Progesterone and 15-Keto-13,14-Dihydroprostaglandin F2α Levels in Peripheral Circulation After Intrauterine Iodine Infusions in Cows

The effect of intrauterine iodine infusion on estrous cycle length was studied in four cows. The infusions were performed at various times of the estrous cycle: early, middle, late, and during luteolysis. Blood samples were drawn every third hour from the jugular vein. Progesterone and 15-keto-13,14-dihydroprostaglandin F2α (the main metabolite of PGF2α) were measured to monitor luteal activity and prostaglandin release. No release of prostaglandins was observed immediately following intrauterine infusion. Infusion in two cows on day 5 of the estrous cycle resulted in prostaglandin release after 54 and 69 hrs., respectively, followed by luteal regression and the occurrence of estrus at approx. five days after infusion. Infusions performed on days 11 or 12 resulted in prostaglandin release after 147 and 120 hrs., respectively, followed by luteolysis and heat after a 19 day estrous cycle. Infusion in two cows at days 16 and 17 resulted in prostaglandin release after 117 hrs. in both animals. One cycle was prolonged whereas the other cycle was normal in duration. One cow infused on day 20 following the occurrence of the first prostaglandin surge had a cycle length of 26 days, whereas another cow infused on day 20 was not affected because luteolysis was essentially complete by the time of infusion. One animal infused on day 5 did not respond to the iodine infusion. In this animal, however, the corpus luteum was not completely developed prior to the infusion. From this study it can be concluded: 1) intrauterine iodine infusions performed after the development of a progesterone secreting corpus luteum result in prostaglandin release within three to six days with the subsequent occurrence of luteolysis; 2) luteolysis wras in all cases observed in connection with prostaglandin F2α release of the same order of magnitude and duration as during normal luteolysis. kw|Keywords|k]prostaglandin release; k]progesterone; k]cow; k]es trous cycle; k]iodine infusion     

Corpus luteum function and pregnancy rate in lactating dairy cows given human chorionic gonadotropin at middiestrus

The effect of human chorionic gonadotropin (HCG) on corpus luteum (CL) function in lactating dairy cows and its subsequent effect on pregnancy rates achieved with artificial insemination was studied in two parts. In Part 1, four IM injections at intervals of seven days or two IM injections at a 14 day interval of HCG (10,000 IU) were given to two groups of six cows each, groups A and B respectively, beginning on days 9, 10 or 11 of the estrous cycle. Ten control cows were given 10 ml of isotonic saline solution on days 9, 10 or 11. Interestrous intervals were prolonged by an average of 8.1 and 3.4 days for groups A and B respectively over controls. In Part 2, 200 lactating dairy cows on nine farms were assigned, on an alternate basis at insemination, to control or treatment groups to study the effect of CL prolongation via HCG on pregnancy rate. A single injection of HCG (10,000 IU) was given 10, 11 or 12 days after a first or second insemination. Pregnancy rates for control and treated cows were similar (64 101 = 63% and 58 99 = 59% respectively). Interestrous intervals of treated nonpregnant cows were prolonged by approximately five days. Providing additional time for developing embryos to become established in utero by delaying luteolysis did not improve pregnancy rates achieved with artificial insemination in lactating dairy cows.     

Failure of exogenous LH to prevent PGF2alpha-induced luteolysis in beef cows

Henderson and McNatty (Prostaglandins 9:779, 1975) proposed that LH from the preovulatory LH surge attached to receptors on luteal cells and that this attachment might protect the early corpus luteum from PGF2alpha induced luteolysis. To test this hypothesis, experiments were performed on heifers at day 10-12 of the cycle. Both jugular veins were catheterized and infusions of either saline (0.64 ml/min) or LH-NIH-B9 (10 microgram/min; 0.64 ml/min) were given. Saline infusions were from 0-12 h; LH infusions were for 10 h and were preceded by a 2 h saline infusion. All animals were given 25 mg PGF2alpha im at 6 h (6 h into the saline infusion and 4 h into the LH infusion). Blood samples were taken at 0.5 h, 1 h and 4 h intervals from 0-12h, 13-18 h and 12-42 h respectively. Serum was assayed for LH and progesterone by radioimmunoassay methods. Two animals received saline and two received LH in each experiment. Each treatment was replicated 6 times. LH infusion resulted in a mean serum LH of 75 ng/ml compared to 0.90 ng/ml in saline infused animals. This elevation of LH did not alter PGF2alpha induced luteolysis as indicated by decline in serum progesterone. This experiment does not support the hypothesis that the newly formed corpus luteum is resistant to PGF2alpha because of protection afforded by the proestrus LH surge.     

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.     

Profiles of reproductive hormones associated with fertile and nonfertile inseminations of dairy cows

Blood and urine samples were collected at 8-hr intervals prior to ovulation (Day 0), daily through Day 8, and during alternate Days 10 to 18. The purpose was to determine if hormonal profiles differed between fertile and nonfertile inseminations (cases). Among 37 cases involving 30 lactating cows, 32 had a palpable corpus luteum after insemination, and 11 were fertile. Hormonal values among every nonfertile case were outside the limits observed for 10 of 11 fertile cases during one or more segments of the sampling period. A consistent difference among nonfertile cases was delayed estrus and delayed preovulatory increase in LH after progesterone decreased to less than .75 ng/ml. These 14 cases represented 75% of the cases having subnormal progesterone after ovulation. Compared to fertile cases, asynchronies among nonfertile cases involved plasma progesterone, LH and estrogen, and urinary estradiol-17α most frequently prior to Day 0. Examples of abnormal hormonal profiles also included those for ovarian cysts, and early and late regression of the corpus luteum.     

Effect of oxytocin infusion on secretion of progesterone and luteinizing hormone and the concentration of uterine oxytocin receptors during the periovulatory period in cloprostenol-treated ewes.

Oxytocin infusions were initiated on day 10 of the oestrous cycle in ewes, and luteal regression was induced by injection of 100 micrograms cloprostenol on day 12. Blood samples were collected at frequent intervals via an indwelling jugular vein cannula to measure concentrations of progesterone and luteinizing hormone (LH) during the luteal and follicular phases in saline (n = 6) and oxytocin (n = 5) infused animals. The oxytocin infusion maintained peripheral plasma concentrations of 53 +/- 3.2 pg oxytocin ml-1 (mean +/- SEM) compared with values of about 1 pg ml-1 during oestrus in control ewes. Oxytocin infusion had no effect on luteal phase progesterone concentrations, the timing of luteolysis, basal luteinizing hormone (LH) secretion, LH pulse frequency, or the timing or height of the LH surge. Treated ewes came into oestrus significantly earlier than controls (P < 0.05) but ovulated normally. Uterine samples collected 96 h after cloprostenol injection (approximately day 2 of the cycle) showed that oxytocin receptor concentrations were significantly higher in the endometrium in ewes that had been given a 5 day oxytocin infusion than in control animals (556 and 262 fmol mg-1 protein, respectively: geometric means from ANOVA, P < 0.001), whereas myometrial receptor concentrations were not affected (113 and 162 fmol mg-1 protein, respectively). We conclude that the previously reported delay in luteal development caused by oxytocin infusion (Wathes et al., 1991) is not due to the inhibition or delay of ovulation, but must instead occur via a direct influence on the developing corpus luteum.(ABSTRACT TRUNCATED AT 250 WORDS)     

A combined radioimmunoassay and immunocytochemical study of ovarian oxytocin production during the periovulatory period in the ewe

Corpora lutea and follicles were taken from the ovaries of 12 ewes at intervals from the start of luteolysis until 3 days after ovulation. RIA analysis of the tissue oxytocin content showed that luteal oxytocin concentrations declined during luteolysis to reach basal values at about the time of the next ovulation. Oxytocin was first measurable in the walls of 3 out of 6 preovulatory follicles during the LH surge, with a small increase in concentration to 26.1 +/- 6.6 pg/mg before ovulation, and a further increase in the young corpus luteum to concentrations exceeding 1 ng/mg 2-3 days later. After the LH surge, oxytocin was also found in the follicular fluid at a concentration of 3.4 +/- 0.3 ng/ml. Using immunocytochemical techniques, oxytocin and neurophysin were first detected in the follicle wall immediately before ovulation, and were localized in the granulosa cells. After ovulation the stained cells initially formed strands which appeared to break down to clusters and then to individual cells as the corpus luteum matured. The immunocytochemical picture also suggested that neurophysin immunoreactivity increased within a few hours of ovulation but that processing to oxytocin may be delayed. Measurements of circulating oxytocin concentrations revealed a pulsatile release pattern throughout the follicular phase with the height of the pulses decreasing from 25 +/- 5 pg/ml during luteolysis to a minimum of 11 +/- 2 pg/ml during the LH surge.     

Interrelationships between progesterone, 13,14-dihydro-15-keto PGF-2 alpha (PGFM) and LH in cyclic and early pregnant cows

Plasma progesterone and LH secretion patterns were examined in 18 mature dairy cows during the oestrous cycle and after insemination. Blood samples were collected every 15 min for 8 h per day on Days 3, 5, 6, 7, 8, 9, 10, 12, 14, 16, 17, 18, 19, 20 and 21 of the oestrous cycle, then, in the same cows, at the same times during early pregnancy. PGF-2 alpha secretion rates (as determined by plasma PGFM concentrations) were also monitored on Days 14, 16 and the day of, or equivalent to, luteal regression. Mean daily plasma progesterone concentrations were similar until Day 16 in cyclic and pregnant cows, after which values in non-pregnant animals declined. Regression analysis indicated that progesterone concentrations were best described by a quadratic expression with fitted maximum values on Day 13 in non-pregnant animals but values increased linearly over the whole period to Day 21 in pregnant cows. The frequency, amplitude and area under the curve of LH episodes showed no significant differences between cyclic and pregnant animals. In pregnant cows, the amplitude and area under the curve of progesterone episodes increased linearly between Days 8 and 21, although no such increase occurred in cyclic cows. Low-level PGFM episodes were present in cyclic and pregnant cows on Days 14 and 16 after oestrus, and high amplitude episodes occurred in non-pregnant cows during luteal regression. Pregnant cows showed a significant depression of the amplitude, but not the frequency of episodes at the expected time of luteal regression. These results confirm that the corpus luteum of pregnancy secretes an increasing amount of progesterone per se and per unit of LH until at least Day 21 after mating. They further suggest that the corpus luteum of the cyclic cow may experience small episodes of PGF-2 alpha and be subjected to initial degenerative changes by Day 14 after oestrus, some time before the onset of definitive luteolysis.     

The effect of Escherichia coli endotoxin on luteal function in Holstein heifers

This study was undertaken to elucidate the possible role of endotcxin in mediating premature luteolysis in the well- documented phenomenon of short estrous cycles in postpartum dairy cows. Four groups of Holstein heifers (n = 4 to 6 each) received either intrauterine infusion of sterile culture medium (Group I); intrauterine infusion of Escherichia coli (E. coli ) endotoxin (5 mug/kg) in sterile culture medium (Group II); intrauterine administration of 10 ml of a 24-h culture of a strain of E. coli isolated from the uterus of a cow with metritis (approximately 10(9) colony forming units/ml; Group III); or intravenous administration of E. coli endotoxin (5 mug/kg; Group IV) on Day 7-9 of the estrous cycle. Blood samples were collected every 48 h during the pretreatment estrous cycle and up to the administration of the experimental treatment, thereafter 4-h samples were collected for 5 d. Sample collection was then performed every 48 h for the remainder of the treatment cycle and the post treatment cycle. Serum concentrations of progesterone and plasma concentrations of 15-keto-13, 14-dihydroprostaglandin F(2alpha) (PGFM) were determined by radionmmunoassay. Intrauterine infusion of endotoxin had no effect on the cycle length or on hormone concentrations, while infusion of viable E. coli organisms tended to shorten the estrous cycle. Intravenous administration of endotoxin produced a sharp increase in both progesterone and PGFM concentrations, followed by a transient decrease in progesterone concentrations. Cycle length remained unchanged. It was concluded that the intact endometrium prevents the uptake of endotoxin although pathogenic E. coli organisms may disrupt the endometrial integrity sufficiently to shorten the estrous cycle by premature luteolysis. It is postulated that intravenous administration of endotoxin influences luteal function by the activation of the arachidonic acid cascade, by a direct effect on the corpus luteum, or via other mediators.     

Concentrations of oestradiol-17β and progesterone in the plasma of dairy heifers before and after cloprostenol-induced and natural luteolysis and during early pregnancy

Plasma oestradiol-17β and progesterone levels were measured in seven nulliparous, dairy heifers (British Friesian breed) that were administered cloprostenol (a synthetic analogue of prostaglandin F_2α) between days 8 and 14 of the oestrous cycle and inseminated (AI) 72 and 96 h later, and in seven heifers inseminated (AI) at natural oestrus.In both treated and untreated heifers, the beginning of the progesterone fall and the oestradiol-17β rise associated with luteolysis appeared to be synchronous but, whereas the rate of fall in progesterone level was greater for the treated heifers, that of the oestradiol-17β rise did not differ between treated and untreated heifers. Mean pre-ovulatory peaks of oestradiol-17β were 8 pg/ml and 10 pg/ml for treated and untreated heifers respectively.A post-ovulatory peak of oestradiol-17β in plasma 5–6 days after the pre-ovulatory peak occurred in all heifers whether or not conception had taken place. It is suggested that 7 days after the initiation of oestradiol-17β secretion by the pre-ovulatory follicle, another follicle begins to mature and secrete oestradiol-17β and that the progress of the latter towards full maturation and potential ovulation is stopped by rising progesterone levels from the corpus luteum; as a result in normal, non-pregnant cattle an interval of about 21 days elapses before another ovulation (of another follicle) takes place. In the event of premature luteolysis (in the present study induced between the 8th and 14th day) there is no evidence that the timing of this luteolysis influences the time taken for a follicle to enter the final stages of pre-ovulatory maturation, when increasing amounts of oestradiol-17β are secreted. Thus the interval between ovulations may not be less than 7 days but, depending on corpus luteum survival, may vary between 7 and 21 days.In one heifer after natural luteolysis a normal plasma oestradiol-17β peak followed but this was not associated with ovulation and corpus luteum formation. The second oestradiol-17β peak 6 days after the first, however, evidently assumed the ovulatory role; presumably the secreting follicle concerned, not being subject to inhibition by progesterone rising to luteal levels, matured fully and ovulated. Thus the second, normally post-ovulatory, oestradiol-17β peak in cattle can, in the event of failure of ovulation at the normal time, itself assume the ovulatory function, the oestrous cycle length then being about 28 days.     

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

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

Effect of treatment with estradiol valerate on endocrine changes and ovarian follicle populations in dairy cows

A linear-array ultrasound instrument was used to monitor the dynamics of follicular cyst formation following estradiol valerate (EV) administration in postpartum dairy cattle. Twelve cyclic cows were given two intramuscular (i.m.) injections of prostaglandin and F(2alpha) (PGF(2alpha)) 12 d apart to synchronize estrus. On Day 16 (Day 0 = day of estrus) six cows received 10 mg of EV in 1 ml sesame oil; the remaining six cows were treated with 1 ml sesame oil. The ovaries of all cows were scanned rectally each morning from Day 9 until 14 or 30 d post treatment. Plasma concentrations of luteinizing hormone (LH) and progesterone (P(4)) were also determined as objective indices of treatment effects. Day 0 to 16 ultrasound pictures of the ovaries of both control and treated cows were characterized by the presence of a corpus luteum (CL; 19 to 38 mm), several small follicles (<5 mm) and a medium-sized follicle (6 to 28 mm). Following treatment in control cows, the CL regressed gradually, and a preovulatory follicle was identifiable by Day 17 to 18, it increased in size and reached a maximum of 28 to 30 mm by Day 20 after ovulation and was identifiable throughout the rest of the cycle. Administration of 0 mg of EV resulted in a rapid reduction in the size of the CL. Growth of a large follicle was observed in all treated animals around Days 16 to 20, but having reached a maximum diameter of 12 to 24 mm it regressed without resulting in ovulation. Subsequent ultrasound pictures of EV-treated cows were characterized by the absence of a new CL and the presence of medium-sized persistent follicles. Estradiol valerate treatment induced early luteolysis (43 +/- 05 h post EV vs 101 +/- 22 h) and an LH surge (41 +/- 11 h vs 125 +/- 17 h).     

An intra-corpus luteum site for the luteolytic action of prostaglandin F2 alpha in the rhesus monkey

It has not been possible to demonstrate prostaglandin F2 alpha (PGF2 alpha) participation in primate luteolysis under conditions of systemic administration or of acute intraluteal injection. These study designs were hampered by the short biological half-life in the first instance and brevity of administration in the latter. In this study, luteolysis has resulted from chronic, intraluteal delivery of PGF2 alpha. Using the Alzet osmotic pump-cannula system, normally cycling rhesus monkeys were continuously infused, until menses occurred, with PGF2 alpha (10 ng/1/hr) directly into the corpus luteum (CL, n = 6), into the stroma of the ovary not bearing the corpus luteum (NCL, n = 3), or subcutaneously (SC, n = 5). An additional 5 monkeys received vehicle (V) into the corpus luteum. All experiments commenced 5-7 days after the preovulatory estradiol surge. Luteal function was assessed by the daily measurements of plasma progesterone, estradiol, and LH. Intraluteal PGF2 alpha caused premature functional luteolysis in all monkeys, as reflected by a highly significant decline in circulating progesterone and estradiol and the early onset of menstruation, when compared to the other groups. V, NCL, and SC infusions had no effect on either circulating steroid levels or luteal phase lengths. None of the experimental groups showed any change in plasma LH concentrations. These are the first data to indicate that PGF2 alpha can induce functional luteolysis in the primate, and the site of action appears to be the corpus luteum.     

Resumption of follicular activity in the early post-partum period of dairy cows

Lactating Friesian dairy cows (2nd-4th parity) which calved in spring (N = 7) or autumn (N = 15) were used. Their ovaries were examined by ultrasound scanning and blood samples were obtained daily for progesterone and oestradiol concentrations from the 5th day after calving until the first post-partum ovulation occurred. Five autumn-calving cows selected at random were bled every 15 min over a 6-h period on 1 day each week for 4 weeks after calving to assess the patterns of LH secretion. Follicular development during the post-partum anoestrous period was characterized by the growth and regression of small (less than or equal to 4 mm) and medium-sized (5-9 mm) follicles, until a dominant follicle (greater than 10 mm) was detected. The first detected dominant follicle ovulated in 14 cows, became cystic in 4 cows (all in autumn), and failed to ovulate in 1 cow. It was not possible to detect a dominant follicle in 3 cows due to scanning difficulties. The post-partum interval to detection of the first dominant follicle (mean +/- s.d.) was shorter (P less than 0.05) in autumn (6.8 +/- 1.8 days) than in spring (20 +/- 10.1 days). However, there was no significant difference between the respective intervals to first ovulation (autumn 27.4 +/- 25.9 and spring 27.3 +/- 18.9 days). Autumn-calved cows which had cysts had longer (P less than 0.001) intervals to first ovulation (58.2 +/- 23.5 days) than did normal cows (12.0 +/- 2.5 days). All cows with cysts had twin ovulations at their first post-partum ovulation. A pulsatile pattern of LH secretion was detected in the first week post-partum and LH pulse frequency was 2-3 per 6-h period in Weeks 1 and 2 post partum and increased to 5-7 pulses per 6-h period in the presence of a dominant or cystic follicle. Concentrations of progesterone in plasma during post-partum anoestrus were usually low (less than 0.2 ng/ml); oestradiol concentrations were also low (less than 5 pg/ml), but higher values (5-110 pg/ml) were observed in cows that had a dominant or a cystic follicle.     

Effects of intrauterine infection on the function of the corpora lutea formed after first postpartum ovulations in dairy cows

This study was carried out to determine the relationship between postpartum intrauterine infections, endocrine patterns and the function of corpora lutea formed following the first postpartum ovulations in dairy cows. Blood samples were collected daily starting from the day of parturition until 30 d after parturition or until the second postpartum estrus, whichever occurred first. Sera were assayed for progesterone (P(4)), prostaglandin F(2alpha) metabolite (PGFM), and luteinizing hormone (LH) concentrations. Palpations per rectum and real-time ultrasound scanning of the reproductive tracts were carried out in all cows once every 4 d for 1 mo, starting from Day 4 after parturition. In addition, endometrial swabs were collected aseptically from each cow once every 4 d during the first month postpartum. The swabs were cultured for aerobic and anaerobic bacteria. Twelve cows (60%) exhibited short estrous cycles (SC; 6 to 14 d long) following first postpartum ovulations. The mean preovulatory LH surges and LH patterns during the first postpartum cycles were similar in both groups, leading us to believe that lack of luteotrophic stimulation was not a factor in the occurrence of SC. Bacterial isolations were frequent in SC cows. The occurrence of moderate to heavy bacterial growth patterns and the repeated isolations of the similar organisms during postpartum suggests the persistence of uterine infections in SC cows. Increases in PGFM concentrations prior to luteolysis in SC cows were associated with moderate to heavy infection. Thus, postpartum uterine infections do not appear to affect ovulations, but prostaglandin (PGF(2alpha)) released in response to uterine infection may contribute to early demise of the corpus luteum formed after the first postpartum ovulation.     

Sequential Hormonal Changes in the Postpartum Dairy Cow

Peripheral plasma levels of 15-keto-13,14-dihydro-PGF2α, progesterone, Cortisol, LH and prolactin were studied in 6 primiparous postpartum dairy cows. The cows were followed by hormone measurements and clinical examinations from parturition until pregnancy was established. Blood was collected 3 times per day. The cervix, uterus and ovaries were examined by rectal palpation at 6–10 days intervals. The cows were observed for signs of oestrus twice daily and were additionally teased with a bull to provoke standing heat. Four cows had a normal parturition and dropped their fetal membranes shortly afterwards. (NR group). The remaining 2 retained their fetal membranes for more than 24 h following parturition (RFM group). One out of 6 cows showed standing oestrus at the first ovulation, 4 animals were in oestrus at the second ovulation and all cows showed signs of oestrus at the third ovulation. Although the length of the first luteal phase varied from 9 to 22 days a corpus luteum was in all cases palpated. The secretion of progesterone during the first luteal phase was terminated by a PGF2α release. A significant difference in 15-keto-13,14-dihydro-PGF2α levels between the 2 groups was found on days 0–4 (2.39 vs 6.87 nmol/1 at Ρ < 0.06). Postpartum prostaglandin F2α release as reflected by the level of 15-keto-13,14-dihydro-PGF2α lasted shorter in the NR group than in the RFM group (15–17 vs 21 days). Significant positive correlations between 15-keto-13,14-dihydro-PGF2α and Cortisol as well as between prolactin and Cortisol during the first 24 days postpartum were noted only in cows having normal parturition. The most pronounced daily prolactin variations occurred during the second luteal phase (NR group), when a significant difference between the times 8.00, 12.00 and 15.00 was recorded (14.7, 31.5 and 19.7 μg/l, respectively). Moreover, a partial negative correlation between log value of prolactin and arithmetical value of LH was found in these cows only during the first luteal phase after parturition.     

Conception rates and serum progesterone concentration in dairy cattle administered gonadotropin releasing hormone 5 days after artificial insemination

The objectives of this study were to determine the effect of administration of exogenous GnRH 5days after artificial insemination (AI) on ovarian structures, serum progesterone concentration, and conception rates in lactating dairy cows. In experiment 1, 23 Holstein cows were synchronized using the Ovsynch protocol. Five days after AI (day 0) cows were assigned randomly to receive either saline (saline; n=11) or 100microg GnRH (GnRH; n=12). To examine ovarian structures, ultrasonography was performed on day 1 and every other day beginning on day 5 until day 13. On days 5 and 13 blood samples were obtained to measure serum progesterone concentrations. All cows in the GnRH-treated group developed an accessory corpus luteum (CL), whereas cows in the saline group did not. Mean serum progesterone concentrations did not differ between GnRH and saline groups on day 5 (1.64+/-0.46ng/ml versus 2.04+/-0.48ng/ml). On day 13 serum progesterone concentrations were greater (P<0.05) in the GnRH group compared with saline (5.22+/-0.46ng/ml versus 3.36+/-0.48ng/ml). In experiment 2, 542 lactating cows, at two different commercial dairies, were used to test the effect of administering GnRH 5 days after AI on conception rates. Cows were synchronized and detected for estrus according to tail chalk removal. Cows detected in estrus received AI within 1h after detection of estrus. Five days after AI, cows were assigned randomly to receive either GnRH (n=266) or saline (n=276). Pregnancy status was determined by palpation per rectum of uterine contents approximately 40 days after AI. There was no effect of farm on conception rate. There was no effect of treatment as conception rates did not differ between GnRH and saline groups (26.7% GnRH versus 24.3% saline). Regardless of treatment, days in milk, parity, milk yield, and number of services had no effect on the odds ratio of pregnancy. In summary, the results of this study indicated that GnRH administered 5 days after AI increased serum progesterone by developing an accessory CL but did not improve conception rates in dairy cattle.     

Failure of exogenous LH to prevent PGF2α-induced luteolysis in beef cows

Henderson and McNatty (Prostaglandins 9:779, 1975) proposed that LH from the preovulatory LH surge attached to receptors on luteal cells and that this attachment might protect the early corpus luteum from PGF_2α induced luteolysis. To test this hypothesis, experiments were performed on heifers at day 10–12 of the cycle. Both jugular veins were catheterized and infusions of either saline (0.64 ml/min) or LH-NIH-B9 (10 μg/min; 0.64 ml/min) were given. Saline infusions were from 0–12 h; LH infusions were for 10 h and were preceded by a 2 h saline infusion. All animals were given 25 mg PGF_2α im at 6 h (6 h into the saline infusion and 4 h into the LH infusion). Blood samples were taken at 0.5 h, 1 h and 4 h intervals from 0–12, 13–18 h and 22–24 h respectively. Serum was assayed for LH and progesterone by radioimmunoassay methods. Two animals received saline and two received LH in each experiment. Eact treatment was replicated 6 times. LH infusion resulted in a mean serum LH of 57 ng/ml compared to 0.90 ng/ml in saline infused animals. This elevation of LH did not alter PGF_2α induced luteolysis as indicated by decline in serum progesterone. This experiment does not support the hypothesis that the newly formed corpus luteum is resistant to PGF_2α because of protection afforded by the protestrus LH surge.     

Superovulation of dairy cows with purified FSH supplemented with defined amounts of LH

This study investigated the effects of a purified follicle stimulating hormone (FSH) preparation supplemented with three different amounts of bovine luteinizing hormone (bLH) and a commercially available FSH with a high LH contamination on superovulatory response, plasma LH and milk progesterone levels in dairy cows. A total of 112 lactating Holstein-Friesian crossbred dairy cows were used for these experiments; the cows were randomly assigned to treatment groups consisting of purified porcine FSH (pFSH) supplemented with bLH. Group 1 was given 0.052 IU LH 40 mg armour units (AU) FSH (n = 6); Group 2 was given 0.069 IU LH (n = 32); Group 3 received 0.423 IU LH (n = 34); while Group 4 cows (n = 36) were superovulated with a commercially available FSH-P((R)). This compound appeared to contain 8.5 IU LH 40 mg AU FSH according to bioassay measurement. All animals received a total of 40 mg AU FSH at a constant dose twice daily over a 4-d period. Levels of milk progesterone and plasma LH were determined during the course of superovulatory treatment. The Group 1 treatment did not reveal multiple follicular growth, and no embryos were obtained. Superovulation of Group 3 cows resulted in significantly (P<0.05) more corpora lutea (CL; 12.6+/-1.1) and fertilized ova (5.1+/-1.3) compared with Groups 2 and 4 (10.1+/-0.9 and 2.6+/-0.6, 9.0+/-0.9 and 2.7+/-0.5, respectively). Due to a high percentage of degenerated embryos (33%) Group 3 yielded only one more transferable embryo than Groups 2 and 4. Among groups, LH levels differed in the period prior to induction of luteolysis and were similar thereafter. The progesterone pattern following FSH LH administration reflected the amount of LH supplementation. Milk progesterone levels on the day prior to embryo collection were correlated to the number of CLs and recovered embryos. It is concluded that under the conditions of our experiment superovulation with 0.423 IU LH 40 mg AU FSH may yield a significantly improved superovulatory response in dairy cows. It is further suggested that LH supplementation exerts its effects mainly on follicular and oocyte maturation during the period prior to luteolysis.     

Follicular dynamics during postpartum anestrus and the first estrous cycle in suckled or non-suckled Brahman (Bos indicus) cows

Brahman (Bos indicus) cows, were selected at 28+/-10 days after calving and analyzed by real time rectal ultrasonography three times a week, in order to evaluate and compare follicular and corpus luteum development during postpartum (PP) anestrus and the first PP estrous cycle under sylvopastoril conditions. Suckling (S, n=11) or non-suckling (NS, n=5) cows were evaluated in a zone of tropical dry forest (450m of altitude, mean temperature=27 degrees C, annual rainfall=1000mm). Estrous detection was performed twice daily by direct observation. Progesterone was quantified using RIA. From 28+/-10 days postcalving to resumption of estrous cycles, there were no differences (P>0.05) between NS and S cows for diameter of the dominant or first subordinate follicle, follicular growth rate, or interdominance interval. Silent ovulation, corpus luteum formation and subsequent progesterone concentrations ranging from 0.3 to 9. 7ng/ml, were found in both groups. The first calving to ovulation and calving to standing estrus intervals were shorter (P<0.01) in NS (34.8+/-5.81 and 41.2+/-9.03 days) than in S (65+/-4.82 and 81+/-6. 21 days) cows. Follicular development and progesterone concentrations during the first PP estrous cycle did not differ (P>0. 05) between NS and S cows. These results suggest that Brahman cows could have an early PP resumption of follicular recruitment if fed under sylvopastoril system conditions. However, non-suckled cows did have an earlier standing estrus and ovulation than did suckled cows.