Maintenance of pregnancy in postpartum beef cows that have short-lived corpora lutea.

The first two experiments examined the role of the uterus in low pregnancy rates of beef cows induced to ovulate by early weaning. At 20 to 25 d postpartum, one-half of the cows in Exp. 1 and 2 received a s.c. implant containing 6 mg of norgestomet (NOR) for 9 d (NOR-pretreated) and the remaining cows were untreated controls (CON). Lengths of first postpartum luteal phase after weaning of calves at d 7 after implant insertion were expected to be normal in NOR-pretreated and short in CON cows. In Exp. 1, cows of both groups received an implant containing 3 mg of NOR at d 4 after first estrus and a silastic implant with 15 or 25 mg of NOR at d 7 after first estrus. At 7 d after first estrus, two embryos were transferred into the uterus of each cow and pregnancy was diagnosed by ultrasonography at d 35. Blood samples were collected daily from onset of treatment to d 8 after estrus and then every other day to d 24. Only 4 of 22 cows were pregnant at d 35, concentrations of estradiol (E2) were elevated after luteolysis, and large follicles were present at d 35. In Exp. 2, all cows were injected with 100 mg of progesterone (P4) twice daily from d 4 to 35 after first estrus. Embryos were transferred, pregnancy was diagnosed, and blood samples were collected as in Exp. 1, except blood sampling was continued to d 34.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of the previously gravid uterine horn and postpartum interval on follicular diameter and conception rate in beef cows treated with estradiol benzoate and progesterone

An experiment was performed to evaluate the effect of the side of ovulation with respect to the previously gravid uterine horn on fertility of cows inseminated at one of two periods postpartum. All cows were treated with an intravaginal progesterone insert for 7 d and received estradiol benzoate (2 mg, i.m.) at the time of device insertion, prostaglandin F2alpha (25 mg, i.m.) at the time of device removal, and estradiol benzoate (1 mg, i.m.) 30 h after device removal. All cows were inseminated 28 to 30 h after the second treatment with estradiol benzoate, regardless of observed estrus. Cows treated in Period 1 received inserts at 16 to 20 d postpartum and were inseminated at 25 to 29 d postpartum. Cows treated in Period 2 received inserts at 26 to 30 d postpartum and were inseminated at 35 to 39 d postpartum. Diameter of the largest follicle at insert removal was greater in cows treated in Period 2 (10.1 +/- 0.3; mm +/- SEM) than in cows treated in Period 1 (9.1 +/- 0.3; P < .05). Diameter did not differ with the side of ovulation in respect to the previously gravid uterine horn. Diameter was greater in cows 5 to 9 (10.3 +/- 0.3) than in cows 3 to 4 (9.0 +/- 0.3) or 10 to 13 (9.4 +/- 0.6) yr of age (P < .01). The proportion of cows that ovulated from the ovary contralateral to the previously gravid uterine horn was greater (P < .05) than of those that ovulated from the ipsilateral ovary, and the incidence of ovulation was reduced in cows 3 to 4 yr of age (P < .01). Conception rate tended to be greater for ovulation from the ipsilateral compared with the contralateral ovary, relative to the previously gravid uterine horn (P < .10) and for ovulation from the right than the left ovary (P < .06). Conception rate was less if cows ovulated a follicle that was < 9 mm than a follicle > or = 9 mm in diameter at insert removal (P < .01) and was greater in cows inseminated in June than in April or May (P < .05). In conclusion, in cows in which estrus was synchronized at 25 to 39 d postpartum, ovulation from either the ovary ipsilateral to the previously gravid uterine horn, or the right ovary, tended to increase fertility.     

A simple pharmacokinetic model linking plasma progesterone concentrations with the hormone released from bovine intravaginal inserts

On the basis of pharmacokinetic modeling, this study provides some insights into predicting in vivo plasma progesterone concentrations when using bovine intravaginal inserts for systemic progesterone delivery. More significantly, this contribution is the first attempt to build a simple pharmacokinetic model that links plasma progesterone concentrations with the hormone released from bovine intravaginal inserts. After evaluating three rival pharmacokinetic models and considering some phenomena involved in the intravaginal administration of progesterone, a primary pharmacokinetic model having a good data fitting capability with only two adjustable parameters is proposed to the above mentioned task. Kinetic parameters are given for lactating Holstein dairy cows with two levels of daily milk yields; and non-pregnant, non-lactating Holstein-Friesian cattle. Model predictions indicate the occurrence of a preferential distribution of the intravaginally administered progesterone dose through a first uterine pass effect.     

Synchronisation of oestrus and reproductive performance of dairy cows following administration of oestradiol benzoate or gonadotrophin releasing hormone during a synchronised pro-oestrus

OBJECTIVE: To compare the use of gonadotrophin releasing hormone (GnRH) and oestradiol benzoate (ODB) administered following a synchronised pro-oestrus on reproductive performance of lactating dairy cows and the submission rates of non-pregnant cows following resynchronisation. DESIGN: Cohort study. PROCEDURE: Lactating Holstein cows enrolled in a controlled breeding program were first treated with an intravaginal progesterone releasing insert (IVP4) for 8 days, 2.0 mg of ODB intramuscular (i.m.) at device insertion (Day 0), an analogue of PGF2alpha at device removal and either 1.0 mg of ODB i.m., 24 h after device removal (ODB group, n = 242), or 0.25 mg of a GnRH agonist (GnRH group, n = 152) injected i.m. approximately 34 h after device removal. Every cow was artificially inseminated between 49 and 56 h after removal of its insert (Day 10). Cows detected in oestrus 1 day after artificial insemination (AI) that were not detected in oestrus on the previous day were re-inseminated on that day. All cows treated on Day 0 were resynchronised for reinsemination by insertion of a used IVP4 device on Day 23. Oestradiol benzoate at a dose of 1.0 mg was administered i.m. at the time of device insertion. Inserts were removed 8 days later (Day 31) and 1.0 mg of ODB was injected i.m. 24 h later. Those cows detected in oestrus between Days 31 and 35 were artificially inseminated. On Day 46 these cows were resynchronised for a third round of AI by insertion of an IVP4 device, used previously to synchronise cows for the first and second rounds of AI, and administration of 1.0 mg of ODB i.m.. Eight days later inserts were removed. Cows detected in oestrus between Days 54 and 58 were artificially inseminated. Bulls were run with the herd between rounds of AI and removed after 21 weeks of mating. RESULTS: Treatment with ODB or GnRH at the first synchronised pro-oestrus did not significantly alter the reproductive performance over three rounds of AI or over a 21-week breeding period. Treatment also did not alter submission rates at the second round of AI or the proportion of non-pregnant and non-return cows ('phantom' cows) detected and did not result in significant differences in concentrations of progesterone in plasma 10 and 18 days after removal of inserts at the first round of AI. Treatment with GnRH reduced the proportion of cows detected in oestrus at the first round of AI (36.2 vs 97.5%; P < 0.001). CONCLUSION: Administration of GnRH compared to ODB at a synchronised pro-oestrus results in similar reproductive performance. Treatment with GnRH reduced the proportion of cows detected in oestrus following treatment. This may offer advantages to the way AI is managed by enabling insemination at a fixed-time and removing the need for the detection of oestrus.     

Clinical response of ovarian cysts in dairy cows after PRID treatment

We investigated the therapeutic effects of a progesterone releasing intravaginal device (PRID) on cystic ovarian disease (COD) and reproduction performance of cows. The possible influence of PRID on metabolic and/or health status was also examined. A total of 40 Holstein-Friesian cattle, with ovarian cystic structures, > or =2.5 cm in diameter, persisting for more than 7-14 days, without a corpus luteum (CL) were used for the study. PRID or placebos were inserted into the vagina for 12 days. Five animals lost the intravaginal device before removal and one was culled. Based on plasma progesterone concentration on the day of treatment, 20 (17 PRID and 3 placebos) of the remaining 34 cows had follicular cysts (progesterone < or =1 ng/ml) and 14 (10 PRID and 4 placebos) had luteal cysts (progesterone >1 ng/m l). Fourteen (82%) of the PRID-treated follicular cystic cows responded with formation of a CL within 14 days after treatment, and an overall conception rate of 53.8%. Likewise, 70% of the treated luteal cystic cows responded with CL formation and 71.4% conception rate. No significant differences were observed in hematocrit (Ht), white blood cell count and serum levels of glucose, blood urea nitrogen, aspartate aminotransferase, and alanine aminotransferase, between the day of PRID insertion and removal, in animals with follicular and luteal cysts. PRID treatment resulted in ovulation 2-4 days later and formation of a CL in cows that recovered.     

The effect of progesterone alone or in combination with estradiol on follicular dynamics, gonadotropin profiles, and estrus in beef cows following calf isolation and restricted suckling

The effects of calf isolation and restricted suckling on LH pulse characteristics and interval to first ovulation (postpartum interval) were studied in 52 multiparous beef cows, with or without exogenous progesterone. At 30 d postpartum, cows were randomly allocated to one of four treatments (n = 13/treatment): 1) Ad lib, ad libitum access of cows to calves; 2) CI/RS, calf isolation/restricted suckling, where suckling was restricted to once daily; 3) CI/RS+P4, same as CI/RS but cows received an intravaginal progesterone-releasing device at calf isolation for 6 d; or 4) CI/RS+P4+E2, as CI/RS+P4 but the intravaginal progesterone-releasing device had a 10-mg estradiol capsule attached. Daily ovarian scanning and twice-daily blood sampling were performed from d 25 postpartum until the day of second ovulation. A random sample of cows from each treatment (n = 31 in total) were blood-sampled at 15-min intervals for 10 h on d 29, 32, 35, and 38. Ovulatory response to treatment was regarded as ovulation of either the dominant follicle growing at d 30 or the subsequent DF. There was a treatment x day effect (P = .09) on LH pulse frequency, but neither progesterone (CI/ RS+P4) nor progesterone and estradiol (CI/RS+P4+E2) treatment suppressed the calf isolation/restricted suckling-induced increase in LH pulse frequency. The estradiol capsule (CI/RS+P4+E2) delivered sufficient estradiol to delay new follicle wave emergence (treatment x stage; P < .001) and the associated preemergence increase in concentrations of FSH (treatment, P < .05) in cows treated at the postselection stage of follicle wave development, prolonging dominance of the dominant follicle present at treatment initiation (P < .001). The number of cows that ovulated in response to treatment was greater (P < .001) in cows with calf isolation/restricted suckling than in cows suckled ad libitum. Hence, cows assigned to the Ad lib treatment had a longer postpartum interval (P < .001) than cows of the other treatments. Exogenous progesterone treatment increased the frequency of cows exhibiting clinical signs of estrus at first ovulation (P < .001) and reduced the frequency of short estrous cycles (P < .001). We conclude that, in beef cows with calves, a 6-d progesterone treatment does not suppress the calf isolation/restricted suckling-induced increase in LH pulse frequency. Hence, on progesterone withdrawal, the LH pulse frequency is sufficient to stimulate first ovulation, accompanied by overt estrous expression and elimination of a short estrous cycle in most cows.     

Influence of estradiol,progesterone, and nutrition on concentrations of gonadotropins and GnRH receptors,and abundance of mRNA for GnRH receptors and gonadotropin subunits in pituitary glands of beef cows

Nutritionally induced anovulatory cows (n = 28) were used to determine the effect of steroids on regulation of synthesis and secretion of gonadotropins. Anovulatory cows were ovariectomized and received intravaginal inserts containing estradiol (E2), progesterone (P4), E2 and P4 (E2P4), or a sham intravaginal insert (C) for 7 d. Concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were quantified in serum and E2 and P4 were quantified in plasma. Cows were exsanguinated within 1 to 2 h after removal of intravaginal inserts and pituitary glands were collected and stored at -80 degrees C until messenger ribonucleic acid (mRNA) for gonadotropin-releasing hormone receptor (GnRH-R) and gonadotropin subunits, pituitary content of GnRH-R, and LH and FSH were quantified. Pituitary glands from five proestrous cows were harvested to compare gonadotropin characteristics between ovariectomized, anovulatory cows and intact cows. Plasma concentrations of E2 were greater (P < 0.05) in E2-treated cows than in sham-treated cows. Concentrations of P4 were greater (P < 0.05) in cows treated with P4 than in sham-treated cows. Mean serum concentrations of LH and FSH were not significantly influenced by steroid treatments. However, frequency of LH pulses of ovariectomized, nutritionally induced anovulatory cows was increased (P < 0.05) by treatment with E2 and amplitude of LH pulses was greater (P < 0.05) in cows treated with E2 or P4 than in cows treated with E2P4 or sham-treated. Quantity of mRNA for LHbeta in the pituitary gland was greater when cows were treated with P4. Concentrations of LH in the pituitary gland were not affected by steroid treatments; however, pituitary concentrations of FSH were less (P < 0.1) in E2 cows than in sham-treated cows. The number of GnRH-R was increased (P < 0.05) in cows treated with E2, but P4 treatment did not influence the number of GnRH-R. Abundance of mRNA for GnRH-R, common alpha-subunit, and FSHbeta were not affected by treatments. Pituitary concentrations of LH were greater (P < 0.05) and concentrations of FSH were less (P < 0.05) in proestrous cows than in ovariectomized, anovulatory cows treated with or without steroids. Abundance of mRNA for GnRH-R, common alpha-subunit, LHbeta and FSHbeta were similar for proestrous and anovulatory cows. We conclude that treatment of nutritionally induced anovulatory cows with progesterone and estradiol may cause pulsatile secretion of LH.     

Efficacy of intravaginal progesterone administration combined with prostaglandin f(2alpha) for cystic ovarian disease in Japanese Black cows

The objective of this study was to investigate cystic ovarian disease (COD) in commercial Japanese Black cows and to evaluate the efficacy of 7-day insertion of an intravaginal progesterone insert (CIDR) combined with prostaglandin F(2alpha) (PGF(2alpha)) at CIDR removal. Experiment 1 was conducted to group cysts into 4 patterns based on alteration of plasma progesterone (P(4)) concentrations on d -7 and d 0 (=CIDR insertion) with 1.0 ng/ml as the cut-off level by ultrasonographic examination of 28 cows with COD that were >or=40 days postpartum and anoestrous after calving. In Experiment 2, a total of 55 cows under the same conditions as in Experiment 1 were utilized, and the same regimen as in Experiment 1 was performed without 7 days of pre-observation before treatment. As a result, 92.9% of CLs on d 21 were highly formed in Experiment 1 and 83.6% were highly formed in Experiment 2. The conception rates within 60 days after CIDR removal were also satisfactory high and were 71.4% and 54.5%, respectively. There were no differences in any overall reproductive parameters between Experiments 1 and 2 (P>0.05). The average days between CIDR removal and conception were 24.4 +/- 5.3 and 24.0 +/- 6.5 days, respectively (P>0.05); therefore, the conception dates of the cows in Experiment 2 were at least 7 days earlier compared with Experiment 1. In conclusion, treatment with a CIDR and PGF(2alpha) against COD could minimize the risk of incorrect treatment and provide sufficient reproductive performance in Japanese Black cows.     

Response of postpartum beef cows to exogenous progestogens and gonadotropin releasing hormone

Plasma progesterone (P4) profile and estrous detection were used during three experiments to evaluate the effects of exogenous progestogens on the life span of gonadotropin releasing hormone (GnRH)-induced corpora lutea (CL) in postpartum (pp) beef cows. Experiment 1 utilized primiparous fall-calving cows (n = 28, trial 1); and spring-calving cows (n = 29, trial 2). On d 18 to 27 pp (d 0) all cows received intravaginal devices containing either P4 or no P4 (NP) for 5 d. On d 5 the devices were removed and calves were either removed (CR) or were present (CP) with half of the cows within steroid group. At 50 h after device removal, 500 micrograms of GnRH was given (iv) to all cows, and weaned calves were reunited with their dams. The induced CL had a normal life span (greater than 16 d) in 17 and 86% (trial 1) and 8 and 79% (trial 2) of NP and P4 cows, respectively. Calf removal did not affect (P greater than .10) the life span of the CL. In Exp. 2, spring-calving multiparous cows (d 18 to 24 pp; d 0) received either no P4 (NP; n = 19), P4 for 6 d via intravaginal devices (P4H; n = 19) or a single im injection of 300 mg P4 (P4 IM; n = 18). At 48 h after device removal or at 8 d after the injection of P4, half of the cows within steroid group received either 500 micrograms GnRH or saline. Corpora lutea had a normal life span in 0, 11, and 80% of NP, P4 IM and P4H cows, respectively, that received GnRH and in 22% of P4-saline cows. In Exp. 3, fall-calving multiparous and primiparous cows (d 25 to 31 pp) received either no progestogen (NP; n = 20), P4 via intravaginal devices for 5 d (P4H; n = 21) or melengestrol acetate (MGA; .5 mg.head-1.d-1 for 5 d orally, n = 15). At 48 d after device removal or at 72 h after the last MGA feeding, all cows received 500 micrograms GnRH. Progesterone post-GnRH injection was increased (greater than 1 ng/ml) at d 7 in 64, 100 and 100%, and remained elevated at d 14 in 11, 46 and 100% of NP, MGA and P4H cows, respectively. For all experiments plasma P4 was increased (range 2 to 5 ng/ml) when the devices containing P4 were in place, then decreased (less than 1 ng/ml) by 48 to 50 h after device removal.(ABSTRACT TRUNCATED AT 400 WORDS)     

Optimizing the use of sex-sorted sperm in timed artificial insemination programs for suckled beef cows

Three experiments were designed to evaluate methods to optimize the use of sex-sorted sperm in timed AI (TAI) programs for suckled beef cows. In all 3 experiments, suckled Bos indicus cows were synchronized using an intravaginal progesterone (P4) device during 8 d and a 2.0-mg injection of intramuscular estradiol benzoate (EB) at device insertion. The females received PG and eCG (300 IU) at P4 device removal and 1.0 mg of EB 24 h later. The cows were inseminated 60 to 64 h after P4 device withdrawal. All cows had their ovaries scanned by transrectal ultrasound at TAI to indentify and to measure the largest follicle (LF) present. In Exp. 1, a total of 853 cows had their LF classified as <9 mm or ≥9 mm at the time of TAI; these cows were then randomly assigned to 4 groups according to their LF diameter (<9 mm or ≥9 mm) and the type of sperm used (sex-sorted or non-sex-sorted). There was an interaction (P = 0.02) between the type of sperm and LF diameter beginning at TAI[non-sex-sorted ≥9 mm = 58.9%a (126/214); non-sex-sorted <9 mm = 49.5%b (106/214);sex-sorted ≥9 mm = 56.8%ab (134/236); and sex-sorted <9 mm = 31.2%c (59/189), a≠b≠c = P < 0.05]. In Exp. 2, suckled cows (n = 491) were classified immediately before TAI as having displayed estrus or not (estrus or no estrus) between P4 device removal and TAI. These cows were randomly assigned to 4 groups according to the occurrence of estrus and the type of sperm (sex-sorted or non-sex-sorted). There were effects of the occurrence of estrus (P = 0.0003) and the type of sperm (P = 0.05) on pregnancy per AI [P/AI; no estrus, non-sex-sorted = 43.6% (27/62); estrus, non-sex-sorted = 58.5%; (107/183); no estrus, sex-sorted = 33.9% (21/62), and estrus, sex-sorted = 50.0% (92/184)]; however, no interaction between the occurrence of estrus and type of sperm was observed (P = 0.87). In Exp. 3, a total of 200 suckled cows presenting LF ≥9 mm at TAI were randomly assigned to receive sex-sorted sperm deposited into the uterine body (n = 100) or into the uterine horn ipsilateral to the recorded LF (n = 100). No effect of deeper AI on P/AI was found (P = 0.57). Therefore, the LF diameter at TAI and the occurrence of estrus can be used as selection criteria to identify cows with greater odds of pregnancy to receive sex-sorted sperm in TAI programs. In addition, performing TAI with sex-sorted sperm deeper into the uterus did not alter the pregnancy results.     

The therapeutic effects of a progesterone-releasing intravaginal device (PRID) with attached estradiol capsule on ovarian quiescence and cystic ovarian disease in postpartum dairy cows

The objective of this study was to evaluate the effects of a progesterone-releasing intravaginal device (PRID) containing an estradiol benzoate capsule on ovarian dysfunction, including ovarian quiescence, follicular cyst (FC) and luteal cyst or cystic corpus luteum (LC/CCL), in postpartum dairy cows. These ovarian dysfunctions were examined by palpation per rectum relative to plasma progesterone status. The results of clinical examination and hormone assay determined ovarian quiescence in 13 cows, FC in 15 cows and LC/CCL in 7 cows. These cows were treated with PRID for 12 d and then clinical examination was performed. After PRID removal, the proportion of cows exhibiting estrous signs within 7 d and confirmed formation of CL within 7-14 d (markedly effective) were 69.2 % (n=9) for ovarian quiescence, 46.7 % (n=7) for FC, and 28.6 % (2 cows) for LC/CCL. Two cows (15.4 %) in ovarian quiescence, 5 cows (33.3%) with FC and 4 cows (57.1 %) with LC/CCL did not exhibit estrous signs but were recognized as having formed CL within 12-16 d after removal of PRID (effective). These results suggest that treatments of PRID with estradiol benzoate for 12 d have therapeutic efficacy on ovarian dysfunction including ovarian quiescence, FC and LC/CCL in postpartum dairy cows.     

GnRH treatment at CIDR insertion influences ovarian follicular dynamics in Japanese black cows

Ovarian follicular dynamics and estrous synchronization after Gonadotropin-releasing hormone (GnRH) treatment at Controlled Internal Drug Releasing device (CIDR) insertion were investigated in Japanese Black cows. CIDR was inserted for eight cows at 7 days after estrus. Cows were allocated to either Group A: 8-day CIDR insertion with GnRH treatment on d 0 (n=4, d 0=CIDR insertion) or Group B: 8-day CIDR insertion (n=4). Both groups were injected with prostaglandin F2alpha (PGF2alpha) on d 7. Ultrasonography and blood sampling were performed twice daily. Intensive sampling was performed every 15 min for 8 hr to determine the pulsatile release of LH on d -1, d 5 and d 10. Three of four cows showed intermediate ovulation within 2 days after GnRH treatment during CIDR insertion in Group A, whereas no ovulation was found in Group B. Three of four cows in Group A and all four cows in Group B ovulated after CIDR removal. Plasma progesterone concentrations from d 3 to d 7 in three intermediate ovulatory cows in Group A (8.4 +/- 1.6 ng/ml) was significantly higher than those in Group B (4.1 +/- 1.2 ng/ml; 4 cows) during CIDR insertion (P<0.01). Interval to estrus and ovulation after CIDR removal was observed at 60.0 +/- 12.0 hr and 76.0 +/- 6.9 hr in three cows in Group A, and 75.0 +/- 15.1 hr and 93.0 +/- 20.5 hr in Group B, respectively. There was a significant increase in LH pulse frequency on d 10 compared on d -1 or d 5 in both groups (P<0.05), in addition those on d 10 in Group A tended to be higher than in Group B. As a result, GnRH treatment at CIDR insertion at 7 days after estrus induced intermediate ovulation with formation of corpus luteum (CL) and rather synchronized emergence of ovulatory follicle during CIDR insertion. These induced CL increased plasma progesterone concentrations and contributed to precise synchronization.     

Effects of suckling on oestradiol benzoate induced LH release in post-partum, ovariectomised cows

Twelve suckling and 12 non-lactating post-partum cows were treated with a progesterone-releasing pessary for 7 or 14 d followed by an injection of 500 micrograms oestradiol benzoate (ODB) 24 h after pessary removal or no injection. Suckling had no significant effect on plasma progesterone concentrations (plasma P4) or plasma luteinising hormone concentrations (plasma LH) during pessary insertion. After pessary removal plasma LH rose in response to ODB. The extent of the rise was similar in suckling and non-lactating cows treated with progesterone for 14 d but was significantly lower in non-lactating than suckling cows (P less than 0.025) treated for 7 d. These findings indicate that suckling increases the responsiveness of plasma LH in post-partum cows following progesterone and oestrogen treatment compared to non-lactating cows and that extended treatment with progesterone can remove this difference.     

The reproductive performance of dairy cows with anovulatory anoestrus that were injected with either gonadotrophin-releasing hormone or oestradiol benzoate as part of a re-treatment process after insemination

This experiment compared the reproductive performance of synchronised anoestrous dairy cows that were treated initially with a combination of progesterone and oestradiol benzoate and then with either gonadotrophin-releasing hormone (GnRH) or oestradiol benzoate to resynchronise returns to service. It was hypothesised that injecting anoestrous dairy cows with GnRH 12-15 days after insemination and coinciding with the time of insertion of a controlled intravaginal progesterone-releasing (CIDR) device would increase conception rates to the preceding 1st insemination compared with oestradiol benzoate treated cows; both GnRH and oestradiol benzoate would resynchronising the returns to service of those cows that did not conceive to the preceding insemination. Groups of cows in 11 herds were presented for a veterinary examination after they had not been seen in oestrus postpartum. Those cows diagnosed with anovulatory anoestrus (n = 1112) by manual rectal palpation and/or ultrasonography were enrolled in the trial. Each enrolled cow was injected with 2 mg oestradiol benzoate i.m. on Day -10, (where Day 0 was the 1st day of the planned insemination) concurrently with vaginal insertion of a CIDR device. The device inserted was withdrawn on Day -2 and then each cow injected i.m. with 1 mg of oestradiol benzoate on Day -1 unless it was in oestrus. Observation for oestrus preceded each insemination. Every cow that had been inseminated on Days -1,0,1 or 2 was presented for treatment for resynchrony on Day 14 (n = 891). They were divided into 2 groups; those with an even number were each injected i.m. with 250 microg of a GnRH agonist (Treatment group n = 477); each of the cows with an odd number injected i.m. with 1 mg of oestradiol benzoate (control group, n = 414). Each GnRH or oestradiol benzoate injection preceded reinsertion of a CIDR device previously inserted from Days -10 to -2. It was withdrawn on Day 22, 24 hours before injecting 1 mg oestradiol benzoate. Cows observed in oestrus were submitted for a 2nd insemination. Every enrolled cow still present in the herd was pregnancy tested by palpation of uterine contents per rectum about 6 weeks later and again at the end of a herd's seasonal breeding programme. The alternative use of GnRH instead of oestradiol benzoate did not affect the percentage of cows conceiving within 3 days of the mating start date (MSD) (35.6 % vs 35.3 %, P = 0.90), resubmission rates for a 2nd insemination among cows not pregnant to the 1st insemination (81.6 % vs 83.5 %, P = 0.41), 6-week pregnancy rate (59.3 % vs 60.6 %, P = 0.65), 21-weekpregnancy rate (86.6 vs 85.0, P = 0.36), mean interval from MSD to conception (32.5 +/- 1.8 days vs 29.9 +/- 1.8 days, P = 0.26) or conception rate of cows reinseminated by Day 28 (43.3 % vs 38.8 %, P = 0.39). When GnRH was compared with oestradiol benzoate, it did not increase conception rates to the 1st service; it was as effective as oestradiol benzoate in synchronising returns to service in previously treated anoestrous cows that did not conceive to the 1st service. Its use affected neither conception rates to the preceding 1st inseminations nor to the following 2nd inseminations.     

Hormonal, estrual, ovulatory and milk traits in postpartum dairy cows following multiple daily injections of oxytocin

Release of oxytocin at suckling or milking may delay onset of estrous cycles in postpartum cows. Twenty lactating Holsteins of mixed parity were given 100 mU oxytocin iv (n = 10) or 2 ml saline (control; n = 10) via jugular catheters at 0530, 0930, 1730 and 2130 daily from calving (d o) until 28 d postpartum. All cows were milked twice daily at 0130 and 1330. Blood was collected thrice weekly (Monday, Wednesday, Friday at 0530) for 12 wk and analyzed by radioimmunoassay for progesterone and 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM) in serum. On d 12, blood was collected every 15 min for 6 h via jugular catheters and concentrations of luteinizing hormone (LH), cortisol and PGFM were determined. Rate of involution of the reproductive tract was estimated twice weekly by palpation per rectum. Overall mean, baseline concentrations, number of pulses/6 h, and pulse duration of LH on d 12 were similar among treatment groups. However, oxytocin seemed to reduce (P less than .10) pulse amplitude of LH in multiparous cows (.4 +/- .2 vs .8 +/- .1 ng/ml), but not in primiparous cows. Concentrations of cortisol and PGFM in serum on d 12 were unaffected by treatment. The average intervals from calving to first ovulation, based on changes of progesterone in serum and the intervals to first estrus, were similar between treatment groups. Rates of involution of the cervix and uterus also were similar between treatments. Milk yield, percent protein in milk and somatic cell counts did not differ between treatment groups. However, percent fat in milk tended to be higher (P less than .10) in cows given oxytocin than in controls (3.99 +/- .22 vs 3.68 +/- .21). These data indicate that multiple daily injections of oxytocin did not affect: 1) length of anestrus and anovulation in postpartum dairy cows, 2) LH release and 3) rates of cervical and uterine involution.     

Impact of ovarian and uterine conditions on some diagnostic tests output of endometritis in postpartum high-yielding dairy cows

The effect of ovarian predominating structures and uterine condition on the result of some diagnostic tools for the evaluation of endometritis was studied in postpartum (pp) Holstein-Friesian dairy cows (n = 58). Endometrial cytology (EC) and the evaluation of vaginal mucus by vaginoscopy or Metricheck were performed weekly from week 3 to 7 pp. The ovarian studies involved the predominating structures including cystic follicles with plasma progesterone (P(4) ; more or <1 ng/ml; >23 mm), corpus luteum (CL), pre-ovulatory follicles (10-23 mm) and small follicles (<10 mm). The uterine conditions comprised uterine involution, tonicity and fluid in uterus (FIU) regarding echogenicity extent. During week 5, the percentage of polymorphonuclear neutrophils (PMN%) was higher (p < 0.05) in animals with pre-ovulatory follicles (mean ± SEM, 26.3 ± 7.6%) than animals having CL (11.0 ± 3.6%). In cystic ovaries, during week 5, PMN% was higher (p < 0.05) in follicular cysts with low progesterone (P(4) < 1 ng/ml; 9.3 ± 2.6%) than those with high P(4) (P(4) ≥ 1 ng/ml; 1.5 ± 1.1%). Moreover, PMN% was higher (p < 0.01) in animals with non-involuted uterus (11.5 ± 7.4%) than those with involuted uterus (2.7 ± 0.6%) during week 7 pp. The animals that had abnormal mucus determined by Metricheck was higher in animals with atonic uterus than those with tonic uterus during week 6 (82.6% vs 51.5%; p < 0.05) and 7 (71.4% vs 25.7%; p < 0.01) pp. In addition, by vaginoscopy, the prevalence of animals with abnormal discharge showing small follicles (100%, 5/5) during week 3 pp and pre-ovulatory follicles (40.0%, 8/20) during week 5 pp was higher (p < 0.05) when compared to those having CL during week 3 (33.3%; 1/3) and week 5 pp (7.7%; 2/26), respectively. In conclusion, endometrial cytology, Metricheck and vaginoscopy were influenced by the predominating various ovarian structures and uterine condition in early pp high-yielding dairy cows.     

Supplementing treated anoestrous dairy cows with progesterone does not increase conception rates

AIM: To determine whether conception rates of anoestrous dairy cows treated with progesterone and oestradiol benzoate (ODB) could be increased by treating them with additional progesterone following insemination at the induced oestrus. METHODS: Cows which had not been detected in oestrus for at least 21 days after calving in 18 herds were confirmed anovulatory anoestrus (AA) by veterinary examination, due to the absence of a detectable corpus luteum in the ovaries. All cows were treated with intra-vaginal progesterone (CIDR insert) for 6 days and injected with 1 mg ODB 24 h after insert removal (Day 0). Only cows which were seen in oestrus on Days 0, 1 or 2 were enrolled in the trial. These cows were either treated with a second CIDR insert on Day 8, for 7 days (P4+; n=422), or remained untreated (Control; n=756). Milk progesterone concentrations were measured in a subset of enrolled cows (n=669) on Day 8 to determine the proportion of cows that ovulated following the induced oestrus. RESULTS: Conception rates to first insemination were similar in P4+ and Control cows (40.3% and 37.2%, p=0.59). Of cows which had milk progesterone concentrations measured on Day 8, 78.6% displayed oestrus and ovulated, (range: 53.8% to 94.6% among herds). Of the cows that ovulated, conception rate to first insemination was 46.8% and 43.5% in P4+ and Control cows, respectively (p=0.86). CONCLUSION: Conception rates to first insemination in AA cows treated with progesterone and ODB were not increased by progesterone supplementation using CIDR inserts following insemination. KEY WORDS: dairy cattle, postpartum anoestrus, reproduction, progesterone treatment, CIDR insert.     

Ovarian and endocrine responses associated with the treatment of cystic ovarian follicles in dairy cows with gonadotropin releasing hormone and prostaglandin F2alpha, with or without exogenous progesterone

The objectives of this observational study were to document ovarian and endocrine responses associated with the treatment of cystic ovarian follicles (COFs) in dairy cows, using gonadotropin releasing hormone (GnRH) and prostaglandin F2alpha (PGF) with or without exogenous progesterone. A secondary objective was to determine pregnancy establishment following synchronization of ovulation and timed insemination in cows diagnosed with COFs. In trial I, 18 Holstein cows diagnosed with COFs received 2 injections of 100 microg GnRH, 9 d apart, with 25 mg PGF given 7 d after the 1st GnRH. A new follicle developed in all 18 cows after the 1st GnRH, and 83% of cows ovulated following the 2nd GnRH. Cows were inseminated 16 h after the 2nd GnRH. Of the 17 cows available for pregnancy diagnosis, 7 were confirmed pregnant. In trial II, 8 cows with COFs received GnRH and an intravaginal progesterone device (CIDR) concurrently, then PGF 7 d later. The CIDR was removed 2 d after PGF administration. Plasma estradiol concentrations declined following CIDR insertion. In all cows, a new follicle developed following GnRH treatment; estradiol-surge and estrus occurred spontaneously after CIDR-removal. Seven of 8 cows ovulated the new follicle. In dairy cows diagnosed with COFs, treatment with GnRH followed by PGF 7 d later, with or without exogenous progesterone, resulted in the recruitment of a healthy new follicle; synchronization of ovulation and timed insemination resulted in a 41% pregnancy rate.     

Administration of progesterone to cows with ovarian follicular cysts results in a reduction in mean LH and LH pulse frequency and initiates ovulatory follicular growth

Cows with ovarian follicular cysts were treated with progesterone to determine whether a reduction in LH concentrations and initiation of ovulatory follicular waves would occur. Cysts were diagnosed using transrectal ultrasonography when single follicular structures > 20 mm or multiple structures > 15 mm in diameter were present for 7 d in the presence of low progesterone concentrations. Three groups were studied: 1) cows with normal estrous cycles (CYC, n = 8); 2) cows with untreated cysts (CYST, n = 7); and 3) cows with cysts treated with two progesterone-releasing intravaginal devices (PRID, n = 8) for 9 d. Ovaries were examined with transrectal ultrasonography, and blood samples were collected daily for analysis of progesterone and FSH. Serial blood samples for determination of mean LH and LH pulse frequency were collected on d 0 (CYST and PRID cows only), 1, 5, 9, and 10. Progesterone concentrations were higher in PRID cows than in CYST cows throughout the PRID treatment period (P < .002). On d 0, LH pulse frequency was similar (P = .10) in PRID (6.6+/-.6 pulses/8 h) and CYST cows (5.1+/-.6 pulses/8 h), but mean LH tended to be higher (P = .054) on d 0 in PRID cows (2.5+/-.2 ng/mL) than in CYST cows (1.9+/-.2 ng/mL). Mean LH and LH pulse frequency decreased (P < .002) by d 1 in PRID cows (1.1+/-.2 ng/mL, 1.8+/-.6 pulses/8 h) compared with CYST cows (2.1+/-.2 ng/mL, 5.6+/-.6 pulses/8 h) and remained lower throughout most of the experimental period. The FSH concentrations were higher (P < .01) in PRID cows than in CYC and CYST cows on d 3 and 4. The increase in FSH concentrations preceded emergence of the PRID-induced follicular wave. All PRID cows and four of seven CYST cows initiated new follicular waves during the period of PRID treatment. Follicular waves were initiated later (P < .05) in CYST cows (d 5.2+/-1.7) and PRID cows (d 5.5+/-.6) than in CYC cows (d 1.8+/-.3). Cysts were smaller (P < .01) at the end of the treatment period in PRID cows compared with CYST cows. No CYST cows ovulated, but all PRID cows ovulated newly developed follicles 3 or 4 d after PRID removal. Treatment with exogenous progesterone reduced LH in cows with cysts, and this was followed by development of normal ovulatory follicles.     

Influence of experimental intrauterine infusion of Arcanobacterium pyogenes solution on ovarian activity in cycling cows

To examine the ovarian response to Arcanobacterium pyogenes (A. pyogenes) in uterus, bacterial solution was infused into the uteri of cows, and the follicle and corpus luteum (CL) development were monitored with a real-time ultrasound instrument. In addition, the plasma concentrations of progesterone (P(4)) and 13, 14-dihydro-15-keto-PGF(2alpha) (PGFM) were determined. A 10 ml bacterial solution that contained A. pyogenes (8 to 15 x 10(8) CFU/ml) was infused into the uterus of eight cows transcervically three days after natural ovulation. As a control, sterile physiological saline was infused into 4 other cows. The dominant follicle developed normally in 8 cows after bacteria inoculation. In 4 of these 8 cows, the developing CL regressed, and the first wave dominant follicles, which normally become atretic, ovulated after the inoculation. In the remaining 4 cows, the CL did not regress. The PGFM concentration increased transiently in all 8 cows after the infusion. The mean PGFM concentration of the cows with a regressed CL was significantly lower (P<0.01) than that of the cows whose CLs did not regress. In the control cows, there was no regression of developing CLs, no ovulation of first wave dominant follicles and no transient increase in PGFM after the infusion of sterile physiological saline. These results show that infusion of A. pyogenes into the uterus did not affect folliculogenesis and might have induced PGF(2alpha) production from the uterus.