Endocrine pathogenesis of postweaning anestrus in swine: response of the persistently anestrous sow to hormonal stimuli

The endocrine function of the individual components of the hypothalamo-hypophyseal-ovarian axis of the postweaning anestrous sow was evaluated by monitoring the sow's response to exogenous estradiol, gonadotropin releasing hormone (GnRH), and gonadotropins. Sows (4 to 6/group) not returning to estrus by 42.8 +/- 3.1 days after weaning were assigned to 1 of the following treatments: 10 micrograms of estradiol benzoate (EB)/kg of body weight; 200 micrograms of GnRH, 1,000 IU of pregnant mare's serum gonadotropin (PMSG); 1,000 IU of human chorionic gonadotropin (HCG); or 4 ml of saline solution plus 2 ml of corn oil. A preovulatory-like surge of luteinizing hormone [(LH) greater than 12 hours in duration] was observed in all weaned sows treated with EB. All EB-treated sows exhibited estrus and ovulated but none conceived. Sows given GnRH had transiently increased (less than 3 hours) LH concentrations that were not associated with estrus or ovulation. Treatment with PMSG caused an increase in peripheral concentrations of 17 beta-estradiol that was followed by an LH surge, estrus, ovulation, and conception. Treatment with HCG caused an increase in circulating concentrations of 17 beta-estradiol that was accompanied by a surge of LH in some sows and ovulation in all sows. Not all sows treated with HCG exhibited estrous behavior, but conception occurred in 2 of 3 sows that were mated at estrus. None of the sows treated with saline plus corn oil had consistent changes in circulatory concentrations of 17 beta-estradiol or LH and none exhibited estrus or ovulated.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of plasma progesterone concentrations on LH release and ovulation in beef cattle given GnRH

The effects of plasma progesterone concentrations on LH release and ovulation in beef cattle given 100 microg of GnRH im were determined in three experiments. In Experiment 1, heifers were given GnRH 3, 6 or 9 days after ovulation; 8/9, 5/9 and 2/9 ovulated (P<0.02). Mean plasma concentrations of progesterone were lowest (P<0.01) and of LH were highest (P<0.03) in heifers treated 3 days after ovulation. In Experiment 2, heifers received no treatment (Control) or one or two previously used CIDR inserts (Low-P4 and High-P4 groups, respectively) on Day 4 (estrus=Day 0). On Day 5, the Low-P4 group received prostaglandin F(2alpha) (PGF) twice, 12 h apart and on Day 6, all heifers received GnRH. Compared to heifers in the Control and Low-P4 groups, heifers in the High-P4 group had higher (P<0.01) plasma progesterone concentrations on Day 6 (3.0+/-0.3, 3.0+/-0.3 and 5.7+/-0.4 ng/ml, respectively; mean+/-S.E.M.) and a lower (P<0.01) incidence of GnRH-induced ovulation (10/10, 9/10 and 3/10). In Experiment 3, 4-6 days after ovulation, 20 beef heifers and 20 suckled beef cows were given a once-used CIDR, the two largest follicles were ablated, and the cattle were allocated to receive either PGF (repeated 12h later) or no additional treatment (Low-P4 and High-P4, respectively). All cattle received GnRH 6-8 days after follicular ablation. There was no difference between heifers and cows for ovulatory response (77.7 and 78.9%, P<0.9) or the GnRH-induced LH surge (P<0.3). However, the Low-P4 group had a higher (P<0.01) ovulatory response (94.7% versus 61.1%) and a greater LH surge of longer duration (P<0.001). In conclusion, although high plasma progesterone concentrations reduced both GnRH-induced increases in plasma LH concentrations and ovulatory responses in beef cattle, the hypothesis that heifers were more sensitive than cows to the suppressive effects of progesterone was not supported.     

Appearance of the uterine specific proteins following induction of ovulation in prepubertal gilts.

The appearance of the uterine specific proteins following induction of ovulation in prepubertal gilts is described. 12 gilts each at 3, 4, and 5 months of age were allotted randomly to 1 of 2 treatment groups prior to induction of ovulation: 1) saline treated and 2) human chorionic gonadotropin (HCG) treated (400 IU daily from Days 12 through 16 of the induced cycle). Ovulation was induced with HCG following treatment with pregnant mare serum gonadotropin and the day of ovulation was designated as Day 1. All the gilts were laparotomized and uteri infused with phosphate-buffered saline on Day 16 to obtain uterine protein secretions. Plasma progesterone levels on Day 11 and observations made at laparotomy indicated that only 1 gilt 3 months of age failed to ovulate. The number of corpora lutea, plasma progesterone, total recoverable uterine protein, and uterine specific protein on Day 16 were markedly affected by the age of the gilt. These same characteristics, except uterine specific protein, were additionally affected by HCG treatment. Total recoverable uterine protein and uterine specific protein in saline and HCG-treated gilts at 3, 4, and 5 months of age were 6.3 and 1.5, 10.4 and 2.8; 38.8 and 15.2, 51.6 and 15.9; 20.4 and 7.7, 47.8 and 14.6 mg, respectively. Polyacrylamide slab gel electrophoresis showed that HCG-treated gilts at 3 months of age and both saline- and HCG-treated gilts at 4 and 5 months of age generally produced the purple basic protein and the complete profile of the low molecular weight acidic proteins during the induced cycle.     

Characteristics of developing prolonged dominant follicles in cattle

In cattle, sub-luteal circulating progesterone induces an increase in the frequency of LH pulses, prolonged growth of the dominant follicle, increased peripheral estradiol and reduced fertility. The objective of this study was to examine the earliest stages of development of prolonged dominant follicles, to gain insight into the etiology of this aberrant condition. Heifers were treated with an intravaginal progesterone-releasing device (CIDR) from Day 4-8 post-estrus and PGF2alpha was injected on Day 6 and again 12h later (early prolonged dominant group). Follicular phase (CIDR: Day 4-6, with PGF2alpha) and luteal phase (CIDR: Day 4-8, without PGF2alpha) groups served as controls. As expected, peripheral progesterone in heifers of the early prolonged dominant group was intermediate between luteal and follicular phase groups after luteal regression (P<0.05). On Day 7, the frequency of LH pulses was higher in heifers of the follicular phase and early prolonged dominant groups than the luteal phase group (P<0.05). Dominant follicles (n = 4 per group) were collected by ovariectomy on Day 8 and were similar in size among groups (P>0.05). Estradiol and androstenedione concentrations in the follicular fluid at ovariectomy were higher in the follicular phase and early prolonged dominant groups versus the luteal phase group (P<0.01), whereas progesterone did not differ among groups (P>0.05). Granulosa cells and theca interna isolated from dominant follicles were incubated for 3h with or without gonadotropins or frozen for later analysis of mRNA for steroidogenic enzymes. Luteinizing doses (128 ng/ml) of LH and FSH increased secretion of progesterone (P<0.05) but did not affect secretion of estradiol by granulosa cells in all groups. Low (2 or 4 ng/ml) and luteinizing doses of LH increased secretion of androstenedione by theca interna to a similar extent among groups. Expression of mRNA for P450 side chain cleavage (P450scc), 3beta-hydroxysteroid dehydrogenase (3beta-HSD), P450 aromatase (aromatase) and Steroidogenic Acute Regulatory (StAR) protein by granulosa cells did not differ among groups (P>0.05). Levels of mRNA for P450scc, 3beta-HSD, 17alpha-hydroxylase (17alpha-OH) and StAR protein in theca interna were similar in the follicular phase and early prolonged dominant groups (P>0.05), but lower in the luteal phase group (P<0.05-0.1). In summary, the premature follicular luteinization observed in previous studies after prolonged periods of sub-luteal progesterone was absent in early prolonged dominant follicles, exposed to sub-luteal progesterone for 36 h, and their characteristics resembled those of control follicles during the follicular phase.     

Characteristics of protein-protease inhibitors, 17-beta estradiol and progesterone in the blood of heifers returning to estrus following LH/FSH-RH treatment]

Tests were performed to study the inhibitive capacity of protein-protease inhibitors, 17-beta estradiol, and progesterone in the blood serum of 21 heifers returning to oestrus which had been treated intramuscularly with 400 microgram LH/FSH-RH. Four selected heifers were examined as to the relationships between the concentrations of 17-beta estradiol, progesterone and the inhibitive capacity of the serum. Heifer no. 1 had low concentrations of 17-beta estradiol and higher concentrations of progesterone (average levels 7.8 and 657.4 pg ml-1 of serum). On the other hand, heifer no. 4 was found to have higher concentration of 17-beta estradiol and lower values of progesterone (average levels 28.3 and 325 pg ml-1 of serum). Having shown heat, the heifers were fertilized and remained in calf. Heifer no. 2 with a low concentration of progesterone (292 pg ml-1 of serum) and 17-beta estradiol (12.4 pg ml-1 of serum), as well as heifer no. 3 with a high concentration of both (830 pg ml-1 of serum and 22.7 pg ml-1 of serum, respectively) showed no heat and they were not mated. Heifers with significant differences in the concentrations of progesterone and 17-beta estradiol and with low values of protein-protease inhibitory capacities remained in-calf, whereas heifers with lower progesterone and 17-beta estradiol values, or adversely, with higher inhibition values, showed no heat. Synthetic LH/FSH-RH was found not to increase the amount of protein-protease inhibitors in heifer serum, as distinct from the cervical mucus of breeding cows treated with gestagens in heat synchronization.     

Pulsatile or continuous infusion of luteinizing hormone-releasing hormone and hormonal concentrations in prepubertal beef heifers

An experiment was conducted to determine if exogenous luteinizing hormone-releasing hormone (LHRH) administered iv intermittently as pulses (P) or by continuous sc infusion (I) using osmotic minipumps could sustain pulsatile LH release and induce estrous cyclicity in prepubertal heifers. Prepubertal heifers were assigned randomly to: 1) receive pulses of LHRH (n = 6; 2.5 micrograms LHRH/2 h for 72 h), 2) be infused with LHRH (n = 11; 1.25 micrograms LHRH/h for 72 h), or 3) serve as controls (n = 16). Blood was collected at 20-min intervals for 8 h (0900 to 1700 h) from six heifers in each group on d 1, 2, 3 (during treatment), and on d 4 (during 8 h after terminating LHRH treatments). Heifers given LHRH had higher (P less than .01) LH concentrations than controls. Preovulatory-like LH surges occurred in three I, two P and no control heifers during treatment. Pulse frequencies of LH (no. LH pulses/8 h) were greater (P less than .001) for P heifers than for I and control heifers due to pulsatile LHRH treatment. Serum estradiol was higher (P less than .01) during treatment for LHRH-treated heifers than for controls. Serum follicle-stimulating hormone, cortisol, and progesterone were unchanged during treatment. High levels of cortisol on d 1 declined (P less than .001) to baseline by d 2. Characteristic progesterone rises or short luteal phases occurred within 10 d of treatment initiation in more (P less than .05) LHRH-treated heifers (I = 45%, P = 33%) than controls (6%), although days to first observed estrus and first ovulation were unaffected by treatments. Although both continuous and pulsatile administration of LHRH successfully induced LH and estradiol release as well as preovulatory-like LH surges in some heifers, earlier initiation of estrous cycles was not achieved. Estrous cycles appeared to be delayed by exposure to continuous LHRH infusions during the peripubertal period.     

Energy balance and body condition influence luteal function in Holstein heifers

A factorial experiment was conducted to determine influence of energy balance (EB) and body condition (BC) on luteal function in heifers. Heifers with moderate (MBC) or fat (FBC) BC were fed individually to sustain positive EB (PEB) or to cause negative EB (NEB). Intake of feed was measured daily and body weight weekly. Progesterone was quantified daily in serum for 3.5 estrous cycles. On days 9, 10, or 11 after fourth estrus, blood was sampled every 15 min for 12 hr to quantify luteinizing hormone (LH), growth hormone (GH), insulin and non-esterified fatty acids (NEFA). The next day, luteal cells were incubated and proportions of small to large cells were determined. After fourth estrus, area of progesterone profiles in serum for 10 days postestrus was reduced in all heifers relative to MBC-PEB heifers. But, luteal weight from FBC-PEB and MBC-NEB heifers was less than MBC-PEB heifers and FBC-NEB heifers were intermediate. Secretion of progesterone in vitro was increased by LH for PEB but not NEB heifers. MBC-NEB heifers had increased ratios of small to large luteal cells. Independent of BC, NEB decreased concentrations of insulin and increased GH and NEFA. Secretion of progesterone was not associated with LH, GH or insulin, but was correlated negatively with NEFA. We conclude that reduced concentrations of progesterone in serum of FBC-PEB and MBC-NEB heifers is due to impaired luteal development. But, reduced concentrations of progesterone in serum of NEB heifers is due also to reduced basal (MBC) and LH-induced (MBC and FBC) secretion of progesterone by luteal cells. Body condition at onset of NEB may determine when effects of NEB on progesterone are detected.     

Estrus synchronization and pregnancy rates in beef cattle given CIDR-B, prostaglandin and estradiol, or GnRH

Two experiments were conducted to determine estrous response and pregnancy rate in beef cattle given a controlled internal drug release (CIDR-B) device plus prostaglandin F2 alpha (PGF) at CIDR-B removal, and estradiol or gonadotropin releasing hormone (GnRH). In Experiment I, crossbred beef heifers received a CIDR-B device and 1 mg estradiol benzoate (EB), plus 100 mg progesterone (E + P group; n = 41), 100 micrograms gonadotropin releasing hormone (GnRH group; n = 42), or no further treatment (Control group; n = 42), on Day 0. On Day 7, CIDR-B devices were removed and heifers were treated with PGF. Heifers in the E + P group were given 1 mg EB, 24 h after PGF, and then inseminated 30 h later. Heifers in the GnRH group were given 100 micrograms GnRH, 54 h after PGF, and concurrently inseminated. Control heifers were inseminated 12 h after onset of estrus. The estrous rate was lower (P < 0.01) in the GnRH group (55%) than in either the E + P (100%) or Control (83%) groups. The mean interval from CIDR-B removal to estrus was shorter (P < 0.01) and less variable (P < 0.01) in the E + P group than in the GnRH or Control groups. Pregnancy rate in the E + P group (76%) was higher (P < 0.01) than in the GnRH (48%) or Control (38%) groups. In Experiment II, 84 cows were treated similarly to the E + P group in Experiment I. Cows received 100 mg progesterone and either 1 mg EB or 5 mg estradiol-17 beta (E-17 beta) on Day 0 and either 1 mg of EB or 1 mg of E-17 beta on Day 8 (24 h after CIDR-B removal), in a 2 x 2 factorial design, and were inseminated 30 h later. There were no differences among groups for estrous rates or conception rates. The mean interval from CIDR-B removal to estrus was 44.2 h, s = 11.2. Conception rates were 67%, 62%, 52%, and 71% in Groups E-17 beta/E-17 beta, E-17 beta/EB, EB/E-17 beta, and EB/EB, respectively. In cattle given a CIDR-B device and estradiol plus progesterone, treatment with either EB or E-17 beta effectively synchronized estrus and resulted in acceptable conception rates to fixed-time artificial insemination.     

Induction of precocious puberty in heifers III: hastened reduction of estradiol negative feedback on secretion of luteinizing hormone

Precocious puberty (<300 d of age) can be induced in beef heifers by early weaning and feeding a high-concentrate diet. The objective of this experiment was to determine whether precocious puberty occurs as a result of a hastened reduction of estradiol negative feedback on secretion of LH. Thirty crossbred Angus and Simmental heifers were weaned at 83 +/- 2 d of age and 114 +/- 3 kg of BW, blocked by BW, and randomly assigned to receive a high-concentrate (60% corn; H) or control (30% corn; C) diet and to receive ovariectomy (OVX), OVX plus an estradiol implant (OVXE), or to remain intact (INT). Residual ovarian tissue after OVX necessitated withdrawal of 6 heifers during the course of the experiment, resulting in the following treatment groups: OVX-C, n = 3; OVX-H, n = 5; OVXE-C, n = 4; OVXE-H, n = 2; INT-C, n = 5; INT-H, n = 5. To determine concentrations of progesterone and estradiol, blood samples were collected weekly beginning at a mean age of 160 d. To characterize LH concentrations, serial blood samples were collected at 12-min intervals for 12 h at mean ages of 119, 149, 188, 217, 246, 281, 323, 365, 407, and 449 d. By a mean age of 202 d, heifers fed the H diet were heavier (P < 0.05) than those fed the C diet. Heifers in the INT-H treatment attained puberty earlier (P < 0.05) than in the INT-C treatment (275 +/- 30 vs. 385 +/- 14 d of age, respectively). Overall mean concentrations of estradiol did not differ between OVXE-H and OVXE-C, between INT-H and INT-C, or between OVXE and INT treatments. The OVX treatments exhibited greater LH pulse frequency than the OVXE and INT treatments by the first serial blood collection (treatment x age, P < 0.05). The frequency of LH pulses was greater (P < 0.05) in the INT-H than the INT-C treatment by a mean age of 246 d and was greater (P < 0.05) in the OVXE-H than the OVXE-C treatment by a mean age of 281 d. In the OVXE-H treatment, LH secretion increased and subsequently "escaped" from estradiol negative feedback (detection of > or = 1 LH pulse/h) earlier (P < 0.05) than in the OVXE-C treatment (307 +/- 30 and 420 +/- 21 d of age, respectively). It is concluded that advancing the reduction of estradiol negative feedback on secretion of LH is the mechanism by which early weaning and feeding a high-concentrate diet results in precocious puberty in heifers.     

Progesterone and Luteinizing hormone secretion patterns in early pregnant gilts

We studied luteinizing hormone (LH) pulsatility and episodic progesterone release of the corpus luteum (CL) on Day 11 and Day 21 in inseminated gilts and aimed to establish a relationship between these two hormones. Blood was collected at 15-min intervals for 12 hr on Days 11, 16 and 21 from a vena cava caudalis catheter. At euthanasia, eight gilts were pregnant and six gilts were not pregnant. Progesterone parameters (basal, mean, pulse frequency and pulse amplitude) did not differ between pregnant and non-pregnant gilts on Day 11, LH pulse frequency and amplitude tended to differ (p = .07 and p = .079). In pregnant gilts, basal and mean progesterone, progesterone pulse amplitude and frequency declined significantly from Day 11 to Day 21 (p < .05). A significant decline was also seen in the LH pulse amplitude from Day 11 to Day 21 (p < .05). None of the LH pulses was followed by a progesterone pulse within 1 hr on Day 21. On Day 11 and Day 21 appeared a synchronicity in the LH pulse pattern, as there were two or three LH pulses in 12 hr and these LH pulses appeared in the same time window. We conclude that on Day 11 and Day 21 of pregnancy in gilts, progesterone pulses do not follow an LH pulse within one hour. Further we demonstrated that the successful or not successful formation of a CL of pregnancy is independent of progesterone release on Day 11 after insemination. We confirmed the decline of progesterone from Day 11 to Day 21 in the vena cava caudalis and could demonstrate that this decline is partly due to lower progesterone pulse amplitude and frequency and that the decline occurs simultaneously with a decline in LH pulse amplitude.     

Arginine vasopressin stimulation of prostaglandin F2 alpha release in heifers during the estrous cycle

The effect of arginine vasopressin on the stimulation of prostaglandin F2 alpha (PGF2 alpha) release has been examined in vivo. Fifty-eight heifers received one intravenous injection of 10 IU arginine vasopressin on either Day 0 (n = 14), Day 6 (n = 12), Day 13 (n = 14) and Day 18 or 19 or 20 (Day 18-20, n = 18) after the onset of oestrus (Day 0) to determine the effect of arginine vasopressin at different times of the oestrous cycle. Frequent blood samples were taken before and after arginine vasopressin injection for the measurement of 13,14-dihydro-15-keto-PGF2 alpha (PGFM) by radioimmunoassay (RIA). Blood samples for progesterone determinations were taken 2 hr before and 24 hr after arginine vasopressin to monitor luteal function. The data show that arginine vasopressin causes an increase (P less than 0.005) in PGFM concentrations only at Day 18-20 of the cycle in 67% of the experimental heifers.     

On the negative feedback regulation of gonadotropins in castrate and intact cattle with comparison of two FSH radioimmunoassays

Two homologous radioimmunoassays for bovine follicle stimulating hormone (bFSH) were utilized in comparing the differential regulation of FSH and luteinizing hormone (LH) in response to ovariectomy or administration of gonadal steroids in cattle. There appeared to be significant LH cross-reactivity in one of the bFSH systems (bFSH-HS-2-17), but not in the other (bFSH-BP3). Concentrations of FSH in plasma measured by these two systems suggested both qualitative and quantitative differences. Following ovariectomy in heifers, LH concentrations in plasma were increased by 7.5 h, while FSH (measured in the bFSH-BP3 system) was not significantly elevated until 18 h. Administration of 200 micrograms of estradiol-17 beta to ovariectomized heifers inhibited levels of FSH in plasma but large doses of testosterone (100 mg), androstenedione (400 mg) and dihydrotestosterone (800 mg) had no effect. Similarly, LH was not affected by the androgens, while estradiol induced LH surges, leading to increased mean LH concentrations. In contrast to the results in heifers, LH concentrations in plasma from steers were inhibited by administration of androgens as well as by estradiol. In steers, FSH (bFSH-BP3) was marginally inhibited by estradiol and not at all by the androgens. Differences in the secretory patterns of FSH and LH also occurred in intact heifers during the estrous cycle. The 72-h period preceding estrus (follicular phase) was characterized by rapidly declining serum progesterone concentrations, followed by concurrent increases in both LH and estradiol. The circulating levels of bFSH (BP3) tended to decline during this interval. Overall, during the estrous cycle, progesterone levels were positively correlated with bFSH-BP3 (r = .37) and negatively correlated with LH (r = -.39). The gonadotropins were not significantly related (r = -.15). These relationships are consistent with the concept that LH controls the final stages of follicular development in cattle and that FSH may exert only a permissive effect.     

Effect of constant infusion of oxytocin on luteal lifespan and oxytocin-induced release of prostaglandin F2α in heifers

Two experiments were conducted to determine whether constant infusion of oxytocin would prolong the luteal phase and inhibit uterine prostaglandin F2 alpha (PGF2 alpha) secretion in heifers. In Experiment 1, twelve heifers, treated with saline (SAL) or oxytocin (OXY) via jugular cannulae infusions (INF) or osmotic minipumps (OMP), were allotted at estrus into four treatment groups (n = 3). Treatments were: SAL-INF, SAL-OMP, OXY-INF and OXY-OMP. Physiological saline or oxytocin was given from Days 10 to 23 (Day 0 = estrus) of the estrous cycle. Method of treatment (jugular cannula infusion or osmotic minipump) had no effect (P greater than 0.05) on estrous cycle length or pattern of secretion of progesterone; therefore, data were pooled. Estrous cycle lengths were extended (P less than 0.01) for heifers which received oxytocin (25.3 +/- 0.4 d) compared to saline (20.5 +/- 0.4 d). Luteolysis did not occur in oxytocin-treated heifers until after treatment ceased. Experiment 2 was designed and conducted identically to Experiment 1 with the addition of a "challenge" injection of oxytocin (100 IU oxytocin, i.v.) given on Day 16 of the estrous cycle. Treatment of heifers with oxytocin extended (P less than 0.05) estrous cycle length by an average of 3 d compared to heifers treated with saline. The "challenge" injection induced (P less than 0.05) secretion of PGF2 alpha (as measured by the stable PGF2 alpha metabolite, 15-keto-13,14-dihydro-PGF2 alpha) in saline-treated but not oxytocin-treated heifers. In both Experiment 1 and 2, serum concentrations of FSH were elevated (P less than 0.05) in oxytocin-treated heifers. No increase was observed for LH or prolactin. The rise in estradiol-17 beta at luteolysis was not affected (P greater than 0.10) by treatment. In summary, constant infusion of oxytocin extended luteal lifespan, prolonged secretion of progesterone, and inhibited oxytocin-induced secretion of PGF2 alpha. Constant infusion of oxytocin did not affect serum concentrations of estradiol-17 beta, LH or prolactin; however, serum concentrations of FSH were elevated during the oxytocin treatment period.     

Characterization of ovine follicles destined to form subfunctional corpora lutea

A study was done to test whether ovulatory follicles destined to form subfunctional corpora lutea differed from normal ovulatory follicles in steroidogenic function. Twenty-five ewes were treated with prostaglandin F2 alpha on d 11 of the estrous cycle, then unilaterally ovariectomized before (n = 13) or after (n = 12) the surge of luteinizing hormone (LH) at the induced estrus to collect "control" follicles, which would have produced normal corpora lutea. In 15 ewes, the second ovary was removed 63 to 84 h later to collect "treated" follicles before (n = 7) or after (n = 8) the second expected surge of LH. Five ewes (control) were allowed to ovulate from the remaining ovary at first estrus and another five (treated) at the second estrus (3 to 4 d later). Treated ewes had lower serum progesterone than control ewes during the ensuing cycle (P less than .05). Treated follicles contained less estradiol in the theca (4.4 +/- .6 vs 10.0 +/- 2.5 ng; P less than .05), less androstenedione (.1 +/- .1 vs 1.0 +/- .2 ng) and estradiol (.5 +/- .1 vs 2.9 +/- 2.2 ng) in the granulosa (P less than .05) and less progesterone in the follicular fluid (.8 +/- .4 vs 3.3 +/- .8 ng; P less than .05) than control follicles, when removed before the surge of LH. Follicles removed after the surge of LH did not differ. In conclusion, ovulatory follicles with low steroidogenic function became corpora lutea that secreted lower-than-normal quantities of progesterone.     

Estrus synchronization and conception rate after a progesterone releasing intravaginal device (PRID) treatment from the early luteal phase in heifers

The objective of the present study was to evaluate estrus synchronization and conception rate after progesterone releasing intravaginal device (PRID) treatment from the early luteal phase in the presence or absence of estradiol benzoate (EB) in heifers. Heifers (n=11) were assigned randomly to two treatments; insertion of a PRID containing 1.55 g progesterone with a capsule attached including 10 mg EB (P+EB; n=6) and the PRID withdrawn the EB capsule (P-EB; n=5). The PRID was inserted into the vagina on Day 2 of the estrous cycle (Day 0 was the day of ovulation) and was left for 12 days. The proportion of heifers exhibiting standing estrus within 3 days after PRID removal was 83.3% (5/6) for the P+EB group, and 80.0% (4/5) for the P-EB group, respectively. Conception rate by artificial insemination on synchronized estrus was 80.0% (4/5) in the P+EB group, and 100% (4/4) in the P-EB treatment group, respectively. These results suggest that a PRID treatment from 2 days after ovulation for 12 days in the presence or absence of EB has an effect on the synchronization of estrus and produces a beneficial conception rate in heifers.     

Endocrine changes during restoration of estrous cycles following induction of anestrus by restricted nutrient intake in beef heifers

The working hypotheses in this experiment were: that ovarian estradiol would inhibit luteinizing hormone (LH) secretion in heifers that were anestrus as a result of restricted dietary energy intake and the responsiveness of LH secretion to estradiol negative feedback would decrease during the period when restoration of estrous cycles occurred following feeding of diets adequate in energy. Fifteen heifers weighing 341 +/- 12 (mean +/- SE) kg were fed a diet containing 50% of the energy required for maintenance until 40 to 50 d following cessation of estrous cycles. Heifers were assigned to intact control (C, n = 5), ovariectomized (OVX, n = 5) or ovariectomized-estradiol-17 beta-implanted (OVX + E2, n = 5) treatments. Heifers were subsequently provided a high-energy (HE) diet until termination of the study. Progesterone concentrations indicating cessation of corpus luteum function were detected after heifers had lost 71 +/- 8 kg body weight over 186 +/- 28 d. Control heifers re-initiated estrous cycles as indicated by increased progesterone concentrations in serum at 49 +/- 9 d after initiation of feeding the HE diet (360 +/- 18 kg body weight). Initiation of pulsatile LH secretion was observed in heifers by d 12 following OVX. Estradiol suppressed LH secretion in OVX + E2 heifers during the period of nutritional anestrus in C heifers. Suppressive effects of E2 on LH secretion continued in OVX heifers after C heifers had initiated corpus luteum function. Therefore, the working hypothesis that LH secretion is inhibited by E2 in the nutritionally anestrous heifer is accepted but responsiveness to estradiol does not subside with re-initiation of estrous cycles, thus this working hypothesis is rejected.     

提高奶牛受胎率的卵泡和黄体活动调控研究

在夏季和秋季对经产荷斯坦奶牛进行定时排卵-输精程序（TAI）处理并在输精后进行HCG处理（第5d和第10d分别肌注HCG2000IU／头），另于冬季单独对自然发情人工授精母牛进行输精后HCG处理。从各季节HCG处理组及其对照组中抽取部分奶牛，在输精后第0d（输精当天）、7d、14d、和21d分别采血，以测定孕酮和雌二醇水平。所有试验牛在输精后45-60d进行直肠妊娠检查以确定是否受胎。结果显示：夏季TAI＋HCG组、TAI组、对照组母牛输精后的受胎率差异不显著（p〉0．05），分别为26．67％（4／15）、16．67％（2／12）和10．71％（3／28）。秋季TAI＋HCG组、TAI组、对照组情期受胎率差异也不显著（p〉0．05），分别为：55．00％（11／20）、42．11％（8／19）和31．43％（11／35）。冬季HCG组和对照组的受胎率分别为63．16％（12／19）和45．00％（9／20）（P〉0．05）。在夏秋冬三个季节，输精后HCG处理相应显著提高第7d、14d和21d血液中孕酮水平（p〈0．05），但稍稍降低雌二醇水平（p〉0．05），显著降低E2／P比值（p〈0．05）。与受胎率的差异相关，秋冬季各组牛输精后血浆P水平高于（p〉0．05）或（极）显著高于（p〈0．01或p〈0．05）夏季相应组；血浆E2水平则稍高于（p〉0．05）夏季相应组。以上结果说明，TAI单独处理可以将受胎率提高约10～15个百分点；输精后单独HCG处理可以将爱胎率又提高约10-15个百分点，这主要是促进黄体功能的结果，而且提高的幅度在秋冬季要高于夏季。     

Effect of human chorionic gonadotropin on induced ovine corpora lutea during the anestrous season

Experiments were conducted to determine the effect of additional gonadotropic support on induced corpora lutea of anestrous ewes. In one series of experiments, ewes were superovulated and half the ewes received an i.v. injection of 500 IU human chorionic gonadotropin (hCG) on day 5 after ovulation. Corpora lutea were collected from both groups on day 10 after ovulation. Dissociated corpora lutea collected from ewes which received additional hCG contained proportionately more large luteal cells than did those from control ewes (P<.05). In neither cell type was content of receptors for luteinizing hormone (LH) or secretion of progesterone in response to LH affected by an additional injection of hCG. Large cells from anestrous ewes produced more progesterone in response to LH (P<.05) than did large cells from similarly treated ewes during the breeding season. Small cells collected during either season responded similarly to LH. In another series of experiments, anestrous ewes were induced to ovulate and were exposed to fertile rams. Half the ewes received an i.v. injection of 500 IU hCG on day 5 after ovulation. Serum content of progesterone was higher on day 10 in ewes which received hCG 5 days earlier than in control ewes, although progesterone levels declined to generally nondetectable levels in nonpregnant ewes of both groups by day 16. Pregnancy rates in the two groups were not different. We concluded that additional gonadotropic support affects the morphology and function of corpora lutea from anestrous ewes and may be useful for enhancing fertility during the nonbreeding season.     

Intravaginal progesterone devices in synchronization protocols for artificial insemination in beef heifers

Two experiments were designed to investigate the administration of intravaginal progesterone in protocols for oestrus and ovulation synchronization in beef heifers. In Experiment 1, cyclic Black Angus heifers (n = 20) received an Ovsynch protocol and were randomly assigned to receive (CIDR‐Ovsynch) or not (Ovsynch) a progesterone device between Days 0 and 7. Treatment with a controlled internal drug release (CIDR) device significantly increased the size of the dominant follicle prior to ovulation (12.8 ± 0.4 CIDR‐Ovsynch vs 11.4 ± 0.4 Ovsynch) (p < 0.02). Plasma progesterone concentrations throughout the experiment were affected by the interaction between group and day effects (p < 0.004). In Experiment 2, cyclic Polled Hereford heifers (n = 382) were randomly assigned to one of the six treatment groups (3 × 2 factorial design) to receive a CIDR, a used bovine intravaginal device (DIB), or a medroxiprogesterone acetate (MAP) sponge and GnRH analogues (lecirelin or buserelin). All heifers received oestradiol benzoate plus one of the devices on Day 0 and PGF on Day 7 pm (device withdrawal). Heifers were detected in oestrus 36 h after PGF and inseminated 8–12 h later, while the remainder received GnRH 48 h after PGF and were inseminated on Day 10 (60 h). The number of heifers detected in oestrus on Day 8 and conception rate to AI on Day 9 were higher (p < 0.01) in the used‐DIB than in the CIDR or MAP groups, while the opposite occurred with the pregnancy rate to FTAI on Day 10 (p < 0.01). There was no effect of progesterone source, GnRH analogue or their interaction on overall pregnancy rates (64.9%). Progesterone treatment of heifers during an Ovsynch protocol resulted in a larger pre‐ovulatory follicle in beef heifers. Progesterone content of intravaginal devices in synchronization protocols is important for the timing of AI, as the use of low‐progesterone devices can shorten the interval to oestrus.     

Synchronization of estrus in beef cattle with norgestomet and estradiol valerate

Fifty-six cows received a norgestomet implant and an injection of norgestomet and estradiol valerate; half (n = 28) received 500 IU equine chorionic gonadotrophin (eCG) at implant removal, 9 d later. A third group (n = 25) received 2 doses of cloprostenol (500 micrograms) 11 d apart. Estrous rate was higher (P < 0.05) for cows given norgestomet and estradiol plus 500 IU eCG (75.0%) than for those receiving cloprostenol (44.0%); for those receiving norgestomet and estradiol alone, it was intermediate (67.8%). Pregnancy rates to artificial insemination (after estrus or timed) were higher (P < 0.05) for cows given norgestomet and estradiol than for those given cloprostenol (23 of 28, 82.1% vs 13 of 25, 52.0%), and intermediate (67.8%) for those given norgestomet and estradiol plus eCG. In a second experiment, for heifers treated with norgestomet and estradiol plus eCG (n = 15) or with 2 doses of cloprostenol (n = 16), estrous rates were 66.7% vs 56.2% (P > 0.5), ovulation rates were 100.0% vs 81.2% (P = 0.08), intervals from implant removal or cloprostenol treatment to estrus were 48.0 +/- 4.4 hours vs 61.3 +/- 7.0 hours (P = 0.12) and to ovulation were 70.4 +/- 4.4 hours vs 93.2 +/- 7.5 hours (P < 0.01), respectively; pregnancy rates were 41.7 and 35.7%, respectively (P > 0.5). Norgestomet and estradiol were as good as (heifers) or superior to (cows) a 2-dose cloprostenol regimen. In cows given norgestomet and estradiol, injecting eCG at implant removal did not significantly improve estrous or pregnancy rates.