Effects of prepartum lipid supplementation on FSH superstimulation and transferable embryo recovery in multiparous beef cows.

The objective of this experiment was to determine the effect of prepartum lipid supplementation on the number and quality of embryos recovered following ovarian super-ovulation in postpartum suckled beef cows. Mature cows (n = 40) were assigned to one of two treatments (lipid versus. no lipid) and supplemented for approximately 40 days prior to calving. Supplements provided to cows were isocaloric and isonitrogenous. The treatment group was fed 1.6 kg hd(-1) per day of whole soybeans (WSB; 19.8% ether extract, and 41.8% crude protein) and the control group received a supplement consisting of 1.8 kg hd(-1) day of a soybean meal and soy-hull combination (SBS; 2.15% EE and 36.81% CP). Cows were synchronized using a GnRH [Cystorelin((R)) 100 microg im]-GnRH-PGF(2alpha) [Lutalyse 25 mg im] protocol. Cows were administered two injections of GnRH seven days apart and PG seven days after the second GnRH injection. Twenty-eight cows (WSB, n = 15; SBS, n = 13) responded to estrus synchronization and were superstimulated. Super-ovulation was initiated on day 8-10 of the synchronized cycle by twice-daily injections of pFSH (Pluset) over four days in decreasing doses using a total of 608.4 IU per cow. Prostaglandin F(2alpha) was administered 96 and 108 h after super-stimulation was initiated with FSH. Days postpartum (WSB = 59 days; SBS = 57 days) at initiation of FSH treatments were similar (P > 0.10) for both treatments. Cows were monitored for estrus activity by the HeatWatch Estrus Detection System. Twenty-seven cows (WSB, n = 15; SBS, n = 12) exhibited estrus after FSH and inseminated at 0, 12, and 24 h after the onset of estrus with 1, 2, and 1 units of semen, respectively. Embryos were recovered and evaluated 7-8 days later. Only cows that responded to FSH and that were inseminated were used for statistical analysis. Data were analyzed using the General Linear Models Procedure of SAS. Body condition scores did not differ (P > 0.10) between treatments when cows were evaluated at the initiation of the experiment, two weeks prior to calving, and at initiation of superovulation with FSH. Estrous cyclicity prior to the initiation of estrus synchronization did not differ (P > 0.10) between treatments. There was no difference (P > 0.10) between treatments in recovery of total embryos (WSB, 14.7 +/- 3.5; SBS, 17.5 +/- 3.0), transferable embryos (WSB, 10.3 +/- 2.5; SBS, 13.6 +/- 2.6), degenerate embryos (WSB, 3.3 +/- 1.1; SBS, 1.6 +/- 1.7) or unfertilized ova (WSB, 1.1 +/- 0.5; SBS, 2.3 +/- 1.2). Cows that were supplemented with whole soybeans prior to parturition failed to produce an increased total number of ova or transferable embryos following super-ovulation.

A comparison of progestin-based protocols to synchronize ovulation and facilitate fixed-time artificial insemination in postpartum beef cows.

The experimental objective was to compare pregnancy rates after fixed-time AI in postpartum suckled beef cows following administration of two progestin-based protocols to synchronize ovulation. Cows (n = 424) at three locations (n = 208, 122, and 92 per location) were stratified by age, BCS, and days postpartum (DPP) and assigned randomly to one of the two treatment protocols. The MGA Select-treated cows (MGA Select; n = 213) were fed melengestrol acetate (MGA, 0.5 mg x cow(-1) x d(-1)) for 14 d and carrier for 8 d, and then GnRH (100 microg i.m. Cystorelin; d 26) was injected 12 d after MGA withdrawal, and PG (25 mg i.m. Lutalyse) was administered 7 d after GnRH. Cows assigned to the 7-11 Synch protocol (7-11 Synch; n = 209) were fed carrier for 15 d and MGA for 7 d, and then injected with PG on d 22 (d 7 of MGA), GnRH on d 26, and PG again on d 33. Artificial insemination was performed at fixed times for cows in both treatments at 60 or 72 h after d 33 PG for 7-11 Synch and MGA Select groups, respectively. All cows were injected with GnRH (100 microg of i.m. Cystorelin) at AI. There was no treatment x location interaction for age (P = 0.90), BCS (P = 0.64), or DPP (P = 0.93), and the results were therefore pooled for the respective treatments (age [7-11 Synch, 5.5 +/- 0.2; MGA Select, 5.5 +/- 0.2], BCS [7-11 Synch, 5.7 +/- 0.1; MGA Select, 5.6 +/- 0.1], and DPP [7-11 Synch, 41.1 +/- 1.1; MGA Select, 42.1 +/- 1.1]). Blood samples were collected 8 and 1 d before MGA or carrier to determine pretreatment estrous cyclicity (progesterone >or=1 ng/mL; 7-11 Synch, 59/209 [28%]; MGA Select, 54/213 [25%]; P = 0.50) and again on d 33 PG to evaluate treatment response as a percentage of cows with progesterone concentrations in serum >or=1ng/mL (7-11 Synch, 184/209 [88%]; MGA Select, 177/213 [83%]; P = 0.15). Pregnancy rates resulting from fixed-time AI did not differ (P = 0.25) between treatments (7-11 Synch, 128/209 [61%]; MGA Select, 142/213 [67%]), nor did pregnancy rates (P = 0.77) at the end of the breeding season (7-11 Synch, 198/208 [95%]; MGA Select, 204/213 [96%]). These data indicate that pregnancy rates were comparable after fixed-time AI, following administration of the 7-11 Synch and MGA Select protocols. Both protocols provide opportunities for beef producers to use AI and eliminate the need to detect estrus.

Estrus synchronization in beef heifers with progestin-based protocols. I. Differences in response based on pubertal status at the initiation of treatment.

Two progestin-based protocols for estrus synchronization in replacement beef heifers were compared on the basis of estrous response, interval to and synchrony of estrus, and pregnancy rate. The objective was to determine, whether addition of GnRH to a melengestrol acetate (MGA)-prostaglandin F2alpha (PGF2alpha) estrus synchronization protocol would improve synchrony of estrus without compromising fertility in yearling beef heifers. Heifers at two locations (Location 1, n = 60 and Location 2, n = 64) were assigned randomly to one of two treatments by breed and pubertal status. Heifers were defined as, pubertal when concentrations of progesterone in serum were elevated (> or = 1 ng/mL) in either one of two samples obtained 10 and 1 day prior to treatment initiation. Prior to MGA administration, 18/60 (30%) and 36/64 (56%) of the heifers at Locations 1 and 2, respectively, were pubertal. Heifers in both treatments were fed MGA (0.5 mg/head/day in 1.8 kg/head/day supplement) for 14 days followed by 25 mg of PGF2alpha i.m. (MGA-PGF2alpha) 19 days after MGA withdrawal (Day 33 of treatment). One-half of the heifers at each location received 100 microg of GnRH i.m. 12 days after MGA withdrawal (Day 26 of treatment; MGA Select). The control group received only MGA-PGF2alpha. Heifers were observed for signs of behavioral estrus continuously during daylight hours for 7 days beginning on the day PGF2alpha was administered. Heifers were inseminated 12 h after observed estrus. There was a treatment by location by pubertal status interaction (P < 0.05) for interval to estrus. Compared to the respective control treatment at each location, prepubertal heifers assigned to the MGA Select protocol at Location 1 had longer intervals to estrus, whereas at Location 2, prepubertal heifers assigned to the MGA-PGF2alpha protocol had longer intervals to estrus. The higher number of pubertal heifers at Location 2 was associated with a reduced variance in the interval to estrus among MGA Select treated heifers. Total estrous response and synchronized conception rates were similar between treatments at both locations. These data suggest that addition of GnRH to the MGA-PGF2alpha protocol may improve synchrony of estrus, however, the degree of synchrony may be influenced by pubertal status of heifers at the time treatments are imposed. Further studies are needed to define production systems in which the MGA Select protocol is warranted for use in beef heifers.

Regulation of frequency of luteinizing hormone pulses by magnitude of acute change in circulating concentration of progesterone of female cattle.

The hypothesis was the greater the magnitude of acute increase in circulating concentration of progesterone of female cattle, the greater the acute inhibitory effect on frequency of pulsatile LH release. From Day 0 to 4 of the treatment period, females without functional corpora lutea were treated with varying doses of progesterone to result in varying concentrations of progesterone within the typical physiological range in blood. From Day 4 to 7, cattle were treated with a single dose of progesterone to achieve a similar circulating concentration of progesterone among all females in the study. Therefore, from Day 0 to 4 relative to Day 4 to 7 of the treatment period, females had a: (1) large (3.1 ng/ml), (2) moderate (2.5 ng/ml), or (3) small (0.5 ng/ml) increase in concentration of progesterone in blood. Frequency of LH pulses was greater (P <0.10) in females with the greatest magnitude of change in concentration of progesterone during the first 24 h following the change in concentration as compared with females with the moderate or small of change in concentration of progesterone suggesting our working hypothesis should be rejected. The greater the magnitude of acute change in concentration of progesterone, however, the longer time required for re-initiation of release of LH pulses of the amplitude of pulses that preceded the change in concentration of progesterone.

A comparison of progestin-based protocols to synchronize estrus in postpartum beef cows.

Two progestin-based protocols for estrus synchronization in postpartum beef cows were compared following treatment administration on the basis of estrous response, interval to and synchrony of estrus, and pregnancy. Cows were assigned to one of the two treatment protocols by age, body condition score (BCS), and days postpartum (DPP). The MGA Select-treated cows (MGA Select; n = 109) were fed melengestrol acetate (MGA; 0.5mg x cow-1 x d(-1)) for 14 d, fed carrier for 8 d, GnRH (100 microg of Cystorelin) was injected i.m. 12 d after MGA withdrawal, and PG (25 mg of Lutalyse) was administered i.m. 7 d after GnRH. Cows assigned to the 7-11 Synch protocol (7-11 Synch; n = 111) were fed carrier for 15 d, fed MGA for 7 d, injected with PG on d 22 (d 7 of MGA), injected with GnRH on d 26, and injected with PG on d 33. Mean BCS (4.8 +/- 0.1, MGA Select; 4.7 +/- 0.1, 7-11 Synch) and DPP (40 +/- 1, MGA Select; 40 +/- 1, 7-11 Synch) did not differ between treatments. Blood samples were collected 8 d and 1 d before feeding of MGA or carrier to determine the pretreatment estrous cyclicity (progesterone > or = 1 ng/mL; 10/109 [9%], MGA Select; 12/111 [11%], 7-11 Synch), and again at PG on d 33 to evaluate treatment response (81/109 [74%], MGA Select; 84/111 (76%), 7-11 Synch). Serum concentrations of progesterone at PG on d 33 differed (P < 0.01) between treatments (3.3 +/- 0.3 ng/mL [MGA Select] vs. 1.7 +/- 0.1 ng/mL [7-11 Synch]). HeatWatch was used for 6 d after PG on d 33 to detect estrus, and AI was performed 12 h after the onset of estrus. Estrous response did not differ between treatments (100/109 [92%], MGA Select; 101/111 [91%], 7-11 Synch). Mean interval to estrus (65 +/- 2.7 h, MGA Select; 52 +/- 1.8 h, 7-11 Synch) and synchrony of estrus differed (P < 0.01) between treatments. Synchronized conception and pregnancy rates (61/100 [61%], 61/109 [56%], MGA Select; 71/101 [70%], 71/111 [64%], 7-11 Synch), and final pregnancy rates (94/109 [86%], MGA Select; 99/110 [90%], 7-11 Synch) did not differ between treatments. In summary, estrous response and fertility did not differ among cows assigned to the MGA Select or 7-11 Synch protocols. Synchrony of estrus, defined as the variance in the interval to estrus from PG, however, was improved following treatment with the 7-11 Synch protocol.

Follicular dynamics and steroid profiles in cows during and after treatment with progestin-based protocols for synchronization of estrus.

Two progestin-based protocols for the synchronization of estrus in beef cows were compared. Cyclic, nonlactating, crossbred, beef cows were assigned by age and body condition score to one of two treatments. Cows assigned to the MGA Select protocol were fed melengestrol acetate (MGA; 0.5 mg x cow(-1) x (-1)) for 14 d, GnRH was administered (100 microg i.m. of Cystorelin) 12 d after MGA withdrawal, and PGF2alpha (25 mg of i.m. Lutalyse) was administered 7 d after GnRH. Cows assigned to the 7-11 Synch protocol were fed MGA for 7 d and were injected with PG on d 7 of MGA, GnRH on d 11, and PG on d 18. Transrectal ultrasonography was performed daily to monitor follicular dynamics from the beginning of MGA feeding through ovulation after the synchronized estrus. All cows exhibited estrus in response to PG. Mean interval to estrus was shorter (P < 0.01) for 7-11 Synch-treated cows (56 +/- 1.5 h) than for cows assigned to the MGA Select protocol (73 +/- 4.7 h). Mean interval from estrus to ovulation did not differ between treatments (P > 0.10). Variances for interval to estrus differed (P < 0.01) between treatments. Mean follicular diameter at GnRH injection, PG injection, and estrus did not differ (P > 0.10) between treatments. Relative to MGA Select, serum estradiol-17beta concentrations were higher (P < 0.01) for 7-11 Synch 2 d and 1 d before, on the day of GnRH injection, in addition to 4 d after GnRH, and 24 h after PG. Mean progesterone concentrations were greater (P < 0.01) for MGA Select cows from 4 d before to 7 d after GnRH. Forty-four percent of the variation in interval to estrus between treatments was explained by differences in estradiol-17beta concentrations 24 h after PG. This study suggests that follicular competence is likely related to steroidogenic capacity of the follicle and the endocrine environment under which growth and subsequent ovulation of the dominant follicle occurs.

Fixed-time artificial insemination of postpartum beef cows at 72 or 80 h after treatment with the MGA Select protocol.

The objective was to determine the appropriate timing of fixed-time artificial insemination (AI) following administration of the MGA Select protocol. Cows at two locations (Location 1, n=114; Location 2, n=97 ) were assigned to fixed-time AI at 72 or 80 h by age, body condition score (BCS), days postpartum (DPP), AI technician, and sire. All cows were synchronized with the MGA Select protocol, consisting of oral administration of melengestrol acetate (MGA; 0.5mg/hd per day) for 14 days, GnRH (Cysotrelin, 100 microg, i.m.; Day 26) 12 days after MGA withdrawal, followed in 7 days with PGF(2alpha) (PG; Lutalyse, 25mg i.m.; Day 33). Cows were inseminated at 72 h ( n=108 ) or 80 h ( n=103 ) after PG and GnRH (100 microg) was given at insemination. Location was not significant and, therefore, was removed from the model. Mean BCS ( 5.2+/-0.1, 72 h; 5.3+/-0.1, 80 h) and DPP ( 34+/-2, 72 h; 35+/-2, 80 h) did not differ ( P>0.1 ) between treatments. Serum progesterone concentrations 7 and 1 day prior to MGA were used to determine pre-treatment cyclicity: cows with at least one sample with progesterone > or =1 ng/ml were defined as cyclic (33/108, 31%, 72 h, versus 32/103, 31%, 80 h; P>0.1). Cows with serum progesterone concentrations > or =1 ng/ml on the day of PG were defined as responding to the synchronization protocol (74/108 (69%), 72 h versus 69/103 (67%), 80 h; P>0.1 ). Although pregnancy rates were higher ( P<0.05 ) for cows inseminated at 72 h (69/108, 64%) versus 80 h (52/103, 50%) after PG, pregnancy rates at the end of the breeding season did not differ ( P>0.1 ) between treatments (98/108 (91%), 72 h; 88/103 (85%), 80 h). In conclusion, pregnancy rates were higher when postpartum beef cows synchronized with the MGA Select protocol were inseminated at 72 h versus 80 h after PG.

Frequency of luteinizing hormone pulses in cattle influences duration of persistence of dominant ovarian follicles, follicular fluid concentrations of steroids, and activity of insulin-like growth factor binding proteins.

The objectives of the present study were to determine how varying frequency of LH pulses as controlled by varying treatments with progesterone (P4) in cattle would affect: (1) concentration of steroid hormones and activity of insulin-like growth factor binding proteins (IGFBPs) in the ovarian follicular fluid and blood plasma, and (2) duration of persistence of largest ovarian follicles. There were four treatment groups (n=7 per group) and a control group (n=5) of mature, non-lactating beef cows. Treatments were: (1) two progesterone releasing intravaginal devices (PRIDs) for 16 days (2PRID); (2) a half PRID for 16 days (0.5PRID); (3) two PRIDs for 8 days, then a half PRID for 8 days (2-0.5PRID); or (4) a half PRID for 8 days, then two PRIDs for 8 days (0.5-2PRID). Treatment was initiated on the fifth day of the estrous cycle, which was designated as Day 0, and continued for 16 days. All P4-treated females were administered prostaglandin F2alpha on Day 0 and 1 to regress their corpora lutea. Frequency of LH pulses was greater during treatment with the smaller dose of P4 compared with treatment with the larger dose of P4 and the control group. Ovarian follicles were classified into five categories based on ultrasonographic observations: growing (G); atretic (A); growing dominant (GD); growing persistent (GP); or atretic persistent (AP). At ovariectomy on Day 16, the largest and second largest follicles collected were re-classified into five categories based on follicular concentration of steroids. Classification of the largest follicle collected on Day 16 was influenced by treatment (P<0.005), with the 2PRID group having A follicles, the 2-0.5PRID group GP follicles, the 0.5-2PRID group AP follicles, and the 0.5PRID group GD and GP follicles. Concentrations of 17beta-estradiol (E2) were greatest in GD and GP follicles (P<0.05). There was less (P<0.05) activity of IGFBP-2 in GD follicles and less (P<0.05) activity of IGFBP-3 in GD and GP follicles than other follicles. Activity of IGFBP-4 and -5 was greater (P<0.05) in A and AP follicles than G, GD, and GP follicles. Maintenance of a frequent release of LH pulses over a 16-day period did not result in maintenance of persistent follicles throughout this period indicating that duration of dominance of these follicles is finite even when there is frequent release of LH pulses. Follicular atresia is associated with greater activity of IGFBP-2, -4, -5, and greater concentrations of P4 in follicles, whereas growing dominant and persistent follicles contained greater concentrations of E2, androstenedione (A4), and less IGFBP-2 activity than follicles of other classes. Follicle classifications based on ultrasonography or follicular concentration of steroids did differ (P<0.05) for the largest follicles from the 2PRID group. Two follicles in this group appeared as GD follicles by ultrasonography, but these were atretic based on follicular steroid contents. Objective 1 of the present study yielded the conclusion that concentrations of steroid hormones in follicular fluid and blood plasma could be predictably controlled by regulating the frequency of LH pulses with varying doses of P4. Objective 2 yielded the conclusion that maintain frequent release of LH pulses over a 16-day period could not maintain persistent follicles throughout this period, indicating that duration of dominance of these follicles is finite even when there is frequent release of LH pulses. Follicular atresia in the present study was associated with increased follicular fluid activity of IGFBP-2, -4, -5, and P4, whereas growing dominant and persistent follicles contained greater concentrations of E2, A4, and less IGFBP-2 activity than follicles of other classes.

Synchronisation of oestrus in dairy cows using prostaglandin F2alpha, gonadotrophin-releasing hormone, and oestradiol cypionate.

An oestrous synchronisation protocol was developed for use in lactating dairy cows using PGF(2alpha), GnRH, and oestradiol cypionate (ECP). In experiment 1, lactating dairy cows received two injections of PGF(2alpha) (on days 0 and 11) (PP; n=10) or two injections of PGF(2alpha) (days 0 and 11) and 100 microg of GnRH on day 3 (PGP; n=10). In experiment 2, cows were treated with PGP (n=7), or PGP and 1 mg of ECP at the same time (PGPE(0); n=7) or 1 day after the second PGF(2alpha) injection (PGPE(1); n=7). In experiment 3, 101 lactating dairy cows in a commercial herd were assigned to one of three treatments; PP, PGP, or PGPE(1). Follicular growth was measured by ultrasound in experiments 1 and 2. Every cow (experiments 1, 2, and 3) was blood sampled at selected intervals for progesterone and oestradiol assays and inseminated at oestrus. In experiment 1, a higher percentage of GnRH-treated cows ovulated after the first PGF(2alpha) injection (90% versus 50%; P<0.05). The GnRH-treated cows tended to have a larger dominant follicle present at the time of the second PGF(2alpha) injection (16.5+/-0.5 mm versus 15.0+/-0.7 mm; P<0.10). The percentage of cows that ovulated after the second PGF(2alpha) injection was similar (60%). In experiment 2, cows treated with ECP had higher peak preovulatory concentrations of oestradiol in plasma (6.99+/-0.63 versus 3.63+/-0.63; P<0.01) following the second PGF(2alpha) injection and a higher percentage ovulated (86% versus 43%; P<0.05). A higher percentage of PGPE(1)-treated cows in experiment 3 were observed in standing oestrus and ovulated after the second PGF(2alpha) injection (standing oestrus, 26.4, 34.3, and 62.6%, P<0.01; ovulated, 56, 63, and 78%, P<0.05; PP, PGP, and PGPE(1), respectively). In conclusion, the PGP protocol increased the number of cows that ovulated after the first PGF(2alpha) injection and produced a more mature dominant follicle at the time of the second PGF(2alpha) injection. Adding ECP to PGP (PGPE(1)) enhanced the expression of oestrus and increased ovulation percentage. The combination of PGP and ECP is potentially a new method to routinely synchronise oestrus and ovulation in dairy cows.

Effect of an orally active progestin on follicular dynamics in cycling and anestrous postpartum beef cows.

Although treatment of cycling cows with low concentrations of melengesterol acetate (MGA) results in formation of persistent follicles, in the absence of corpora lutea, it is not known whether persistent follicles form in anestrous cows in response to a similar treatment. The objective of this experiment was to determine the effect of long-term MGA treatment (14 d) on follicular dynamics and the secretion of estradiol in anestrous postpartum beef cows. Treatment groups (replicated over 2 yr) included the following: anestrous control (AC; n = 11), anestrous MGA (AM; n = 16), and cycling MGA (CM; positive control; n = 16). Angus-crossbred cows were assigned to treatment by age, cow body condition, and days postpartum. Cows were fed carrier (AC group) or 0.5 mg MGA x animal(-1) x d(-1) (AM and CM groups) for 14 d beginning approximately 38 d postpartum. Cows allotted to the CM group were injected with PGF2alpha, on the first day of MGA treatment to induce luteolysis. The preceding treatment (CM) results in formation of persistent follicles and secretion of elevated concentrations of estradiol. Ovaries of each cow were examined daily by transrectal ultrasonography beginning 5 to 7 d preceding the initiation of feeding MGA or carrier and continued until ovulation or 7 d following MGA feeding. There was no difference among groups in the stage of follicular wave or diameter of the largest follicle at the start of carrier or MGA feeding. The length of the follicular wave present at the start of MGA feeding was greater (P < 0.01) for cows in the CM (14.5 d, yr 1; 18.3 d, yr 2) group compared to the AM (9.4 d, yr 1; 7.9 d, yr 2) or AC (9.7 d, yr 1; 10.7 d, yr 2) groups. Maximum follicular diameter over both years was greater (P < 0.01) for the CM (20.6 mm) group than the AM (15.1 mm) or AC (16.4 mm) groups. Circulating concentrations of estradiol were also increased (P < 0.05) in the CM group compared to the AM or AC groups. However, MGA appeared to have no effect (P > 0.05) on the number of follicles recruited, growth rate of the dominant follicle during the first 6 d oftreatment, or growth rate to the maximum follicular diameter. In summary, MGA treatment did not increase the duration ot the follicular wave, maximum follicular diameter, or secretion of estradiol in anestrous postpartum cows, nor did MGA affect the number of follicles recruited or growth rate of dominant follicles in cycling or anestrous animals.

Effect of short-term calf removal at three stages of a follicular wave on fate of a dominant follicle in postpartum beef cows.

The hypothesis that ovulation in response to short-term (48 h) calf removal (CR) is dependent on the developmental stage of the dominant follicle was tested in two studies. The objective of Exp. 1 was to characterize the fate of a dominant follicle following 48-h CR on d 2, 4, or 8 of a postpartum follicular wave. Ovaries of 61 beef cows were examined daily by transrectal ultrasonography starting at d 20 to 21 postpartum. Treatments were no CR (n = 14) and CR on d 2 (n = 12), 4 (n = 16), or 8 (n = 10) of first detected follicular wave. Percentage of cows that ovulated a dominant follicle following treatment was not different among groups (P = 0.62). Maximum size of dominant follicles was larger in cows that ovulated (P = 0.002) than in cows that did not ovulate. The objectives of Exp. 2 were 1) to determine whether a follicular wave could be synchronized in anestrous cows following injection of 1 mg of estradiol benzoate (EB) and 200 mg of progesterone (P4; EB + P4); 2) to characterize the fate of dominant follicles following 48-h CR at three stages of a synchronized follicular wave; and 3) to determine whether estrous cycles of normal length followed ovulation in cows pretreated with EB + P4. Ovaries of 50 anestrous beef cows were examined daily as in Exp. 1. Treatments were sesame oil (SO) injected (i.m.) on d 25 postpartum and no CR (n = 9); EB + P4 and no CR (n = 9); EB + P4 and CR on 6 (n = 12), 8 (n = 9), or 12 (n = 11) d after injection. The EB and P4 injections were given on d 25 postpartum. Variability in day of emergence of subsequent follicular waves was lower in cows receiving EB + P4 than in SO-injected cows (P < 0.05). The percentage of cows that ovulated was not different (P = 0.16), but CR increased the percentage of cows that ovulated when groups that received EB + P4 were compared to the EB + P4 group that did not have CR (53.1 vs 11.1%, respectively; P < 0.05). Maximum diameter of dominant follicles was larger (P = 0.05) in ovulatory follicles. The luteal phase was longer (P < 0.03) in cows receiving EB + P4 injection (10.6 +/- 1.2 d) than in cows receiving SO (4.4 +/- 2.2 d). In summary, the maximum size of ovulatory follicles was greater than that of nonovulatory follicles, the ovulatory response of postpartum anestrous cows was maintained through d 8 of a follicular wave, synchronization of follicular waves was accomplished in postpartum cows using EB + P4, and the subsequent luteal phase length was increased in animals that were administered EB + P4.

Interaction of endophyte-infected fescue and heat stress on ovarian function in the beef heifer.

The objective of the experiment was to examine the interaction of endophyte-infected tall fescue and environmental temperature on follicular and luteal development and function in beef heifers. Heifers were fed endophyte-free or endophyte-infected tall fescue seed at thermoneutral or heat stress temperatures (n = 6/treatment) 4 wk before and 3 wk after synchronized ovulation. All heifers were subjected to thermoneutral conditions (19 degrees C, 50% relative humidity) from Days -7 to -2; temperature increased incrementally from Days -1 to 0 and cycled between 25 degrees C and 31 degrees C between Days 1 and 20 for heat-stressed heifers. Serum was collected and ovaries monitored every other day after induced luteolysis between Days 1 and 23 or until ovulation. Size and location of follicles >4 mm and corpora lutea were recorded. Serum concentrations of prolactin were reduced in heat-stressed heifers fed infected seed and both heat stress and infected seed decreased total cholesterol. Rectal temperature and respiration rate were greatest in heifers fed the infected seed when exposed to maximal temperatures. Heat stress led to reduced diameter of the corpus luteum and serum progesterone compared with thermoneutral conditions. Progesterone was reduced more so in heifers fed infected seed. The combination of infected seed and heat stress was associated with reduced diameter of the preovulatory dominant follicle, and consumption of infected seed led to fewer large follicles during the estrous cycle. Both stressors led to reduced serum estradiol. Impaired follicle function may explain reduced pregnancy rates commonly observed in heifers grazing infected tall fescue pasture.

Role of ascorbic acid in promoting follicle integrity and survival in intact mouse ovarian follicles in vitro.

Ascorbic acid has three known functions: it is necessary for collagen synthesis, promotes steroidogenesis and acts as an antioxidant. Within the ovary, most studies have concentrated on the role of ascorbic acid in luteal formation and regression and little is known about the function of this vitamin in follicular growth and development. Follicular growth and development were investigated in this study using an individual follicle culture system that allows the growth of follicles from the late preantral stage to Graafian morphology. Follicles were isolated from prepubertal mice and cultured for 6 days. Control media contained serum and human recombinant FSH. Further groups of follicles were cultured in the same media but with the addition of ascorbic acid at concentrations of either 28 or 280 micromol l(-1). Addition of ascorbic acid at the higher concentration significantly increased the percentage of follicles that maintained basement membrane integrity throughout culture (P < 0.001). Ascorbic acid had no effect on the growth of the follicles or on oestradiol production. Metalloproteinase 2 activity tended to increase at the higher concentration of ascorbic acid and there was a significant concomitant increase in the activity of tissue inhibitor of metalloproteinase 1 (P < 0.01). Follicles cultured without the addition of serum but with FSH and selenium in the culture media underwent apoptosis. Addition of ascorbic acid to follicles cultured under serum-free conditions significantly reduced apoptosis (P < 0.05). From these data it is concluded that ascorbic acid is necessary for remodelling the basement membrane during follicular growth and that the ability of follicles to uptake ascorbic acid confers an advantage in terms of granulosa cell survival.

Development of an estrus synchronization protocol for beef cattle with short-term feeding of melengestrol acetate: 7-11 synch.

An estrus synchronization protocol (7-11 Synch) was developed to synchronize the first follicular wave and timing of ovulation in postpartum beef cows. In Exp. 1, follicular development and timing of ovulation in response to the following protocol were evaluated. Beef heifers (n = 12) and cows (n = 6), at random stages of the estrous cycle, were fed melengestrol acetate (MGA; .5 mg x animal(-1) x d(-1)) for 7 d and injected with PGF2alpha (PG; 25 mg) on the last day of MGA. A second injection of PG was administered 11 d after cessation of MGA. After the second injection of PG, estrus was synchronized in 6/12 heifers and 3/6 cows. The interval to estrus in heifers and cows was 54 and 64 h, respectively (P > .10). All animals exhibiting estrus ovulated first-wave follicles. Animals that failed to respond to the second injection of PG were in estrus later than 6 d after cessation of MGA and had corpora lutea that were unresponsive to the injection of PG. Based on the variation in interval to estrus following the first PG injection on the last day of MGA feeding in Exp. 1, an injection of GnRH (100 microg) was added to the protocol 4 d after the cessation of MGA to ensure ovulation or luteinization of dominant follicles and synchronization of first-wave follicular development. This revised protocol was termed "7-11 Synch." In Exp. 2, two estrus synchronization protocols were compared. Multiparous beef cows were stratified by breed and postpartum interval and randomly assigned to the 7-11 Synch (n = 44) or Select Synch protocols (GnRH injection followed by PG injection 7 d later; n = 45). Timing of estrus after the last PG injection (0 h) ranged from 42 to 102 h in the 7-11 Synch group and -30 to 114 h in the Select Synch group. Eight cows (18%) in the Select Synch group exhibited estrus 30 h before to 18 h after PG. Synchronized estrus peaked between 42 and 66 h after the last PG injection, and a maximum number of cows were in estrus at 54 h for both treatment groups. Synchrony of estrus from 42 to 66 h was greater (P < .05) in 7-11 Synch (91%: 41/44) than in Select Synch cows (69%: 31/45). Artificial insemination pregnancy rate from 42 to 66 h was greater (P < .05) in the 7-11 Synch group (66%: 29/44) than in the Select Synch group (40%: 18/45). In summary, the 7-11 Synch protocol improved synchrony of estrus without reducing fertility. This protocol has potential future application for fixed-time AI in beef cattle production systems.

Corpus luteum development and function in cattle with episodic release of luteinizing hormone pulses inhibited in the follicular and early luteal phases of the estrous cycle.

The influence of episodic LH pulses before and subsequent to ovulation on size and function of the corpus luteum (CL) in cattle was examined. Treatments were 1) control; 2) LHRH antagonist starting 2 days before the preovulatory LH surge (Antagonist [Ant] -2); 3) LHRH antagonist at initiation of the preovulatory LH surge (Ant 0); and 4) LHRH antagonist starting 2 days after the preovulatory LH surge (Ant 2). Treatments with an LHRH antagonist were continued until 7 days after the preovulatory surge. Diameter of the CL and concentrations of progesterone were monitored during the luteal phase that ensued after treatment. Maximum average diameters of CL were 9.5, 17.5, 21.6, and 28.8 mm for females from the Ant -2, Ant 0, Ant 2, and control groups, respectively (P < 0. 01). Compared with those in control animals, concentrations of progesterone in plasma were less (P < 0.01) in animals in which release of LH pulses was inhibited by treatment with antagonist. Arbitrary units under the curve for concentrations of progesterone during the luteal phase of the estrous cycle for Ant -2, Ant 0, Ant 2, and control groups were 19.6, 41.6, 43.6, and 142.2, respectively. There was no difference in circulating concentrations of progesterone (P > 0.1) among antagonist-treated groups. In conclusion, episodic release of LH pulses before, during, and after the time of the preovulatory surge of LH may stimulate development and function of the CL in cattle.

Estrus synchronization of beef cattle with a combination of melengestrol acetate and an injection of progesterone and 17beta-estradiol.

Our hypothesis was that estrus synchronization in beef cattle using melengestrol acetate (MGA) and an injection of progesterone (P4) and 17beta-estradiol (E2) to regress dominant ovarian follicles would improve pregnancy rate (number conceived/number in group) to AI compared with feeding only MGA or injecting PGF2alpha. During 2 yr, peripubertal heifers (n = 52) and cows (n = 327) received either 1) MGA for 18 d (d 0 = 1st d of MGA) plus an injection of P4 and E2 in sesame oil (vehicle) on d 11 to regress persistent ovarian follicles (MGA+P4), 2) MGA for 18 d plus vehicle on d 11 (MGA), or 3) two injections of PGF2alpha 10 d apart (d 7 and 17, PG). Concentration of P4 was assessed in blood samples obtained on d 0, 7, and 17 to indicate estrual status (anestrual or estrual) during treatment to induce estrus synchrony. Observations for detection of estrus occurred every 6 h for 180 h following treatment cessation. Females showing estrus were inseminated 6 to 12 h after estrus detection. Conception to AI was determined by ultrasonography 35 to 40 d later. Conception rate was greater (P < .05) in females in the PG than in those in the MGA group but did not differ from conception rate of females in the MGA+P4 group. Among anestrual females, estrus synchrony rates were greatest (P < .10) among females treated with MGA+P4. Among females that were estrual before treatment cessation, estrus synchrony rates were greater (P < .10) among females treated with MGA+P4 or PG than among those given MGA. Pregnancy rates were greater (P < .05) among females that were anestrual before treatment cessation and treated with MGA or MGA+P4 than among those treated with PG. Estrus synchronization using MGA+P4 and E2 differentially improves estrus synchronization and pregnancy rates among anestrual and estrual beef cattle while maintaining conception rates similar to those of PGF2alpha-treated females.

Regulation of ovarian extracellular matrix remodelling by metalloproteinases and their tissue inhibitors: effects on follicular development, ovulation and luteal function.

In most organs, remodelling of tissues after morphogenesis is minimal; however, normal ovarian function depends upon cyclical remodelling of the extracellular matrix (ECM). The ECM has a profound effect on cellular functions and probably plays an important role in the processes of follicular development and atresia, ovulation, and development, maintenance and regression of corpora lutea. Matrix metalloproteinases (MMPs; collagenases, gelatinases, stromelysins and membrane-type MMPs) cleave specific components of the ECM and are inhibited by tissue inhibitors of metalloproteinases (TIMPs). MMPs have been detected at all stages of follicular development and probably modulate follicular expansion or atresia within the ovarian stroma. In addition, increased MMP activity appears to be required for ovulation since follicular rupture occurred in the absence of plasminogen activator activity and inhibitors of MMPs blocked follicular rupture. Development and luteolysis of the corpus luteum are accompanied by extensive remodelling of the ECM. Differentiation and regression of luteal cells are associated with construction and degradation of ECM, respectively. There is increasing evidence that ECM components enhance luteinization; whereas loss of ECM results in luteal cell death. Ovine large luteal cells may be the primary type of cell responsible for controlling the extent of remodelling of luteal ECM since they produce TIMP-1, TIMP-2 and plasminogen activator inhibitor 1. The ratio of active MMPs to TIMPs may be important in maintaining an ECM microenvironment conducive to the differentiation of follicular-derived cells into luteal cells, and maintenance of the phenotype of luteal cells.

Concentrations of gonadotropins, estradiol and progesterone in sows selected on an index of ovulation rate and embryo survival.

The objective of this study was to determine concentrations of follicle stimulating hormone (FSH), luteinizing hormone (LH), progesterone (P4) and 17beta-estradiol (E2) in sows from a line selected on an index which emphasized ovulation rate (Select) and from a control line. A further classification of the sows in each line was made according to the estimated number of ovulations during an estrous cycle. Sows in the Select line were ranked into a high (HI) or low group (LI) when their estimated number of ovulations were 25 or more and 14 to 15, respectively. Sows of the control line were classified into groups as high (HC) or low (LC) when the estimated values for ovulation rate were 14-15 and 8-9 ovulations, respectively. Blood samples were collected every 12 h during a complete estrous cycle and samples were analyzed for concentrations of FSH and LH. Samples collected every 24 h were assayed for P4 and E2. Mean concentrations of FSH, LH, P4 and E2 did not differ (P > 0.10) between lines or between HI and LI or HC and LC groups. Selection of pigs for ovulation rate and embryonal survival did not affect concentrations of FSH, LH, P4 and E2 in sows during the estrous cycle.

Prolonged increased concentrations of 17beta-estradiol associated with development of persistent ovarian follicles do not influence conception rates in beef cattle.

Objectives were to evaluate conception rates and time to estrus following cessation of treatments designed to either cause prolonged elevated concentrations of 17beta-estradiol associated with development of persistent ovarian follicles or to inhibit elevated concentrations of 17beta-estradiol and development of persistent ovarian follicles. Beef heifers (n = 80) and 2-yr-old nonlactating cows (n = 39) were stratified by age, blocked by estrual status (previously exhibited estrus or anestrus) and assigned to receive either 1) four norgestomet implants (4 Norg; n = 59) for 9 d (d 0 = treatment initiation) or 2) one norgestomet implant from d 0 to 7 and three additional norgestomet implants from d 7 to 9 (1 + 3 Norg; n = 60). All animals received PGF2alpha on d 0 to lyse corpora lutea. All implants were removed on d 9 followed by estrus detection every 6 h for 7 d following implant removal. Females exhibiting estrus were artificially inseminated 6 to 12 h after detection of estrus. A treatment x day interaction (P < .01) for concentrations of 17beta-estradiol from d 0 to 9 of the experiment with elevated 17beta-estradiol occurring in females treated with 1 + 3 Norg implants. The interval from treatment withdrawal to estrus was longer (P < .01) in females treated with 1 + 3 Norg (105 h) than in those treated with 4 Norg (61 h). Synchrony of estrus among anestrous females was greater (P < .10) in females treated with 4 Norg (97%) than in females treated with 1 + 3 Norg (67%) but was similar in estrual females. Conception rates (number conceiving to AI/number bred by AI) did not differ (4 Norg = 67%; 1 + 3 Norg = 72%; P > .10). Pregnancy rates (number conceiving to AI/number in treatment group) also did not differ between treatment groups of either estrual or anestrous females. Conception rates are not compromised in females that develop persistent ovarian follicles and have prolonged elevated concentrations of 17beta-estradiol when persistent ovarian follicles are not allowed to ovulate.

Development of a persistent ovarian follicle and associated elevated concentrations of 17beta-estradiol preceding ovulation does not alter the pregnancy rate after embryo transfer in cattle.

It was hypothesized that prolonged elevation in 17beta-estradiol (E(2)) preceding ovulation as a result of a persistent ovarian follicle would have a detrimental effect on pregnancy rate after Day 7 (behavioral estrus = Day 0) of the estrous cycle. Cows were either treated with exogenous progesterone (P(4)) for 10 d or remained untreated (CON; n = 76). Cows were treated with 1 of 2 doses of P(4) from Day 6 to 16 which was intended to result in either elevated E(2) (EE(2); n = 76) or normal E(2) (NE(2); n = 76) concentration in the circulation. At the initiation of P(4) treatment, cows received prostaglandin F(2alpha) (PGF(2alpha)) to eliminate the endogenous source of P(4). On Day 16, the exogenous source of P(4) was removed from treated cows, while cows in the CON group received PGF(2alpha). A single embryo was transferred into each cow 7 days after observation of behavioral estrus. Blood samples were taken on alternating days during the treatment period to determine concentrations of P(4) and E(2). The pregnancy rate was determined by ultrasonographic examination 25 to 32 d after embryo transfer. There was a treatment-by-day interaction (P < 0.0001) on E(2) concentrations in the plasma during the 10-d treatment period. Cows in the EE(2) group had a higher concentration of E(2) by Day 8 (6.1 +/- 0.5 pg/ml) and this concentration remained elevated until PRID removal compared with that of cows in the NE(2) (2 +/- 0.2 pg/ml) and CON (2.0 +/- 0.3 pg/ml) groups, which had concentrations of E(2) similar to those at the initiation of treatment. Pregnancy rates after embryo transfer did not differ (P = 0.56; X(2) = 1.1) among cows in the EE(2) (30.7%), NE(2) (36.2%) and CON (32.9%) groups. Prolonged elevation of E(2) concentrations associated with the development of a persistent ovarian follicle preceding ovulation did not affect the pregnancy rate to embryo transfer after Day 7 of the estrous cycle in cows.