Controlled Internal Drug Release Insert Removal Research Articles

15 Influence of early progesterone removal on follicular development, estrus expression, and pregnancy rates in presynchronized beef cows

Abstract The objective of this study was to evaluate the impact of early progesterone removal on pregnancy rates to fixed-time artificial insemination (FTAI) in presynchronized beef cows. Postpartum beef cows (n = 922; body condition score = 4.9 ± 0.77; days postpartum = 77.3 ± 20.77 d) from 7 locations were enrolled in a completely randomized design. Within location, cows were stratified by parity and days postpartum before being randomly assigned to 1 of 2 treatments: 1) 7&7 Synch: Cows received a controlled internal drug release insert (CIDR) and a 25-mg injection of prostaglandin F2α (PG1; d 0). Cows received 100 μg of GnRH on d 7 and the CIDR was removed 7 d later, concurrently with a second prostaglandin F2α (PG2) injection (d 14). Cows received a second injection of GnRH and were FTAI 60-66 h after PG2 (d 17); 2) 7&6 Synch: Cows received the same treatment as 7&7 Synch cows; however, CIDR removal and PG2 were performed on d 13, and cows were FTAI 60-66 h after CIDR removal. Ovarian ultrasonography was performed to determine follicle diameter and CL presence at PG2 and FTAI in a subset of cows (n = 40). Estrus detection patches were utilized to evaluate estrus expression between PG2 and FTAI. Pregnancy diagnosis was performed 28-35 d after FTAI. Cows exposed to the 7&7 Synch tended to have larger follicle diameter at PG2 compared with 7&6 Synch cows (P = 0.09); however, there were no differences in follicle diameter at FTAI. Cows that expressed estrus tended to have larger follicle diameter at FTAI compared with cows that failed to express estrus (P = 0.10). Follicular growth between PG2 and FTAI, percentage of cows with a corpus luteum at PG2, and ovulation rate were not impacted by treatment (P ≥ 0.56). No differences were determined between treatments for estrus expression (7&7 Synch: 77.1 ± 5.91; 7&6 Synch: 74.6 ± 6.28; P = 0.50) or pregnancy rates to FTAI (7&7 Synch: 60.1 ± 4.32; 7&6 Synch: 58.2 ± 4.26; P = 0.69). In conclusion, early removal of progesterone did not impact pregnancy rates in presynchronized beef cows. Thus, the 7&6 Synch protocol described herein may be used as an alternative to the conventional 7&7 Synch protocol without compromising fertility.

Association of gastrointestinal parasite burden, serum cytokines and hormones concentrations, and pregnancy in Angus-cross beef cows

The objective was to elucidate the relationships among gastrointestinal (GI) parasite load, serum cytokines (Th 1 – Interleukin (IL) 2, Interferon (IFN) γ and Tumor necrosis factor (TNF) α; Th 2- IL4, IL6, and IL10) levels, hormones (progesterone, cortisol, 8-epi-prostaglandin F2 alpha (isoprostane), prolactin, substance-p, and prostaglandin F metabolites) concentrations, and pregnancy in beef cattle. Angus-cross beef cows (n = 700; age, 3–8 y) were blocked by age and body condition score (BCS, 1–9), and were randomly assigned to treatment (n = 350, TRT, 50 mg of eprinomectin/50 kg BW, im) or control (n = 350, CON, no treatment) on Day -30. Cows were synchronized using Controlled Internal Drug Release insert (CIDR) + CO-Synch protocol and artificially inseminated at a fixed time on Day 0 (66 h after CIDR removal). Fecal samples were collected to determine fecal egg count per gram (FEG, McMaster method) on Days -30, -23, -16, -7, 7, 0, 16 and 23, and blood samples were collected on Days -7, 0, 7, 16 and 23. Serum cytokines were determined on Days -7, 0, 7, 16 and 23, and circulating hormones were measured on Day 16. BCS were recorded on Day 16 following artificial insemination (AI), and pregnancy status was diagnosed on Day 30 and 60. Pregnancy/AI varied among treatment groups on Day 30 [TRT, 62.0% (217/350); CON, 54.9% (192/350) (P = 0.05)] and Day 60 [TRT, 60.9% (213/350); CON, 51.7% (181/350) (P < 0.05)]. Pregnancy loss between 30 and 60 days for TRT and CON groups were 1.8% (4/217) and 5.7% (11/192), respectively (P < 0.05). The BCS on Day 16 did not differ among treatment groups (P> 0.1). Four groups of 40 cows were selected based on their pregnancy status and treatment: pregnant, TRT; non-pregnant, TRT; pregnant, CON; and non-pregnant, CON to compare the mean FEG, cytokines, and hormones levels. The FEG and cytokine concentrations were significantly (P < 0.05) influenced by treatment, pregnancy status, day, treatment by pregnancy status, and treatment by day. Day 16 hormone concentrations were considerably influenced by treatment, pregnancy status, and treatment by pregnancy. Although FEG on Day -30 did not differ among the groups (P> 0.1), it was lower in treated, pregnant cows compared with cows in other three groups from Day -23 onwards (P < 0.05). Overall and pairwise comparisons showed that serum concentrations of Type 1 cytokines, IL2, IFNγ, and TNFα were lower (P < 0.05) from gestational Day 7 onwards in treated, pregnant cows compared with cows in other three groups. In contrast, serum concentrations of Type 2 cytokines, IL4, IL6 and IL10 were greater (P < 0.05) from gestational Day 7 onwards in treated, pregnant cows compared with cows in other groups. Serum concentrations of progesterone was greater and other hormones were lower for pregnant cows in TRT group compared to cows in other groups on gestational Day 16. In conclusion, GI parasite load was reduced; Th 1 cytokines levels were decreased; Th 2 cytokines concentrations were increased; progesterone level was increased; and cortisol, substance-P, prolactin, isoprostane, and PGFM were decreased in pregnant, TRT cows. These changes also resulted in an increase in P/AI. It is plausible that direct and bidirectional host-parasite interactions mediated by cytokines and hormones may have promoted maternal tolerance of an immunologically diverse conceptus and the establishment of pregnancy.

Reproductive performance of replacement dairy heifers submitted to first service with programs that favor insemination at detected estrus, timed artificial insemination, or a combination of both

Our objective was to compare the insemination dynamics and time to pregnancy for up to 100 d after the beginning of the artificial insemination period (AIP) for heifers managed with first artificial insemination (AI) service programs that relied primarily on insemination at detected estrus (AIE) after PGF2α treatments, timed artificial insemination (TAI), or a combination of both. Holstein heifers were randomly assigned to receive first AI service with sex-selected semen after 368 ± 10 d of age with (1) AIE after synchronization of estrus with up to 3 PGF2α treatments every 14 d starting on the first day of the AIP (PGF+AIE; n = 317). Heifers not AIE up to 9 d after the third PGF2α received a 5-d Cosynch protocol with progesterone supplementation [GnRH + controlled internal drug release insert (CIDR)-5 d-CIDR removal and PGF2α-3 d-GnRH and TAI] before TAI. Heifers detected in estrus from CIDR removal and PGF2α until the day before TAI received AIE with no GnRH treatment; (2) 2 PGF2α treatments 14 d apart with the second treatment at the beginning of the AIP (PGF+TAI; n = 334). Heifers received AIE for up to 9 d after the second PGF2α treatment. Heifers not AIE received TAI after the 5-d Cosynch protocol and (3) TAI after the 5-d Cosynch protocol (ALL-TAI; n = 315). Heifers failing to conceive to a previous AI received a subsequent AI with conventional semen at detected estrus or TAI after the 5-d Cosynch protocol. Binomial outcomes were analyzed by logistic regression, whereas time to AI and pregnancy were analyzed with Cox's regression. The hazard of first AI up to 45 d of the AIP was greater for ALL-TAI than for PGF+AIE [hazard ratio (HR) = 1.72; 95% confidence interval (CI) =1.45 to 2.03] and PGF+TAI (HR = 1.51; 95% CI = 1.28 to 1.77), but similar for PGF+AIE and PGF+TAI (HR = 1.14; 95% CI = 0.97 to 1.33). A greater proportion of heifers received AIE in PGF+AIE (98.7%) than in PGF+TAI (78.5%). Overall, first service pregnancy per AI did not differ (PGF+AIE = 42.0%; PGF+TAI = 47.3%, ALL-TAI = 43.8%). Time to pregnancy was reduced for ALL-TAI compared with PGF+AIE (HR = 1.20, 95% CI = 1.02 to 1.42), but was similar to that of PGF+TAI (HR = 1.13, 95% CI = 0.96 to 1.33). Time to pregnancy did not differ for PGF+AIE and PGF+TAI (HR = 1.07, 95% CI = 0.91 to 1.25). Median days to pregnancy were 27, 23, and 21 for heifers in PGF+AIE, PGF+TAI, and ALL-TAI, respectively. We concluded that an ALL-TAI program for first service reduced time to pregnancy, albeit a relatively small reduction, when compared with a program that relied primarily on AIE after induction of estrus with PGF2α treatments. The program that combined synchronization of estrus and TAI (PGF+TAI) resulted in similar time to pregnancy than the predominant TAI and predominant AIE programs.

Fertility after two doses of PGF2α concurrently or at 6-hour interval on the day of CIDR removal in 5-day CO-Synch progesterone-based synchronization protocols in beef heifers

Timed artificial insemination protocols in beef cattle are designed to synchronize ovulation in a greater proportion of females while simultaneously achieving acceptable pregnancy rates and a concise calving season. Protocols achieving such goals reduce time and labor associated with estrus detection and make advanced reproductive technologies implementable for beef producers. The objective of the study was to determine the effect of three different PGF2α (PGF) dosage schemes on artificial insemination (AI) pregnancy rates in beef heifers. We hypothesized that two doses of PGF administered concurrently at the time of controlled internal drug release (CIDR) removal would attain similar pregnancy rates compared with two doses given 6-hours apart—one at CIDR removal and the next 6 hours later in the 5-day CO-Synch progesterone-based synchronization protocol. Angus heifers (n = 875) at six locations in Washington, Idaho, and Oregon states were included in this study. Heifers within locations were assigned a body condition score (BCS). All heifers received a CIDR (1.38 g of progesterone) and 100 μg IM of GnRH on Day 0. The CIDRs were removed on Day 5, heifers were randomly allocated to one of three protocol groups: 1PGF (n = 291), received 25 mg IM of dinoprost (PGF); 2CO-PGF (n = 291), received 50 mg IM of dinoprost at CIDR removal, 2PGF (n = 293), received 25 mg IM of dinoprost at CIDR removal, and an additional 25 mg IM of dinoprost 6 hours later. Each heifer was given GnRH (100 μg, IM) and artificially inseminated at 56 hours after CIDR removal. Heifers were examined for pregnancy status between 50 and 70 days after AI to determine time of conception. A mixed-model procedure (PROC GLIMMIX of SAS) was used to evaluate the effect of treatments (1PGF, 2CO-PGF, and 2PGF) on AI pregnancy rates. Models included were treatments, BCS categories (≤5 and >5), and treatment by BCS category interaction. Location (state), handling facilities, handlers, inseminators, and AI sires were included as a random effect in the model. The 2PGF group had greater AI pregnancy rate of 63.6% (185/291), compared with the 2CO-PGF group at 51.9% (151/291) and 1PGF group at 54.9% (161/293; P < 0.001). An AI pregnancy rate of 50% (104/208) was observed for heifers with BCS less than or equal to 5 versus 58.9% (393/667) for heifers with BCS greater than 5 (P < 0.05). Location did not influence the AI pregnancy rate (P > 0.1). In conclusion, beef heifers received two 25-mg doses of PGF at 6-hour interval on Day 5 at CIDR insert removal in a 5-day CO-Synch + CIDR synchronization protocols achieved greater pregnancy compared with heifers received 50 mg of PGF concurrently at CIDR removal.

Evaluation of treatment with a 5-day versus 7-day controlled internal drug-release insert on reproductive outcomes of beef heifers using a modified timed–artificial insemination protocol

The objective of this experiment was to determine the effect of reducing the duration of controlled internal drug-release insert (CIDR) exposure in a CIDR-based, fixed-time-AI protocol on pregnancy per AI (P/AI) and pregnancy rates in beef heifers. The experiment was conducted in 4 consecutive years. British crossbred heifers (yr 1, n = 82; yr 2, n = 68; yr 3, n = 86; yr 4, n = 53) were stratified by BW and age and were assigned randomly to 1 of 2 treatments: (1) CIDR7 heifers (n = 145) received a CIDR insert for 7 d; and (2) CIDR5 heifers (n = 144) received a CIDR insert for 5 d. Heifers in CIDR7 and CIDR5 were given prostaglandin F2α (25 mg) upon CIDR removal followed by gonadotropin-releasing hormone (75 μg) and fixed-time AI 56 to 59 h after CIDR removal. Estrual behavior was monitored following CIDR removal. In all 4 yr, heifers received AI by a single inseminator. Fourteen days after AI, heifers were exposed to bulls for 40 d. Blood samples were collected on the day of CIDR insertion. Pregnancy status was determined by ultrasonography at d 35 and d 67 to 82 after AI. Data were analyzed by logistic regression and ANOVA. Percentage of heifers detected in estrus was different (P 0.2) between treatments. At CIDR insertion, mean serum progesterone concentrations were similar between CIDR7 and CIDR5, and there was no effect of treatment-by-year interaction on progesterone concentrations (P > 0.2). There was a treatmentby-year interaction effect on P/AI (P < 0.05). For CIDR7 and CIDR5, in yr 1, P/AI was 39 and 65.8% (P < 0.05), respectively; in yr 2, P/AI was 64.7and 41.1% (P < 0.05), in yr 3, P/AI was 59 and 73.8% (P = 0.09), and in yr 4, P/AI was 50 and 70.4% (P = 0.25). Overall, P/AI tended (P = 0.07) to be greater for CIDR5 compared with CIDR7 (63.8 vs. 53%). Regardless of treatment, heifers detected in estrus had a greater (P < 0.01) P/AI compared with those not detected in estrus (65.9 vs 47.9%). The final pregnancy rates of heifers were unaffected by the treatment protocols or year (92 and 95.5% for CIDR7 and CIDR5, respectively). The P/AI results from this study suggest that reducing the duration of CIDR treatment from 7 to 5 d in a CIDR-based and single-prostaglandin-injection, fixed-time-AI protocol may improve P/AI in beef heifers.

Technical note: Physiological response of beef heifers after receiving a reused controlled internal drug release insert processed with different heat-treating methods1

Eighty-one prepubertal beef heifers were used to evaluate effects of used controlled internal drug release (CIDR) insert heating methods on concentrations of progesterone after CIDR insert reinsertion. Heifers were stratified by weight and birth date and then assigned to receive a new CIDR insert (New; n = 10) or 1 of 8 used (7 d prior use) CIDR insert treatments: 1) no processing (Used; n = 10), 2) autoclaved (Autoclaved; n = 8), 3) processed in dishwasher (Dishwasher; n = 8), 4) processed in microwave for 30 s (Microwave; n = 10), 5) processed in toaster oven (Oven; n = 9), 6) processed in clothes dryer (Dryer; n = 10), 7) processed in boiling water (Boiled; n = 8), or 8) stored outdoors for 60 d (Outside; n = 8). Used CIDR inserts were processed at 121°C for 30 min for autoclaved and oven treatments, at 121°C for boiled treatment, and for 30 min for dryer and dishwasher treatments. Blood samples were collected on d -10, immediately before CIDR insert insertion (d 0), 3 h after CIDR insert insertion (3 h), daily while CIDR insert was in place (d 1 to 11), and 24 h after CIDR insert removal (d 12) for analysis of concentrations of progesterone. Subjective color scores (1 = bright white to 5 = completely stained yellow/red) were assigned to each CIDR insert after d 11. A treatment × time interaction (P < 0.0001) was present for concentrations of progesterone. Concentrations of progesterone were similar (P > 0.10) for heifers receiving a used CIDR insert compared with heifers receiving CIDR inserts processed in a dishwasher, microwave, oven, dryer, or boiling water (collectively reported as "Processed"). However, heifers receiving autoclaved CIDR inserts had greater (P < 0.05) concentrations of progesterone from h 3 to d 3 but similar (P > 0.10) concentrations of progesterone from d 4 to d 11 compared with heifers receiving used or processed CIDR inserts. From d 1 to 11 heifers receiving outside CIDR inserts had decreased (P < 0.05) concentrations of progesterone compared with all other treatments. Heifers receiving autoclaved CIDR inserts had greater (P < 0.05) concentrations of progesterone compared with all other treatments at 3 h and 1 d, whereas heifers receiving new CIDR inserts had greater (P < 0.05) concentrations of progesterone from d 6 to 11 compared with all other treatments. Outside CIDR inserts were more discolored (P < 0.001) compared with all other treatments. Processing used CIDR inserts with a dishwasher, microwave, oven, clothes dryer, boiling water, or full environmental exposure did not result in a pattern of concentrations of progesterone similar to that of autoclaved or new CIDR inserts.

Altered progesterone concentrations by hormonal manipulations before a fixed-time artificial insemination CO-Synch + CIDR program in suckled beef cows

We hypothesized that pregnancy outcomes may be improved by inducing luteal regression, ovulation, or both (i.e., altering progesterone status) before initiating a timed–artificial insemination (TAI) program in suckled beef cows. This hypothesis was tested in two experiments in which cows were treated with either PGF2α (PG) or PG + GnRH before initiating a TAI program to increase the proportion of cows starting the program in a theoretical marginal (<1 ng/mL; experiment 1) or elevated (≥1 ng/mL; experiment 2) progesterone environment, respectively. The control was a standard CO-Synch + controlled internal drug release (CIDR) program employed in suckled beef cows (100 μg GnRH intramuscularly [IM] [GnRH-1] and insertion of a progesterone-impregnated intravaginal CIDR insert on study Day −10, 25 mg PG and CIDR insert removal on study Day −3, and 100 μg GnRH IM [GnRH-2] and TAI on study Day 0). In both experiments, blood was collected before each injection for later progesterone analyses. In experiment 1, cows at nine locations (n = 1537) were assigned to either: (1) control or (2) PrePG (same as control with a PG injection on study Day −13). The PrePG cows had larger (P < 0.05) follicles on study Day −10 and more (P < 0.05) ovulated after GnRH-1 compared with control cows (60.6% vs. 36.5%), but pregnancy per TAI was not altered (55.5% vs. 52.2%, respectively). In experiment 2, cows (n = 803) at four locations were assigned to: (1) control or (2) PrePGG (same as control with PG injection on study Day −20 and GnRH injection on study Day −17). Although pregnancy per TAI did not differ between control and PrePGG cows (44.0% vs. 44.4%, respectively), cows with body condition score greater than 5.0 or 77 or more days postpartum at TAI were more (P < 0.05) likely to become pregnant than thinner cows or those with fewer days postpartum. Presynchronized cows in both experiments were more (P < 0.05) likely than controls to have luteolysis after initial PG injections and reduced (P < 0.05) serum progesterone; moreover, treatments altered the proportion of cows and pregnancy per TAI of cows in various progesterone categories before the onset of the TAI protocol. In combined data from both experiments, cows classified as anestrous before the study but with elevated progesterone on Day −10 had increased (P < 0.05) pregnancy outcomes compared with anestrous cows with low progesterone concentrations. Progesterone concentration had no effect on pregnancy outcome of cycling cows. In summary, luteal regression and ovulation were enhanced and progesterone concentrations were altered by presynchronization treatments before the 7-day CO-Synch + CIDR program, but pregnancy per TAI was not improved.

Effect of the first GnRH and two doses of PGF2α in a 5-day progesterone-based CO-Synch protocol on heifer pregnancy

The objectives were (1) to determine the effects of gonadorelin hydrochloride (GnRH) injection at controlled internal drug release (CIDR) insertion on Day 0 and the number of PGF2α doses at CIDR removal on Day 5 in a 5-day CO-Synch + CIDR program on pregnancy rate (PR) to artificial insemination (AI) in heifers; (2) to examine how the effect of systemic concentration of progesterone and size of follicles influenced treatment outcome. Angus cross beef heifers (n = 1018) at eight locations and Holstein dairy heifers (n = 1137) at 15 locations were included in this study. On Day 0, heifers were body condition scored (BCS), and received a CIDR. Within farms, heifers were randomly divided into two groups: at the time of CIDR insertion, the GnRH group received 100 μg of GnRH and No-GnRH group received none. On Day 5, all heifers received 25 mg of PGF2α at the time of CIDR insert removal. The GnRH and No-GnRH groups were further divided into 1PGF and 2PGF groups. The heifers in 2PGF group received a second dose of PGF2α 6 hours after the administration of the first dose. Beef heifers underwent AI at 56 hours and dairy heifers at 72 hours after CIDR removal and received 100 μg of GnRH at the time of AI. Pregnancy was determined approximately at 35 and/or 70 days after AI. Controlling for herd effect (P < 0.06), the treatments had significant effect on AI pregnancy in beef heifers (P = 0.03). The AI-PRs were 50.3%, 50.2%, 59.7%, and 58.3% for No-GnRH + PGF + GnRH, No-GnRH + 2PGF + GnRH, GnRH + PGF + GnRH, and GnRH + 2PGF + GnRH groups, respectively. The AI-PRs were ranged from 50% to 62.4% between herds. Controlling for herd effects (P < 0.01) and for BCS (P < 0.05), the AI pregnancy was not different among the treatment groups in dairy heifers (P > 0.05). The AI-PRs were 51.2%, 51.9%, 53.9%, and 54.5% for No-GnRH + PGF + GnRH, No-GnRH + 2PGF + GnRH, GnRH + PGF + GnRH, and GnRH + 2PGF + GnRH groups, respectively. The AI-PR varied among locations from 48.3% to 75.0%. The AI-PR was 43.5%, 50.4%, and 64.2% for 2.5 or less, 2.75 to 3.5, and greater than 3.5 BCS categories. Numerically higher AI-PRs were observed in beef and dairy heifers that exhibited high progesterone concentrations at the time of CIDR insertion (>1 ng/mL, with a CL). In addition, numerically higher AI-PRs were also observed in heifers receiving CIDR + GnRH with both high and low progesterone concentration (<1 ng/mL) initially compared with heifers receiving a CIDR only with low progesterone. In dairy heifers, there were no differences in the pregnancy loss between 35 and 70 days post-AI among the treatment groups (P > 0.1). In conclusion, GnRH administration at the time of CIDR insertion is advantageous in beef heifers, but not in dairy heifers, to improve AI-PR in the 5-day CIDR + CO-Synch protocol. In addition, in this study, both dairy heifers that received either one or two PGF2α doses at CIDR removal resulted in similar AI-PR in this study regardless of whether they received GnRH at CIDR insertion.

Evaluation of reproductive and economic outcomes of dairy heifers inseminated at induced estrus or at fixed time after a 5-day or 7-day progesterone insert-based ovulation synchronization protocol

The objectives of the current experiment were to evaluate the reproductive performance and economic outcome of 3 synchronization strategies for first artificial insemination (AI) of dairy heifers. Holstein heifers from 2 herds (site A, California, n=415; site B, Idaho, n=425) were assigned to 1 of 3 treatments. Heifers assigned to the AI on estrus (AIE) treatment received an injection of 25mg of PGF2α at enrollment (d 0) and every 11 d thereafter until AI occurred. Heifers assigned to the CIDR5 treatment received a controlled internal drug release insert (CIDR) containing 1.38g of progesterone, which was removed 5 d later concomitantly with an injection of 25mg of PGF2α, and received fixed-time AI (TAI) concomitantly with an injection of 100μg of GnRH 53 to 60 h later. Heifers assigned to the CIDR7 treatment received a CIDR insert, which was removed 7 d later concomitantly with an injection of 25mg of PGF2α, and received TAI concomitantly with an injection of 100μg of GnRH 53 to 60 h later. Heifers were observed for estrus and inseminated up to 98 and 73 d after enrollment in sites A and B, respectively. Thereafter, heifers were moved to pens with bulls and considered failure to conceive to AI if still not pregnant at the end of the observation period. Economic outcomes were based on cost of synchronization protocol (CIDR treatment=$11, PGF2α or GnRH treatments=$2.5/treatment, estrous detection=$0.80/heifer per day), rearing cost ($2.75/heifer per day), and economic loss if a heifer did not conceive to first AI ($150). Input cost of the reproductive programs=synchronization protocol cost + semen cost + rearing cost + replacement cost. Pregnancy per AI (P/AI) 38±3 d after first AI was greatest for AIE heifers (61.1%) followed by CIDR5 (44.8%) and CIDR7 (35.7%) heifers. Furthermore, P/AI 73±7 d after first AI was greatest for AIE (58.8%) and tended to be greater for CIDR5 (42%) than for CIDR7 (34.1%) heifers. The percentage of heifers that had spontaneous luteolysis from CIDR insertion to CIDR removal was reduced for CIDR5 compared with CIDR7 (13.8 vs 31.8%). Pregnancy rate was greatest for AIE heifers but did not differ between CIDR5 [adjusted hazard ratio (95% confidence interval)=0.75 (0.63, 0.90)] and CIDR7 [adjusted hazard ratio (95% confidence interval)=0.65 (0.54, 0.77)] heifers. Consequently, rearing cost and input cost of AIE heifers ($67.1±4.4 and −$107.1±7.0, respectively) were reduced compared with CIDR5 ($86.9±5.1 and −$143.4±8.1, respectively) and CIDR7 ($98.3±5.1 and −$156.5±8.2, respectively) heifers, but no differences were observed between CIDR5 and CIDR7 heifers.

Evaluation of the 5-day versus a modified 7-day CIDR breeding program in dairy heifers

Dairy heifers were used to compared the effects of two timed AI + controlled internal drug release (CIDR) protocols (5-day vs. a modified 7-day) on: (1) luteal regression to initiate a new ovarian follicular wave; (2) ovarian response to the initial GnRH injection; and (3) pregnancy outcomes. Holstein heifers (N = 543) were assigned randomly to two treatments: (1) 25 mg PGF2α (im) and a CIDR insert on Day −7 followed by 100 μg of GnRH (GnRH-1) on Day −5 and 25 mg PGF2α (im) at CIDR insert removal (7-day [7D]) on Day 0; or (2) 100 μg GnRH (GnRH-1) and insertion of a CIDR on Day −5 and 25 mg PGF2α (im) at CIDR removal (5-day [5D]) on Day 0. Insemination with frozen-thawed conventional or gender-biased semen occurred after detected estrus from Days 0 to 2 or by appointment at 72 h after PGF2α when a second 100-μg dose of GnRH was given. Blood was collected on Days −7, −5, 0, and 3 to determine concentrations of progesterone and incidence of luteolysis. Ovaries were scanned on Days −5 and 0. Luteolysis in the 7D treatment by 48 h after the initial PGF2α was greater (P < 0.01) than what occurred spontaneously in the 5D treatment (36.2% vs. 19.7%, respectively). Incidence of ovulation after GnRH-1 on Day −5 was greater (P < 0.05) for 7D than for 5D heifers, but the proportion of heifers with an induced CL on Day 0 did not differ between treatments. Heifers inseminated after detected estrus (166/543, 30.6%) on Days 0, 1, and 2 had greater (P < 0.05) pregnancy per AI (P/AI) at 32 days post AI than after timed AI (38.2% vs. 28.3%) on Day 3. Pregnancy P/AI, however, was greater (P < 0.05) for 7D heifers inseminated at estrus (46.5%) than for 7D heifers receiving the timed AI (26.8%) and differed (P < 0.05) from all 5D heifers regardless of insemination time at estrus (30.5%) or at timed AI at 72 h (29.9%). At the Florida location in which conventional and sexed semen were used during two breeding clusters, P/AI using sexed semen (43.9%, N = 56) did not differ from that of conventional semen (21.2%, N = 50). Remaining replicates of sexed semen produced similar P/AI at the other two locations (sexed = 27.6%, N = 71; and sexed = 31.9%, N = 215). We concluded that the modified 7-day CO-Synch + CIDR program produced more P/AI in heifers inseminated at estrus than a standard 5-day CO-Synch + CIDR program, but when timed AI occurred at 72 h after PGF2α and CIDR insert removal, P/AI did not differ between programs.

Artificial insemination at 56 h after intravaginal progesterone device removal improved AI pregnancy rate in beef heifers synchronized with five-day CO-Synch + controlled internal drug release (CIDR) protocol

The objective was to determine whether timed artificial insemination (TAI) 56 h after removal of a Controlled Internal Drug Release (CIDR, 1.38 g of progesterone) insert would improve AI pregnancy rate in beef heifers compared to TAI 72 h after CIDR insert removal in a 5-days CO-Synch + CIDR protocol. Angus cross beef heifers (n = 1098) at nine locations [WA (5 locations; n = 634), ID (2 locations; n = 211), VA (one location; n = 193) and WY (one location; n = 60)] were included in this study. All heifers were given a body condition score (BCS; 1-emaciated; 9-obese), and received a CIDR insert and 100 μg of gonadorelin hydrochloride (GnRH) on Day 0. The CIDR insert was removed and two doses of 25 mg of dinoprost (PGF2α) were given, first dose at CIDR insert removal and second dose 6 h later, on Day 5. A subset of heifers (n = 629) received an estrus detector aid at CIDR removal. After CIDR removal, heifers were observed thrice daily for estrus and estrus detector aid status until they were inseminated. Within farm, heifers were randomly allocated to two groups and were inseminated either at 56 h (n = 554) or at 72 h (n = 544) after CIDR removal. All heifers were given 100 μg of GnRH at AI. Insemination 56 h after CIDR insert removal improved AI pregnancy rate compared to insemination 72 h (66.2 vs. 55.9%; P < 0.001; 1 – β = 0.94). Locations, BCS categories (≤ 6 vs. > 6) and location by treatment and BCS by treatment interactions did not influence AI pregnancy rate (P > 0.1). The AI pregnancy rates for heifers with BCS ≤ 6 and > 6 were 61.8 and 60.1%, respectively (P > 0.1). The AI pregnancy rates among locations varied from 54.9 to 69.2% (P > 0.1). The AI pregnancy rate for heifers observed in estrus at or before AI was not different compared to heifers not observed in estrus [(65.4% (302/462) vs. 52.7% (88/167); P > 0.05)]. In conclusion, heifers inseminated 56 h after CIDR insert removal in a 5-days CO-Synch + CIDR protocol had, on average, 10.3% higher AI pregnancy rate compared to heifers inseminated 72 h after CIDR insert removal.

Comparison of the CIDR Select and 5-day Select Synch + CIDR protocols that included limited estrus detection and timed insemination for synchronizing estrus in beef heifers

The objective of this study was to compare AI pregnancy rates between the 5-d Select Synch + CIDR (5dCIDR; CIDR = controlled internal drug-release insert) and CIDR Select (CS) programs in replacement beef heifers with a limited period of estrus detection and AI followed by a timed insemination (TAI) of heifers not exhibiting estrus. Crossbred beef heifers (n = 318) were equally and randomly assigned to either the 5dCIDR or CS treatment. The 5dCIDR treatment consisted of gonadotropin-releasing hormone (GnRH) at CIDR insertion, followed 5 d later by CIDR removal and administration of two 25-mg doses of prostaglandin F2α (PGF) administered approximately 12 h apart. The CS treatment consisted of insertion of a CIDR for 14 d, an injection of GnRH 9 d after CIDR removal, and administration of PGF 7 d following GnRH. In both treatments, heifers were detected for estrus for 52 h following PGF, and those exhibiting estrus were inseminated based on the AM/ PM rule. Heifers not observed in estrus were TAI 72 h after PGF, concurrent with GnRH administration. Ten days following TAI, bulls were placed with the heifers for approximately 45 d. Estrus response tended (P = 0.07) to be greater in the CS treatment than in the 5dCIDR treatment. However, conception rate of heifers exhibiting estrus, TAI conception rate, overall AI pregnancy rate, and breeding season pregnancy rate did not differ (P > 0.10) between treatments. In conclusion, 5dCIDR and CS protocols were equally effective at synchronizing estrus in replacement beef heifers.

Evaluation of the 5- vs. 7-day CIDR program in dairyheifers before timed artificial insemination (2011)

Our objectives were to determine: (1) the effectiveness of an injection of PGF2α to regress the corpus luteum before initiating an timed artificial insemination (TAI) program, (2) ovulation response to gonadotropin-releasing hormone (GnRH), and (3) pregnancy outcomes in dairy heifers inseminated with conventional and gender-biased semen. Heifers (n = 545) from 3 locations (Florida, Kansas, and Mississippi) were assigned randomly to 1 of 2 treatments: (1) 25-mg prostaglandin F2α (PGF2α) injection and controlled internal drug release (CIDR) insert on day −7 followed by 100 μg of GnRH administered on day −5, and a 25-mg PGF2α injection at CIDR insert removal (7D) on day 0; or (2) 100 μg of GnRH and insertion of previously used autoclaved CIDR on day −5 and a 25-mg PGF2α injection at CIDR removal (5D) on day 0. Artificial insemination occurred after detected estrus from days 0 to 3. Those heifers not detected in estrus were inseminated on day 3 (72 hours after PGF2α) and given a second 100-μg dose of GnRH (72 hours after CIDR removal). Blood collected on days −7 and −5 was assayed to determine concentrations of progesterone and presence of a CL (progesterone ≥1 ng/mL) on d −7. Blood progesterone concentrations on days 0 and 3 were used to determine if luteolysis occurred in all heifers. Pregnancy was determined on days 32 and 60 and intervening pregnancy loss was calculated. Of those heifers in the 7D treatment having progesterone ≥1 ng/mL on day −7, the proportion having progesterone <1 ng/mL 2 days later (luteolysis) was greater (P < 0.05) than that in the 5D treatment (43.0 vs. 22.9%), respectively. A treatment by location interaction was detected for pregnancies per AI. The Kansas location had no detectable treatment differences. In contrast, the 7D treatment produced more (P < 0.05) pregnancies in the first replicate of the Florida location and at the Mississippi location. We concluded that the 5D protocol was not more effective in producing acceptable luteolysis, pregnancy, and ovulation rates compared with the modified 7D protocol.

Insemination timing affects pregnancy rates in beef cows treated with CO-Synch protocol including an intravaginal progesterone insert

Our objective was to determine the optimal time to artificially inseminate lactating beef cows (Bos taurus typicus) after using the standard CO-Synch protocol that also included a progesterone-releasing, intravaginal controlled internal drug release (CIDR) insert. Cows (N = 605) at three locations were inseminated at four different times after CIDR insert removal and the prostaglandin F 2α administration of the CO-Synch + CIDR protocol: 48, 56, 64, or 72 h. Blood samples were collected 9 to 10 d before and on the day of CIDR insertion. Based on elevated (>1 ng/mL) serum progesterone concentrations, 60% of 605 cows had previously ovulated (were cycling) at the initiation of the study, with a range of 39.6% to 67.9% among locations (P < 0.05). Age of cow, body condition score, and days postpartum affected (P ≤ 0.05) cycling status before ovulation was synchronized. Averaged across treatments, pregnancy rate to artificial insemination (AI) at Day 32 was affected (P ≤ 0.05) by pretreatment cycling status and body condition. Younger cows (≤3 yr) tended to have greater AI pregnancy rates when inseminated at 56 h, whereas older cows had similar pregnancy rates when inseminated at 56 h or later (timing of AI by age interaction; P = 0.085). Pregnancy loss between Days 32 and 63 was greatest (quadratic effect; P < 0.05) when cows were inseminated at 48 and 72 h. In summary, insemination times at or after 56 h improved AI pregnancy rates when using the CO-Synch + CIDR program. Further work is warranted to examine age effects on timing of AI in the CO-Synch + CIDR program.

Fixed-time AI pregnancy rate following insemination with frozen-thawed or fresh-extended semen in progesterone supplemented CO-Synch protocol in beef cows

The objective of this study was to compare fixed-time AI pregnancy rate in Angus crossbred beef cows inseminated with frozen-thawed or fresh-extended semen. Two ejaculates from each of two Angus bulls were collected by artificial vagina and pooled for each bull. The pooled semen from each bull was divided into two aliquots; Aliquot 1 was extended using Caprogen ® (LIC, Hamilton, New Zealand) to a concentration of 3 × 10 6 sperm/straw and Aliquot 2 was extended using egg-yolk–glycerol extender to a concentration of 20 × 10 6 sperm/straw. Semen extended with Caprogen ® was maintained at ambient temperature and semen extended with egg-yolk–glycerol extender was frozen and maintained at −196 °C until insemination. In each of two breeding seasons (Fall 2007 and Spring 2008), Angus-crossbeef cows ( N = 1455) at 12 locations were randomly assigned within location to semen type [Fresh ( N = 736) vs. Frozen ( N = 719)] and sire [1 ( N = 731) vs. 2 ( N = 724)]. All cows were synchronized with 100 μg of GnRH im and a progesterone Controlled Internal Drug Release insert (CIDR) on Day 0, and on Day 7, 25 mg of PGF2 α im and CIDR removal. All cows received 100 μg of GnRH im and were inseminated at a fixed-time on Day 10, 66 h after CIDR removal. Timed-AI pregnancy rates were influenced by season ( P < 0.05), cows detected in estrus prior to and at AI ( P < 0.001), and dam age ( P < 0.01). Pregnancy rates were not affected by semen type (Fresh = 51.5% vs. Frozen = 50.4%; P = 0.66) and there were no significant interactions of semen type by estrus expression, semen type by sire, or semen type by season ( P > 0.1). In conclusion, commercial beef cows inseminated with fresh-extended semen (3 × 10 6 sperm/straw) yielded comparable pregnancy rates to conventional frozen-thawed semen in a progesterone supplemented, CO-Synch fixed-time AI synchronization protocol and may provide an alternate to frozen semen for more efficient utilization of superior genetics.

Evaluation of human chorionic gonadotropin as a replacement for GnRH in an ovulation synchronization protocol before fixed-time insemination (2007)

Two experiments were conducted to evaluate the difference between gonadotropinreleasing hormone (GnRH) and human chorionic gonadotropin (hCG) given at the beginning of a timed AI protocol and their effects on fertility. In Experiment 1, beef cows (n = 672) at six different locations were assigned randomly to treatments based on age, body condition, and days postpartum. On day −10, cattle were treated with GnRH or hCG and a progesterone-releasing controlled internal drug release (CIDR) insert was placed in the vagina. An injection of PGF2αwas given and CIDR inserts were removed on day −3. Cows were inseminated at one fixed timed at 62 hr (day 0) after CIDR insert removal. Pregnancy was diagnosed at 33 days (range of 32 to 35) after insemination to determine pregnancy rates. For cows that were pregnant after the first insemination, a second pregnancy diagnosis was conducted 35 days (range of 33 to 37) after the first diagnosis to determine pregnancy survival. Pregnancy rates were reduced by the hCG injection compared with the GnRH injection (39.1 vs. 53.5%). In Experiment 2, cattle were assigned randomly to three treatments, balanced evenly across the two treatments (GnRH vs. hCG) applied in Experiment 1. Cows were injected with GnRH, hCG, or saline seven days before the first pregnancy diagnosis of cows inseminated in Experiment 1. At the time of pregnancy diagnosis, cattle found not pregnant (n = 328) were given PGF2αand inseminated 56 hours later. A second pregnancy diagnosis was conducted 35 days (range of 33 to 37) after the second insemination to determine pregnancy rate at the second AI. Injections of GnRH, hCG, or saline had no effect on pregnancy rates of cows already pregnant to the first insemination. Pregnancy rates after second insemination in cows given an injection of hCG or GnRH, however, tended to be reduced. Percentage of cows pregnant after two timed inseminations exceeded 60% without any need to detect estrus.

Treatment of Cycling and Noncycling Lactating Dairy Cows with Progesterone During Ovsynch

Our objective was to determine whether progesterone (P4) supplementation during an Ovsynch protocol would enhance fertility in lactating dairy cows. Lactating dairy cows (n = 634) at 6 locations were assigned randomly within lactation number and stage of lactation to receive the Ovsynch protocol [OVS; synchronization of ovulation by injecting GnRH 7 d before and 48h after PGF2α, followed by one fixed-time AI (TAI) 16 to 20h after the second GnRH injection] or Ovsynch plus a controlled internal drug release (CIDR) P4-releasing insert for 7 d, beginning at the first GnRH injection (OVS + CIDR). Blood was sampled to quantify P4 10 d before the first GnRH injection, immediately before the first GnRH injection, at the time of CIDR removal, before the PGF2α injection (1 to 2h after CIDR insert removal), and 48h after the PGF2α injection to determine cyclicity status before initiation of treatment, luteal status at the PGF2α injection, and incidence of luteal regression. Overall, conception rates at 28 (40 vs. 50%) and 56 d (33 vs. 38%) after TAI differed between OVS and OVS + CIDR, respectively; but a treatment×location interaction was detected. Compared with OVS, pregnancy outcomes were more positive for OVS + CIDR cows at 4 of 6 locations 28 d after TAI and at 3 of 6 locations 56 d after TAI. An interaction of luteal status (high vs. low) before CIDR insert removal and PGF2α injection with pretreatment cycling status indicated that cows having low P4 at PGF2α injection benefited most from P4 supplementation (OVS + CIDR = 36% vs. OVS = 18%), regardless of pretreatment cycling status. Pregnancy loss between 28 and 56 d after TAI was greater for noncycling cows (31%) compared with cycling cows (16%). Pregnancy loss for cows receiving P4 (21%) did not differ from that for cows not receiving P4 (21%). Supplementation of P4, pretreatment cycling status, and luteal status before PGF2α injection altered follicular diameters at the time of the second GnRH injection, but were unrelated to pregnancy outcomes. Incidence of multiple ovulation was greater in noncycling than in cycling cows. Further, cows having multiple ovulations had improved pregnancy outcomes at 28 and 56 d after TAI. In summary, a CIDR insert during the Ovsynch protocol increased fertility in lactating cows having low serum P4 before PGF2α injection. Improved pregnancy outcomes were observed at some, but not all locations.

Expression of low-affinity p75 nerve growth factor receptor in developing mouse nasal region

We hypothesized that pregnancy outcomes may be improved by inducing luteal regression, ovulation, or both (i.e., altering progesterone status) before initiating a timed–artificial insemination (TAI) program in suckled beef cows. This hypothesis was tested in two experiments in which cows were treated with either PGF2α (PG) or PG + GnRH before initiating a TAI program to increase the proportion of cows starting the program in a theoretical marginal (<1 ng/mL; experiment 1) or elevated (≥1 ng/mL; experiment 2) progesterone environment, respectively. The control was a standard CO-Synch + controlled internal drug release (CIDR) program employed in suckled beef cows (100 μg GnRH intramuscularly [IM] [GnRH-1] and insertion of a progesterone-impregnated intravaginal CIDR insert on study Day −10, 25 mg PG and CIDR insert removal on study Day −3, and 100 μg GnRH IM [GnRH-2] and TAI on study Day 0). In both experiments, blood was collected before each injection for later progesterone analyses. In experiment 1, cows at nine locations (n = 1537) were assigned to either: (1) control or (2) PrePG (same as control with a PG injection on study Day −13). The PrePG cows had larger (P < 0.05) follicles on study Day −10 and more (P < 0.05) ovulated after GnRH-1 compared with control cows (60.6% vs. 36.5%), but pregnancy per TAI was not altered (55.5% vs. 52.2%, respectively). In experiment 2, cows (n = 803) at four locations were assigned to: (1) control or (2) PrePGG (same as control with PG injection on study Day −20 and GnRH injection on study Day −17). Although pregnancy per TAI did not differ between control and PrePGG cows (44.0% vs. 44.4%, respectively), cows with body condition score greater than 5.0 or 77 or more days postpartum at TAI were more (P < 0.05) likely to become pregnant than thinner cows or those with fewer days postpartum. Presynchronized cows in both experiments were more (P < 0.05) likely than controls to have luteolysis after initial PG injections and reduced (P < 0.05) serum progesterone; moreover, treatments altered the proportion of cows and pregnancy per TAI of cows in various progesterone categories before the onset of the TAI protocol. In combined data from both experiments, cows classified as anestrous before the study but with elevated progesterone on Day −10 had increased (P < 0.05) pregnancy outcomes compared with anestrous cows with low progesterone concentrations. Progesterone concentration had no effect on pregnancy outcome of cycling cows. In summary, luteal regression and ovulation were enhanced and progesterone concentrations were altered by presynchronization treatments before the 7-day CO-Synch + CIDR program, but pregnancy per TAI was not improved.

Presence of oxytocin receptor mRNA in GnRH-1 cells

We hypothesized that pregnancy outcomes may be improved by inducing luteal regression, ovulation, or both (i.e., altering progesterone status) before initiating a timed–artificial insemination (TAI) program in suckled beef cows. This hypothesis was tested in two experiments in which cows were treated with either PGF2α (PG) or PG + GnRH before initiating a TAI program to increase the proportion of cows starting the program in a theoretical marginal (<1 ng/mL; experiment 1) or elevated (≥1 ng/mL; experiment 2) progesterone environment, respectively. The control was a standard CO-Synch + controlled internal drug release (CIDR) program employed in suckled beef cows (100 μg GnRH intramuscularly [IM] [GnRH-1] and insertion of a progesterone-impregnated intravaginal CIDR insert on study Day −10, 25 mg PG and CIDR insert removal on study Day −3, and 100 μg GnRH IM [GnRH-2] and TAI on study Day 0). In both experiments, blood was collected before each injection for later progesterone analyses. In experiment 1, cows at nine locations (n = 1537) were assigned to either: (1) control or (2) PrePG (same as control with a PG injection on study Day −13). The PrePG cows had larger (P < 0.05) follicles on study Day −10 and more (P < 0.05) ovulated after GnRH-1 compared with control cows (60.6% vs. 36.5%), but pregnancy per TAI was not altered (55.5% vs. 52.2%, respectively). In experiment 2, cows (n = 803) at four locations were assigned to: (1) control or (2) PrePGG (same as control with PG injection on study Day −20 and GnRH injection on study Day −17). Although pregnancy per TAI did not differ between control and PrePGG cows (44.0% vs. 44.4%, respectively), cows with body condition score greater than 5.0 or 77 or more days postpartum at TAI were more (P < 0.05) likely to become pregnant than thinner cows or those with fewer days postpartum. Presynchronized cows in both experiments were more (P < 0.05) likely than controls to have luteolysis after initial PG injections and reduced (P < 0.05) serum progesterone; moreover, treatments altered the proportion of cows and pregnancy per TAI of cows in various progesterone categories before the onset of the TAI protocol. In combined data from both experiments, cows classified as anestrous before the study but with elevated progesterone on Day −10 had increased (P < 0.05) pregnancy outcomes compared with anestrous cows with low progesterone concentrations. Progesterone concentration had no effect on pregnancy outcome of cycling cows. In summary, luteal regression and ovulation were enhanced and progesterone concentrations were altered by presynchronization treatments before the 7-day CO-Synch + CIDR program, but pregnancy per TAI was not improved.