Changes in ovarian function in mature beef cows grazing endophyte infected tall fescue

The objective was to examine follicular and luteal development and function in mature, lactating beef cows grazing endophyte free (E-) or endophyte infected (E+) tall fescue during the early postpartum period. Angus, Hereford, and Angus x Hereford cows were exposed to pasture for 37-39 days before synchronized estrus. Serum concentrations of prolactin were evaluated during the luteal phase before the synchronized estrus. Every Monday, Wednesday, and Friday for one estrous cycle ovaries were monitored by transrectal ultrasonography and blood was collected for determination of serum concentrations of progesterone and estradiol in cows that responded to synchronization. Signs of fescue toxicosis in E+ cows included decreased serum concentrations of prolactin (84.9+/-13.6 pg/ml versus 32.3+/-12.0 pg/ml; P < 0.009) measured during the luteal phase (day 37 of grazing) and decreased body condition of cows and weight of cows and calves (P < 0.001). Neither serum concentrations of progesterone or estradiol, nor diameter of the CL differed between treatments. Diameter of the largest follicle tended to be smaller for cows grazing E+ fescue, especially between days 8 and 12 of the estrous cycle (P < 0.08). Numbers of class 1 (3-5 mm) and class 3 (>10 mm) follicles were similar (P > 0.05) between treatments, but number of class 2 (6-9 mm) follicles was reduced in E+ cows for most of the cycle (days 10 through 20; P < 0.03). Length of synchronized estrous cycle, days open, calving interval, and pregnancy rate at 30, 45, 60, and 90 days post-breeding was similar (P > 0.05) among treatment groups. Even though follicular dynamics (diameter of the largest follicle and number of class 2 follicles) were altered in cows grazing E+ tall fescue, follicular function was apparently not affected by ergot alkaloids.     

Prostaglandin F2 alpha-induced release of oxytocin from bovine corpora lutea in vitro

Two experiments were conducted to study the in vitro effects of prostaglandins F2 alpha (PGF2 alpha), E2 (PGE2), and luteinizing hormone (LH) on oxytocin (OT) release from bovine luteal tissue. Luteal concentration of OT at different stages of the estrous cycle was also determined. In Experiment 1, sixteen beef heifers were assigned randomly in equal numbers (N = 4) to be killed on Days 4, 8, 12, and 16 of the estrous cycle (Day 0 = day of estrus). Corpora lutea were collected, an aliquot of each was removed for determination of initial OT concentration, and the remainder was sliced and incubated with vehicle (control) or with PGF2 alpha (10 ng/ml), PGE2 (10 ng/ml), or LH (5 ng/ml). Luteal tissue from heifers on Day 4 was sufficient only for determination of initial OT levels. Luteal OT concentrations (ng/g) increased from 414 +/- 84 on Day 4 to 2019 +/- 330 on Day 8 and then declined to 589 +/- 101 on Day 12 and 81 +/- 5 on Day 16. Prostaglandin F2 alpha induced a significant in vitro release of luteal OT (ng.g-1.2h-1) on Day 8 (2257 +/- 167 vs. control 1702 +/- 126) but not on Days 12 or 16 of the cycle. Prostaglandin E2 and LH did not affect OT release at any stage of the cycle studied. In Experiment 2, six heifers were used to investigate the in vitro dose-response relationship of 10, 20, and 40 ng PGF2 alpha/ml of medium on OT release from Day 8 luteal tissue.(ABSTRACT TRUNCATED AT 250 WORDS)     

The synchrony of prostaglandin-induced estrus in cows was reduced by pretreatment with hCG

The induction of optimal synchrony of estrus in cows requires synchronization of luteolysis and of the waves of follicular growth (follicular waves). The aim of this study was to determine whether hormonal treatments aimed at synchronizing follicular waves improved the synchrony of prostaglandin (PG)-induced estrus. In Experiment 1, cows were treated on Day 5 of the estrous cycle with saline in Group 1 (n = 25; 16 ml, i.v., 12 h apart), with hCG in Group 2 (n = 27; 3000 IU, i.v.), or with hCG and bovine follicular fluid (bFF) in Group 3 (n = 21; 16 ml, i.v., 12 h apart). On Day 12, all cows were treated with prostaglandin (PG; 500 micrograms cloprostenol, i.m.). In Experiment 2, cows were treated on Day 5 of the estrous cycle with saline (3 ml, i.m.) in Group 1 (n = 22) or with hCG (3000 IU, i.v.) in Group 2 (n = 20) and Group 3 (n = 22). On Day 12, the cows were treated with PG (500 micrograms in Groups 1 and 2; 1000 micrograms in Group 3). Blood samples for progesterone (P4) determination were collected on Day 12 (Experiment 1) or on Days 12 and 14 (Experiment 2). Cows were fitted with heat mount detectors and observed twice a day for signs of estrus. Four cows in Experiment 1 (1 cow each from Groups 1 and 2; 2 cows from Group 3) had plasma P4 concentrations below 1 ng/ml on Day 12 and were excluded from the analyses. In Experiment 1, cows treated with hCG or hCG + bFF had a more variable (P = 0.0007, P = 0.0005) day of occurrence of and a longer interval to estrus (5.9 +/- 0.7 d, P = 0.003 and 6.2 +/- 0.8 d, P = 0.005) than saline-treated cows (3.4 +/- 0.4 d). The plasma P4 concentrations on Day 12 were higher (P < 0.0001) in hCG- and in hCG + bFF-treated cows than in saline-treated cows (9.4 +/- 0.75 and 8.5 +/- 0.75 vs 4.1 +/- 0.27 ng/ml), but there was no correlation (P > 0.05) between plasma P4 concentrations and the interval to estrus. In Experiment 2, cows treated with hCG/500PG and hCG/1000PG had a more variable (P = 0.0007, P = 0.002) day of occurrence of and a longer interval to estrus (4.2 +/- 0.4 d, P = 0.04; 4.1 +/- 0.4 d, P = 0.03) than saline/500PG-treated cows (3.2 +/- 0.1 d). The concentrations of plasma P4 on Days 12 and 14 of both hCG/500PG- and hCG/1000PG-treated cows were higher (P < 0.05) than in saline/500PG-treated cows (7.3 +/- 0.64, 0.7 +/- 0.08 and 7.7 +/- 0.49, 0.7 +/- 0.06 vs 5.3 +/- 0.37, 0.5 +/- 0.03 ng/ml). The concentrations of plasma P4 on Days 12 or 14 and the interval to estrus were not correlated (P > 0.05) in any treatment group. The concentrations of plasma P4 on Days 12 and 14 of hCG/500PG- or hCG/1000PG-treated cows were correlated (r = 0.65, P < 0.05; r = 0.50, P < 0.05). This study indicated that treatment of cows with hCG on Day 5 of the estrous cycle reduced the synchrony of PG-induced estrus and that this reduction was not due to the failure of luteal regression.     

Effect of estrus synchronization with Syncro-Mate-B((R)) on serum luteinizing hormone, progesterone and conception rate in Brahman heifers

Twenty-two estrous cyclic, 2-yr-old Brahman heifers were randomly assigned to receive either estrus synchronization with Syncro-Mate-B((R)) (SMB; 11) or no treatment (Control; 11). Blood samples were collected via tail vessel puncture at onset of estrus and daily thereafter until Day 11 after estrus. Blood samples were also collected from five SMB and five Control heifers at 0, 4, 8 and 12 h after the onset of estrus. All samples were processed to yield serum and stored at -20 degrees C until radioimmunoassay. Heifers were inseminated by one technician using semen from a single ejaculate of a Brahman bull 12 h after the onset of estrus. All SMB heifers exhibited estrus within 72 h of implant removal. All heifers had corpora lutea (CL) detected by rectal examination 8 to 12 d following estrus. Serum luteinizing hormone (LH) was not affected by treatment, time (4 - h intervals) or an interaction of treatment by time (P > 0.10). Independent analysis with h indicated that at h 12, SMB (2.2 +/- 0.06 ng/ml) had lower LH than did control heifers (8.9 +/- 2.1 ng/ml). Serum progesterone increased from Day 1 through Day 12 in all heifers, which is indicative of functional CL. Serum progesterone was affected by treatment (P < 0.0001) and time (d intervals; P < 0.10). Progesterone elevation was lower (P < 0.05) and area under the progesterone curve was lower (P < 0.03) in SMB (5.6 +/- 0.5 ng/ml, 32.0 +/- 4.5 units, respectively) when compared with control heifers (7.0 +/- 4 ng/ml, 43.7 +/- 2.4 units, respectively). Conception rate was lower (P < 0.01) in SMB heifers (2 of 11) than in control heifers (8 of 11). The lowered conception rate in SMB treated Brahman heifers may be due to altered timing of LH release following estrus, resulting in an altered time of ovulation.     

Relationships between insulin and glucose metabolism and pituitary-ovarian functions in fasted heifers

The effects of fasting between Days 8 and 16 of the estrous cycle on plasma concentrations of luteinizing hormone (LH), progesterone, cortisol, glucose and insulin were determined in 4 fasted and 4 control heifers during an estrous cycle of fasting and in the subsequent cycle after fasting. Cortisol levels were unaffected by fasting. Concentrations of insulin and glucose, however, were decreased (p less than 0.05) by 12 and 36 h, respectively, after fasting was begun and did not return to control values until 12 h (insulin) and 4 to 7 days (glucose) after fasting ended. Concentrations of progesterone were greater (p less than 0.05) in fasted than in control heifers from Day 10 to 15 of the estrous cycle during fasting, while LH levels were lower (p less than 0.01) in fasted than in control heifers during the last 24 h of fasting. Concentrations of LH increased (p less than 0.01) abruptly in fasted heifers in the first 4 h after they were refed on Day 16 of the fasted cycle. Concentrations (means +/- SEM) of LH also were greater (p less than 0.05) in fasted (11.2 +/- 2.6 ng/ml) than in control (4.7 +/- 1.2 ng/ml) heifers during estrus of the cycle after fasting; this elevated LH was preceded by a rebound response in insulin levels in the fasted-refed heifers, with insulin increasing from 176 +/- 35 pg/ml to 1302 +/- 280 pg/ml between refeeding and estrus of the cycle after fasting. Concentrations of LH, glucose and insulin were similar in both groups after Day 2 of the postfasting cycle. Concentrations of progesterone in two fasted heifers and controls were similar during the cycle after fasting, whereas concentrations in the other fasted heifers were less than 1 ng/ml until Day 10, indicating delayed ovulation and (or) reduced luteal function. Thus, aberrant pituitary and luteal functions in fasted heifers were associated with concurrent fasting-induced changes in insulin and glucose metabolism.     

Peripheral inhibin levels in relation to climatic variations and stage of estrous cycle in buffalo (Bubalus bubalis )

The present study investigated the peripheral plasma inhibin levels in relation to 1) the stage of estrous cycle and the effect of climatic variations. Blood samples were collected from cyclic buffalo (n=5) once daily for 32 consecutive days during the tropical hot humid (summer) and cold (winter) seasons. Estrus was recorded by parading a vasectomized bull as well as by plasma progesterone determination. In the winter season, peripheral inhibin concentrations which were lowest (0.35 +/- 0.02 ng/ml) during the mid-luteal phase of estrous cycle (Day 6 to Day 14, Day 0 = day of estrus) increased significantly (P < 0.02) to 0.47 +/- 0.04 ng/ml during the late luteal phase (Day -4 to Day -2) and then further to 0.52 +/- 0.03 ng/ml (P< 0.02) during the periestrus phase (Day -1 to Day 1). Inhibin concentrations then decreased significantly (P < 0.02) to 0.40 +/- 0.03 ng/ml during the early luteal phase (Day 2 to Day 5). In the summer season the differences in peripheral inhibin concentrations among different phases of estrous cycle were found to be nonsignificant. A comparison of the circulating inhibin concentrations between the two seasons indicated that inhibin concentrations were significantly higher in the late luteal phase (P < 0.01) and periestrus phase (P < 0.05) during the winter season compared with corresponding periods during the summer season. The present study suggests that peripheral inhibin concentrations change in the estrous cycle during cooler breeding season and that environmental heat stress can cause a reduction in peripheral inhibin concentrations.     

Effect of estrous cycle stage at Syncro-Mate B treatment on conception and time to estrus in cattle

Two trials involving 85 heifers and 67 cows were conducted to determine the effect of estrous cycle stage at the time of Syncro-Mate-B((R)) (SMB) treatment on interval to estrus following implant removal and on conception rate at the synchronized estrus. In Trial 1, 57 beef and 28 dairy heifers were treated with SMB on each representative day of a 22-d estrous cycle (estrus = Day 0). Beef heifers were artificially inseminated approximately 48 h after implant removal, whereas dairy heifers were inseminated 0 to 12 h after detection of estrus. Inseminations were scored by the inseminator according to their difficulty. Interval to the onset of estrus was not different between heifers treated early ( Day 11) in the cycle (35.2 +/- 7.2 h). Conception rate at the synchronized estrus was slightly higher in early-cycle heifers (22 47 = 47% ) compared to late-cycle heifers (14 38 = 37% , P = 0.2). Heifers that were difficult to inseminate had lower (P < 0.01) conception rates (2 11 = 18% ) at the synchronized estrus than heifers considered normal (21 51 = 41% ) or easier than normal to inseminate (13 23 = 57% ). In Trial 2, of the 131 beef cows synchronized, 67 that were estimated to be either early or late in the estrous cycle by progesterone analysis were utilized. Cows were treated with SMB and inseminated without regard to estrus 48-h after implant removal. Inseminations were scored as in Trial 1. Calves were separated from cows from the time of implant removal to insemination. Conception rate was higher (P < 0.05) in cows treated with SMB early ( Day 11, 16 35 = 46% ). Cows that were difficult to inseminate had a lower (P < 0.01) conception rate (0 8 = 0% ) than cows that were normal (43 94 = 46% ) or easier than normal to inseminate (13 29 = 45% ).     

Plasma progesterone levels during the estrous cycle of Holstein and Brahman cows, Carora type and cross-bred heifers

Daily plasma progesterone (P(4)) was determined during one estrous cycle of 19 cows and 18 heifers of four different breeds: Holstein (H), Brahman (B), Carora-type (C) and crossbred (CB) females. Estrus detection was made by visual observation and using a teaser bull with a chin-ball marker. The P(4) profiles showed no differences among groups. In Group 1 (H), P(4) levels ranged from 0.5 ng/ml plasma on the day of estrus (Day 0) to 5.1 ng/ml at the luteal phase peak (Day 13). In Group 2 (B), P(4) levels ranged from 0.5 ng/ml on Day 0 to 9.2 ng/ml on Day 13. In Groups 3 (C) and 4 (CB), P(4) levels ranged from 0.5 ng/ml, on Day 0, to 13.7 ng/ml on Day 12 and 8.8 ng/ml on Day 13. These last two groups were moved to the same location and then compared. It was found that P(4) concentrations were significantly higher (P < 0.025) in Group 3 between Days 7 and 14 of the estrous cycle. In all groups, P(4) levels were lower than 1 ng/ml one day before the next estrus, and levels of 0.4, 0.5, 0.4 and 0.4 ng/ml were obtained the day of estrus in Groups 1 to 4, respectively. Results indicated that the pattern of P(4) for each one of the groups was similar to those reported by other investigators.     

Concentrations of 13, 14-dihydro-15-keto-prostaglandin F(2)alpha, estradiol-17beta and progesterone during the peripubertal period in heifers

Twenty prepubertal Holstein heifers were utilized to assess plasma 13, 14-dihydro-15-keto-prostaglandin F(2)alpha (PGFM), serum progesterone (P(4)) and estradiol-17beta (E(2)) concentrations as well as the E(2):P(4) ratio during the onset of puberty in cattle. All animals were maintained as a group along with a sterile marker bull to assist in the detection of estrus. Upon detection of the first estrus (Day=O), daily blood samples were collected from a jugular vein until the heifers had completed 3 estrous cycles. The average body weight and age at first estrus were 247.6+/-4.8 kg and 304.0+/-7.5 days, respectively. Frequency of abnormal length estrous cycles was greater (P<0.02) during the first (40%) and second (35%) cycles than during the third estrous cycle (0%). All heifers had normal cycle lengths (18 to 24 days) by the third estrous cycle. Serum P(4) was greater during the third cycle (P<0.05) from Day 10 to Day 4 before the next estrus compared with the same period of the first estrous cycle. Serum E(2) did not peak until the day of estrus in the first cycle, whereas E(2) reached a maximal level 2 days before estrus in the third estrous cycle. Serum E(2) was higher (P<0.0001) 2 days before estrus in the third cycle than in the first estrous cycle. Plasma PGFM reached maximum concentrations 3 days before estrus in the third cycle compared with 1 day before estrus at the end of first estrous cycle. As estrus approached during the third cycle, PGFM rose 1 day before E(2) rose and P(4) declined, while the rise in PGFM and E(2) occurred simultaneously, with P(4) declining at the end of the first estrous cycle. During diestrus, the E(2):P(4) ratio was lower (P<0.07) in the third cycle than in the first, but it was higher (P<0.04) at estrus and 1 day before in the third estrous cycle. These data reveal a high incidence of abnormal length estrous cycles during the first two estrous cycles of the peripubertal period, and demonstrate anomalies in uterine and ovarian endocrine activity during the peripubertal period in cattle.     

Influence of stage of the estrous cycle on endogenous opioid modulation of luteinizing hormone, prolactin, and cortisol secretion in the gilt

Fourteen gilts that had displayed one or more estrous cycles of 18-22 days (onset of estrus = Day 0) and four ovariectomized (OVX) gilts were treated with naloxone (NAL), an opiate antagonist, at 1 mg/kg body weight in saline i.v. Intact gilts were treated during either the luteal phase (L, Day 10-11; n = 7), early follicular phase (EF, Day 15-17; n = 3), or late follicular phase (LF, Day 18-19; n = 4) of the estrous cycle. Blood was collected at 15-min intervals for 2 h before and 4 h after NAL treatment. Serum luteinizing hormone (LH) concentrations for L gilts averaged 0.65 +/- 0.04 ng/ml during the pretreatment period and increased to an average of 1.3 +/- 0.1 ng/ml (p less than 0.05) during the first 60 min after NAL treatment. Serum prolactin (PRL) concentrations for L gilts averaged 4.8 +/- 0.2 ng/ml during the pretreatment period and increased to an average of 6.3 +/- 0.3 ng/ml (p less than 0.05) during the first 60 min after NAL treatment. Serum PRL concentrations averaged 8.6 +/- 0.7 ng/ml and 7.6 +/- 0.6 ng/ml in EF and LF gilts, respectively, prior to NAL treatment, and decreased (p less than 0.05) to an average of 4.1 +/- 0.2 ng/ml and 5.6 +/- 0.4 ng/ml in EF and LF gilts, respectively, during the fourth h after NAL. Naloxone treatment failed to alter serum LH concentrations in EF, LF, or OVX gilts and PRL concentrations in OVX gilts.(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

Inhibition of estrus and corpora lutea function with Norgestomet

Forty-five nonpregnant, nonlactating, Angus and Brangus cows were utilized to determine how long a Norgestomet ear implant would inhibit estrus when administered at various stages of an estrous cycle. All cows completed a nontreated estrous cycle to ensure normal cyclicity. At the second observed estrus (estrus = Day 1), cows were randomly allotted to be treated at metestrus (Day 3 or Day 4, n = 15); at diestrus (Day 9 or Day 10, n = 14); or at proestrus (Day 15 or Day 16, n = 16). All cows received a 2-ml intramuscular injection of 3 mg of Norgestomet accompanied by a 6-mg Norgestomet ear implant, which remained in situ for 21 days, or until individual cows were observed in estrus. Estrus was inhibited for a mean (+/- SEM) of 18.7 +/- 0.7, 19.9 +/- 0.8, and 17.0 +/- 0.8 days, respectively, when cows were treated at metestrus, diestrus, and proestrus (metestrus and diestrus vs proestrus; P < 0.05). Estrus was inhibited for an entire 21-day implantation period in 27, 50, and 38% of cows treated at metestrus, diestrus, and proestrus, respectively (P > 0.10). Norgestomet inhibited estrus in all cows for 11, 17, and 11 days after implantation when treatment was initiated at metestrus, diestrus, and proestrus, respectively (P > 0.10). These data indicate that a 6-mg Norgestomet ear implant effectively inhibits estrus in all cows for a maximum of 11 days, with some cows exhibiting estrus by Day 12 with the Norgestomet implant in situ.     

Induction of a new follicular wave in holstein heifers synchronized with norgestomet

Treatments with progestin to synchronize the bovine estrous cycle in the absence of the corpus luteum, induces persistence of a dominant follicle and a reduction of fertility at doses commonly utilized. The objective of the present research was to induce a new wave of ovarian follicular development in heifers in which stage of the estrous cycle was synchronized with norgestomet. Holstein heifers (n=30) were used, in which estrus was synchronized using two doses of PGF2alpha i.m. (25 mg each) 11 days apart. Six days after estrus (day 0=day of estrus) heifers received a norgestomet implant (6 mg of norgestomet). On day 12, heifers were injected with 25 mg of PGF2alpha i.m. and assigned to treatments (T1 to T4) as follows: treatment 1, heifers received a second norgestomet implant (T1: N+N, n=6), treatment 2, received 100 microg of GnRH i.m. (T2: N+GnRH, n=6), treatment 3, 200 mg of progesterone i.m. (T3: N+P4, n=6), treatment 4, control treatment with saline solution i.m. (T4: N+SS); in the four treatments (T1 to T4) implants were removed on day 14. For treatment 5, heifers received 100 microg of GnRH i.m. on day 9 and 25 mg of PGF2alpha i.m. (T5: N+GnRH+PGF2alpha) at the time of implant removal (day 16). Ovarian evaluations using ultrasonographic techniques were performed every 48 h from days 3 to 11 and every 24 h from days 11 to 21. Blood samples were collected every 48 h to analyze for progesterone concentration. A new wave of ovarian follicular development was induced in 3/6, 6/6, 3/6, 1/6 and 6/6, and onset of estrus in 6/6, 0/6, 6/6, 6/6 and 6/6 for T1, T2, T3, T4 and T5, respectively. Heifers from T1, T3 and T4 that ovulated from a persistent follicle, showed estrus 37.5 +/- 12.10 h after implant removal and heifers that developed a new wave of ovarian follicular development showed it at 120.28 +/- 22.81 h (P<0.01). Ovulation occurred at 5.92 +/- 1.72 and 2.22 +/- 1.00 days (P<0.01), respectively. Progesterone concentration was <1 ng/ml from days 7 to 15 in T1, T2 and T4; for T3 progesterone concentration was 2.25 +/- 0.50 ng/ml on day 13 and decreased on day 15 to 0.34 +/- 0.12 ng/ml (P<0.01). For T5, progesterone concentration was 1.66 +/- 0.58 ng/ml on day 15. The more desirable results were obtained with T5, in which 100% of heifers had a new wave of ovarian follicular development induced, with onset of estrus and ovulation synchronized in a short time period.     

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.     

Radioimmunoassay of a bovine pregnancy-associated glycoprotein in serum: its application for pregnancy diagnosis.

A sensitive and specific double-antibody RIA for a bovine pregnancy-associated glycoprotein (bPAG) is described. The limit of detection was 0.2 ng/ml. The assay was specific for bPAG in that pituitary and placental gonadotropic hormones and other placental or serum proteins assayed in serial dilutions did not cross-react. The RIA allowed measurement of bPAG in placental extracts, fetal serum, fetal fluids, and serum or plasma of pregnant cows. About 20% of unbred heifers and nonpregnant cows had detectable levels ranging from 0.30 +/- 0.09 to 0.50 +/- 0.17 ng/ml (mean +/- SD), and 15% of bull sera showed higher concentrations (3.01 +/- 1.73 ng/ml) of bPAG or bPAG-like protein. Variations among animals was observed in fetal serum bPAG concentrations. Bovine PAG was detected in maternal peripheral blood at Day 22 of pregnancy (mean +/- SD, 0.38 +/- 0.13 ng/ml) in some animals and at Day 30 in all pregnant cows. Peripheral serum bPAG levels increased progressively to 3.60 +/- 1.73 ng/ml (mean +/- SD) at Day 30 of pregnancy, to 24.53 +/- 8.81 ng/ml at Day 120, and to 1551.91 +/- 589.68 ng/ml at Day 270. Peak concentration of bPAG was 2462.42 +/- 1017.88 ng/ml and it occurred 1-5 days prior to parturition. After delivery, bPAG concentrations decreased steadily to 499.63 +/- 267.20 ng/ml at Day 14 postpartum (pp), 10.12 +/- 7.84 ng/ml at Day 60 pp, and 1.44 +/- 1.08 ng/ml at Day 90 pp. The undetectable concentration (less than 0.20 ng/ml) was reached by Day 100 +/- 20 pp. An investigation undertaken in Holstein heifers, Holstein cows, and Hereford cows used as recipients for purebred Holstein embryos supplied evidence of the influence of breed of recipient and sex of fetuses on peripheral concentrations of bPAG. A herd of 430 Holstein-Friesian heifers that had received transferred embryos were bled at Day 35 postestrus (pe) for measurement of bPAG. The bPAG was detected in 287 of 430 serum samples analyzed. By rectal palpation performed at Day 45 pe, 267 heifers with detectable levels of bPAG at Day 35 pe were confirmed to be pregnant as were 3 of 143 heifers previously diagnosed as not pregnant by RIA. These results suggest that detection of this placental-specific antigen in the serum could be used as a specific serological method for early pregnancy diagnosis in cattle from 28 days after breeding.     

Interrelationships of hormonal and ovarian responses in superovulated heifers pretreated with FSH-P at the beginning of the estrous cycle

The objective of this study was to determine the relationships between follicle stimulating hormone, (FSH), estradiol (E(2)), and progesterone (P(4)) concentrations in peripheral blood samples and the follicular dynamics prior to and during superovulation in heifers pretreated with FSH-P (10 mg, i.m.) (FSH-P-primed; n=9) or not (saline-primed; n=9) on Day 3 (Day 0 = estrus) of the estrous cycle. On Day 10, all heifers were superovulated with FSH-P (27.7 mg i.m.) in declining dosages over 5 days. Prior to and during superovulation, blood samples were collected one to five times daily, and the follicular dynamics were monitored daily by ultrasonography. Prior to superovulation, profiles of P(4) and E(2) did not differ (P>1) between the saline- and FSH-P-primed heifers. The FSH concentrations in saline-primed heifers decreased from 0.43 +/- 0.05 ng/ml to 0.30 +/- 0.04 ng/ml between Days 3 and 7 and then increased progressively to 0.59 +/- 0.04 ng/ml on Day 10. In contrast (P<0.002), FSH concentrations in the FSH-P-primed heifers remained constant between Days 3 and 10 and averaged 0.41 +/- 0.03 ng/ml. Higher increases in E(2) during superovulation (maximum values, 100 vs 46 pg/ml) and in P(4) after superovulation (maximum values, 39 vs 22 ng/ml) in the saline-than in the FSH-P-primed heifers reflected the greater increase in the number of follicles (>10 mm) and in the number of corpora lutea (CL) in the saline-primed heifers. Prior to the preovulatory luteinizing hormone (LH) peak during superovulation, there was a parallel (P>0.1) decrease in FSH concentrations in the saline- and FSH-P-primed groups. Within heifers partial correlations indicated that E(2) was correlated positively with the number of follicles (>/= 7 mm) and the size of the largest follicle during superovulation (r=0.54 to 0.81; P<0.01). Negative correlations were detected (P<0.01) between FSH and the number of follicles >/=7 mm prior to (r=-0.26) and during superovulation (r=-0.37). The results cofirm earlier reports indicating that priming with FSH-P decreases the superovulatory response in cattle. Interrelationships of hormonal and ovarian responses support the concept that the presence of large dominant follicles prior to superovulation limits the superovulatory response.     

Follicular dynamics and concentrations of steroids and gonadotropins in lactating cows and nulliparous heifers

Differences in follicular development and circulating hormone concentrations, between lactating cows and nulliparous heifers, that may relate to differences in fertility between the groups, were examined. Multiparous, cyclic, lactating Holstein cows (n=19) and cyclic heifers (n=20) were examined in the winter, during one estrous cycle. The examinations included ultrasound monitoring and daily blood sampling. Distributions of two-wave and three-wave cycles were similar in the two groups: 79 and 21% in cows, 70 and 30% in heifers, respectively. Cycle lengths were shorter by 2.6 days in heifers than in cows, and in two-wave than in three-wave cycles. The ovulatory follicle was smaller in heifers than in cows (13.0+/-0.3 mm versus 16.5+/-0.05 mm). The greater numbers of large follicles in cows than in heifers corresponded well to the higher concentrations of FSH in cows. The duration of dominance of the ovulatory follicle tended to be longer in cows than in heifers. Estradiol concentrations around estrus and the preovulatory LH surge were higher in heifers than in cows (20 versus 9 ng/ml). Progesterone concentrations were higher in heifers than in cows from Day 3 to Day 16 of the cycle. Circulating progesterone did not differ between two-wave and three-wave cycles. The results revealed differences in ovarian follicular dynamics, and in plasma concentrations of steroids and gonadotropins; these may account for the differences in fertility between nulliparous heifers and multiparous lactating cows.     

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.     

Short estrous cycles and estrous signs after premature ovulations induced with cloprostenol and gonadotropin-releasing hormone in cyclic dairy cows

The aim of the present study was to confirm earlier findings, obtained with a small number of animals, that gonadotropin-releasing hormone (GnRH) can shorten corpus luteum functional life when it is administered 24 h after cloprostenol (PG) treatments given 7-9 days after estrus. In addition, the effects of two treatments, PG alone or PG + GnRH given before mid-diestrus, on signs of estrus were studied. Sixty cows in farm conditions were used in the experiment. Eight days after natural estrus, they were given an intramuscularly (i.m.) treatment of cloprostenol (0.5 mg). The animals were then divided into two groups. One group (n = 25) received an i.m. treatment of gonadorelin (0.1 mg) 24 h after the PG treatment (PG + GnRH group), while another group (n = 35) served as controls without any further treatment (PG group). Estrous signs were recorded. Progesterone concentrations were measured from samples of whole milk. No short cycles were observed in the PG group, whereas 33% of the cows in the PG + GnRH group exhibited premature luteal regression (P < 0.05). Cloprostenol treatment on Day 8 had no effect on the intensity of the estrous signs. Instead, GnRH treatment 24 h after PG treatment weakened the estrous signs significantly (P < 0.01). It is concluded that GnRH administration 24 h after a PG treatment given 8 days after estrus can cause short estrous cycles in some cows on an individual basis.     

Intrauterine infusion of prostaglandin E2 and subsequent luteal function in cattle.

The objective was to evaluate the effect of intrauterine infusion of prostaglandin E2 (PGE2) on luteal function in cattle. Heifers and cows were randomly assigned after two normal estrous cycles to either PGE2 or control treatment groups. Females in Treatment A were infused with 1 mg of PGE2 once daily into the uterine horn ipsilateral to the corpus luteum between days 7-10 of the estrous cycle with a 0.25 ml plastic semen straw and an artificial insemination pipette. Females in Treatment B were similarly infused with 1 mg of PGE2 once daily in 20 ml of a carrier vehicle via a catheter on days 10 and 11 of the estrous cycle. Control animals were infused with the carrier vehicle using either a semen straw (Treatment C) or via a catheter (Treatment D) on the same days of the estrous cycle. Blood samples were collected daily to monitor plasma progesterone concentrations during the treatment period. Females infused with PGE2 on days 7-10 of the estrous cycle returned to estrus in a mean of 23.5 days (range 22-25 days) and were similar (P > 0.05) to those infused on days 10 and 11 which returned to estrus in 23.5 days (range 22-25 days). Animals similarly infused with carrier vehicle on the same days of the estrous cycle returned to standing estrus in 20.2 days (range 17-23 days). Plasma progesterone concentrations indicated an extended period of elevated progesterone concentrations in PGE2-treated animals compared with control animals. These results indicate that short term administration of PGE2 early in the estrous cycle may result in extended luteal maintenance.