Effect of peripheral concentrations of progesterone on follicular growth and fertility in ewes

The effects of progesterone (P₄) on follicular growth and fertility in ewes were examined. In Experiment 1, 22 ewes received either one or three packets of P₄ (5 g/packed) or an empty packet subcutaneously (sc) from Days 5 to 15 of the estrous cycle (estrus = Day 0). On Day 6, P₄-treated ewes received 12.5 mg of prostaglandin F₂α. Follicles ⩾3 mm in diameter were observed via transrectal ultrasonography daily from Day 4 through estrus, corpora lutea (CL) were observed 5 to 7 d after estrus. Ewes with low (LOW; ⩽1 ng/ml; n = 5), intermediate (MED; > 1 and <2 ng/ml; n = 10), or normal (NOR; ⩾2 ng/ml; n = 7) P₄ in jugular plasma on Days 7 through 15 differed in follicular development. The largest follicle at estrus was larger in ewes with LOW vs. MED and NOR P₄ (7.8 ± 0.3 vs. 6.9 ± 0.2 mm; P < 0.05). Treatments differed in proportions of multiple-ovulating ewes, in which the oldest ovulatory follicle was first observed before Day 10 (LOW: 3 of 3, MED: 6 of 10, NOR: 0 of 5, respectively; P < 0.05). Estradiol was higher early in the treatment period in LOW ewes than in MED and NOR ewes (day × treatment; P < 0.05). In Experiment 2, ewes received 5 mg of P₄ in corn oil (low progesterone [LP]; n = 51) or 2 ml of corn oil (CON; n = 49) sc every 12 hr on Days 6 through 14 of the estrous cycle before mating. LP ewes received 15 mg of prostaglandin F₂α on Day 6. Mean serum P₄ on Days 7 through 15 was 0.6 ± 0.1 ng/ml in LP and 1.9 ± 0.1 ng/ml in CON ewes. Eleven LP and 12 CON ewes were scanned daily from Day 4 through mating, and in all ewes (n = 93), CL were counted 10 d after mating and embryos were counted at 25, 40, and 60 d of gestation. In multiple-ovulating ewes, day of cycle of appearance was earlier for the oldest (Day 6.1 ± 0.8 vs. 10.4 ± 0.8) but not second oldest (Day 11.7 ± 1.0 vs. 12.2 ± 0.9) ovulatory follicles in LP compared with CON ewes. The conception rate was lower in LP (72%) than in CON ewes (98%; P < 0.01). However, numbers of CL 10 d after mating, and in pregnant ewes, numbers of embryos 25 d after mating and lambs born, did not differ with treatment. In summary, low P₄ increased the size of the largest follicles and the age of the oldest ovulatory follicles. Embryos resulting from the ovulation of older and younger follicles in the same ewe did not differ in their ability to survive.     

Gonadotropin concentrations, follicular development, and luteal function in pituitary stalk-transfected ewes treated with bovine follicular fluid.

Two experiments, each arranged as a 2 x 2 factorial, were conducted in ewes to examine direct effects of bovine follicular fluid (bFF) on follicular development and luteal function and to further characterize follicular development and luteal function after pituitary stalk transection (SS). In Exp. 1, ewes were sham-operated or SS on d 6 of an estrous cycle and received 5 ml of saline or bFF three times daily on d 5 through 11 of the same cycle. In Exp. 2, all ewes were SS on d 6 of an estrous cycle and treated with saline or bFF three times daily on d 5 through 11 and with ovine FSH (60 micrograms; NIADDK-oFSH-16) or saline (1.2 ml) from d 7 to 11. In Exp. 2, ewes were ovariectomized on d 11 to assess effects of treatments on follicular development and luteal function. In both experiments, concentrations (ng/ml) of FSH on d 7 were suppressed (P less than or equal to .005) by bFF compared with saline (.50 +/- .17 vs 1.63 +/- .15) and remained suppressed (P less than or equal to .005) through d 11 (.46 +/- .12 vs 1.54 +/- .12). Replacement therapy (oFSH) restored concentrations of FSH. Concentrations of LH were not affected by bFF but were elevated (P less than or equal to .05) 1 d after SS (d 7; .88 +/- .09 vs .56 +/- .09) and remained elevated (P less than or equal to .05; 1.31 +/- .20 vs .65 +/- .11) from d 6 through 11. Concentrations of progesterone were unaffected by SS.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of exogenous estradiol and progesterone on plasma concentrations of leptin in ewes in non-breeding season

Estradiol and progesterone may play a role in controlling leptin secretion by utilizing their receptors in adipocytes and the genomic mechanisms of the leptin gene. This study was conducted to evaluate the effects of exogenous sex steroids on the blood leptin concentrations in ewes in the non-breeding season. Multiparous ewes were fed to maintenance level for their live weights. Blood samples were collected at 12-h intervals from Days -3 to -1 to determine the basal leptin levels (pre-injection period). From Day 0 to Day 5 (injection period), blood sampling continued at 12-h intervals, and the ewes were injected intramuscularly at 24-h intervals with oil, 50 mg progesterone in oil, 1 mg of estradiol in oil, or both steroids in oil. Leptin was measured using a sensitive and specific radioimmunoassay based on recombinant bovine leptin. Overall, plasma concentrations of leptin were not affected by any of the steroid treatments, and there were no differences in the value of leptin between the pre-injection and injection periods among the 4 groups. Therefore, the exogenous estrogen and progesterone used in this study do not have a strong effect on the blood leptin concentrations of ewes in the non-breeding season.     

高精料日粮对湖羊发情、孕酮水平及黄体PLINs基因表达的影响

高精料日粮会引起瘤胃代谢机能异常,引发机体内分泌紊乱。本试验旨在研究高精料日粮对湖羊发情、孕酮水平及黄体组织围脂滴蛋白(PLINs)家族成员mRNA表达的影响。试验选用5月龄雌性湖羊20只,分为低精料组(LC,精粗比2∶8,n=10)和高精料组(HC,精粗比6∶4,n=10)。于正试期第20 d进行同期发情处理,在正试期的第58天采集卵巢,分离黄体,采集血液检测孕酮(P_4)和脂多糖(LPS)的含量,统计平均日增重(ADG)及干物质采食量(DMI)。应用实时定量PCR检测黄体PLINs基因的表达量,并对LPS浓度、卵巢重、黄体数量、孕酮水平及PLINs基因表达量进行相关性分析。结果显示,高精料组ADG显著高于低精料组(P0.05),DMI显著低于低精料组(P0.05)。高精料组血清LPS浓度显著高于低精料组(P0.05)。高精料和低精料组母羊自然发情率分别为30%和70%(P=0.089)。2组母羊黄体期血清P_4浓度、卵巢重和黄体数量均无显著差异(P0.05)。PLINs家族成员mRNA在黄体组织中均有不同程度的表达,其中PLIN2 mRNA表达量最高,且低精料组PLIN2 mRNA表达量显著高于高精料组(P0.05),而PLIN3、PLIN4和PLIN5的mRNA表达量则显著低于高精料组(P0.05)。LPS浓度与PLIN2 mRNA表达量呈显著负相关(P0.05),与PLIN3 mRNA表达量呈显著正相关(P0.05)。PLIN3 mRNA表达量与PLIN4、PLIN5呈显著正相关(P0.05),与PLIN2呈显著负相关(P0.05)。结果表明,高精料日粮饲喂使湖羊血清LPS浓度升高,可能不利于母羊的发情。黄体组织PLINs基因在高精料组和低精料组之间的表达差异,提示PLINs基因表达可能受LPS及饲料中精粗比的影响,同时为进一步研究营养与繁殖的关系提供数据支撑。     

Effects of repeated hCG injections on reproductive efficiency in mares

Thirty reproductively sound mares were divided into treatment and control groups. In the treatment group, consisting of 14 mares, 2500 I.U. of human chorionic gonadotropin (hCG) was administered intravenously during estrus, in the presence of a 35 mm follicle over five successive cycles in 1987, and at least two cycles in 1988. Beginning with the second cycle of treatment in 1988, these mares were bred to a fertile stallion. The control group, consisting of 16 mares, was followed for two to five cycles in either the 1987 or 1988 season and six of these mares were bred to fertile stallions. Throughout the study period, blood was collected from the mares in the treatment group for analysis of anti-hCG antibodies and cross reactivity of the antibody to purified equine lutenizing hormone (eLH) and equine chorionic gonadotropin (eCG).In 1987, after the first three injections of hCG, mean duration of estrus in treated mares tended to be shorter than in control mares (P<.10). After all five hCG injections in 1987, mean ovulation time for treated mares was shorter than in control mares (P<0.01). However, after two to five hCG injections in 1987, seven treated mares (50%) had some individual ovulation times that did not differ from the control mares.Initially, following the first three injections of hCG in 1988, mean duration of estrus tended to be shorter (P<0.1) in treated mares compared to control mares. A reduction in mean ovulation time was observed after the first two hCG injections of 1988 (P<0.01). However, after one to four hCG injections in 1988, eight treated mares (57.1%) had some individual ovulation times that did not differ from controls.In 1987, all 14 treated mares developed significant levels of antibodies to hCG after one to four injections, and again in 1988, were positive for anti-hCG antibodies after one to three injections. However, no correlation was observed between magnitude of the immune response and duration of ovulation time or pregnancy rate. In cross reactivity studies, no significant binding of plasma anti-hCG antibodies to either eLH or eCG was observed in vitro.Overall, pregnancy and foaling rates of treated (85.7%) and control mares (83.3%) did not differ. Additionally, no difference was observed in number of inseminations per estrus between treated and control mares. In this study, with successive injections of hCG, the expected shortened time to ovulation was not elicited consistently in all mares. However, mares continued to ovulate, conceive and foal in the presence of significant levels of anti-hCG antibodies.     

Leptin infusion during the early luteal phase in ewes does not affect progesterone production

Infusion of leptin during the ovine follicular phase has been shown to increase progesterone secretion during the subsequent luteal phase. In this study, we have assessed the effects of infusing leptin during the early luteal phase. Infusion of leptin (2.5 microg/h) into the ovarian artery of ewes with ovarian autotransplants (n=5) on day 3 of the luteal phase for 12h did not affect progesterone estradiol or LH concentrations compared to control ewes (n=5). These results suggest no direct effect of leptin on ovarian function at this stage of the estrous cycle.     

Effects of charcoal-extracted, bovine follicular fluid on gonadotropin concentrations, the onset of estrus and luteal function in heifers

A study was conducted to determine the effect of charcoal-extracted, bovine follicular fluid (CFF) on plasma follicle stimulating hormone (FSH) and luteinizing hormone (LH) concentrations, the interval from luteolysis to estrus, and subsequent luteal function in heifers. Fifteen Angus, Simmental and Hereford heifers were allotted by age, weight and breed to a control (C, n = 8) or a CFF (n = 7) group. Heifers received injections of saline or CFF (iv, 8 ml/injection) every 12 h from d 1 (d 0 estrus) through d 5 of the estrous cycle. On d 6, each heifer was injected (im) with 25 mg of prostaglandin F2 alpha (PGF2 alpha). Blood samples were collected every 12 h by venipuncture starting just before the first saline or CFF injection and continuing until estrus. Thereafter, blood samples were collected every other day during the subsequent estrous cycle and assayed for FSH, LH, estradiol-17 beta and progesterone by radioimmunoassay. Injections of CFF had no effect (P greater than .05) on circulating FSH or LH concentrations from d 1 to 5 relative to the C group; however, there was a transient rise (P less than .05) in FSH concentrations 24 h following cessation of CFF injections. This transient rise in FSH was not immediately followed by an increase in plasma estradiol-17 beta concentrations. Although CFF injections did not interfere with PGF2 alpha-induced luteolysis, the interval from PGF2 alpha injection to estrus was delayed (P less than .05) by 5 d in the CFF group compared with the C group.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of different concentrations of Chir98014 as an activator of Wnt/beta-catenin signaling pathway on oocyte in-vitro maturation and subsequent embryonic development in Sanjabi ewes

The present study was conducted to investigate the effects of the activator factor of the WNT pathway, chir98014, leading to the in vitro sheep oocyte maturation medium, on the cumulus cell development, different nuclear maturation stages and the following process of embryonic development. Experiments included (a) addition of different concentrations (0, 0.1, 0.5, 1 µm) of chir98014 to the maturation medium and evaluation of the cumulus cell expansion, (b) addition of different concentrations of chir98014 to the maturation medium and investigation of different nuclear maturation stages, (c) addition of different concentrations of chir98014 to the maturation medium and examination of the subsequent embryonic maturation process and (d) addition of different concentrations of chir98014 to the embryonic development culture medium (the first 48 hr) and investigation of the subsequent embryonic development process. The extracted data were analysed using the SPSS software, considering the significance level of p < .05 and making the mean comparisons. The results showed that the addition of the 0.1 µM concentration of chir98014 to the maturation medium had no significant effects on the oocyte maturation and embryo development post-fertilization but it enhanced the Cumulus-oocyte complexes (COCs) expansion. In the fourth experiment, the low concentration of chir98014 in the embryo culture media improved the embryo development process, whereas the high one had a detrimental effect on it, as compared to the control group. Thus, the presence of the lower concentrations of this compound in the embryonic culture medium had favourable effects on the development of embryos.     

Effects of progestagens on follicular growth and oocyte developmental competence in FSH-treated ewes

Previous research has reported evidence for negative effects of progestagens on follicular growth and oocyte competence. In the present study, negative effects of progestagens on follicular growth and oocyte developmental competence were assessed. During the breeding season, 20 Sarda ewes were treated with two doses of cloprostenol, 10 days apart, to assure the presence of a corpus luteum (CL). On day 5 after the second cloprostenol dose, 10 ewes were treated with a progestagen sponge while 10 females remained untreated. Starting on day 7 after the second cloprostenol dose, all the ewes were treated with 6 equal doses of 24 I.U. of FSH (Ovagen™, ICP, NZ), every 12 h. The number of follicles ≥2 mm in diameter increased (P < 0.0005) in all the ewes from 24 h before to 60 h after the first FSH dose (from 12.8 ± 1.1 to 23.4 ± 1.3 in treated and from 12 ± 0.6 to 22 ± 1.2 in untreated ewes, n.s.). There were no significant differences in follicle dynamics between groups, but concentrations of estradiol in control ewes were higher than in the progestagen group (P < 0.05). Twelve hours after the last FSH dose, oocytes were collected by ovum pick-up. Recovery rates were lower for progestagen-treated ewes (71.1 versus 83%; P < 0.001). After IVP procedure, cleavage rate was also lower in the progestagen group (39.1 versus 82.6%; P < 0.001). Furthermore, blastocysts output revealed that oocyte developmental competence was lower in progestagen group (17.3 versus 30.4%; P = 0.245), although differences were not significant. These results suggest deleterious effects from progestagen on oocyte developmental competence and set the basis for new protocols for in vitro embryo production.     

Effects of melatonin and progesterone administered to ewes in spring and summer

Melatonin (MEL) was evaluated for effects on LH, prolactin (PRL) and fertility in spring (Exp. 1, 2) and summer (Exp. 3 to 5). In Exp. 1, 17 ovariectomized ewes bearing estradiol implants were fed 3 mg MEL or vehicle for 44 d beginning May 1. Melatonin decreased (P less than .001) PRL levels but had no effect on LH secretion and response to GnRH. In Exp. 2, 12 ewes each received a 40-d MEL ear implant or a sham implant on March 31. Progesterone-releasing pessaries (CIDR) were applied for 12 d and were withdrawn concomitant with ram joining on May 7. Neither treatment stimulated follicular development or induced estrus or ovulation. Exp. 3 and 4 were contemporary 2 x 2 factorial trials with 24 ewes at each of two locations. Melatonin implants were administered on June 29 and CIDR on July 22. The CIDR were removed and rams (Exp. 3, vasectomized; Exp. 4, fertile) were joined on August 3. Days from introduction of rams to estrus were reduced (P less than .05) by CIDR but not by MEL. All ewes lambed in Exp. 4, and days to estrus and conception were reduced (P less than .001) by CIDR but not by MEL. Exp. 5 was designed like Exp. 4 except that MEL implants were inserted June 20 and rams were joined August 8. Intervals from introduction of rams to estrus were reduced (P less than .01) by both MEL and CIDR treatments.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of GnRH or progesterone treatment on day 5 post-AI on plasma progesterone, luteal blood flow and leucocyte counts during the luteal phase in dairy cows

This study was designed to test the effects of progesterone or GnRH treatment on day 5 post-AI on fertility and luteal function in dairy cows and heifers. Five days after AI, 32 animals were randomly assigned to a control, intravaginal progesterone for 14 days progesterone releasing intravaginal device (PRID) or GnRH treatment group. On days 5, 7, 12, 14, 17 and 19 post-AI, each animal underwent colour Doppler ultrasonography of the corpus luteum and blood samples were collected for cell counts and plasma progesterone determination. Through general linear model repeated measures analysis of variance, significant effects were observed of treatment, parity, inseminating bull, reduced vascularization of the CL and pregnancy on plasma progesterone concentrations, whereas mean plasma progesterone and time luteal phase day, and treatment and plasma progesterone concentration on day 5 post-AI were found to, respectively, affect neutrophil and lymphocyte counts throughout the luteal phase. Moreover, two binary logistic regression analyses were performed. Based on the odds ratio, the likelihood of pregnancy by days 26-32 post-AI was 23.4 times higher in animals with high mean progesterone levels throughout the study period, compared with animals with low mean progesterone. The likelihood of reduced CL vascularization was 14 times higher in animals treated with PRID, compared with control and GnRH-treated animals. In conclusion, our results indicate that treatment on day 5 post-AI with PRID reduced subsequent CL vascularization, whereas GnRH treatment increased plasma progesterone concentrations on day 12 post-AI, although an effect was identified of the inseminating bull on plasma progesterone levels. Pregnant animals showed higher mean plasma progesterone concentrations than in nonpregnant ones and heifers higher than in lactating cows, whereas blood cell counts differed depending on the treatment and on the mean plasma progesterone concentration on day 5 post-AI.     

Effect of ammonia‐generating diet on ovine serum and follicular fluid ammonia and urea levels,serum oestrogen and progesterone concentrations and granulosa cell functions

This study was undertaken to elucidate the effect of ammonia‐generating diet on serum and follicular fluid ammonia and urea levels, serum oestrogen and progesterone concentrations and granulosa cell growth and secretion parameters in ewes (Ovis aries). Ewes were fed with 14% CP diet (control) or ammonia‐generating diet or ammonia‐generating diet plus soluble sugar. The serum and follicular fluid ammonia and urea level, serum oestrogen and progesterone levels and granulosa cell (obtained from ovaries of slaughtered ewes) growth parameters and secretory activities were estimated. Ammonia‐generating diet (high‐protein diet) increased the serum ammonia and urea concentration. Supplementation of soluble sugar significantly reduced the ammonia concentration in serum with comparable levels as in control group; however, the urea level in the same group was higher than that observed in control group. Supplementation of soluble sugar significantly reduced the follicular fluid ammonia concentration; however, the level was significantly higher compared to control group. Supplementation of soluble sugar brought down the follicular fluid urea level comparable to that observed in control group. Oestrogen and progesterone levels remained unchanged in ewes fed with different types of diet. Oestrogen and progesterone secretion were significantly lowered from granulosa cells recovered from ewes fed with high ammonia‐generating diet. Low metabolic activity and high incidence of apoptosis were observed in granulosa cells obtained from ovaries of ewes fed with ammonia‐generating diet.     

Follicle stimulating hormone pattern and luteal function in ewes receiving bovine follicular fluid during three stages of the estrous cycle

The objectives of this study were to determine 1) the ability of charcoal-extracted bovine follicular fluid (bFF) to suppress endogenous follicle stimulating hormone (FSH) at various stages of the estrous cycle and 2) the effects of suppression of FSH on luteal function and lengths of the current and subsequent estrous cycles. Twenty-six mature ewes were assigned randomly to receive 5 ml of either bFF or saline, subcutaneously, at 8-h intervals on d 1 through 5 (bFF n = 6; saline n = 3), d 6 through 10 (bFF n = 6; saline n = 3) or d 11 through 15 (bFF n = 6; saline n = 2) of the estrous cycle (d 0 = estrus). Blood was collected daily beginning at estrus and continued until the third estrus (two estrous cycles) or 40 d; more frequent samples were collected 2 h prior to initiation of treatment (0600), hourly for the first 8 h of treatment, then every 4 h until 0800 on the first day after treatment, and finally at 1600 and 2400 on that day. Plasma concentrations of FSH were lower (P less than .001) in bFF-treated than in saline-treated ewes. Treatment with bFF reduced (P less than .05) plasma concentrations of progesterone during the current but not during the subsequent estrous cycle. Treatment with bFF did not affect plasma concentrations of estradiol-17 beta. Administration of bFF on d 11 through 15 of the estrous cycle lengthened the interval from the decline in progesterone to estrus and the inter-estrous interval by approximately 3 and 4 d, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of exogenous progesterone and/or prolactin on Haemonchus contortus infections in ovariectomized ewes

Reproduction can alter the course of ovine nematodiasis; fecal nematode egg concentrations often increase near lambing and throughout lactation, a phenomenon referred to as the periparturient rise. To identify the host mechanisms that might link these disparate events, i.e. lactation and the fecundity of gastrointestinal trichostrongyles, ovariectomized ewes were injected daily with progesterone and/or prolactin, or saline. Progesterone treatment commenced 20 days before inoculation with Haemonchus contortus and prolactin was administered throughout the 30-day infection period. Ovariectomized ewes receiving prolactin during the experimental infection maintained higher daily fecal egg concentrations than the progesterone, progesterone/prolactin, or the control treatment groups. However, ovariectomized ewes that received 20 days of pre-inoculation exposure to progesterone, as well as prolactin during the infection, had greater numbers of nematodes that were larger than the other treatment groups. Thus, the sequential delivery of these hormones that are associated with the reproductive cycle in ewes produced some of the same results that occur during periparturient rise.