Comparison of plasma concentrations of estradiol-17β and progesterone, and conception in dairy cows with cystic ovarian diseases between Ovsynch and Ovsynch plus CIDR timed AI protocols

The objectives of this study were 1) to determine the effects of adding a CIDR to the Ovsynch protocol on plasma concentrations of estradiol-17β and progesterone and conception in dairy cows with cystic ovarian diseases and 2) to examine associations among the estradiol-17β and progesterone concentrations and conception. Cows were diagnosed as having cystic ovarian diseases if they were found to have a cystic follicle (diameter ≥25 mm) without a corpus luteum by two palpations per rectum with an interval for 7 to 14 days. They were treated with either the Ovsynch (GnRH on Day 0, PGF(2α) on Day 7 and GnRH on Day 9, with AI on Day 10; n=15) or Ovsynch+CIDR protocol (Ovsynch protocol plus a CIDR from Day 0 to Day 7; n=23). Plasma estradiol-17β concentrations were determined on Days 0, 7 and 9, and plasma progesterone concentrations were determined on Days 0, 7, 9 and 17. The plasma estradiol-17β and progesterone concentrations at all of the days examined and conception rates did not differ significantly between the two timed AI protocols. The progesterone concentrations on Day 17 and conception rates were lower (P<0.05) for cows with low concentrations of estradiol-17β (<2 pg/ml) on Day 9 than for cows with high concentrations of estradiol-17β (≥2 pg/ml). The present study suggests that, in dairy cows with cystic ovarian diseases, addition of a CIDR to the Ovsynch protocol had no remarkable effects on plasma estradiol-17β and progesterone concentrations during and after the treatments or on conception after timed AI. This study indicates that the low plasma estradiol-17β concentration at the second administration of GnRH in the protocols can be a predictor for impaired luteal formation and lower likelihood of pregnancy in dairy cows with cystic ovarian diseases.     

Efficiency of fecal steroid hormone measurement for assessing reproductive function in the Hokkaido brown bear (Ursus arctos yesoensis)

The present study aimed to establish simple systems for measuring fecal steroid hormones in order to monitor the reproductive profiles of captive Hokkaido brown bears. The efficiency of fecal sample processing at the steps of dehydration and extraction and the correlation between steroid concentrations in matched fecal and blood samples were studied. Then, monthly changes in fecal estradiol-17 beta and progesterone in female bears, and testosterone in male bears were examined. The procedure was finalized as follows. Fecal samples were dried at 100 degrees C for 3 hr and extracted with diethyl ether. The diethyl ether in the extracts was evaporated and residues were reconstituted in ethanol for the assays. Hormone concentrations were quantified using enzyme immunoassays. Concentrations of progesterone and testosterone in fecal and plasma samples were correlated in the systems. The changes in fecal progesterone and testosterone concentrations were similar to those in serum concentrations of bears as reported previously. In contrast, fecal estradiol concentrations did not correlate with plasma levels probably because of the time lag in excretion. However, the changes in estradiol-17 beta concentrations in feces in the present study were similar to those reported in serum. In conclusion, fecal progesterone and testosterone assay systems appear practical for monitoring ovarian and testicular activities without immobilization, though methodological improvements and further validation may be required. For the fecal estradiol-17 beta assay, there is a need to solve the problem of excretion time lag before the system can be used in the study of reproductive physiology.     

Progesterone and estradiol-17β as a potential method for pregnancy diagnosis in the collared peccary (Pecari tajacu)

In this study, the period of pregnancy of nine collared peccary females has been monitored through the analysis of serum progesterone and estradiol-17β profiles. Serum concentrations of progesterone increased by Day 4 after conception, reaching concentrations of 33.4±5.6ng/mL on Day 10. Between Days 10 and 130 progesterone values were maintained between 20 and 60ng/mL. In the collared peccary, embryonic estradiol synthesis is first observed in the systemic circulation by Day 15 of pregnancy. Between Days 0 and 50 of pregnancy, average estradiol-17β concentrations were between 0 and 30pg/mL. From Day 75 of pregnancy onwards, estradiol concentrations were constantly increasing, reaching maximum concentrations (131.4±40.8pg/mL) on the day of parturition. The combined study of serum progesterone and estradiol-17β concentrations as a potential method for early pregnancy diagnosis presented the best overall accuracy (73%) when the threshold was established at 20ng/mL serum progesterone and 20pg/mL serum estradiol. Nevertheless, the accuracy for diagnosing pregnancy of females at mid and late pregnancy was 78% and 95%, respectively. The analysis of the sexual hormones during pregnancy could be a useful tool as a potential pregnancy diagnosis and an efficient predictor of the day of parturition in the captive collared peccary.     

香猪生殖器官的发育及血浆雌二醇和睾酮含量变化

测试验定了初生、10、20、30、90、150和210日龄香猪公母猪的生殖器官及其血浆雌二醇和睾酮浓度。结果表明,香猪公猪的生殖器官发育较早,母猪生殖器官发育较迟。90日龄时,公猪睾酮水平达3273±1455pg/ml,母猪雌二醇水平也开始上升。发现公猪血浆中查得较母猪高的雌二醇水平。     

Relationship between endogenous estradiol-17 beta and estrous behavior in heifers

Thirty-five Holstein heifers were used to examine the relationship between endogenous estradiol-17 beta and estrous traits. During a non-superovulation period (NSP), estrous cycles were synchronized and during the periovulatory stage blood samples were collected every 6 h for 120 h for subsequent determination of estradiol-17 beta and progesterone. In addition, continuous observation for estrous behavior was performed for 98 h. A gonadotropin-induced superovulation period (SP) was begun 12 d after estrus was detected during NSP. Heifers were injected with FSH twice daily for 4 d and single injections of prostaglandin were given on d 14 and 15. Beginning at d 14, blood samples were collected every 6 h for 120 h for subsequent determination of estradiol-17 beta and progesterone. Continuous observation for estrous behavior was performed for 98 h. Peak estradiol-17 beta was greater during SP than during NSP (49.0 +/- 3.1 vs 12.9 +/- 3.0 pg/ml serum). Thirty-three and 31 of the 35 heifers were in estrus during NSP and SP, respectively; duration of estrus was 2.3 h longer during SP than during NSP. However, number of behavioral interactions during estrus did not differ between NSP and SP. In conclusion, estrous traits were similar, whereas peak estradiol-17 beta concentrations were markedly different between NSP and SP.     

Relationships among dietary lipid intake, serum cholesterol and ovarian function in Holstein heifers

Experiments were performed to determine whether serum concentrations of total cholesterol (TC) undergo cyclic changes during the estrous cycle of dairy heifers and to assess the relationship between serum concentrations of TC and ovarian steroid hormones. The effects of a hypercholesterolemic diet upon luteal progesterone secretion also were determined. Experiment 1 involved five dairy heifers exhibiting normal estrous cycles. Serum concentrations of TC, progesterone, testosterone and estradiol-17 beta were determined in blood samples collected throughout a complete estrous cycle. A transient decline in TC was observed during the luteal phase of all heifers beginning on d 2 and reached a nadir 6 d after estrus. Highest mean concentrations of TC occurred between d -2 and +2 (96.3 +/- 8.2 mg/dl), which were markedly higher (P less than .05) than the lowest mean concentrations (76.3 +/- 10.3 mg/dl) observed on d 6. Concentrations of serum TC were negatively correlated (r = -.40; P less than .01) with progesterone between d 2 and 9. Serum TC was not correlated with testosterone or estradiol-17 beta. In Exp. 2, seven cycling Holstein heifers were fed a control diet for 70 d (Stage I), a diet containing 15% whole sunflower seed as a source of supplemental dietary lipid for 70 d (Stage II) and then the control diet for 70 d (Stage III). Diets were isocaloric and isonitrogenous. All heifers were synchronized with prostaglandin F2 alpha after wk 5 in each of the three feeding stages.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of estradiol cypionate and GnRH treatment on plasma estradiol‐17β concentrations,synchronization of ovulation and on pregnancy rates in suckled beef cows treated with FTAI‐based protocols

Two experiments were conducted to evaluate the effect of different ovulation inducers on E‐17β plasma concentrations, synchronized ovulations and pregnancy rates. In Experiment 1, cows received a progesterone intravaginal device (PID) with 1 g of progesterone (P4) plus 2 mg of estradiol benzoate (EB) (day 0). At PID removal (day 8), cows received 0.150 mg of D‐cloprostenol and were randomly assigned to four treatment groups (n = 10/treatment): Group ECP: 1 mg of estradiol cypionate at PID removal, Group EB: 1 mg of EB 24 hr after PID removal, Group GnRH: 10 μg of GnRH 48 hr after PID removal, Group ECP‐GnRH: 1 mg of ECP at PID removal plus 10 μg of GnRH 48 hr later. Ultrasonographic examinations were performed to detect the dominant follicle and ovulation. GnRH‐treated cows ovulated later (p < .05) compared to ECP‐ and ECP+GnRH‐treated cows. There were effects of treatment, time and their interaction on E‐17β concentrations (p < .05). ECP treatment affected plasma E‐17β concentration, which increased earlier and decreased later compared to treatments without ECP. In Experiment 2, cows received (i) ECP: n = 126; (ii) EB: n = 126; (iii) GnRH: n = 136; (iv) ECP+GnRH: n = 139; FTAI was performed 48–50 hr after PID removal. Pregnancy rates did not differ among ovulation inducers (p > .05; ECP: 54.0%, 68/126; EB: 49.2%, 62/126; GnRH: 40.4%, 55/136; ECP+GnRH: 43.9%, 61/139). In conclusion, ECP administration (ECP and ECP+GnRH treatments) affected E‐17β concentrations, determining its earlier increase and later decrease compared to treatments without ECP (EB and GnRH treatments). ECP+GnRH‐treated cows achieved the best distribution of ovulations without affecting pregnancy rates.     

Plasma concentrations of immunoreactive (ir)-inhibin, gonadotropins and steroid hormones during the ovulatory cycle of the duck

The present study was undertaken to determine changes in circulating levels of immunoreactive (ir)-inhibin, FSH, LH, estradiol-17beta, progesterone, and testosterone during the ovulatory cycle of Shao ducks. Serial blood samples were taken from two groups of laying ducks for measurement of ir-inhibin, gonadotropins, and steroid hormones at 2 h intervals for 24 h. Plasma concentrations of ir-inhibin did not change significantly during the ovulatory cycle. The highest level of plasma ir-inhibin was observed 6 h prior to ovulation, which coincided with a decreased level of plasma FSH. One FSH surge was found 12 h after ovulation. Estradiol-17beta, progesterone, and testosterone were also determined during the ovulatory cycle. Two peak values were detected for estradiol-17beta 8 h before ovulation and 4 h after ovulation, while progesterone started to increase 4 h before ovulation and reached a peak at ovulation. The highest level of plasma testosterone was detected around the time of ovulation. These results suggest that inhibin may be involved in the control of FSH secretion during the ovulatory cycle. In addition, both LH and progesterone are of importance in the ovulation process of Shao ducks.     

Androgens modulate growth hormone-releasing factor-induced GH release from bovine anterior pituitary cells in static culture.

Static primary cultures of bovine anterior pituitary (AP) cells were utilized to study the effect of sex steroids on basal growth hormone (GH) and GH-releasing hormone (GRF)-stimulated release of GH. The AP cells (5 x 10(5) cells/well) were allowed to attach for 72 hr and become confluent before treatments were imposed. Cells were incubated for an additional 24, 48 or 72 hr with either estradiol-17 beta (E2, 10(-11) to 10(-8) M), testosterone (T, 10(-8) to 10(-5) M), dihydrotestosterone (DHT, 10(-9) to 10(-6) M) or 5 alpha-androstane-3 alpha, 17 beta-diol (3 alpha-diol, 10(-11) to 10(-8) M). Media were collected every 24 hr and GH concentrations determined by RIA. Incubation of calf AP cells with gonadal steroids did not affect (P > 0.05) basal GH released at 24, 48, or 72 hr. In another experiment, calf AP cells were incubated with the same concentrations of the steroids for 24 hr, media harvested, cells washed and challenged in serum-free media for 1 hr with bovine GRF 1-44-NH2 (10(-8) M). In non-steroid treated wells, GRF increased (P < 0.05) GH from 58 to 134 ng/ml. Incubation with E2 or 3 alpha-diol did not affect (P > 0.05) GRF-induced GH release; however, preincubation with T (10(-5) M) and DHT (10(-9), 10(-8) and 10(-7) M) increased (P < 0.05) GRF-induced GH release above control concentrations (195, 235, 190 and 185 ng/ml, respectively). At the doses tested, sex steroids did not affect basal release of GH, but androgens increased responsiveness of somatotropes to GRF.     

Effects of the thymic peptide thymulin on in vitro and in vivo testicular steroid concentrations in white composite and Meishan boars.

Immuno-peptides may have positive or negative effects on gonadal steroidogenesis, but few have been tested outside of rodent species or in vivo. In Exp. 1, thymulin, a secreted nonapeptide of the thymus, was incubated (1, 10, 100, or 1,000 ng/mL) with testicular minces (sampled at 3, 6, or 12 h) from Chinese Meishan boars of high gonadotropin/testicular steroidogenic function (n = 8) and White composite boars of European origin (n = 8 ) to test the hypothesis that thymulin could augment hCG stimulation of testicular androgen concentrations. Thymulin alone had few effects on androgen concentrations (testosterone, dehydroepiandrosterone+dehydroepiandrosterone sulfate [DHEA+DHEASO4]) in Meishan boar testicular incubates. In minces from White composites incubated with thymulin, testosterone concentrations were generally below control values (P<.05), but DHEA+DHEASO4 concentrations were unaffected. Thymulin had no effect on estrone concentrations in testicular incubates of White composite boars but stimulated estrone concentrations in Meishan testicular incubates. Thymulin plus hCG increased testosterone (3 and 6 h of incubation; P<.05) but not DHEA+DHEASO4 concentrations in White composite testicular incubates. Thymulin plus hCG did not alter androgen or estrogen concentrations from control values in Meishan testicular incubates. In Exp.2 with a protocol similar to that of Exp. 1 for testicular minces from White composite boars (n = 30), thymulin increased testosterone concentrations during the early incubation period (1 to 3 h; P<.05) and depressed testosterone concentrations at later times (6 h; P<.05). Thymulin synergized with hCG in stimulating increases in testosterone and DHEA+DHEASO4 concentrations (P<.05) but had no effect on estrone concentrations in vitro. Thymulin was tested in vivo in boars from three genetic lines selected for high, medium, or normal circulating LH concentrations (Meishan, select White composites, and control White composites, respectively). Injection of thymulin i.v. (4.4, 44.4, or 444.4 ng/kg BW) generally increased circulating testosterone concentrations (2 to 3 h later; P<.01), but the response was dependent on the boar's general circulating LH concentrations and dose of thymulin. Overall results from these studies support the hypothesis of a thymulin augmentation of LH stimulation of androgen increases in vitro and in vivo in the testis of boars.     

Oxytocin-induced secretion of prostaglandin F2alpha in postpartum beef cows: effects of progesterone and estradiol-17beta treatment

The purpose of the present study was to determine the effect of progesterone or progesterone + estradiol-17beta on oxytocin-induced prostaglandin F2alpha (PGF2alpha) secretion in postpartum beef cows. Thirty-four anestrous postpartum beef cows were ovariectomized (d 32 [Groups 1 to 3] or d 23 [Groups 4 to 6] postpartum [d 0 = parturition]) and allotted to six treatments (Group 1; negative control) to simulate short (Groups 2 through 5) or normal (Group 6) length estrous cycles. Steroid treatments for the respective groups were as follows: Group 1) no estradiol-17beta or progesterone treatment (n = 8; negative control); Group 2) progesterone (d 34 to 40; n = 6); Group 3) estradiol-17beta (d 32 to 33) and progesterone (d 34 to 40; n = 6); Group 4) progesterone (d 23 to 29), no estradiol-17beta (d 32 to 33), and progesterone (d 34 to 40; n = 5); Group 5) progesterone (d 23 to 29), estradiol-17beta (d 32 to 33), and progesterone (d 34 to 40; n = 5); and Group 6) progesterone (d 23 to 29), estradiol-17beta (d 32 to 33), and progesterone (d 34 to 50; n = 4; positive control). Oxytocin (100 IU) was injected (i.v.) at the end of each treatment to test the ability of the postpartum uterus to secrete PGF2alpha as measured by a stable metabolite of PGF2alpha, 15keto-13,14 dihydro-PGF2alpha (PGFM). Peak concentrations ofPGFM (P < 0.08) and total PGFM secreted (area under the curve; P < 0.05) were increased on d 6 following first (Group 2) or second (Group 4) exposure to progesterone and were similar to peak concentrations and total PGFM secreted 16 d following a simulated normal estrous cycle (Group 6). Administration of estradiol-17beta before first progesterone exposure (Group 3) did not reduce peak concentrations of PGFM or total PGFM secreted relative to the preceding groups. Peak concentrations of PGFM (P < 0.08) and total PGFM secreted (P < 0.05) were reduced following a second progesterone exposure, provided that cows were pretreated with estradiol-17beta (Group 5). In summary, oxytocin-induced release of PGFM was inhibited on d 6 following second exposure to progesterone only when cows were pretreated with estradiol-17beta. Therefore, estradiol-17beta and progesterone were both associated with the timing of PGF2, secretion in postpartum cows.     

The relation between ultrasonographic observations in the oviduct and plasma progesterone, luteinizing hormone and estradiol during the egg laying cycle in ostriches

In this study we investigated the temporal relationship between ovulation, egg formation, oviposition and the changes in plasma concentrations of progesterone, luteinizing hormone and estradiol-17beta during the egg laying cycle in farmed ostriches. In 10 egg-producing birds, transcutaneous ultrasound scanning was performed at 3h intervals and blood sampling at hourly intervals during a period of at least 48h (one egg laying cycle). In hens (n=8) that ovulated during the observational period, the ovulated egg was first detected 2h after oviposition; thus, ovulation occurred shortly after oviposition in all birds. During the period between two consecutive ovipositions, the developing egg remained for 9h in the proximal part (infundibulum, magnum or isthmus) and for 39h in the distal part of the oviduct (uterus). In ovulating hens, plasma progesterone concentrations showed a characteristic and consistent profile: from basal levels of around 0.1ng/ml concentrations started to increase 12h before oviposition, reached an average maximum of 3.5ng/ml at 3h before oviposition and returned to basal levels 3h and 30min after oviposition. Changes in plasma luteinizing hormone and estradiol-17beta concentrations showed comparable patterns of elevation and decline relative to the timing of oviposition and ovulation. However, variation in their individual basal concentrations was generally larger and peak values were less conspicuous than those of progesterone. In non-ovulating hens (n=2) neither progesterone, nor luteinizing hormone nor estradiol-17beta showed elevations to peak concentrations before oviposition. These data demonstrate that during the egg laying cycle of ostriches, events such as ovulation, egg development and oviposition evolve according to a rather strict time schedule, and that progesterone, luteinizing hormone and estradiol-17beta reach peak concentrations shortly before ovulation. Additionally, our findings also show that on-farm ultrasound scanning is a useful technique to discriminate between ovulating and non-ovulating hens.     

Efficacy of Long-Acting Formulations of Estradiol or Progesterone Plus Estradiol on Estrous Synchronization in Broodmares

A preliminary trial was performed to evaluate the ability of sustained release preparations of estradiol-17β or progesterone plus estradiol-17β to synchronize estrus in cyclic mares. Group 1 mares were treated with a 50 mg intramuscular (IM) injection of sustained release estradiol-17β, while group 2 mares were treated with estradiol plus 1.5 g of sustained release progesterone. All mares received an IM injection of 10 mg of prostaglandin-F₂α (PGF₂α) 10 days after steroid treatment. Mares were examined by transrectal ultrasonography on Days 1 and 10 of treatment and then at ≤2 day intervals to monitor follicle size. Once a follicle ≥30 mm diameter and uterine edema were detected, 0.5 mg of the GnRH analog histrelin was administered IM. Mares were examined daily thereafter to detect ovulation. Group 1 mares did not exhibit ovulation synchrony (ovulations occurred 12-22 days after steroid treatment), whereas ovulation synchrony was satisfactory in group 2 mares (interval to ovulation being 20.4 ± 1.5 days, range 17-22 days). Using sustained release preparations of progesterone plus estradiol-17β, with PGF₂α administered on Day 10, could eliminate the need for daily injections of steroid preparations in oil when synchronizing estrus and ovulation.     

Ovarian morphology and internal vis-à-vis non internal laying in relation to triacylglycerol,hormones and their receptors concentration around the age of sexual maturity in broiler breeder hens

1. Ovarian morphology, serum hormone concentrations of 17-β-estradiol, progesterone, testosterone, tri-iodothyronine (T3), thyroxine (T4) and triacylglycerol (TAG) were investigated at 23 and 26 weeks of age in broiler breeder hens provided with ad libitum access to feed. Progesterone, oestrogen-β, thyroid-α and -β receptor mRNAs were also quantified in the infundibulum at the same ages.

2. A large variation in the ovarian morphology was observed at 23 weeks of age including hens with undeveloped ovaries, non-laying hens with post ovulatory follicles (POF) and a predominance of non-laying hens without a POF.

3. Serum concentrations of triglyceride, 17-β-estradiol and progesterone at 23 weeks of age were lower in hens with an undeveloped ovary compared with other groups of hens, whereas testosterone, triiodothyronine and thyroxin were higher.

4. At 26 weeks of age, the average number of hierarchical yellow follicles in normal layers was 7.64?±?0·41 whereas in internal layers, the follicular numbers were significantly greater at 8.66?±?0·53. The higher follicular numbers in internal layers were associated with higher serum triglyceride and progesterone concentrations.

5. Oestrogen receptor-β and thyroid receptor-β mRNA was up regulated in the infundibulum of internal layers compared with normal laying hens at 26 weeks of age.     

Effects of calcium soaps of fatty acids on postpartum reproductive function in beef cows.

Twelve multiparous Simmental cows (584 kg) were used to determine the influence of calcium soaps of fatty acids (CSFA) incorporated in a range supplement on postpartum reproductive characteristics. Cows were assigned randomly to receive a control [C; containing grain sorghum (GS) and soybean meal (SBM)] or CSFA-based (containing Megalac [a source of CSFA], GS, and SBM) supplement. Supplements plus prairie hay were individually fed. Diets were isonitrogenous and met the NEm requirement for heavy-milking beef cows in early lactation. Supplement feeding and daily blood collection began at parturition. Calves were removed permanently from cows at 25 +/- 2 d postpartum. Duration of first postpartum estrous cycles was determined by both visual observations and changes in concentrations of progesterone in serum. Concentrations of LH in serum (15-min intervals for 6 h) were determined 12 h before and 48 and 96 h after calf removal. Concentrations of progesterone and estradiol-17 beta in serum were determined daily. Cows receiving CSFA had higher (P = .06) mean concentrations of LH than those receiving C (1.47 vs 1.12 +/- .13 ng/ml). Concentrations of estradiol-17 beta were lower (P less than .02) and serum progesterone were higher (P less than .02) between d 6 and 8 of the induced cycle in CSFA-fed cows. Plasma cholesterol was greater (P less than .01) in cows fed CSFA although plasma triglyceride concentrations were similar between treatments.(ABSTRACT TRUNCATED AT 250 WORDS)     

Excretion patterns of fecal progestagens, androgen and estrogens during pregnancy, parturition and postpartum in okapi (Okapia johnstoni)

The aim of the present study was to establish a simple method to monitor ovarian activity and non-invasively diagnose pregnancy in okapi (Okapia johnstoni). The feces of a female okapi were collected daily or every 3 days for 28 months. Steroids in lyophilized feces were extracted with 80% methanol, and the fecal levels of immunoreactive progestagens (progesterone and pregnanediol-glucuronide), androgen (testosterone), and estrogens (estradiol-17beta and estrone) were determined by enzyme immunoassays with commercially available antisera. Using the progesterone profiles, the durations of the luteal phase, follicular phase, and estrous cycle were determined to be 11.1 +/- 0.4, 5.3 +/- 0.6, and 16.5 +/- 0.7 days (n=22), respectively. Fecal levels of immunoreactive progesterone, pregnanediol glucuronide, and testosterone gradually increased from early pregnancy and peaked several months before parturition. More pregnanediol glucuronide was excreted in feces than progesterone during late pregnancy, but not during the estrous cycle. Although the fecal concentrations of immunoreactive estradiol-17beta and estrone change a little throughout pregnancy and non-pregnancy, they rose sharply and temporarily on the day following parturition. The present study indicates that fecal assays with commercial antisera for progesterone and pregnanediol glucuronide are useful for evaluating luteal activity and diagnosing pregnancy and indicates that estrogens might have some role as a trigger of parturition.     

Endocrine and ovarian changes in response to the ram effect in medroxyprogesterone acetate-primed Corriedale ewes during the breeding and nonbreeding season

Two experiments were performed to determine the endocrine and ovarian changes in medroxyprogesterone acetate (MAP)-primed ewes after ram introduction. Experiment 1 was performed during the mid-breeding season with 71 ewes primed with an intravaginal MAP sponge for 12 days. While the control (C) ewes (n = 35) were in permanent contact with rams, the ram effect (RE) ewes (n = 36) were isolated for 34 days prior to contact with rams. At sponge withdrawal, all ewes were joined with eight sexually experienced marking Corriedale rams and estrus was recorded over the next 4 days. The ovaries were observed by laparoscopy 4-6 days after estrus. Four weeks later, pregnancy was determined by transrectal ultrasonography. In eight ewes from each group, ovaries were ultrasonographically scanned; FSH, LH, and estradiol-17beta were measured every 12 hours until ovulation or 96 hours after estrus. The response to the rams was not affected by the fact that ewes had been kept or not in close contact with males before teasing. No differences were found in FSH, LH, estradiol-17beta concentrations, growth of the ovulatory follicle, onset of estrus, ovulation rate, or pregnancy rate. Experiment 2 was performed with 14 ewes during the nonbreeding season. Ewes were isolated from rams for 1 month, and received a 6-day MAP priming. Ovaries were ultrasonographically scanned every 12 hours, and FSH, LH, estradiol-17beta, and progesterone were measured. Ewes that ovulated and came into estrus had higher FSH and estradiol-17beta levels before introduction of the rams than did ewes that had a silent ovulation. The endocrine pattern of the induced follicular phase of ewes that came into estrus was more similar to a normal follicular phase, than in ewes that had a silent ovulation. The follicle that finally ovulated tended to emerge earlier and in a more synchronized fashion in those ewes that did come into estrus. All ewes that ovulated had an LH surge and reached higher maximum FSH levels than ewes that did not ovulate, none of which had an LH surge. We conclude that (a) the effect of ram introduction in cyclic ewes treated with MAP may vary depending on the time of the breeding season at which teasing is performed; (b) patterns of FSH, and estradiol-17beta concentrations, as indicators of activity of the reproductive axis, may be used to classify depth of anestrus; and (c) the endocrine pattern of the induced follicular phase, which is related to the depth of anestrus, may be reflected in the behavioral responses to MAP priming and the ram effect.     

Pfaffia paniculata-induced changes in plasma estradiol-17beta, progesterone and testosterone levels in mice

The present study undertook chemical analysis of components of Pfaffia paniculata roots. In addition, an animal experiment was conducted in which mice had ad libitum access to water enriched with powdered P. paniculata root for 30 days. Changes in plasma concentrations of estradiol-17beta and progesterone in female mice and of testosterone in male mice were ascertained. The results revealed that P. paniculata roots contain two types of phytosteroids, beta-sitosterol and stigmasterol, in addition to other compounds such as pfaffic acid, allantoin, saponins, beta-sitosteryl-beta-D-glucoside, and stigmasteryl-beta-D-glucoside. Regarding changes in plasma concentrations of hormones, levels of the sex hormones estradiol-17beta, progesterone and testosterone were clearly higher for mice that drank P. paniculata root-enriched water than for mice that drank plain water. Powdered P. paniculata root is easily dissolved in feed or water, and as no adverse reactions were seen in mice within 30 days of oral intake, consumption of P. paniculata for long periods of time appears safe.     

Involvement of high-density lipoprotein in stimulatory effect of hormones supporting function of the bovine corpus luteum

The hypothesis that epinephrine (noradrenaline, NA) enhances utilisation of high-density lipoproteins (HDL) by bovine luteal cells and that this process involves phospholipase (PL) C and protein kinase (PK) C intracellular pathway was tested. Luteal cells from days 2-4, 5-10 or 11-17 of the oestrous cycle were preincubated for 20 h. Subsequently DMEM/Ham's F-12 medium was replaced by fresh medium and the cells were treated for 6 h as follows: In Experiment I with HDL (5-75 micrograms cholesterol per ml), NA, isoprenaline (ISO) or luteinising hormone (LH). In Experiment II cells were incubated for further 24 h in deficient medium (without FCS) and next treated as in Experiment I. In Experiment III cells were stimulated with NA, ISO or LH alone and together with HDL. In Experiment IV cells were treated with PLC inhibitor (U-73122) or with PKC inhibitor (staurosporine) or stimulator (phorbol 12-myristrate 13-acetate) and with either NA, insulin or LH. Only luteal cells from days 5-10 of the cycle responded on HDL and beta-mimetics (P < 0.05). LH stimulated progesterone secretion from the luteal cells during all stages of the cycle (P < 0.001). Cells incubated in deficient medium and supplemented with HDL secreted as much progesterone as those stimulated by LH in all stages of the cycle. Beta-mimetics were unable to enhance the stimulatory effect of HDL. Blockade of PLC had no influence on progesterone secretion from cells treated with either NA or LH, but this did impair the stimulatory effect of insulin (P < 0.05). Similarly, blockade of PKC by staurosporine impaired (P < 0.05) the effect of insulin only but not that observed after LH or NA treatment. We suggest that: (a) noradrenergic stimulation does not enhance utilisation of cholesterol from HDL for progesterone secretion; (b) the fasting of luteal cells seems to activate enzymes responsible for the progesterone synthesis; (c) effect of NA on progesterone secretion from luteal cells does not involve the PLC-PKC pathway.     

Age-related changes of reproductive hormones in young Meishan boars

Meishan pigs are known for their early sexual maturity. On the other hand, they grow slowly. There is no information currently available about the combination of these two characteristics in Meishan pigs. To study the developmental characteristics of Meishan pigs, the plasma concentrations of LH, FSH, inhibin, testosterone, estradiol-17beta, progesterone and insulin-like growth factor-I (IGF-I) in young Meishan boars were determined using RIA and ELISA. Inhibin decreased with age in weeks, while testosterone and estradiol-17 beta increased. Testosterone increased gradually, and an increase in estradiol-17beta occurred after sexual maturity. IGF-I increased before puberty and subsequently decreased just after puberty like a pubertal IGF-1 surge. FSH, LH and progesterone did not change with age. There was no significant correlation among the hormones. During the experimental period, the Meishan boars showed large individual differences. These differences may depend on the fact that Meishan boar reach maturity at 12 weeks of age and continue to grow thereafter.