Cortisol disrupts the ability of estradiol-17β to induce the LH surge in ovariectomized ewes

Stress disrupts the preovulatory luteinizing hormone (LH) surge in females, but the mechanisms are unknown. We tested the hypothesis that cortisol compromises the ability of estrogen to induce a preovulatory-like LH surge in ovariectomized ewes in both the breeding and nonbreeding season. Luteinizing hormone surges were induced in ovariectomized ewes by treatment with progesterone followed by a surge-inducing estradiol-17β (E2) stimulus using a crossover design. The experiment was replicated in the breeding and nonbreeding seasons. Cortisol reduced the incidence of LH surges irrespective of season. Cortisol increased the latency from E2 stimulus to the onset of the surge in the breeding season only and suppressed the LH surge amplitude during both seasons (P < 0.01). We conclude that cortisol can interfere with the LH surge in several ways: delay, blunt, and in extreme cases prevent the E2-induced LH surge. Furthermore, the effect of cortisol to delay the E2-induced LH surge is more pronounced in the breeding season. These results show that cortisol disrupts the positive feedback effect of E2 to trigger an LH surge and suggest the involvement of multiple mechanisms.     

Follicular dynamics and changes in oestradiol‐17β, progesterone and LH profiles following PGF2α induced oestrus in early and late ovulating Beetal goats

This study compares the factors associated with variable interval to oestrus and ovulation between early versus late ovulating goats following PGF_2α administration. The time of ovulation in Beetal goats (n = 38) was monitored through transrectal ultrasound at every 6 hr following a single dose of PGF_2α (experiment 1). Variations in oestrus and ovulation times were further explored through the changes in follicular dynamics, endocrine profiles and behaviour in another set of goats (n = 13) following single PGF_2α given randomly during the luteal phase (experiment 2). The ovulation time varied between 60 and 96 hr, and 57% of ovulations occurred by 72 hr following PGF_2α (experiment 1). Accordingly, the goats (n = 13) in the second experiment were retrospectively divided either into early and/or late ovulating, that is, ≤72 and/or ≥84 hr following PGF_2α. The onset of oestrus, peak estradiol‐17β concentration and LH surge after PGF_2α was first observed in early than late ovulating goats (p < 0.05). The goats ovulating early had larger follicle and smaller CL in diameter at the time of PGF_2α administration than those ovulating late (5.4 ± 0.2 vs. 4.3 ± 0.2 mm and 10 ± 0.6 vs. 11.8 ± 0.3 mm, respectively; p < 0.05). Likewise, plasma progesterone concentration tended to be lower (p = 0.087) in early than late ovulating goats. In conclusion, the size of dominant follicle and CL at the time of PGF_2a determines the interval to ovulation following a single dose of PGF_2a during the luteal phase.     

Altered Expression of 3β‐HSD,CYP17 and 17β‐HSD in the Foetal Porcine Gonads in Response to Anti‐androgen Flutamide Exposure

We investigated whether the limited access to androgens during late prenatal period alters expression of steroidogenic enzymes involved in androgen production: 3β‐hydroxysteroid dehydrogenase/Δ5‐Δ4 isomerase (3β‐HSD), cytochrome P450 17α‐hydroxylase/17,20‐lyase (CYP17) and 17β‐hydroxysteroid dehydrogenase type 1 (17β‐HSD1) or type 3 (17β‐HSD3) in the foetal porcine gonads. Pregnant gilts were injected with anti‐androgen flutamide (for seven days, 50 mg/day/kg bw) or corn oil (control) starting at 83 (GD90) or 101 (GD108) gestational day. To assess 3β‐HSD, CYP17 and 17β‐HSD1 or 17β‐HSD3 expression, real‐time PCR and immunohistochemistry were performed. In testes from flutamide‐treated foetuses, increased 3β‐HSD and CYP17 mRNA expression was observed in the GD90 group, while decreased 3β‐HSD and 17β‐HSD3 mRNA expression and increased CYP17 mRNA expression were found in the GD108 group. CYP17 and 17β‐HSD3 were localized in Leydig cells. Following flutamide administration, the intensity of CYP17 immunostaining was higher in both treated groups, while 17β‐HSD3 intensity was lower in the GD108 group. In ovaries from flutamide‐treated foetuses in the GD90 group, mRNA level for 3β‐HSD was elevated, but it was diminished for CYP17 and 17β‐HSD1. In the GD108 group, flutamide treatment led to lower mRNA level for 3β‐HSD but higher for CYP17. 3β‐HSD was found in granulosa cells, while CYP17 was localized within egg nests and oocytes of forming follicles. Following flutamide treatment, the intensity of 3β‐HSD and CYP17 immunostaining was higher in the GD90 and GD108 groups, respectively. Immunohistochemical staining for 3β‐HSD was restricted to the ovary. Concluding, diminished androgen action in the porcine foetal gonads during late gestation induces changes in steroidogenic enzymes expression, which may led to changes in gonadal function. However, it seems that androgens exert diverse biological effects depending on the gestational period.     

Plasma prolactin, LH, FSH and estrogen excretion patterns in gilts during sexual development

Plasma prolactin (PRL), luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured by radioimmunoassay in groups of eight gilts sampled every 20 min for 6 h at about 2-wk intervals between 15 and 192 d of age. The PRL levels were high at 15 and 28 d, declined at 40 d just after weaning and then rose slowly until 192 d of age. The number of LH pulses during 6 h was higher between 83 and 125 d than at the other periods. Magnitude of LH pulses was highest at 15 d, constant from 54 to 125 d, fell at 137 d and remained low until 192 d. Plasma FSH was high from 15 to 125 d, with a maximum at 54 d. It declined slowly until 168 d and did not change thereafter. Estrogen excretion was estimated from urine excretion of estrone (E1; conjugated plus nonconjugated E1) per 24 h from 40 d until puberty in three gilts and at 156 and 174 d in two other animals. The E1 excretion increased with age and four levels were described before peak values with the onset of first estrus. The first increase in E1 excretion occurred between 68 and 110 d, when antral follicles appeared in the ovaries. It was subsequent to the highest levels of FSH and concomitant with the increased frequency of LH pulses. The drop in levels of both gonadotropins after 125 d probably corresponded to the development of the negative feedback as a result of greater ovarian activity in these gilts.     

Circulating concentrations of 17-estradiol influence pattern of LH in circulation of cows

The objective of the research was to determine the relationship between circulating 17β-estradiol (E2) and secretion of luteinizing hormone (LH) in cows. A second objective was to determine if response to E₂ was influenced by interval between ovariectomy and the start of E₂ treatment. Thirty-one nulliparous cows 3 yr of age were randomly assigned to a 2 × 4 factorial arrangement of treatments. Sixteen cows were ovariectomized at 18 mo of age (long term), and the other 15 cows were ovariectomized at 36 mo of age (short term). At the time of ovariectomy of cows in the short term group, 11 cows in the short term group and 12 cows in the long term group were implanted subcutaneously with 1, 2 or 4 polydimethylsiloxane capsules containing E₂. The other eight cows served as non-implanted controls (n=4-short term, n=4-long term). All cows were fitted with jugular vein catheters on day 29 of treatment, and on day 30 blood samples were collected at 12-min intervals for 6 hr. At the end of 6 hr, luteinizing hormone-releasing hormone (LHRH) was administered and blood sampling continued at 12-min intervals for an additional hour. Serum was analyzed for LH and E₂. Variables of LH secretion analyzed were mean concentration, frequency of pulses, amplitude of pulses and maximum concentration after LHRH. There were no significant interactions for any of the variables of LH among cows ovariectomized for the long and short term. There was a significant linear increase in mean concentration of LH with increased circulating concentration of E₂. Frequency of LH pulses was not affected by circulating concentration of E₂. As circulating concentration of E₂ increased, amplitude of LH pulses increased and response to LHRH increased - resulting in an increase in mean LH. Interval from time of ovariectomy to the start of E₂ treatment only had a minor influence on mean concentration of LH and profile of LH concentrations in circulation.     

Effect of anthelmintic treatment on sexual maturation in prepubertal beef heifers

Heifers treated with ivermectin at weaning have been reported to reach puberty at a younger age and lighter weight than untreated heifers. We tested the hypothesis that heifers administered ivermectin would respond with earlier follicular development and a greater LH response to a 1-mg estradiol-17beta challenge (E2C) than untreated heifers. Fall-born Angus heifers (n = 32) were randomly assigned on 284 +/- 9 d of age (215.5 +/- 20.8 kg) to receive ivermectin (IVR) or albendazole (ALB), IVR + ALB, or to remain as untreated controls (CONT). Each group (n = 8) was housed separately in adjacent pens throughout the trial and managed to gain .8 kg/heifer on a ration containing 13.2% CP, 58.8% TDN, and 49.9% DM. The CONT heifers received an additional 2.27 kg/heifer of corn silage and 1.59 kg/heifer of corn daily to maintain ADG at comparable levels. Individual body weight was recorded weekly, and nematode eggs per gram (EPG) of feces were measured every 21 d. Ultrasonography was performed on alternate days starting 2 wk prior to E2C to characterize follicular wave patterns. Follicles were separated into classes (C1 [3 to 5 mm], C2 [6 to 9 mm], and C3 [10 mm]) and sizes (largest [LF], second [SLF], third [TLF], and fourth largest follicles [FLF]). The sizes of the regressing dominant follicle 1 (DF1) and the progressing dominant follicle 2 (DF2) were also determined. Serum concentrations of LH were determined from hourly jugular blood samples collected 8 to 24 h after injection of E2C. The IVR + ALB treatment group had more C3 follicles than ALB and CONT (P < .07). The IVR-treated heifers had larger TLF than ALB and CONT (P < .04). The IVR- and IVR + ALB-treated heifers had larger FLF and DF2 than ALB and CONT (P < .1). Least squares means for DF2 were 9.5 +/- .5, 8.0 +/- .4, 9.5 +/- .3 and 8.3 +/- .3 mm, for IVR, ALB, IVR + ALB and CONT, respectively (P = .02 for treatment effect). The E2C-induced serum LH concentration did not differ with respect to treatment. We conclude that heifers administered IVR display increased follicular development, supporting our earlier investigations regarding reduced age at puberty in heifers treated with IVR near weaning.     

Effect of estradiol‐17β during in vitro growth culture on the growth,maturation, cumulus expansion and development of porcine oocytes from early antral follicles

Growing porcine oocytes from early antral follicles can acquire meiotic and developmental competence under suitable culture conditions, but at lower rates compared to full‐grown oocytes. We postulated that estradiol‐17β (E₂) supported the acquisition of meiotic and developmental competence as well as cumulus‐expansion ability during growth culture. Growing oocytes from early antral follicles (1.2 to 1.5 mm in diameter) were grown in vitro for 5 days in a medium containing 0, 10^?7, 10^?6, 10^?5 or 10^?4 mol/L E₂; after in vitro maturation, 35, 58, 47, 74 and 49% of oocytes matured to metaphase II, 25, 79, 77, 90 and 97% acquired cumulus‐expansion ability, and 23, 54, 63, 89 and 64% were fully surrounded by cumulus cells, respectively. Following maturation, electro‐stimulation was applied to the oocytes grown with 10^?5 mol/L E₂. After 6 days of culture, in vitro‐grown oocytes developed to the blastocyst stage at a rate similar to that for full‐grown oocytes (31% and 40%, respectively). Therefore, we suggest that the use of E₂ during growth culture improves the meiotic and developmental competence of oocytes, cumulus‐expansion ability, and cumulus cell attachment to the oocytes.     

α- and β-adrenoceptors in the sheep urinary bladder

A study was undertaken to determine the presence and distribution of alpha- and beta-adrenoceptors in the sheep bladder body and base. In the bladder body, noradrenaline and isoproterenol induced relaxation which was significantly inhibited by propranolol, pafenolol and butoxamine. In the presence of propranolol (10(-5) M), noradrenaline induced a small contraction, as well as phenylephrine, but B-HT 920 failed to cause any effect on the bladder body. In the bladder base, noradrenaline caused a contraction that was significantly inhibited by prazosin but not by yohimbine. Phenylephrine also induced a contractile response in this structure which was inhibited by prazosin. Isoproterenol caused a relaxation that was significantly inhibited by propranolol and pafenolol but not by butoxamine. Relaxation was mediated by both beta 1 and beta 2-adrenoceptors in the detrusor muscle and by beta 1-adrenoceptors in the bladder base. Alpha 1-adrenoceptors contributed to maintain the detrusor tone and contract the bladder base.     

Dietary supplementation of β‐hydroxy‐β‐methylbutyrate in animals – a review

The leucine metabolite β‐hydroxy‐β‐methylbutyrate (HMB) has been studied by many researchers over the last two decades. In particular, the utility of HMB supplementation in animals has been shown in numerous studies, which have demonstrated enhanced body weight gain and carcass yield in slaughter animals; positive immunostimulatory effect; decreased mortality; attenuation of sarcopenia in elderly animals; and potential use in pathological conditions such as glucocorticoid‐induced muscle loss. The aim of this study was to summarize the body of research on HMB supplementation in animals and to examine possible mechanisms of HMB action. Furthermore, while the safety of HMB supplementation in animals is well documented, studies demonstrating efficacy are less clear. The possible reasons for differences in these findings will also be examined.     

奶牛子宫复旧期间FSH和LH动态变化规律研究

子宫复旧是奶牛产后生殖器官恢复的一个重要时期,复旧时间的长短会直接影响产后第一次发情的时间,子宫复旧的正常时间为产后30～46d。本试验为研究促卵泡素(FSH)与促黄体素(LH)在子宫复旧过程中的变化规律,选择了年龄、胎次相近,健康状况良好的产后奶牛15头,应用放射免疫分析法在子宫复旧期间每3天对奶牛血清中FSH与LH进行一次测定,前后持续10次。由试验数据可得出,FSH与LH分泌水平在子宫复旧的早期比较低,FSH浓度到产后第15天达到峰值,LH从产后第21天开始大幅上升,到产后第24天浓度达到峰值,之后二者浓度变动范围不大。     

Effects of β‐carotene‐enriched dry carrots on β‐carotene status and colostral immunoglobulin in β‐carotene‐deficient Japanese Black cows

Data from 18 β‐carotene‐deficient Japanese Black cows were collected to clarify the effects of feeding β‐carotene‐enriched dry carrots on β‐carotene status and colostral immunoglobulin (Ig) in cows. Cows were assigned to control or carrot groups from 3 weeks before the expected calving date to parturition, and supplemental β‐carotene from dry carrots was 138 mg/day in the carrot group. Plasma β‐carotene concentrations in the control and carrot groups at parturition were 95 and 120 μg/dL, and feeding dry carrots slightly improved plasma β‐carotene at parturition. Feeding dry carrots increased colostral IgA concentrations in cows and tended to increase colostral IgG₁, but colostral IgM, IgG₂, β‐carotene and vitamin A were not affected by the treatment. Feeding dry carrots had no effects on plasma IgG₁, IgA and IgM concentrations in cows, but plasma IgG₁ concentrations decreased rapidly from 3 weeks before the expected calving date to parturition. These results indicate that feeding β‐carotene‐enriched dry carrots is effective to enhance colostral IgA and IgG₁ concentrations in β‐carotene‐deficient cows.     

Gonadal regulation of LH secretion in prepubertal bull calves

In the present study we examined the influence of castration and exogenous estradiol on pulsatile LH release during the transition from infancy to the prepubertal period of development. Bull calves were assigned to treatments (N = 5 treatment) at 6 weeks of age. Treatments consisted of intact controls, castrates and castrates receiving estradiol implants. Plasma LH response was monitored over 8 hr periods at 7, 8, 9, 10, 11 and 13 weeks of age. Castration alone did not alter LH concentrations, compared to controls until 10 weeks of age. At 10, 11 and 13 weeks, mean LH concentration and the number of LH pulses/8 hr period were greater (P less than .05) in castrates than in controls. In castrates with estradiol implants, mean LH concentration and the number of LH pulses/8 hr period were suppressed at all ages compared to controls and castrate treatments. These results indicate that LH release is not inhibited by gonadal factors from 6 to 9 weeks of age in the bull calf. However, estradiol negative feedback on LH secretion is evident during this same developmental period.     

Retinol, β-carotene and β-tocopherol concentrations in mare and foal plasma and in colostrum

Fat-soluble vitamins transfer poorly across the diffuse epitheliochorial placenta of the mare, so the neonatal foal is dependent on its supply via colostrum. Concentrations of retinol (RT), β-carotene (BC), and β-tocopherol (AT) were assayed in samples of plasma, colostrum and milk from nine mares of mixed light breeding and their foals at parturition and days 1, 2, and 4 postpartum. Samples were analyzed simultaneously for RT, BC, and AT using a new, less time-consuming reverse phase high-pressure liquid chromatography (HPLC) method. Feeds were analyzed, and mean daily intakes calculated. Mare plasma RT increased from day 1 to day 4 (P = .033), and plasma AT declined linearly from day 0 to day 4 (P = .006). Colostrum concentrations of all vitamins increased from parturition to day 1, and then declined rapidly over the 4 days. Foal plasma BC increased from parturition to day 4 (P = .080), and plasma AT increased from parturition to day 2 (P < .001), and 4 (P = .060). These observations suggest that three times the current recommendation of vitamin A is sufficient for pregnant mares, in keeping with previous studies of growth, pregnancy, and lactation. In contrast, the linear decrease in mare plasma AT suggests that 1.2 times the current recommendation of vitamin E may be less than optimal for pregnant mares.

Introduction

The diffuse epitheliochorial placenta of the mare does not allow fat-soluble vitamins to cross with ease to the fetus,[1] so their status in the neonatal foal is dependent on colostrum. This first milk contains these vitamins, which are necessary for many functions.Vitamin A is required for vision, reproduction, growth, development, and maintenance of epithelial cells, and osteoclast activity in bone formation. In addition, vitamin A deficiencies reduce resistance to disease, can cause neurological degeneration, and congenital defects.[2] Vitamin E, an antioxidant, prevents lipid peroxidation and cell membrane damage. Its deficiency in horses has been associated with a form of muscular dystrophy and impaired immune function. [3]Studies on the mare and foal have described blood concentrations of one or two of these vitamins,[4, 5 and 6] but only one involved all three. [7] In that study, vitamin concentrations were assayed in mare blood and milk, but only once in foal blood at parturition, so data during the periparturant period is incomplete.The objectives of this enumerative study were to determine concentrations of RT, BC, AT in plasma and colostrum, to evaluate colostral transfer of these vitamins, to assess the adequacy of vitamin intakes of mares, and to develop a time-saving single-run HPLC method.

Materials and methods

Animals

Nine healthy aged multiparous mares of mixed light breeding (13.6 ± 1.5 years, 592 ± 20 kg body mass) and their foals were sampled. The institutional animal care and use committee approved the protocol. Mares were kept on meager late-winter mixed grass pasture, with free access to round bale orchard and fescue grass hay, water, and trace mineral/vitamin salt. They were fed 2 kg of concentrate twice daily at 7:00 and 3:00 . The diet met or exceeded current recommendations for pregnant mares at 11 months gestation for energy.[8] Samples of hay and concentrate were submitted for partial proximate analysis to the Virginia Tech Forage Testing Laboratory where standard AOAC [9] methods are used ( Table 1). Samples of feeds were also taken for analysis of RT, BC, and AT ( Table 2).