Short-term relationships between plasma LH, FSH and progesterone concentrations in post-partum dairy cows and the effect of Gn-RH injection

Jugular venous blood samples were obtained from 7 dairy cows every 10 min for 10-19 h during the early- or mid-luteal phase of the oestrous cycle, and each cow was given 1 or 2 i.v. injections of 100 micrograms synthetic Gn-RH. Four of these cows were also sampled in a different cycle with no treatment being administered. Peaks of plasma LH, FHS and progesterone were detected in each animal in the absence of treatment; those of LH and progesterone often occurred in parallel. Injection of Gn-RH was always followed by a significant increase in plasma LH and progesterone concentrations and in most cases by a significant FSH increase. There was a significant temporal relationship between the peaks of all 3 hormones. A further 8 cows were sampled during the first 10 days post partum when the mean plasma progesterone concentration was low. An i.v. injection of 200 micrograms synthetic Gn-RH was given to each animal and this resulted in a significant increase in plasma LH and FSH concentrations, but in only one cow was the Gn-RH injection followed by a significant increase in plasma progesterone concentration. The LH response to Gn-RH injection was significantly less in cows injected on or before Day 5 post partum than in cows injected on Days 7-10 post partum.     

Effect of exogenous progesterone and oestradiol on plasma progesterone concentrations and follicle wave dynamics in anovulatory anoestrous post-partum dairy cattle

The effect of exogenous progesterone (P4) and of oestradiol benzoate (ODB) on plasma progesterone concentration and follicle dynamics was studied in anovulatory anoestrus (AA) post-partum pasture-fed dairy cattle. Cows (n=32) were defined AA based on not detecting a corpus luteum upon transrectal ultrasonography of the ovaries. Cows were randomly assigned to treatment with an intravaginal P4-releasing device containing 1.56 g of P4 (1Q; Cuemate, Pfizer Animal Health, Auckland, NZ; n=11) or with modified devices with double (2Q; n=11) or triple (3Q; n=10) the normal P4 dose for 8 days. Half of each group received 2 mg ODB at device insertion (Day 0) while the other half did not receive ODB at this time. All cows were treated with 1 mg ODB 1 day after intravaginal device removal (Day 9). Ultrasonography occurred daily until either ovulation or Day 15 whichever occurred sooner. Blood samples were drawn on Days 0, 1, 3, 5, 7, 8, 9, 15 and 22 for plasma P4 determination. Increasing P4 dose was associated with an increase in plasma P4 concentration during the time of device insertion (P <0.05). The highest P4 dose was associated with a delay in emergence of, but a shorter interval from emergence to maximum diameter and ovulation of, the subsequent dominant follicle (DF2) compared to the lowest P4 dose. Treatment with ODB resulted in a delay in emergence of DF2 (4.2 (0.4) versus 2.0 (0.4) days (S.E.M.) to emergence for ODB versus no-ODB; P=0.01), a smaller maximum diameter of DF2 (15.2 (0.5) versus 17.9 (0.6)mm (S.E.M.) for ODB versus no-ODB; P <0.01), a shorter interval to maximum DF2 diameter (5.0 (0.3) versus 6.8 (0.3) days (S.E.M.) for ODB versus no-ODB; P=0.03), a shorter interval from DF2 emergence to ovulation (6.3 (0.4) versus 8.5 (0.4) days (S.E.M.) for ODB versus no-ODB; P=0.02) and a tendency for a lower average plasma P4 concentration post-ovulation (i.e. average of Days 15 and 22; 2.5 (0.4) versus 3.4 (0.4) ng/ml plasma P4 for ODB versus no-ODB, respectively; P=0.08). The DF present at device insertion, was still present at device removal in three (9%) cows of which two were treated with 1Q + no-ODB and one with 3Q + ODB. It is concluded that increasing P4 dose and ODB treatment are associated with a delay in subsequent follicle wave emergence and more rapid follicle growth. Oestradiol benzoate treatment also tends to reduce the plasma P4 concentration in the subsequent luteal phase in post-partum, anoestrous dairy cattle.     

Plasma progesterone and gonadotrophin concentrations and ovarian activity in post-partum dairy cows

Plasma progesterone concentrations in jugular vein blood samples collected every other day after calving from 13 Friesian dairy cows indicated that ovarian cyclic activity was initiated by 16.6 +/- 1.1 (s.e.m.) days post partum, except for 1 cow which did not resume cyclic activity until Day 98 post partum. Rectal palpation of the ovaries indicated that a developing follicle was recognizable at a mean time of 15.7 +/- 2.0 days after calving. During the first oestrous cycle after parturition there was a significantly shorter period when plasma progesterone levels were elevated than during the next 2 cycles. Concentrations of progesterone, LH, FSH and prolactin were determined for 4 cows, in blood samples taken every 6 h from 2 to 36 days post partum. Tonic LH release was lower during the first 10 days than subsequently, but the lack of change in pattern for FSH suggests dissimilar control mechanisms for these hormones during this time. Three cows showed evidence of a resumption of ovarian cyclicity during the sampling period: in 2 there was an initial LH surge of a magnitude which would normally give rise to ovulation, followed 4 days later by an increase in plasma progesterone lasting only 5 and 9 days. This progesterone was considered to be of follicular origin. A second LH surge was followed by the presence of a corpus luteum.     

Effect of endogenous and exogenous progesterone on the oestradiol-induced LH surge in dairy cows

Four cows released an LH surge after 1.0 mg oestradiol benzoate administered i.m. during the post-partum anoestrous period with continuing low plasma progesterone. A similar response occurred in the early follicular phase when plasma progesterone concentration at the time of injection was less than 0.5 ng/ml. Cows treated with a progesterone-releasing intravaginal device (PRID) for 8 days were injected with cloprostenol on the 5th day to remove any endogenous source of progesterone. Oestradiol was injected on the 7th day when the plasma progesterone concentration from the PRID was between 0.7 and 1.5 ng/ml. No LH surge occurred. Similarly, oestradiol benzoate injected in the luteal phase of 3 cows (0.9-2.1 ng progesterone/ml plasma) did not provoke an LH surge. An oestradiol challenge given to 3 cows 6 days after ovariectomy induced a normal LH surge in each cow. However, when oestradiol treatment was repeated on the 7th day of PRID treatment, none released LH. It is concluded that ovaries are not necessary for progesterone to inhibit the release of LH, and cows with plasma progesterone concentrations greater than 0.5 ng/ml, whether endogenous or exogenous, did not release LH in response to oestradiol.     

Preovulatory LH profiles of superovulated cows and progesterone concentrations at embryo recovery

Forty-two Holstein cows were randomly assigned to three superovulatory treatment groups of 14 cows each. Cows in Group I received follicle stimulating hormone (FSH; 50 mg i.m.); those in Group II received FSH (50. mg i.m.) along with GnRH (250 ug in 2 % carboxymethylcellulose s.c.) on the day of estrus; and cows in Group III were infused FSH (49 mg) via osmotic pump implants. FSH was administered over a 5-d period for cows in Groups I and II (twice daily in declining doses). Cows in Group III received FSH over a 7-d period (constantly at a rate of 7 mg/day). All cows received 25 mg PGF(2)alpha (prostaglandin F(2)alpha) 48 hours after initiation of the FSH treatment. Blood samples were collected from seven cows from each group at 2 hour intervals on the fifth day of superovulation for serum luteinizing hormone (LH) concentration analysis by radioimmunoassay, and blood samples were collected from all cows on the day of embryo recovery for plasma progesterone determination. The LH profile was not altered (P>0.05) by either GnRH administration or by the constant infusion of FSH as compared to FSH treatment alone. Plasma progesterone concentrations were highly correlated with the number of corpora lutea (CL) palpated (r=0.92; P<0.01) and with the number of ova and/or embryos recovered (r=0.88; P<0.01). The accuracy of predicting the number of recoverable ova and/or embryos by the concentration of plasma progesterone was 86%.     

Seasonal variations in post-partum plasma progesterone levels and conception in primiparous and multiparous dairy cows

Progesterone concentrations in peripheral plasma of 18 primiparous and 34 multiparous dairy cows were determined by radioimmunoassay every 4 days, from 10 days post partum until the next conception. The interval from parturition to the first progesterone peak (greater than 4 ng/ml plasma) was significantly longer (P less than 4 ng/ml plasma) was significantly longer (P less than 0.01) in primiparous than in multiparous cows. The progesterone concentrations on Days 4-15 of the oestrous cycle were significantly lower (P less than 0.025) during the summer than in cycles occurring in winter. The conception rate during the summer was lower than in winter. In cows inseminated in summer, conception was related to the shape of the progesterone curve in the cycle preceding insemination.     

The effect of active immunization against progesterone on plasma concentrations of total and free progesterone, estradiol-17beta and LH in the cyclic ewe

Nine mature cyclic ewes were actively immunized against progesterone which was rendered immunogenic by conjugation to bovine serum albumin (BSA). Seven control ewes were immunized with BSA. In ewes immunized against progesterone, the concentration of total plasma progesterone increased to 24.3 ng/ml vs 2.8 ng/ml in control animals (P<0.001). However, immunization did not affect the plasma levels of free, unbound progesterone. The correlation coefficient between total plasma progesterone concentrations on Days 4 to 11 of the estrous cycle and antibody titer was r=0.983. Estradiol-17beta concentrations in immunized ewes were higher than in controls on Days 6 to 15 of the estrous cycle (P approximately 0.05). Frequent sampling for LH over a 6-h period on Days 2, 5, 8, 11 and 14 of the cycle revealed no significant differences in the frequency and amplitude of LH pulses between immunized and control ewes. The immunized ewes had estrous cycles of normal length and maintained normal pregnancies. It is suggested that the immunized cyclic ewe is capable of maintaining adequate levels of free progesterone by greatly increasing progesterone synthesis, thus neutralizing the effect of the antibodies.     

The effect of progesterone implants on ovulation and plasma levels of LH, FSH and progesterone in anoestrous ewes.

The effects of 100-mg progesterone implants in anoestrous ewes on plasma progesterone and gonadotrophin levels are described. Implant removal resulted in a surge of plasma gonadotrophins and ovulation, but there was no evidence of behavioural oestrus in 90% of the ewes. These results are discussed.     

Effect of exogenous LH on plasma concentrations of progesterone and oestradiol in relation to the cessation of egg laying induced by different moulting methods

Artificially induced cessation of egg laying caused regression of the reproductive tract in hens, as well as changes in circulating concentrations of sex steroids. Hens were bled at several stages during and after artificial moult induced by fasting or overfeeding a diet low in calcium or high in zinc. Hens received a single injection of 200 i.u. of horse LH at Day 0, 7, 21, 35 and 77 (Exp. 1) or Day 0, 8, 23, 35 and 71 (Exp. 2) after start of the treatment to induce moult. Blood samples were taken before and 20, 40 and 60 min (Exp. 1) or 15, 30 and 45 min (Exp. 2) after LH injection. Hens which were fasted or given the diet high in zinc had low plasma progesterone concentrations and the response to LH was reduced or delayed. In hens fed low calcium the reduction in plasma progesterone was less pronounced and the responsiveness to LH was more or less maintained. Conversely, there was no response of oestradiol to LH in laying hens. However, oestradiol concentrations increased in moulting hens after LH injection, due to the high oestradiol secretion from the small white follicles, since all yolky follicles were atretic.     

Hormone responses to exogenous GnRH pulses in post-partum dairy cows

Nine Friesian dairy cows were treated with 2.5 micrograms GnRH i.v. at 2-h intervals for 48 h commencing between Days 3 and 8 post partum. Hormone concentrations were measured in jugular venous plasma. An episodic pattern of LH release was induced in all animals and there was no significant change in amplitude during treatment. However, cows treated between Days 7 and 8 ('late') showed higher LH episode peaks than did those treated between Days 3 and 6 ('early'). Plasma FSH concentrations showed a less clear episodic pattern in response to GnRH injection. The mean height of FSH responses to GnRH tended to be higher in the 'early' group than in the 'late' group, as did mean FSH concentrations during the pretreatment sampling period. Although clear episodic changes were not observed, GnRH treatment induced a rapid sustained rise in plasma oestradiol-17 beta concentrations, indicating the responsiveness of ovarian follicles to gonadotrophin stimulation early in the post-partum period. There was no difference in oestradiol-17 beta concentrations between the 'early' and 'late' groups during the treatment period. Only one cow exhibited preovulatory-type LH, FSH and oestradiol-17 beta surges during the 96-h post-treatment sampling period. It is concluded that: (1) responsiveness to GnRH pulses increases significantly and FSH responsiveness tends to decrease with time post partum, (2) ovarian follicles are able to secrete oestradiol-17 beta in response to GnRH-induced LH and FSH release during the early post-partum period and there is no time-dependent change in responsiveness; and (3) the lack of preovulatory surges, except in one cow, may reflect a temporary defect in the positive-feedback mechanism by which high concentrations of oestradiol-17 beta induce preovulatory gonadotrophin release.     

Effect of induced suprabasal progesterone levels around estrus on plasma concentrations of progesterone, estradiol-17beta and LH in heifers

A controlled study was carried out to investigate the effects of suprabasal plasma progesterone concentrations on blood plasma patterns of progesterone, LH and estradiol-17beta around estrus. Heifers were assigned to receive subcutaneous silicone implants containing 2.5 g (n=4), 5 g (n=4), 6 g (n=3), 7.5 g (n=3) or 10 g (n=4) of progesterone, or implants without hormone (controls, n=5). The implants were inserted on Day 8 of the cycle (Day 0=ovulation) and left in place for 17 d. The time of ovulation was determined by ultrasound scanning. Blood was collected daily from Days 0 to 14 and at 2 to 4-h intervals from Days 15 to 27. Control heifers had the lowest progesterone concentrations on Days 20.5 to 21 (0.5 +/- 0.1 nmol L(-1)); a similar pattern was observed in heifers treated with 2.5 and 5 g of progesterone. In the same period, mean progesterone concentrations in the heifers treated with 6, 7.5 and 10 g were larger (P < 0.05) than in the controls, remaining between 1 and 2.4 nmol L(-1) until implant removal. A preovulatory estradiol increase started on Days 16.4 to 18.4 in all the animals. In the controls and in heifers treated with 2.5 and 5 g of progesterone, estradiol peaked and was followed by the onset of an LH surge. In the remaining treatments, estradiol release was prolonged and increased (P < 0.05), while the LH peak was delayed (P < 0.05) until the end of the increase in estradiol concentration. The estrous cycle was consequently extended (P < 0.05). In all heifers, onset of the LH surge occurred when progesterone reached 0.4 to 1.2 nmol L(-1). The induction of suprabasal levels of progesterone after spontaneous luteolysis caused endocrine asynchronies similar to those observed in cases of repeat breeding. It is suggested that suprabasal concentrations of progesterone around estrus may be a cause of disturbances oestrus/ovulation.     

The effect of “Estrumate” followed by progesterone coils on oestrus synchronization and conception of post-partum beef and dairy cows

To synchronize oestrus, a single dose of 0.5 mg of the prostaglandin F_2α analogue cloprosterol (Estrumate) was administered to 200 post-partum beef and dairy cows. Six, 9 or 13 days following the injection of Estrumate a progesterone-releasing intravaginal device (PRID) was inserted for 12 to 16 days. Among the synchronization treatments investigated, the most promising one was to administer 0.5 mg of Estrumate followed 13 days later by a PRID inserted for 12 days. Following the use of this treatment 87% of post-partum suckling beef and dairy cows manifested oestrus within the first 3 days after PRID removal; 80% appeared in heat between approximately 36 and 72 h following PRID removal. Conception rates for untreated controls, cows inseminated once at 55 h, and cows inseminated twice at 48 and 72 h following PRID removal, were 52, 50 and 48% respectively.     

Radioimmunoassay of FSH in the plasma of post-partum dairy cows

A method for measurement of bovine plasma FSH has been established with an interassay coefficient of variation of 15.5% over the workable range of the assay. Compared to values at the end of pregnancy (42-122 ng/ml), FSH concentrations were greater (P less than 0.01) between 0 and 20 days post partum in 3 cows.     

The effects of prostacyclin (PGI2) and 6-keto-PGF1 alpha on bovine plasma progesterone and LH concentrations

Injections of 1 mg PGI2 directly into the bovine corpus luteum significantly increased peripheral plasma progesterone concentrations within 5 min. Concentrations were higher in the PGI2-treated heifers than in saline-injected controls between 5 and 150 min and at 3.5, 4, 5, and 7 h post-treatment. Levels tended to remain elevated through 14 h. Saline and 6-keto-PGF1 alpha were without effect on plasma progesterone levels. The luteotrophic effect of PGI2 was not due to alterations in circulating LH concentrations. An in vitro experiment assessed the effects of either PGI2 alone or in combination with LH on progesterone production by dispersed luteal cells. Progesterone accumulation over 2 h for control, 5 ng LH, 1 microgram PGI2, 10 micrograms PGI2, and 10 micrograms PGI2 plus 5 ng LH averaged 99 +/- 42, 353 +/- 70, 152 +/- 35, 252 +/- 45, and 287 +/- 66 ng/ml (n = 4), respectively. Thus PGI2 has luteotrophic effects on the bovine CL both in vivo and in vitro.     

Plasma,milk and faecal progesterone concentrations during the oestrous cycle of lactating dairy cows with different milk yields

The hypotheses tested in this study were that neither average progesterone (P4) concentrations in plasma and milk nor average progesterone metabolites concentrations in faeces would differ during an oestrous cycle in two groups of cows with differing daily milk yields. High producing (HP = 8) and low producing (LP = 8) dairy cows were selected randomly for the study. Their oestrous cycles were initially synchronised using P4 and prostaglandin F2alpha. Chromic oxide capsules were administered twice daily to measure total faecal output. Samples of blood, faeces and milk were taken daily throughout one oestrous cycle, plasma and milk P4, and faecal progesterone metabolites (FP4M) assayed. The average daily milk yields in the two groups were 30.8 and 21.9l per day, respectively (P < 0.0001), although daily faecal output was similar in both the groups (HP, 7.7 versus LP, 6.9 kg DM; P = 0.24). Mean plasma and milk P4 concentrations were similar in both the groups (plasma P4, 4.12 versus 4.05 ng/ml; P = 0.3; milk P4, 8.2 versus 8.3; P = 0.9) during dioestrus. Average daily excretion of P4 to the milk was greater in HP than LP cows (252 versus 185 microg, P = 0.04). Neither concentration nor the daily yield of FP4Ms was affected by level of milk yield (concentration: 12.2 versus 11.5 microg/g; daily yield: 89.1 versus 82.9 mg per day; P > 0.05). These data showed that the concentrations of P4 in plasma and milk, and the concentrations and daily yields of FP4M were not affected by the level of daily milk yields which differed by about 41% of the LP average of 21.9l.     

The effect of ACTH administration on plasma testosterone, dihydrotestosterone and serum LH concentrations in normal men

Plasma cortisol (F), testosterone (T), dihydrotestosterone (DHT) and serum luteinizing hormone (LH) concentrations were measured in 8 normal young men at 8 AM on two control days. Exogenous adrenocorticotrophin (ACTH) 20 U.S.P. units per m² of body surface area every 12 or 6 hours was administered intramuscularly for 4 days. Twenty-four hours after starting ACTH administration, the plasma T and DHT concentrations were significantly lower than those of the control days on a paired t test. No significant change in serum LH concentration could be demonstrated. Similar results were observed after 48, 72 and 96 hours of ACTH stimulation.     

Ultradian, circadian and seasonal variations of plasma progesterone and LH concentrations during the luteal phase

The circadian variations in plasma progesterone (P) and LH concentrations were investigated in six women, aged 23-40 years. All were studied in the mid-luteal phase (7 +/- 2 days after LH mid-cycle surge). Experiments were conducted in autumn and in spring. Blood samples were obtained every 15 min for 24 hr. Plasma P and LH concentrations were measured by RIA. Each subject's time-series was analysed using three methods; visual inspection (chronogram), spectral analysis to estimate component periods of rhythms (tau) and cosinor analysis to quantify the rhythms parameters. Marked temporal variations in plasma P concentration were observed in each subject. The maximal variations over a 24-hr period, ranged between 13-58.5 mmol/l. Differences related to sampling time were statistically validated by ANOVA (p less than 0.00001). Significant harmonic periods were detected by spectral analysis but differed among subjects. In all subjects but one, a circadian rhythm was detected. The acrophase location was similar (about 0700 hr) in the four subjects studied in autumn, but ranged from 1940 to 0320 hr in those studied in spring. An ultradian rhythm with tau = 8 hr was also validated in six time-series with similar acrophases (about 0200, 1000, and 1800 hr). Cosinor analysis of pooled data revealed that the 24-hr, 12-hr, and 8-hr rhythms were statistically significant (p = 0.001) in autumn. algebraic sum of these three cosine functions yielded a circadian waveform with peak-times occurring near 0300 and 1130 hr and a trough-time about 2200 hr. In spring, the circadian pattern appeared quite different, and peak-times were found near 0700 and 2000 hr, and trough-times near 0300 and 1500 hr. Furthermore, the 24-hr mean of P was higher in autumn (28.9 +/- 0.4 nmol/l) than in spring (17.2 +/- 0.4 nmol/l), p from ANOVA less than 0.00001. The evidence for a similar circadian LH pattern is not as strong. Seasonal, circadian and ultradian rhythms characterize the physiologic time structure of plasma P concentration in mid-luteal phase.