Resynchronization of previously timed-inseminated beef heifers with progestins

The objective was to determine the efficacy of a previously used CIDR or melengestrol acetate (MGA; 0.5mg/head/day) for resynchronization of estrus in beef heifers not pregnant to timed-AI (TAI). In three experiments and a field trial, heifers were reinseminated 6-12 h after first detection of estrus. Pregnancy diagnosis was done from approximately 25-43 days after either TAI or reinsemination. In Experiment 1, 79 heifers received a once-used CIDR from 13 to 20 days after TAI and 80 heifers were untreated controls. For these two groups, there were 34 and 35 heifers, respectively, not pregnant to TAI; median +/- S.E. intervals from TAI to onset of estrus were 22 +/- 0.2 days versus 20 +/- 0.6 days (P < 0.001); estrus rates were 70.6% versus 85.7% (P = 0.1); conception rates were 62.5% versus 76.7% (P < 0.3); and pregnancy rates were 44.1% versus 65.7% (P = 0.07), for CIDR and untreated (control) groups, respectively. In Experiment 2, heifers (n = 651) were TAI (Day 0) and 13 days later randomly assigned to one of seven groups (n = 93 per group) to receive a once-used CIDR (three groups; Days 13-20), MGA (three groups; Days 13-19), or no treatment (control group). Groups given a CIDR or MGA also received: no further treatment (CIDR or MGA alone); 1.5mg estradiol-17beta (E-17beta) and 50 mg progesterone (P4) in 2 mL canola oil on Day 13; or E-17beta and P4 on Day 13 and 0.5 mg E-17beta on Day 21 (24 h after CIDR removal or 48 h after the last feeding of MGA). Pregnancy rate to TAI was lowest (P < 0.05) for the group given a CIDR plus E-17beta and P4 on Day 13 and E-17beta on Day 21. Variability in return to estrus was greater (P < 0.001) in the control and MGA groups than in CIDR groups. Conception and pregnancy rates in heifers given a CIDR (65.1 and 61.4%) were higher (P<0.01) than those fed MGA (49.6 and 40.4%), but not different from controls (62.2 and 54.9%, respectively). In Experiment 3, 616 heifers received a once- or twice-used CIDR for 7 days, beginning 13+/-1 days after TAI, with or without a concurrent injection of 150 mg of P4 (2 x 2 factorial design). Pregnancy rate to TAI was 47.2%. In heifers that returned to estrus, there was no significant difference between a once- or twice-used CIDR for rates of estrus (68.8%, P < 0.3), conception (65.9%, P < 0.6) and pregnancy (45.3%, P < 0.8). Injecting progesterone at CIDR insertion increased the median interval from CIDR removal to onset of estrus (P < 0.05) and reduced rates of estrus (63.8% versus 73.8%, P<0.05), conception (60.5% versus 70.6%, P = 0.1) and pregnancy (38.6% versus 52.2%, P < 0.02). In a field trial, 983 heifers received a once-used CIDR for 7 days, beginning 13 +/- 1 days after TAI. Pregnancy rate to TAI was 55.2%. The median (and mode) of the interval from CIDR removal to estrus was 2.5 days. Estrus, conception and pregnancy rates were 78.2, 70.3 and 55.0% (overall pregnancy rate to TAI and rebreeding, 78.7%). In summary, a once- or twice-used CIDR for 7 days, starting 13 +/- 1 days after TAI resulted in the majority of nonpregnant heifers detected in estrus over a 4-day interval, with acceptable conception rates; however, injecting progesterone at CIDR insertion significantly reduced both estrus and pregnancy rates, and estradiol treatment after CIDR removal was associated with a decreased pregnancy rate to TAI. Fertility was higher in heifers resynchronized with a once-used CIDR than with MGA.     

Estrogen concentrations in milk at estrus and ovulation in dairy cows

The aim of the present study was two-fold. First, to characterize the secretory profiles of oestradiol-17beta and progesterone in relation to the structural changes observed by ultrasonography during follicular dynamics in non-ovulating llamas. Second, to evaluate the effect of exogenous progesterone on follicular activity, in terms of follicle development and hormone production. In experiment one, six adult non-pregnant, non-lactating llamas were examined daily by rectal palpation and transrectal ultrasonography during 70 days. On day 54, intravaginal devices containing 0.33 g of progesterone (CIDR) were inserted and left in the vagina during 16 days. The mean duration of a follicular wave was 22.6+/-2.5 days. The follicular growth phase (follicles growing from 3mm to maximum size) averaged 9.2+/-2.8 days, the mature phase (follicles around maximum size) 5.2+/-1.4 days and regression phase (follicles with decreasing size) 8.2+/-2.2 days. Oestradiol-17beta plasma concentrations exhibited a similar wave pattern (P<0.05). In addition, oestradiol-17beta peak plasma concentrations (46.9+/-3.3 pmoll(-1)) were attained approximately 12 days after the beginning of the growing phase in connection with maximum follicle size (11.8+/-1.6mm). After CIDR insertion, a rapid increase in plasma progesterone concentrations was observed, with peak concentrations attained on day 1 after insertion. Thereafter, concentrations decreased gradually. Mean follicle size steadily decreased from the day of CIDR insertion to day 11 post-insertion (10.3+/-1.6 and 3.3+/-0.8mm, respectively). In order to investigate the effect of follicle size at CIDR insertion on the outcome of progesterone treatment, experiment two was designed. Sixteen adult non-pregnant and non-lactating llamas were divided into four groups according to follicle development at the time of CIDR insertion (group I: follicles < or =6 mm; group II: follicles between 6 and 9 mm; group III: follicles between 10 and 14 mm and group IV, regressing follicles). In groups II, III and IV, a significant decrease in follicle size was observed after the insertion of the CIDR device. In group I, no further development of dominant follicles was observed until the device was withdrawn. In all cases, the smallest diameter was registered between days 5 and 7 after the beginning of treatment. In conclusion, a detailed characterization of follicular waves using ultrasound and hormone determinations simultaneously in non-ovulating llamas and after the insertion of progesterone releasing devices, is presented.     

Resynchronization of ovulation and timed insemination in lactating dairy cows III. Administration of GnRH 23 days post AI and ultrasonography for nonpregnancy diagnosis on day 30

The objective was to compare pregnancy rates to resynchronization and timed AI (TAI) protocols in lactating dairy cows that received GnRH at 23 d and were diagnosed not pregnant at 30 d after the pre-enrollment AI. Nonpregnant cows (624) at ultrasonography on day 30 (study day 0) were classified as diestrus (74.8%), metestrus (5.6%) and without a CL (19.5%). Cows in diestrus were assigned either to the GnRH group (PGF2alpha on day 0, GnRH on day 2 and TAI 16 h later, n = 238) or the estradiol cypionate (ECP) group (PGF2alpha on day 0, ECP on day 1, and TAI 36 h later, n = 229). Cows in metestrus were assigned to the Modified Heatsynch Group (GnRH on day 0, PGF(2alpha) on day 7, ECP on day 8 and TAI on day 9, n = 35). Cows without a CL (n = 122) were classified either as proestrus (10.6%), ovarian cysts (7.5%) or anestrus (1.4%), and assigned to factorial treatments (i.e., use of GnRH versus CIDR) to either the GnRH group (GnRH on day 0, PGF2alpha on day 7, GnRH on day 9 and TAI 16 h later, n = 28), the CIDR group (CIDR insert from days 0 to 7, PGF2alpha on day 7, GnRH on day 9 and TAI 16 h later, n = 34), the GnRH + CIDR group (GnRH on day 0, CIDR insert from days 0 to 7, PGF2alpha on day 7, GnRH on day 9 and TAI 16h later, n = 32), and the control group (PGF2alpha on day 7, GnRH on day 9 and TAI 16 h later, n = 28). For cows without a CL, plasma P4 concentrations were determined on days 0, 7, 10 and 17 and ovarian structures determined on days 0, 7 and 17. Pregnancy rates were evaluated at 30, 55 and 90 d after the resynchronized AI. For cows in diestrus, there were no differences in pregnancy rates on days 30, 55 and 90 for cows in the GnRH (27.5, 26.5 and 24.2%) or ECP (29.1, 25.5 and 24.1%) groups. In addition, there were no differences in pregnancy losses between days 30 and 55 and 55 and 90 between the GnRH (7.0 and 8.6%) and ECP (9.8 and 5.4%) groups. For cows without a CL, GnRH on day 0 increased the proportion of cows with a CL on days 7 and 17 and plasma P4 concentration on day 17 in cows with ovarian cysts but not for cows in proestrus. The CIDR insert increased pregnancy rate in cows with ovarian cysts but reduced pregnancy rate for cows in proestrus.     

Body condition influences maintenance of a persistent first wave dominant follicle in dairy cattle

The objective of this study was to determine whether plasma concentrations of progesterone (P4) from a controlled internal drug releasing (CIDR) device (approximately 2 ng/ml) were adequate to sustain a persistent first wave dominant follicle (FWDF) in low body condition (LBC, body condition score [BCS] 1 = lean, 5 = fat [2.3 +/- 0.72, n = 4]) compared with high body condition (HBC, BCS = 4.4 +/- 0.12, n = 4) nonlactating dairy cows. On Day 7 of the estrous cycle (Day 0 = estrus), cows were treated with PGF2 alpha (25 mg i.m. Lutalyse, P.M., and Day 8 A.M.) and a used CIDR device containing P4 (1.2 g) was inserted into the vagina until ovulation or Day 16. Plasma was collected for P4 and estradiol (E2) analyses from Day 5 to Day 18 (or ovulation), and ovarian follicles were monitored daily by ultrasonography. Mean concentrations of plasma P4 were greater in HBC than LBC cows between Days 5 and 7 (4.6 > 3.4 +/- 0.37 ng/ml; P < 0.04). All LBC cows maintained the first wave dominant follicle and ovulated after removal of the CIDR device (18.3 +/- 0.3 d, n = 3; Cow 4 lost the CIDR device on Day 11 and ovulated on Day 15), whereas in the HBC cows ovulation occurred during the period of CIDR exposure (11.3 +/- 0.3 d; n = 3; a fourth cow developed a luteinized first wave dominant follicle that did not ovulate during the experimental protocol on Day 19). Mean day of estrus was 17 +/- 0.4 for LBC (n = 3) and 10 +/- 0.4 for HBC (n = 3) cows. Sustained concentrations of plasma E2 (12.9 +/- 2.8 pg/ml; Days 8 to 17) in LBC cows reflected presence of an active persistent first wave dominant follicle. The differential effect of BCS on concentrations of plasma P4 (y = ng/ml) was reflected by the difference (P < 0.01) in regressions: yLBC = 19.9 - 3.49x + 0.166x2 vs yHBC = 37.3 - 7.04x + 0.340x2 (x = day of cycle, Days 7 to 12). Although P4 concentration was greater for HBC cows prior to Day 8, a greater clearance of plasma P4 released from the CIDR device in the absence of a CL altered follicular dynamics, leading to premature ovulation in the HBC cows. A greater basal concentration of P4 was sustained in LBC cows that permitted maintenance of a persistent first wave dominant follicle.     

Follicular wave emergence, luteal function and synchrony of ovulation following GnRH or estradiol benzoate in a CIDR-treated, lactating Holstein cows

This study evaluated the effect of GnRH or estradiol benzoate (EB) on follicular wave emergence and progesterone concentrations, and following a second injection of GnRH, synchrony of ovulation, and pregnancy rates in a controlled internal drug release (CIDR)-based timed AI (TAI) protocol in lactating Holstein cows. Cows received a CIDR device without hormone (controls), with an injection of 100 microg GnRH or with an injection of 4 mg EB. Thereafter, all received PGF(2 alpha) at the time of CIDR removal on Day 7, GnRH on Day 9, and TAI 16 h later. Follicular wave emergence occurred within 7 days in 19/20 GnRH-treated, 14/20 EB-treated and 5/20 control cows (P < 0.05). The interval to wave emergence was the shorter and less variable (P < 0.01) in the GnRH group (2.9 +/- 0.2 days) than in the EB (4.7 +/- 0.5 days) or control (4.8 +/- 1.0 days) groups. Serum progesterone concentrations from Days 4 to 7 were higher (P < 0.01) in the GnRH-treated cows that ovulated than in those that did not ovulate, or in control and EB-treated cows. The diameters of dominant follicle on Day 7 differed among groups (P < 0.01), and the diameters of the preovulatory follicle on Day 9 were larger (P < 0.01) in the control and GnRH groups than in the EB group. The proportion of cows with synchronized ovulations did not differ among groups, but pregnancy rate to TAI was higher (P < 0.05) in the GnRH group (65%; 13/20) than in the control (30%; 6/20) or EB (35%; 7/20) groups. Results suggest that GnRH treatment of CIDR-treated lactating Holstein cows will result in synchronous follicular wave emergence, large preovulatory follicles and synchronous ovulation, resulting in an acceptable pregnancy rates to TAI.     

Influence of a CIDR prior to bull breeding on pregnancy rates and subsequent calving distribution

We determined whether insertion of a CIDR for 7 days prior to the breeding season enhanced pregnancy rates and altered the date of conception in suckled beef cows mated naturally. Suckled beef cows (n=2033) from 15 locations were randomly assigned to one of two treatments: (1) cows received a CIDR 7 days prior to the breeding season for 7 days (CIDR; n=999); (2) cows received no treatment (Control; n=1034). On the first day of the breeding season bulls were introduced to herds at a rate of 15-25 cows per yearling bull or 20-30 cows per mature bull. Pregnancy status and the date of conception were determined via transrectal ultrasonography at 56 and 120 days after initiation of the breeding season. Overall pregnancy rates ranged from 59.3 to 98.9% among the 15 locations. The percentage of cows becoming pregnant during the first 30 days of the breeding season was similar between CIDR (68.2%) and Control (66.7%) cows, and overall pregnancy rates were similar between CIDR (88.9%) and Control (88.6%) cows. The average day of conception after initiation of the breeding season was shorter (P<0.01) for CIDR (20.1+/-0.8 days) compared to Control cows (23.2+/-0.8 days). Of cows conceiving during the breeding season, more (P<0.05) CIDR cows (35.9%) conceived during the first 10 days of the breeding season than Control cows (30.8%). Neither body condition score and nor parity affected pregnancy rates or days to conception, whereas pregnancy rates and days to conception were affected (P<0.01) by location and days postpartum. Days to conception were greater for cows that calved within 40 days (31.6+/-1.2 days) of initiation of the breeding season compared to cows calving between 40 and 50 days (25.3+/-1.2 days) prior to initiation of the breeding season, which were greater than those cows calving between 50-60 days (20.0+/-0.8 days) and 60-70 days (21.3+/-1.0 days) prior to initiation of the breeding season. Cows calving greater than 70 days (17.3+/-1.5 days) from initiation of the breeding season had the shortest interval to conception. We concluded that insertion of a CIDR prior to the breeding season failed to increase overall pregnancy rates, but did influence the average day of conception.     

Comparison of the effect of estradiol benzoate plus progesterone and GnRH on the follicular wave emergence and subsequent follicular development in CIDR-treated, lactating dairy cows with follicular cysts

This study examined the effect of estradiol benzoate (EB) plus progesterone (P4) as compared with GnRH on follicular wave emergence and follicular development, and synchrony of ovulation and pregnancy rates following a second injection of GnRH in a controlled internal drug release (CIDR)-based timed AI (TAI) protocol in lactating dairy cows with follicular cysts. Lactating dairy cows diagnosed with follicular cysts received a CIDR device, with an injection of 2mg EB plus 50mg P4 (EB+P4 group) or with an injection of 100 microg GnRH (GnRH group) at the beginning of the experiment (day 0). Thereafter, all received PGF(2alpha) at the time of CIDR removal on day 7, GnRH on day 9, and TAI 16 h later. Follicular wave emergence occurred within 7 days in 12/15 EB plus P4-treated and 14/15 GnRH-treated cows (P>0.05). The interval to wave emergence was longer in the EB+P4 group (4.8+/-0.4 days) than in the GnRH group (2.0+/-0.2 days). The mean diameters of preovulatory follicles and the proportion of cows with preovulatory follicles greater than 12 mm on day 9 did not differ between groups (P>0.05). The proportion of cows with synchronized ovulations by 40 h after the GnRH injection on day 11 and pregnancy rates to TAI did not differ between the EB+P4 (13/15 and 36.7%) and the GnRH (14/15 and 53.3%) groups, respectively. Results suggest that a single treatment with EB plus P4 as compared with GnRH simultaneously with CIDR insertion in lactating dairy cows with follicular cysts will result in relatively asynchronous emergence of a new follicular wave, but subsequently similar sizes of preovulatory follicles and synchronous ovulation, resulting in similar pregnancy rates to TAI.     

The effect of subluteal levels of exogenous progesterone on follicular dynamics and endocrine patterns during early luteal phase of the ewe

Nineteen Corriedale ewes were treated with an im dose of a PGF2alpha during the luteal phase to synchronize estrus. After ovulation had been detected by using ultrasonography (Day 0); the ewes were randomly assigned to 2 different groups. In 11 ewes a CIDR, which had previously been used for 10 d, was inserted on the fourth day after ovulation. The ewes then received a dose of PGF2alpha on Day 5 to induce luteolysis. The CIDR remained in place until the end of the experiment (Day 9). Control ewes (n = 8) received no treatment. Blood samples were taken daily for estradiol, progesterone and FSH determinations. In the untreated ewes, 2 follicular waves were detected in all of the animals throughout the monitoring period, with a mean wave interval of 4.5 d. The total number of follicles which were > or =2 mm decreased from Day 0 to Day 4 (8.8+/-1.0 to 5.3+/-0.6; P< or =0.05) and then increased at Day 7 (7.5+/-0.9; P< or =0.05). The growth profiles of both the largest and the second largest follicles of Wave 1 showed significant divergence, while no divergence was observed in Wave 2. Serum estradiol concentrations decreased significantly from the day before to the day of ovulation and then increased again during the growing phase of the largest follicle of Wave 1. Concentrations of FSH were high on the day of emergence of both waves, but while a significant decline was observed after emergence in Wave 1, the levels remained high in Wave 2. In 8 of the 11 treated ewes, the largest follicle of Wave 1 was still present on the ninth day after ovulation (persistent follicle). In the other 3 ewes, the largest follicle of Wave 1 was already regressing on the day that the treatment was administered, and the largest follicle that was present on Day 9 originated from Wave 2 (nonpersistent follicle). In persistent follicle ewes, the largest follicle of Wave 1 prolonged its lifespan significantly, attaining the maximum diameter (Day 8.1+/-0.8) later than in untreated (Day 3.0+/-0.4) and nonpersisted follicle ewes (Day 2.0+/-0.6). The total number of follicles decreased in persistent follicle ewes between Day 0 and Day 4 (7.9+/-1.5 to 4.5+/-0.5, respectively; P< or =0.05) and remained low until the end of the experiment. Progesterone concentrations (nmol/L) between Days 6 and 9 were significantly different between untreated and persistent follicle ewes (12.8+/-1.0 vs. 9.4+/-1.0, P< or =0.02). The present study confirms that the largest follicle of Wave 1 is dominant in the ewe and that subluteal progesterone concentrations can prolong its lifespan and extend this dominance.     

Length of progesterone exposure needed to resolve large follicle anovular condition in dairy cows

Hypothalamic unresponsiveness to an estradiol surge appears to be an underlying cause of large follicle anovular condition (follicular cysts), but progesterone exposure for 7 days resolves this condition. In this study, dairy cows with induced (Experiment 1) or naturally occurring (Experiment 2) follicular cysts were treated for different times with progesterone. In Experiment 1, 16 of 26 cows (62%) were induced into anovulation by causing a GnRH/LH surge when no ovulatory follicle was on the ovary. Anovular cows (n = 16) were assigned to one of four treatment groups ( 0, 1, 3, or 7 days of progesterone treatment) using an intravaginal, progesterone-releasing implant (CIDR). All anovular cows had low circulating progesterone concentrations before controlled internal drug releasing (CIDR) and greater concentrations that reached steady state (1.3 +/- 0.1 ng/mL progesterone) by 3 h after CIDR insertion. Circulating progesterone decreased to basal concentrations by 4 h after CIDR removal. Cows were treated with 5mg estradiol benzoate (EB) 12 h after CIDR removal. None (n = 4) of the control cows (0 day) had an LH surge after EB. All of the 3 days (5/5) and 7 days (4/4) CIDR-treated cows had an LH surge following EB, but only one of the 1 day (1/3) CIDR-treated cows. Magnitude of the LH peak was similar in the 3 and 7 days cows. All cows treated for 7 days ovulated (4/4), whereas, ovulation occurred in only 3/5, 1/3, and 0/4 of the cows treated for 3, 1, and 0 day, respectively. The two cows in the 3 days group that did not ovulate had a normal LH surge, but these two cows had a smaller maximal follicle size than cows that ovulated. In Experiment 2, naturally anovular lactating dairy cows (24 of 248) were identified using weekly ultrasonography. All anovular cows grew follicles to >12 mm, with 54% (13 of 24) having follicles larger than ovular size (15-24 mm) and 33% (8 of 24) having follicles that would be considered cystic (>25 mm). Anovular cows were randomly assigned to CIDR treatment for 0, 1, or 3 days. All (7/7) of 3 days, 33% (3/9) of 1 day, and 25% (2/8) of control (0 day) cows ovulated by 1 week after CIDR removal. Thus, 3 days but not 1 day of progesterone exposure appears to be sufficient to reinitiate estradiol responsiveness of the hypothalamus.     

Comparison of follicular dynamics and hormone concentrations between the 7-day and 5-day CO-Synch + CIDR program in primiparous beef cows

The objectives were to compare follicular dynamics, preovulatory estradiol concentrations, and progesterone concentrations between the 7-day (7CO, n = 15) and 5-day (5CO, n = 13) CO-Synch + controlled internal drug release device (CIDR) program in primiparous suckled beef cows. On Day −7 (7CO) or Day −5 (5CO), GnRH (100 μg) was administered (GnRH-1) and a CIDR was inserted. On Day 0, hour 0, CIDR was removed and cows received PGF_2α (25 mg) at hours 0 and 12. Animals were administered GnRH (100 μg, GnRH-2) at either hour 60 (7CO) or 72 (5CO). Follicular growth and ovulation to both GnRH-1 and GnRH-2 were evaluated using ultrasonography. Concentrations of estradiol were determined in blood samples taken at hours 0, 36, 60, and 72 (5CO). Blood samples were collected on Days 5, 8, and 14 for progesterone quantification. Ovulation rate to GnRH-1 did not differ between the 7CO (11/15) and 5CO (8/13) treatments, and for all dependent variables the statistical model included treatment, ovulation to GnRH-1, and their interaction. Diameter (mm) of the ovulatory follicle did not differ between treatments (13.4 ± 0.3) but was greater (P < 0.05) in cows that responded to GnRH-1 (13.8 ± 0.3) than those did not (12.6 ± 0.6). Maximum estradiol concentrations tended (P = 0.06) to be greater in the 5CO (7.3 ± 0.5 pg/mL) than 7CO (6.1 ± 0.7 pg/mL) treatment and tended to be greater (P = 0.08) in cows that responded to GnRH-1 (7.1 ± 0.5 pg/mL) than those did not (5.6 ± 0.9 pg/mL). Three cows in the 7CO treatment failed to develop a CL after GnRH-2. There was a treatment by response to GnRH-1 interaction (P < 0.05) for progesterone concentrations. In cows that did not respond to GnRH-1 in the 7CO treatment, progesterone concentrations were less (P < 0.05) than in those that responded to GnRH-1 in the 7CO treatment and tended (P = 0.09) to be less than in cows in the 5CO treatment that did not respond to GnRH-1. In conclusion, these findings demonstrate that failure to respond to GnRH-1 is detrimental to estradiol and progesterone concentrations with a 7-day interval between GnRH-1 and PGF_2α but of little consequence when this interval is shortened to 5 days.     

Progesterone concentrations, exogenous equine chorionic gonadotropin, and timing of prostaglandin F2α treatment affect fertility in postpuberal Nelore heifers

Two experiments were performed to test the hypothesis that elevated progesterone concentrations impair pregnancy rate to timed artificial insemination (TAI) in postpuberal Nelore heifers. In Experiment 1, postpuberal Nelore heifers (n = 398) received 2 mg estradiol benzoate (EB) and either a new progesterone-releasing intravaginal device containing 1.9 g of progesterone (CIDR) (first use) or a CIDR previously used for 9 d (second use) or for 18 d (third use) on Day 0, 12.5 mg prostaglandin F_2α (PGF_2α) on Day 7, 0.5 mg estradiol cypionate (ECP) and CIDR withdrawal on Day 9, and TAI on Day 11. Largest ovarian follicle diameter was determined on Day 11. The third-use CIDR treatment increased largest ovarian follicle diameter and pregnancy rate. Conception to TAI was reduced in heifers with smaller follicles in the first- and second-use CIDR treatments, but not in the third-use CIDR treatment. In Experiment 2, postpuberal Nelore heifers received the synchronization treatment described in Experiment 1 or received 12.5 mg PGF_2α on Day 9 rather than Day 7. In addition, 50% of heifers received 300 IU equine chorionic gonadotropin (eCG) on Day 9. Heifers were either TAI (Experiment 2a; n = 199) or AI after detection of estrus (Experiment 2b; n = 125 of 202). In Experiment 2a, treatment with eCG increased pregnancy rate to TAI in heifers that received PGF_2α on Day 9 but not on Day 7 and in heifers that received a first-use CIDR but not in heifers that received a third-use CIDR. Treatments did not influence reproductive performance in Experiment 2b. In summary, pregnancy rate to TAI in postpuberal Nelore heifers was optimized when lower concentrations of exogenous progesterone were administered, and eCG treatment was beneficial in heifers expected to have greater progesterone concentrations.     

Effect of early luteolysis in progesterone-based timed AI protocols in Bos indicus, Bos indicus x Bos taurus, and Bos taurus heifers

The objective of this study was to evaluate the effects of treatment with an intravaginal progesterone-releasing device (CIDR) and estradiol benzoate (EB) on follicular dynamics in Bos indicus (n=23), Bos taurus (n=25), and cross-bred (n=23) heifers. To assess the influence of reduced serum progesterone concentrations during 8 days of treatment with a progesterone-releasing device on follicular dynamics, half of the heifers received PGF at CIDR insertion (Day 0; 3 x 2 factorial design). Mean (+/-S.E.M.) serum progesterone concentrations during CIDR treatment varied (P<0.05) among genetic groups: B. indicus (5.4+/-0.1 ng/mL), B. taurus (3.3+/-0.0 ng/mL), and cross-bred (4.3+/-0.1 ng/mL). Maximum diameter of the dominant follicle (DF) was smaller (P<0.01) in B. indicus heifers (9.5+/-0.5 mm) than in cross-bred (12.3+/-0.4 mm) or B. taurus heifers (11.6+/-0.5 mm). B. indicus experienced lower (P<0.01) ovulation rate (39.1%) than did B. taurus (72.7%) and cross-bred (84.0%). Heifers treated with PGF on Day 0 had lower (P<0.05) serum progesterone concentrations during progesterone treatment. The PGF treatment on Day 0 increased (P<0.01) the diameter of the DF (11.9+/-0.4 mm vs. 10.5+/-0.4 mm). Moreover, greater (P=0.02) ovulation rates (78.8 vs. 54.0%) occurred in heifers treated with PGF on Day 0. In summary, B. indicus heifers had greater serum progesterone concentrations, smaller DF diameter, and a lower ovulation rate compared to B. taurus heifers. Prostaglandin treatment on the day of CIDR insertion reduced serum progesterone during treatment, and resulted in increased maximum DF diameter and ovulation rate.     

The effect of a progesterone (P4) intravaginal device (CIDR) on resynchronisation of oestrus and embryonic loss in previously timed inseminated dairy heifers

A study was done to evaluate the effect of using progesterone (P4) intravaginal device (CIDR: controlled internal drug-releasing dispenser) to synchronise the return to oestrus of previously timed inseminated (TAI) dairy heifers, and to evaluate embryo survival and pregnancy rate (PR) in the return to oestrus heifers. At the onset of the artificial insemination (AI) breeding period (day -9), heifers were randomly assigned into two groups (treated group CGPG, n = 79) and (control group GPG, n = 83). Every heifer in both groups was injected with gonadotropin-releasing hormone (GnRH) agonist and prostaglandin F2-alpha (PGF2α) as follows: GnRH on day -9; PGF2α on day -2; GnRH and TAI on day 0. Heifers in both groups received TAI within 30 min after the second GnRH injection. Artificial insemination at first breeding was conducted for all heifers during 55 days from day 0. On day 14 after timed insemination, every heifer in the CGPG group received CIDR device for 6 days. Within 3 days after CIDR removal, more heifers in CGPG group showed oestrus within 1.9 days compared to heifers that showed oestrus within 2.9 days in the control. Within 10 days after CIDR removal, more heifers in the CGPG group showed oestrus within 2.4 days compared to heifers that showed oestrus within 6.7 days in the control. PRs on days 30 and 55 were not different between both groups, while PR on day 55 during September were higher (P = 0.032) in CGPG group (58.0%) than GPG group (37.0%). In addition, PR from first to second AI was higher (P = 0.037) for CGPG group (79.8%) than for GPG group (65.1%) but it was similar after that. Pregnancy losses between days 30 and 55 tended to be lower (P = 0.089) for the CGPG group (12.7%) compared to 25.1% for the GPG group. Interval between first and second AI was lower (P = 0.052) for the CGPG group (27.5 ± 1.6 days) compared to 31.6 ± 1.3 days for heifers in the GPG group but no differences were detected for intervals from second to third AI and from third to fourth AI between the two groups. Number of services per pregnancy was not different between CGPG and GPG groups. Results indicate that the CIDR device improved synchronisation to return to oestrus and increased PR to first AI during high temperature months by reducing embryonic losses.     

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 PGF_2α (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.     

Effects of estradiol cypionate (ECP) on ovarian follicular dynamics,synchrony of ovulation,and fertility in CIDR-based,fixed-time AI programs in beef heifers

Estradiol cypionate (ECP) was used in beef heifers receiving a controlled internal drug release (CIDR; insertion = Day 0) device for fixed-time AI (FTAI) in four experiments. In Experiment 1, heifers (n = 24) received 1mg ECP or 1mg ECP plus 50mg commercial progesterone (CP) preparation i.m. on Day 0. Eight or 9 days later, CIDR were removed, PGF was administered and heifers were allocated to receive 0.5mg ECP i.m. concurrently (ECP0) or 24h later (ECP24). There was no effect of treatment (P = 0.6) on mean (+/-S.E.M.) day of follicular wave emergence (3.9+/-0.4 days). Interval from CIDR removal to ovulation was affected (P<0.05) only by duration of CIDR treatment (88.3+/-3.8h versus 76.4+/-4.1h; 8 days versus 9 days, respectively). In Experiment 2, 58 heifers received 100mg progesterone and either 5mg estradiol-17beta or 1mg ECP i.m. (E-17beta and ECP groups, respectively) on Day 0. Seven (E-17beta group) or 9 days (ECP group) later, CIDR were removed, PGF was administered and heifers received ECP (as in Experiment 1) or 1mg EB 24h after CIDR removal, with FTAI 58-60h after CIDR removal. Follicular wave emergence was later (P<0.02) and more variable (P<0.002) in heifers given ECP than in those given E-17beta (4.1+/-0.4 days versus 3.3+/-0.1 days), but pregnancy rate was unaffected (overall, 69%; P = 0.2). In Experiment 3, 30 heifers received a CIDR device and 5mg E-17beta, with or without 100mg progesterone (P) i.m. on Day 0. On Day 7, CIDR were removed and heifers received ECP as described in Experiment 1 or no estradiol (Control). Intervals from CIDR removal to ovulation were shorter (P<0.05) in ECP0 (81.6+/-5.0h) and ECP24 (86.4+/-3.5h) groups than in the Control group (98.4+/-5.6h). In Experiment 4, heifers (n = 300) received a CIDR device, E-17beta, P, and PGF (as in Experiment 3) and after CIDR removal were allocated to three groups (as in Experiment 2), with FTAI 54-56h (ECP0) or 56-58h (ECP24 and EB24) after CIDR removal. Pregnancy rate did not differ among groups (overall, 63.6%, P = 0.96). In summary, although 1mg ECP (with or without progesterone) was less efficacious than 5mg E-17beta plus 100mg progesterone for synchronizing follicular wave emergence, 0.5mg ECP (at CIDR removal or 24h later) induced a synchronous ovulation with an acceptable pregnancy rate to fixed-time AI.     

Ovarian responses to progesterone and oestradiol benzoate administered intravaginally during dioestrus in cattle

This study examined the effects of administering progesterone and oestradiol benzoate (ODB) during mid-dioestrus, on ovarian follicular dynamics in cattle. Twelve cycling cows were used in a 4 x 4 latin square design, with the 4 treatments being initiated on Day 13 of the cycle (oestrus = Day 0) and comprising intravaginal insertion for 5 days of: (i) a progesterone releasing device (CIDR; 'P4'); (ii) a CIDR device with a gelatin capsule containing 10 mg ODB and 1 g lactose (CIDIROL; 'P4/ODB') attached; (iii) a placebo CIDR device with the 10 mg ODB capsule (ODB); and, (iv) a placebo CIDR device alone (CTRL). The ovaries of each cow were examined daily by transrectal ultrasonography from Day 7 of the cycle until subsequent ovulation. Blood samples were collected daily from Day 11, and at intervals of 2-4 h during the 24 h period either side of treatment initiation. The second dominant follicle (DF2) emerged on Day 10.7 +/- 0.2 (mean +/- SEM), and was 8.5 +/- 0.2 mm in diameter by Day 13. The DF2 developed through to ovulation (2-wave cycles) in half of the animals in the CTRL group; while in the other half of cases, the ovulatory follicle originated from the third follicle wave that emerged on Day 17.2 +/- 0.4. Administration of a CIDR device alone (P4 group) did not alter the 1:1 ratio of 2 and 3-wave cycles, but the third dominant follicle (DF3) in those cows with 3-wave cycles emerged earlier on Day 15.6 +/- 0.2. In contrast, the DF2 of every animal in the ODB and P4/ODB groups became atretic and was replaced by a DF3 which emerged 4.0 +/- 0.3 days later. The effects of ODB on luteal function were limited to an earlier decline in plasma progesterone concentrations from 2 to 4 days after device insertion and a reduction in diameter of the corpus luteum when administered concurrently with progesterone. Intravaginal administration of 10 mg ODB on Day 13 of the oestrous cycle, with or without progesterone, was effective in promoting follicle wave turnover. In the absence of ODB, progesterone administration alone (P4 group) did not alter the ratio of animals with 2 or 3-wave cycles from that observed in animals in the CTRL group, but did advance the timing of subsequent follicle wave emergence in those animals with 3-wave cycles.     

Ability of ram introduction to induce LH secretion, estrus and ovulation in fall-born ewe lambs during anestrus.

The ability of ram introduction (RI) and progesterone pre-treatment to induce increases in LH secretion and ovulation, and the ability of progesterone pre-treatment with or without estrogen to induce estrus and ovulation in fall-born ewe lambs during seasonal anestrus was investigated. In early July, lambs of mixed breeds (41.8+/-0.6 kg and 250.7+/-1.3 days of age) were assigned to receive no treatment (C, n=7), to be introduced to rams (7:1 ewe:ram ratio; R, n=7), to be treated with progesterone (a used CIDR device) for 5 days (P, n=5), to be treated with progesterone and introduced to rams at CIDR removal (PR, n=11), or to receive the latter treatment plus an injection of estradiol benzoate (25 microg, E2beta i.m.) 24 h after CIDR withdrawal/RI (PER, n=11). Blood samples were collected from all lambs every 4h for 60 h beginning at RI/CIDR withdrawal (0 h), to characterize the LH surge profile and in groups R and C every 15 min for 8 h between 12 and 20 h for determination of LH pulse frequencies. Ultrasonographic examinations of the ovaries were conducted at 0, 36 and 60 h. In ram-exposed groups lambs were also observed for raddle marks every 4h from 0 to 60 h. The LH pulse frequency (pulses/8 h) was higher in group R (P<0.01; 7.7+/- 0.5) than group C lambs (2.7+/- 0.8). More lambs in groups exposed to rams than in the C or P groups showed an LH surge (P<0.05; 0, 100, 0, 72.7 and 100%, for C, R, P, PR and PER groups, respectively). Time from RI/CIDR removal to initiation of the LH surge was greater in lambs in the PR (43.5+/- 3.8h) than in the R (32.6+/- 4.6h; P=0.08) or PER (33+/- 1.2h; P<0.01). Diameter of the largest follicle at 0 h (3.2+/- 0.2mm) was not different among groups. Growth rate of the largest follicle between 0 and 36 h was greater (P<0.05) in RI than in C or P groups. Diameter of the largest follicle at 36 h was larger (P<0.05) in lambs in R (5.6+/- 0.2mm) and PR (5.1+/- 0.5mm) than C (4.0+/- 0.6mm) or P (3.8+/- 0.4mm) groups, and in R than PER (4.3+/- 0.4mm) treatment groups. Only lambs in the RI groups ovulated. Among RI groups the percentage of lambs ovulating was greater in the R (P<0.05; 85.7%) than PR (33.3%) groups with an intermediate response observed in lambs in treatment group PER (71.4%). The estrous response in progesterone pre-treated groups was greater (P<0.05) in lambs also treated with estrogen (PER; 81.8%), than in lambs introduced to rams alone (PR; 45.5%). In conclusion, ram introduction by itself, but not progesterone treatment alone, induces increases in LH pulse frequency, follicular development, and ovulation in fall-born ewe lambs during seasonal anestrus, further, P4 pre-treatment and RI when combined with E2 results in a high estrous response.     

Evaluation of an ovarian synchronization scheme for fixed-time artificial insemination in wapiti

The ovarian response to an empirically derived treatment protocol used commercially for fixed-time insemination in wapiti (Cervus elaphus) was evaluated by transrectal ultrasonography in hinds during transition into the ovulatory season. On September 29, hinds (n=7) were given an intravaginal progesterone-releasing device (CIDR-B, 1.9 g of progesterone) or left untreated (controls, n=9). Fourteen days later, hinds in the treated group were given 200 IU eCG and the CIDR was removed. Hinds in the control group ovulated randomly over a 15 day period. In the treated group, five hinds ovulated 3 days after eCG treatment, one ovulated 7 days after treatment, and one failed to ovulate by November 1. All extant dominant follicles ceased growth and/or began to regress within 2 days of CIDR placement. Two waves of follicular development were detected between CIDR insertion and removal; the first emerged 5.1+/-0.5 days after CIDR insertion and the second at 11.0+/-0.7 days. Serum progesterone concentration was 0.6+/-0.5 ng/mL (range 1.0-0.3 ng/mL) before CIDR placement, remained above 6 ng/mL during CIDR placement, and fell to 0.8+/-0.9 ng/mL after CIDR removal. In the control group, maximal luteal-phase progesterone concentration was lower (1.1+/-0.1 ng/mL; P<0.05) and emergence of the first follicular wave was more variable (P=0.05) than in the treated group. The protocol to synchronize ovulation was effective in 5/7 (71%) hinds, and 4/7 (57%) became pregnant and calved. The pregnancy rate (6/9) and calving rate (5/9) was similar in the control group. In conclusion, synchronization with CIDR-B was effective; however, the protocol may be improved by shortening the interval of CIDR placement to < or = 7 days and by reducing the circulating concentrations of progesterone to physiologic concentrations (< 4 ng/mL).     

Effect of estradiol valerate on ovarian follicle dynamics and superovulatory response in progestin-treated cattle

Three experiments evaluated the effects of estradiol valerate (EV) on ovarian follicular and CL dynamics, intervals to estrus and ovulation, and superovulatory response in cattle. Experiment 1 compared the efficacy of two norgestomet ear implants (Crestar and Syncro-Mate B; SMB) for 9 d (with PGF at implant removal), combined with either 5 mg estradiol-17beta and 100 mg progesterone (EP) or 5 mg EV and 3mg norgestomet (EN) im at the time of implant insertion on CL diameter and follicular wave dynamics. Ovaries were monitored by ultrasonography. There was no effect of norgestomet implant. Diameter of the CL decreased following EN treatment (P < 0.01). Mean (+/- S.D.) day of follicular wave emergence (FWE) was earlier (P < 0.0001) and less variable (P < 0.0001) in EP- (3.6 +/- 0.5 d) than in EN- (5.7 +/- 1.5 d) treated heifers. Intervals from implant removal to estrus (P < 0.001) and ovulation (P < 0.01) were shorter in EN- (45.7 +/- 11.7 and 74.3 +/- 12.6 h, respectively) than in EP- (56.4 +/- 14.1 and 83.3 +/- 17.0 h, respectively) treated heifers. Experiment 2 compared the efficacy of EP versus EN in synchronizing FWE for superovulation in SMB-implanted cows. At random stages of the estrous cycle, Holstein cows (n = 78) received two SMB implants (Day 0) and were randomly assigned to receive EN on Day 0 or EP on Day 1. Folltropin-V treatments were initiated on the evening of Day 5, with PGF in the morning and evening of Day 8, when SMB were removed. Cows were inseminated after the onset of estrus and embryos were recovered 7 d later. Non-lactating cows had more CL (16.7 +/- 11.3 versus 8.3 +/- 4.9) and total ova/embryos (14.7 +/- 9.5 versus 7.9 +/- 4.6) than lactating cows (P < 0.05). EP-treated cows tended (P = 0.09) to yield more transferable embryos (5.6 +/- 5.2) than EN-treated cows (4.0 +/- 3.7). Experiment 3 compared the effect of dose of EV on ovarian follicle and CL growth profiles and synchrony of estrus and ovulation in CIDR-treated beef cows (n = 43). At random stages of the estrous cycle (Day 0), cows received a CIDR and no further treatment (Control), or an injection of 1, 2, or 5 mg im of EV. On Day 7, CIDR were removed and cows received PGF. Follicular wave emergence occurred within 7 d in 7/10 Control cows and 31/32 EV-treated cows (P < 0.05). In responding cows, interval from treatment to FWE was longer (P < 0.05) in those treated with 5 mg EV (4.8 +/- 1.2 d) than in those treated with 1 mg (3.2 +/- 0.9 d) or 2 mg (3.4 +/- 0.8 d) EV, while Control cows were intermediate (3.8 +/- 2.0 d). Diameter of the dominant follicle was smaller (P < 0.05) at CIDR removal and tended (P = 0.08) to be smaller just prior to ovulation in the 5 mg EV group (8.5 +/- 2.2 and 13.2 +/- 0.6 mm, respectively) than in the Control (11.8 +/- 4.6 and 15.5 +/- 2.9 mm, respectively) or 1mg EV (11.7 +/- 2.5 and 15.1 +/- 2.2 mm, respectively) groups, with the 2mg EV group (10.7 +/- 1.5 and 14.3 +/- 1.7 mm, respectively) intermediate. Diameter of the dominant follicle at CIDR removal was less variable (P < 0.01) in the 2 and 5mg EV groups than in the Control group, and intermediate in the 1mg EV group. In summary, treatment with 5mg EV resulted in a longer and more variable interval to follicular wave emergence than treatment with 5mg estradiol-17beta, which affected preovulatory dominant follicle size following progestin removal, and may have also affected superstimulatory response in Holstein cows. Additionally, 5 mg EV appeared to induce luteolysis in heifers, reducing the interval to ovulation following norgestomet removal. Conversely, intervals to, and synchrony of, follicular wave emergence, estrus and ovulation following treatment with 1 or 2 mg EV suggested that reduced doses of EV may be more useful for the synchronization of follicular wave emergence in progestogen-treated cattle.     

Ultrasonic identification of follicular populations and return to estrus in early postpartum dairy cows given intravaginal progesterone for 15 days

In an attempt to program ovarian function in the early post partum period, 52 lactating Holstein cows were injected with 25 mg prostaglandin F(2alpha) (PGF) and given a CIDR device containing 1.9 g progesterone for 15 d starting on Day 25 post partum. Ovarian follicles were measured by ultrasound on 0, 5, 10 and 15 d after insertion and on alternate days after CIDR removal until estrus. Not all cows were devoid of corpora lutea (CL) during the CIDR (11, 9 and 8 cows had a CL on Days 5, 10 and 15, respectively). There was a CL by day interaction (P<0.01) for the number of 10- to 15-mm follicles per cow; the average number of large follicles (>15 mm) was twice greater (0.75 vs 0.37) for those cows not having a CL during the period of CIDR exposure. The average size of the largest follicle increased to a maximum of 19.3 +/- 0.7 mm by 15 d after insertion in cows not having a CL. Plasma estradiol increased for 10 d after insertion, then decreased to the end of the CIDR period. After removal of the CIDR, 34 cows ovulated, eight cows developed ovarian follicular cysts, and eight cows did not ovulated by 14 d. Cows becoming cystic or not ovulating had a declining number of follicles during the CIDR compared with those cows ovulating (P<0.07). The diameter of the largest follicle in cystic cows was equivalent to noncystic cows until removal of the CIDR, but then it increased markedly. Interval to estrus was longer in cows having more 6- to 9-mm follicles on Day 15 (day of CIDR removal). These results demonstrate the existence and maintenance of a large dominant follicle after CIDR insertion and PGF injection which was influenced apparently by the presence of a CL. Furthermore, subsequent reproductive responses after the CIDR treatment was a function of follicular populations prior to withdrawal of the CIDR device. This system may be appropriate for the study of factors regulating follicular growth and fertility in domestic cattle.