Effects of cortisol on pregnancy rate and corpus luteum function in heifers: an in vivo study

To determine whether glucocorticoids affect the function of the bovine corpus luteum (CL) during the estrous cycle and early pregnancy, we examined the effects of exogenous cortisol or reduced endogenous cortisol on the secretion of progesterone (P4) and on pregnancy rate. In preliminary experiments, doses of cortisol and metyrapone (an inhibitor of cortisol synthesis) were established (n=33). Cortisol in effective doses of 10 mg blocked tumor necrosis factor-induced prostaglandin F(2α) secretion as measured by its metabolite (PGFM) concentrations in the blood. Metyrapone in effective doses of 500 mg increased the P4 concentration. Thus, both reagents were then intravaginally applied in the chosen doses daily from Day 15 to 18 after estrus (Day 0) in noninseminated heifers (n=18) or after artificial insemination (n=36). Pregnancy was confirmed by transrectal ultrasonography between Days 28-30 after insemination. Plasma concentrations of P4 were lower in cortisol-treated heifers than in control heifers on Days 17 and 18 of the estrous cycle (P<0.05). However, the interestrus intervals were not different between control and cortisol-treated animals (P>0.05). Moreover, metyrapone increased P4 and prolonged the CL lifespan in comparison to control animals (P<0.05). Interestingly, in inseminated heifers, cortisol increased the pregnancy rate (75%) compared with control animals (58%), whereas metyrapone reduced the pregnancy rate to 16.7% (P<0.05). The overall results suggest that cortisol, depending on the physiological status of heifers (pregnant vs. nonpregnant), modulates CL function by influencing P4 secretion. Cortisol may have a positive influence on CL function during early pregnancy, leading to support of embryo implantation and resulting in higher rates of pregnancy in heifers.     

The effect of metritis and subclinical hypocalcemia on uterine involution in dairy cows evaluated by sonomicrometry

The objective of this study was to examine the effects of metritis and subclinical hypocalcemia on reduction of uterine size in dairy cows using ultrasonography and sonomicrometry. Four piezoelectric crystals were implanted via laparotomy into the myometrium of the pregnant uterine horn of 12 pluriparous Holstein Friesian cows 3 weeks before the calculated calving date. Sonometric measurements were conducted daily from 2 days before parturition (= Day 0) until Day 14 after calving and then every other day until Day 28. Distances between adjacent crystals were expressed in relation to reference values obtained before calving. The diameter of the formerly pregnant uterine horn was measured using transrectal B-Mode sonography starting on Day 10. Cows were retrospectively divided into the following groups: cows without metritis (M–; n = 7), cows with metritis (M+; n = 5), cows with normocalcemia (SH–; Ca > 2.0 mmol/l on Days 1 to 3; n = 5) and cows with subclinical hypocalcemia (SH+; Ca < 2.0 mmol/l in at least one sample between Days 1 and 3; n = 7). Metritis did not affect (P > 0.05) sonometric measurements, but the diameter of the formerly pregnant horn was larger (P ≤ 0.05) between Days 15 and 21 in M+ cows than in M‒ cows. Reduction in uterine length in hypocalcemic cows was delayed (P ≤ 0.05) between Days 8 and 21 compared with normocalcemic cows, but the uterine horn diameter was not related to calcium status. In conclusion, both diseases affected reduction of uterine size until Day 28. Cows with metritis had a larger uterine diameter, possibly attributable to accumulation of lochia, and cows with subclinical hypocalcemia had delayed reduction of uterine length, presumably related to reduction of myometrial contractility.     

奶牛奶中孕铜含量变化模式及其对繁殖率的影响

通过３３２头奶牛孕酮含量变化周期的分析,获得孕酮谷点超前、居中和滞后３种模式,其排卵对应有超前、适中和延缓３种。这３种模式的奶牛情期受胎率分别为１９．２％,６３．６％和４１．４％。奶牛配种后,怀孕和未孕对孕酮含量变化有影响：怀孕牛,配种后从孕酮谷点至第１４天孕酮含量有提高（Ｐ＜０．０５）,特别是第１４天提高更显著（Ｐ＜０．０１）;未孕牛,配种后除谷点处孕酮含量较一致外,其余各处平均值都下降（Ｐ＜０．０５）。     

Factors affecting the success of resynchronization protocols with or without progesterone supplementation in dairy cows

The objective of this study was to investigate factors that influence the success of resynchronization protocols for bovines with and without progesterone supplementation. Cow synchronized and not found pregnant were randomly assigned to two resynchronization protocols: ovsynch without progesterone (P4) supplementation (n = 66) or with exogenous P4 administered from Days 0 to 7 (n = 67). Progesterone levels were measured on Days 0 and 7 of these protocols as well as 4 and 5 days post-insemination. Progesterone supplementation raised the P4 levels on Day 7 (p < 0.05), but had no overall effect on resynchronization rates (RRs) or pregnancy per artificial insemination (P/AI). However, cows with Body Condition Score (BCS) > 3.5 had increased P/AI values while cows with BCS < 2.75 had decreased P/AI rates after P4 supplementation. Primiparous cows had higher P4 values on Day 7 than pluriparous animals (p = 0.04) and tended to have higher RRs (p = 0.06). Results of this study indicate that progesterone supplementation in resynchronization protocols has minimal effects on outcomes. Parity had an effect on the levels of circulating progesterone at initiation of the protocol, which in turn influenced the RR.     

Histological Characteristics and Steroid Concentration of Ovarian Follicles at Different Stages of Development in Pregnant and Non-pregnant Dairy Cows

The aim of the study was to investigate the histological characteristics and steroid concentrations in follicular fluid of different populations of follicles at different stages of development, during pregnancy and the oestrous cycle in cows. Follicles from ovaries collected at a slaughterhouse were allocated into three size categories (small, 2–5.9 mm; medium, 6–13.9 mm; and large, 14–20 mm) in pregnant and non-pregnant cows. Slices were stained with HE and PAS for histological analysis. Follicular fluid was pooled according to size and pregnancy status and estradiol, testosterone and progesterone concentrations in follicular fluid were determined by RIA. Characteristics of healthy follicles did not differ, regardless of follicle size or pregnancy status. Total histological atresia was significantly higher in pregnant cows than in non-pregnant cows (p < 0.05). Estradiol increased and testosterone decreased significantly, while follicles increased in size, in both non-pregnant and pregnant cows (p < 0.05). Nonpregnant cows had the highest estradiol values in follicles of all sizes. Medium and large follicles from pregnant cows showed the lowest testosterone concentration (p < 0.05). Progesterone levels increased with follicle size only in non-pregnant animals. In large follicles, progesterone concentration was significantly higher in non-pregnant cows than in pregnant cows (p < 0.05). Considering steroid concentration and histological findings, most large follicles might be atretic during pregnancy in cattle.     

Effects of flunixin meglumine and transportation on establishment of pregnancy in beef cows

Objectives of these studies were to determine the effects of flunixin meglumine (FM) administration on early embryonic mortality and circulating PG and cortisol concentrations in transported and non-transported cows. Cows (n = 483) from 3 locations were used to evaluate the effects of transportation and FM approximately 14 d after AI on the establishment of pregnancy and serum concentrations of progesterone, PGF metabolite (PGFM), and cortisol. Treatments were transport (n = 129), transport + FM (n = 128), no transport (n = 130), and no transport + FM (n = 96). Multiparous cows (n = 224) were used at 2 locations, and nulliparous cows (n = 259) were used at 1 location. The no transport + FM treatment was used at only 2 locations. Flunixin meglumine (approximately 1.1 mg/kg of BW; i.m.) was administered before the cows were separated into transportation groups. Transportation included 4 to 6 h of transportation, without calves, via semitractor trailer. Nontransported cows remained penned, with their calves in adjacent pens, during the same period as the transported cows. Blood samples were collected from all cows before and after treatment and, at 2 locations, approximately 3 h after the onset of treatment. Location affected AI pregnancy rate (P < 0.01). Treatment effects, although not significant (P = 0.16), were of a magnitude to be considered practically important. Cows that received transportation + FM tended (P = 0.07) to have greater AI pregnancy rates (74%) than those that did not receive FM (66%), irrespective of transportation. Cortisol concentration was greater (P < 0.05) for transported cows than for nontransported cows. Cows receiving FM had greater (P < 0.05) AI pregnancy rates than non-FM cows (71 vs. 61%, respectively). Cows receiving transportation had lower (P < 0.01) mean PGFM concentrations than nontransported cows (45.4 vs. 54.6 pg/mL, respectively), and cows receiving FM had lower (P < 0.01) mean PGFM concentrations than non-FM cows (39.4 vs. 60.6, respectively). We conclude that transportation of cows approximately 14 d after AI increased serum cortisol concentrations but did not affect AI pregnancy rates. However, treatment of cows with FM increased AI pregnancy rates, irrespective of whether they were transported.     

Effect of ovulatory follicle size and expression of estrus on progesterone secretion in beef cows

Induced ovulation of small dominant follicles (SF, < 12 mm; CO-Synch protocol) in postpartum beef cows resulted in formation of corpora lutea (CL) that exhibited a delayed rise in progesterone (P4) compared with CL from large dominant follicles (LF, > 12 mm). Experiment 1 characterized P4 concentrations from ovulation to subsequent estrus among GnRH-induced or spontaneously ovulated SF (or= 12 mm) to determine if P4 secretion by CL formed from GnRH-induced SF remains lower postovulation in nonlactating beef cows. Nonlactating beef cows were induced to ovulate 48 h after PGF(2alpha) (CO-Synch; GnRH on d - 9, PGF(2alpha) on d - 2, and GnRH on d 0) or exhibited estrus and spontaneously ovulated after PGF(2alpha). Follicle size was measured at the second GnRH in cows induced to ovulate or approximately 3 h after the onset of estrus for cows that ovulated spontaneously. Cows were classified into 1 of 4 groups: 1) GnRH-induced ovulation-SF (or= 12 mm; Ind-LF; n = 16); 3) spontaneous ovulation-SF (or= 12 mm; Spon-LF; n = 22). Serum concentrations of P4 from d 3 to 15 were reduced in the Ind-SF compared with the Ind-LF (P = 0.05), Spon-SF (P = 0.07), and Spon-LF (P = 0.03). Experiment 2 characterized P4 concentrations (0 to 60 d postAI) among GnRH-induced or spontaneously ovulated SF (or= 13 mm) to determine if P4 secretion by CL formed from GnRH-induced SF remained lower during early gestation. Ovulation was induced with GnRH 48 h after PGF(2) (CO-Synch) or occurred spontaneously, and ovulatory follicle size was measured at AI. Lactating cows were classified into 1 of 3 groups: 1) GnRH-induced ovulation-SF (or= 13 mm; Ind-LF; n = 43); or 3) spontaneous ovulation-LF (>or= 13 mm; Spon-LF; n = 27). The increase in P4 concentrations was greater (P = 0.06) in pregnant (d 2 to 12) compared with nonpregnant cows. Also, the increase in P4 from d 2 to 12 was greater (P = 0.01) in the Ind-LF compared with the Ind-SF groups, but there was no difference (P = 0.94) among groups in P4 from d 14 to 60 in pregnant cows. Follicle size at AI influenced the increase in P4 in cows that failed to conceive (P = 0.007), but not among cows that became pregnant (P = 0.32) to AI. In summary, P4 secretion after GnRH-induced ovulation of SF was decreased from d 2 to 12 compared with that of LF, but was similar among pregnant cows from d 14 to 60 postAI (d 0).     

Endocrine effects of GnRH agonist application to early pregnant gilts

The hypothesis of the present study was that a GnRH agonist application at early pregnancy would alter the pattern of the key reproductive hormones LH and FSH, and subsequently that of estradiol (E2) and especially progesterone (P4), and improve the conditions for embryo survival in early pregnant gilts. Therefore, the endocrine effects of a GnRH agonist (GnRHa) application to gilts (n=11 GnRHa treated, n=9 saline Controls) were studied in blood samples from the Vena cava caudalis. GnRHa injected on Day 12 after insemination induced elevated (P<0.01) LH and FSH levels for at least 180 min. However, subsequent LH concentrations were not altered up to Day 21 of pregnancy. LH pulse number, estimated in 6-h period samples on Days 13, 15 and 17, was not influenced by treatment and pregnancy. LH pulse amplitude was decreased (P<0.05) on Days 13 to 17 in pregnant gilts of both groups, but not in nonpregnant animals. In pregnant GnRHa-treated gilts, the basal LH level was elevated compared with the Controls (P<0.01). Additionally, differences (P<0.05) in basal LH were present between the pregnant and nonpregnant animals. The P4 and E2 secretion pattern was not affected by GnRHa. P4 concentrations increased (P<0.01) from Day 10 to Day 14 regardless of the treatment. P4 revealed a pulse-like pattern, but without a definite relation to the LH pulse characteristics. Also, pregnancy rate (73 vs. 67%) and the number of fetuses (12.8 ± 2.3 vs. 11.6 ± 2.3) were unaffected in the treated and Control gilts, respectively. The present study did not confirm the initial hypothesis that a GnRHa-mediated LH effect could alter ovarian steroid secretion and favorably support early embryo development and pregnancy outcome.     

Pregnancy diagnosis by milk progesterone on days 18, 22, and 24 postbreeding in dairy cows

Milk samples were collected from 91 dairy cows on days 18, 22, and 24 postbreeding. The progesterone values of these samples were assayed, using enzyme-immuno-assay for pregnancy diagnosis. These animals were classified by nonpregnant (low progesterone values, 5 ng/ml or less) and pregnant (high progesterone values, more than 5 ng/ml). The accuracy of diagnosis on days 18, 22, and 24 postbreeding were 56%, 78%, and 79% for pregnant cows and 90%, 93%, and 93% for nonpregnant animals. The sampling date had a significant effect on diagnostic accuracy (P less than 0.05).     

Resynchronization of estrus in cattle of unknown pregnancy status using estrogen,progesterone, or both

Our objective was to develop treatments applied to cattle of unknown pregnancy status that would resynchronize the repeat estrus of nonpregnant females. In Exp. 1, previously inseminated dairy and beef heifers were assigned randomly to each of three treatments 13 d after AI: 1) no treatment (controls; n = 44); 2) 0.5 mg of estradiol cypionate (ECP) i.m. on d 13 and 20 at the time of insertion and removal of a used intravaginal progesterone (P4)-releasing insert (CIDR; P4 + ECP; n = 44); and 3) same as P4 + ECP without injections of ECP (P4; n = 42). The P4 + ECP (>90%) and P4 (>75%) protocols effectively synchronized repeat periods of estrus to 2 d and did not harm established pregnancies. In Exp. 2, treatments similar to those in Exp. 1 were applied to previously inseminated beef heifers (n = 439). Feeding 0.5 mg of melengestrol acetate (MGA) from d 13 to 19 after AI replaced the CIDR as a source of progestin. Of those heifers not pregnant (n = 65) after the initial AI, more than 86% were reinseminated, but conception was decreased (P < 0.05) by 28 to 39% compared with controls. In Exp. 3, previously inseminated lactating beef cows at four locations were assigned within herd to each of three treatments: 1) no treatment (control; n = 307); 2) same as in Exp. 1, but with P4 + 1 mg of estradiol benzoate on d 13 and 20 (P4 + EB; n = 153); and 3) same as in Exp. 1, P4 + ECP (n = 149). Treatments with P4 plus estrogen did not decrease conception rates in pregnant cows at any location, but increased (P < 0.05) the percentage of nonpregnant cows returning to estrus between 19 and 23 d after timed AI from 29% in controls to 86% in P4 + EB and 65% in P4 + ECP cows. Conception rates at the return estrus were not decreased when treatments occurred between d 13 and 20. In Exp. 4, lactating beef cows were assigned as in Exp. 3 to each of three treatments: 1) no treatment (controls; n = 51); 2) P4 + ECP (n = 47), as in Exp. 1; and 3) a single injection of ECP on d 13 (n = 48). Previously established pregnancies were not harmed (P = 0.70), and return rates of nonpregnant cows did not differ (P = 0.78) among treatments. In summary, in both heifers and lactating beef cows, the P4-based resynchronization treatments increased synchronized return rates when estrus detection rates were low, had no negative effects on established pregnancies, and decreased or tended to decrease conception rates at the resynchronized estrus.     

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 reproductive state on pituitary-adrenal activity in the ewe.

Plasma corticosteroid concentrations are altered in pregnancy, during the reproductive cycle and by ovariectomy in many species. This study was designed to examine basal ACTH and cortisol in ewes of four different reproductive statuses: ovariectomized, nonpregnant cycling, nonpregnant noncycling, and pregnant. Blood samples were drawn every 4 hr for 48 hr from ewes during quiet, undisturbed conditions and analyzed for plasma ACTH, cortisol and progesterone concentrations. There were no significant changes in ACTH, cortisol or progesterone over time. Mean progesterone concentrations were significantly greater in the pregnant ewes than in all other ewes, and were greater in cycling ewes than noncycling or ovariectomized ewes. Mean ACTH was significantly greater in pregnant ewes than noncycling ewes, and mean cortisol was significantly greater in cycling ewes than in nonpregnant cycling or noncycling ewes. Ovariectomized ewes also had significantly greater mean cortisol concentrations than cycling ewes. The results demonstrate that there is an increase in basal ACTH and cortisol in ovine pregnancy.     

Relations between plasma IGF-I concentrations during treatment with CIDR-based or Ovsynch protocol for timed AI and conception in early postpartum Japanese black beef cows

We examined the relations between plasma insulin-like growth factor (IGF) -I concentrations during treatment with CIDR-based or Ovsynch protocol for timed AI and conception and plasma steroid concentrations in early postpartum Japanese Black beef cows. Cows in the control group (Ovsynch; n = 21) underwent Ovsynch protocol (GnRH analogue on Day 0, PGF(2alpha) analogue on Day 7, and GnRH analogue on Day 9), with AI on Day 10, approximately 20 h after the second GnRH treatment. Cows in the Ovsynch+CIDR group (n = 22) received Ovsynch protocol plus a CIDR for 7 days (starting on Day 0). Cows in the further treatment group (EB+CIDR+GnRH; n = 22) received 2 mg of estradiol benzoate (EB) on Day 0 in lieu of the first GnRH treatment, followed by the same treatment as in the Ovsynch+CIDR protocol. Plasma IGF-I concentrations were determined on Days -7, 0, 7, 9 and 17. Conception rates were improved in the CIDR-combined groups (both CIDR-treated groups were combined) relative to Ovsynch group (P < 0.05) for cows with low IGF-I concentrations (<1,000 ng/ml) on Days -7, 0, and 7, but improved conception rate produced by the CIDR-based protocols did not occur in cows with a high IGF-I concentration (> or =1,000 ng/ml). Plasma estradiol-17beta concentrations increased from Day 0 to 7 (P < 0.05) and were unchanged from Day 7 to 9 in the Ovsynch group with low IGF-I concentrations on Day 0, while they were unchanged from Day 0 to 7 and increased from Day 7 to 9 (P < 0.05) in the Ovsynch group with high IGF-I concentrations on Day 0 and in the CIDR-combined group. Plasma progesterone concentrations in the Ovsynch group with low IGF-I concentrations on Day 0 were higher on Day 14 than in the Ovsynch group with high IGF-I concentrations on Day 0 and in the CIDR-combined group (P < 0.05). In conclusion, CIDR-based protocols may improve conception relative to Ovsynch in early postpartum beef cows with lower plasma IGF-I concentrations at the start of the protocols. This improvement is probably due to prevention of premature increases of estradiol-17beta and progesterone concentrations, which occurred in cows with low IGF-I concentrations treated with Ovsynch, by the CIDR treatment.     

Influence of a controlled internal drug release after fixed-time artificial insemination on pregnancy rates and returns to estrus of nonpregnant cows

We determined whether an ovulatory estrus could be resynchronized in previously synchronized, AI nonpregnant cows without compromising pregnancy from the previous synchronized ovulation or to those inseminated at the resynchronized estrus. Ovulation was synchronized in 937 suckled beef cows at 6 locations using a CO-Synch + progesterone insert (controlled internal drug release; CIDR) protocol [a 100-microg injection of GnRH at the time of progesterone insert, followed in 7 d by a 25-mg injection of PGF(2alpha) at insert removal; at 60 h after PGF(2alpha), cows received a fixed-time AI (TAI) plus a second injection of GnRH]. After initial TAI, the cows were assigned randomly to 1 of 4 treatments: 1) untreated (control; n = 237); 2) progesterone insert at 5 d after TAI and removed 14 d after TAI (CIDR5-14; n = 234); 3) progesterone insert placed at 14 d after TAI and removed 21 d after TAI (CIDR14-21; n = 232); or 4) progesterone insert at 5 d after TAI and removed 14 d after TAI and then a new CIDR inserted at 14 d and removed 21 d after TAI (CIDR5-21; n = 234). After TAI, cows were observed twice daily until 25 d after TAI for estrus and inseminated according to the AM-PM rule. Pregnancy was determined at 30 and 60 d after TAI to determine conception to the first and second AI. Pregnancy rates to TAI were similar for control (55%), CIDR5-14 (53%), CIDR14-21 (48%), and CIDR5-21 (53%). A greater (P < 0.05) proportion of nonpregnant cows was detected in estrus in the CIDR5-21 (76/110, 69%) and CIDR14-21 (77/120, 64%) treatments than in controls (44/106, 42%) and CIDR5-14 (39/109, 36%) cows. Although overall pregnancy rates after second AI service were similar, combined conception rates of treatments without a CIDR from d 14 to 21 [68.7% (57/83); control and CIDR5-14 treatments] were greater (P = 0.03) than those with a CIDR during that same interval [53.5% (82/153); CIDR5-21 and CIDR14-21 treatments]. We conclude that placement of a progesterone insert 5 d after a TAI did not compromise or enhance pregnancy rates to TAI; however, conception rates of nonpregnant cows inseminated after a detected estrus were compromised when resynchronized with a CIDR from d 5 or 14 until 21 d after TAI.     

Resynchrony of previously anoestrous cows and treatment of cows not detected in oestrus that had a palpable corpus luteum with prostaglandin F(2 alpha)

AIMS: To determine if resynchrony, using a progesterone (P4) / oestradiol benzoate (ODB) regime, of previously treated, anovulatory anoestrous (AA) cows would increase the probability of oestrus, conception and pregnancy compared to no resynchrony. Additionally, the effect of prostaglandin F2alpha (PG) treatment of cows not detected in oestrus, but in which a corpus luteum (CL) was palpable transrectally (NDO/CL+), was compared with no treatment. METHODS: Cows not detected in oestrus by 1 week before the start of the seasonal breeding programme were categorised as AA or NDO/CL+, based on absence or presence of a palpable CL, respectively. All AA cows were treated with an intravaginal device containing 1.36 g P4 (CIDR-B) for a period of 6 days, and with 1 mg ODB by intramuscular (I/M) injection 1 day after the device was removed (Day 0). Half the AA cows that were detected in oestrus between Days 0 and 3 were randomly assigned to be resynchronised by reinsertion of a clean used CIDR-B device on Day 15, for a period of 6 days, followed by an I/M injection of 0.5 mg ODB, 1 day after the device was removed (resynchrony, RS), while the other half were not resynchronised (no-RS). NDO/CL+ cows were alternately assigned to be either treated with 25 mg of the PG, dinoprost tromethamine, on Day 0 or left as untreated controls (Con). RESULTS: Resynchrony increased the percentage of cows detected in oestrus between Days 14 and 28 (212/282 = 75.2% vs 155/285 = 54.4%, for RS vs no-RS, respectively; p<0.001), but had no effect on conception rate to a service within the first 3 days of the mating period (146/397 = 36.8% vs 148/399= 37.1%, for RS vs no-RS cows, respectively; p>0.9), or to a service between Days 14 and 28 (84/159 = 52.8% vs 114/217 = 52.5%, for RS vs no-RS cows, respectively; p>0.9). Resynchrony increased the Day 28 pregnancy rate (264/432 = 61.1% vs 237/435 = 54.5%, for RS vs no-RS cows, respectively; p=0.03), but had no effect on the Day 56 or final pregnancy rates (p>0.1).Prostaglandin treatment of NDO/CL+ cows did not affect the percentage of cows detected in oestrus by Day 7 (43/106 = 40.6% vs 51/101 = 50.5%, for Con vs PG, respectively; p=0.15), or Day 28 (92/106 = 86.8% vs 91/101 = 90.1%, for Con vs PG, respectively; p>0.4). Treatment did not affect the Day 28 pregnancy rate (55/103 = 53.4% vs 54/98 = 55.1%, for Con vs PG, respectively; p>0.9), the Day 56 pregnancy rate (81/103 = 78.6% vs 74/98 = 75.5%, for Con vs PG, respectively; p>0.6), or the final pregnancy rate (98/103 = 95.1% vs 89/97 = 91.8%, for Con vs PG, respectively; p>0.3). CONCLUSIONS: Resynchrony of AA cows treated using the present protocol increased the proportion of non-pregnant cows detected in oestrus between Days 14 and 28 and increased the Day 28 pregnancy rate. This resynchrony protocol may be useful for increasing the proportion of the herd pregnant in the first month of the breeding programme, especially in herds that have a history of a low proportion of non-pregnant cows detected in oestrus between Days 14 and 28. There was no benefit in treating NDO/CL+ cows with PG compared to no treatment. KEYWORDS: Dairy cows, anoestrus, treatment, resynchrony, prostaglandin, progesterone, oestradiol benzoate.     

Involvement of Plasma Progesterone, Oestradiol-17β and Cortisol in Ovulatory Response to Gonadotropin-releasing Hormone in Dairy Cows with Cystic Follicles

The aim of the present study was to examine the plasma concentrations of progesterone, oestradiol-17beta and cortisol in the cows with cystic follicle and to examine its relationship with the ovulatory response to gonadotropin-releasing hormone (GnRH). Eighty-five post-partum Holstein-Friesian cows with cystic follicles regardless of the presence of corpus luteum were studied. Follicular size, presence of corpus luteum and occurrence of ovulation were checked by palpation per rectum. Blood collection and palpation per rectum were conducted on days 0, 7 and 14. Gonadotropin-releasing hormone was administered at day 7. Plasma concentrations of progesterone, oestradiol-17beta and cortisol were determined. Progesterone concentrations of <3.2, 3.2-4.8 and >4.8 nmol/l were defined as low, intermediate and high, respectively. Sixty-three (74.1%) of 85 cows showed low (<3.2 nmol/l =1 ng/ml) progesterone concentrations on day 0. Only 40 (47.1%) of them showed low-low pattern of progesterone at days 0 and 7. In 27 (31.8%) of them, progesterone concentration had increased by day 14. Of 22 cows having high progesterone concentration (>/=4.8 nmol/l) on day 0, corpus luteum was not detected in 18 cows (21.2%). Only in 10 cows, cystic follicle disappeared after GnRH administration. However, only one of 27 cows in which progesterone pattern was low-low-high at days 0, 7 and 14 experienced ovulation of the cystic follicle. Significantly lower oestradiol-17beta concentration was found on day 7 in cows showing a low-low-low pattern than a low-low-high pattern of progesterone (43.0 +/- 4.6 vs 55.8 +/- 2.8 pmol/l, p < 0.05). There was no significant difference in cortisol concentration on any days (days 0, 7 and 14) between cows showing a low-low-low and low-low-high pattern of progesterone. These results suggest that approximately one-fifth of cows diagnosed to have ovarian cysts possess luteal cysts and that a high oestradiol-17beta concentration at the time of GnRH administration is involved in the subsequent ovulation of the follicle, although ovulated follicle may not be cystic.     

Plasma bovine placental lactogen concentration throughout pregnancy in the cow; relationship to stage of pregnancy,fetal mass,number and postpartum milk yield

This study characterized the peripheral plasma placental lactogen (bPL) profile throughout gestation and examined the relationship between the stage of gestation, fetal mass, number, and postpartum lactation with circulating levels of bPL in Holstein cows after nonsurgical embryo transfer. Cows (n = 12) were divided into two groups: Group 1 = single embryo recipient cows (n = 5); Group 2 = twin embryo recipient cows (n = 7). Blood was collected about every third day from Day 0 (Day 0 was defined as the first day of standing estrus), then daily for the last 10 d of gestation, and sampling was stopped 1 d postpartum. The cows were milked twice daily at 0800 and 1800 hr. Two twin-embryo recipient cows had abnormal pregnancies; therefore, their data were excluded from that of the group and reported separately. The time trend concentrations of plasma bPL were significantly affected by the stage of gestation (P < 0.01) but not fetal number (P < 0.21). In both groups bPL levels remained low during the first two trimesters, then increased rapidly (P < 0.01) to peak concentrations between Days 200 and 220, and stabilized at this elevated level until parturition. Postpartum milk yields were indistinguishable between the singleton and twin-bearing cows. Calf birth weight and postpartum lactation were both correlated (P < 0.01) to peripheral bPL concentration in singleton cows, however, this relationship decreased with a subsequent increase in fetal number. Cows giving birth prematurely to stillborn calves or to a schistosomus reflexus calf exhibited a deviating bPL profile. These results indicate that peripheral bPL levels are positively associated with the stage of gestation but not with fetal number. Otherwise, the peripheral pattern of bPL is a valuable index for predicting feto-placental viability.     

In vivo investigation of luteal function in dogs: Effects of cabergoline,a dopamine agonist,and prolactin on progesterone secretion during mid-pregnancy and -diestrus

The role of prolactin on luteal function in dogs was investigated in vivo. The function of prolactin in mid-luteal phase was compared in pregnant and nonpregnant dogs. A dopamine agonist, cabergoline, known for its prolactin secretion inhibitory effects, was injected subcutaneously at a dose of 5 μg/kg body weight in five pregnant and five nonpregnant Beagle bitches. Mean plasma prolactin and progesterone were dramatically suppressed for 4 to 5 days after injection in both groups when compared with control pregnant and non-pregnant animals, whereas no effect on luteinizing hormone (LH) secretion was observed. The decline in plasma progesterone occurred after that in prolactin, suggesting plasma progesterone was impaired by inhibition of prolactin secretion. These results confirm the luteotropic importance of prolactin in pregnant bitches, and also demonstrate its importance in luteal phase of the nonpregnant dog.Second, to demonstrate that the effects of cabergoline were mediated by prolactin inhibition and not by a direct action on the corpus luteum, concomitant administration on Day 30 of cabergoline and prolactin (375 μg IV twice daily on Days 30 and 31) or cabergoline and LH (750 μg IV twice daily on Days 30 and 31) was affected in two groups of five pregnant animals each. Results showed that only prolactin was able to reverse the negative effects of cabergoline on circulating progesterone. This confirms the indirect mode of action of the dopamine agonist, cabergoline on corpus luteum function.Third, further investigation on the precise luteotropic role of prolactin was made by IV injection of 375 μg pure canine prolactin twice daily in five pregnant bitches on Days 30 and 31, and in five pregnant bitches on Days 40 and 41. No direct stimulatory effect of prolactin on plasma progesterone secretion occurred. Nor was there a noticeable effect on plasma LH secretion. These results suggest that prolactin is unable to directly stimulate progesterone secretion by the corpus luteum of pregnancy.The results of this study suggest that prolactin is an essential luteotropin in the dog from mid-luteal phase in both pregnant and nonpregnant animals. However, it appears to act by sustaining corpus luteum lifespan and function rather than by direct stimulatory effects on progesterone secretion.     

Quantitative analyses of insulin-like peptide 3 and sex steroid hormones in dominant follicles and corpora lutea during the estrus cycle and in follicular cysts in beef heifers

Insulin-like peptide 3 (INSL3) and sex steroids were measured in bovine dominant follicles and corpora lutea during the estrus cycle and in follicular cysts. Paired ovaries from beef heifers (n = 47) were classified, by their morphological features, either into four stages of the estrus cycle (Day 1 = day of ovulation, Day 20 = day of estrus) as Stage I (Days 1–4; n = 8), Stage II (Days 5–10; n = 10), Stage III (Days 11–17; n = 8), and Stage IV (Days 18–20; n = 11) or follicular cystic (n = 10). Cysts (n = 15) were subdivided into estrogen-active (n = 7) and estrogen-inactive (n = 8) cysts. INSL3, testosterone, and estradiol-17β concentrations in the dominant follicular fluid of Stage IV were higher than those in Stages II and III (P < 0.05). INSL3 concentrations in the cystic fluid were similar to those in dominant follicles at Stage IV, whereas testosterone and estradiol-17β concentrations were lower in cysts (P < 0.05). INSL3 content per estrogen-inactive cyst was higher than that of Stage IV (P < 0.05). INSL3 and progesterone concentrations in luteal tissue and contents per corpus luteum were higher in Stages II and III (P < 0.05). In conclusion, INSL3 secretion in bovine dominant follicles increased with maturation. Follicular cysts may retain the production of INSL3 during their formation but tend to lose the capacity for testosterone secretion. Estrogen-inactive cysts subjected to advanced atresia may accumulate more INSL3. INSL3 production in bovine corpora lutea is enhanced during maturation.     

Pregnancy Loss in Dairy Cattle: Relationship of Ultrasound,Blood Pregnancy‐Specific Protein B,Progesterone and Production Variables

Objectives were to determine associations between percentage pregnancy loss (PPL) in dairy cattle and: (i) pregnancy diagnosis by ultrasonography; (ii) pregnancy diagnosis by serum pregnancy‐specific protein B (PSPB) concentrations, with or without serum progesterone concentrations; and (iii) production and environmental factors. This study included 149 822 pregnancy diagnoses conducted over 13 years in Holstein‐Friesian cows in Hungarian dairy herds. The following were determined: PPL in cows diagnosed pregnant by transrectal ultrasonography 29–42 days after artificial insemination (AI; n = 11 457); PPL in cows diagnosed pregnant by serum PSPB 29–35 days after AI (n = 138 365); and PPL and its association with serum progesterone concentrations, PSPB and production/environmental variables. The definition of PPL was percentage of cows initially diagnosed pregnant based on ultrasonography or PSPB, but not pregnant when examined by transrectal palpation 60 –70 days after AI. The PPL was lower (p < 0.001) in cows following ultrasonographic vs PSPB diagnosis of pregnancy at 29–35 days (8.1 vs 19.3%, respectively), but was higher in cows following ultrasonographic pregnancy diagnosis on 29–35 vs 36–42 days (8.1 vs 7.1%, respectively, P < 0.05). Furthermore, 72.9% of pregnancies with ultrasound‐detected morphological abnormalities resulted in pregnancy loss. As a subset of PSPB data, a fully quantitative PSPB assay was used for 20 430 samples; PPL in cows with a high PSPB concentration (>1.1 ng/ml) was lowest (15.0%), whereas cows with low concentrations of both PSPB and progesterone (0.6–1.1 and <2 ng/ml, respectively) had the highest PPL (76.3%; p < 0.0001). Furthermore, PPL was higher in cows with advanced parity and with high milk production, when ambient temperatures were high, although body condition score (BCS) had no effect on PPL. Finally, there were no significant associations between serum PSPB and environmental temperatures or number of post‐partum uterine treatments.