Effects of season and estradiol-17 beta on luteinizing hormone release in ovariectomized sows.

This study examined the ability of estradiol-17 beta (E2) to suppress LH release in the sow during different months of the year. Six chronically ovariectomized sows were fitted with vena caval cannulas (d 0) and blood samples were collected at 6-h intervals for 6 d. Sows were treated s.c. with E2 capsules (24 mg of E2/275 kg of BW) at d 3. Additional blood samples were collected at 15-min intervals for 8 h on d 2 and 5. After each 8-h frequent sampling period, sows were treated i.v. with GnRH at .5 microgram/kg of BW, and blood samples were collected at 10-min intervals for 3 h. The protocol was repeated at monthly intervals for 13 mo. Luteinizing hormone concentrations were determined for all serum samples, and E2 concentrations were quantified in samples collected at 6-h intervals. Data were analyzed by split-block analyses of variance. Serum E2 concentrations increased (P less than .001) from 5.0 +/- .3 pg/ml before E2 treatment to 26.0 +/- .2 pg/ml after E2 treatment. The interval from GnRH administration to peak LH concentration was shorter (P less than .001) before E2 treatment than after E2 treatment (28.7 +/- 2.2 vs 71.0 +/- 2.2 min). It was evident that baseline LH, mean LH, pulse frequency, and pulse amplitude and LH release after GnRH administration failed to demonstrate seasonal changes. In summary, LH release was suppressed after treatment with E2 and was affected minimally by month of the year. In addition, E2 inhibitory effects of LH release included hypothalamic and anterior pituitary sites of action.     

The concentration of estradiol-17 beta in bovine semen

This study was conducted to evaluate the influence of age, breed, epididymectomy and semen processing on the concentration of estradiol-17 beta (E2) in bovine semen. Semen was collected either by electroejaculation or with an artificial vagina. Neat semen samples were stored at -20 C until analysis. Processed, frozen semen and an egg yolk-citrate semen extender were obtained from a commercial semen processing firm and stored in liquid nitrogen at -196 C. The concentration of E2 in semen was determined by radioimmunoassay. Semen from mature (greater than 24 mo), fertile Brahman (n = 19), Brangus (n = 16), Charolais (n = 29), Holstein (n = 15) and Santa Gertrudis (n = 25) bulls was analyzed for E2 concentration, and no difference (P greater than .10) between breeds was found. There was no difference (P greater than .10) in seminal E2 concentration between mature, fertile bulls (n = 104) and epididymectomized bulls (n = 22). In semen collected from prepuberal (12 to 16 mo, n = 21), peripuberal (17 to 20 mo, n = 17) and mature (greater than 24 mo, n = 19), Brahman bulls, the mature bulls had a lower (P less than .01) semen E2 concentration than peripuberal and prepuberal bulls. There were no differences (P greater than .10) in seminal E2 concentration among peripuberal Angus (n = 8), Hereford (n = 8) and Brahman (n = 17) bulls.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of exogenous estradiol-17 beta on luteinizing hormone, progesterone, and estradiol-17 beta concentrations before and after prostaglandin F2 alpha-induced termination of pregnancy and pseudopregnancy in gilts.

This study evaluated the influence of exogenous estradiol-17 beta (E2) administration on LH concentrations and the number of animals returning to estrus after the termination of pregnancy or pseudopregnancy in gilts. Gilts were mated (pregnant; n = 11) on the 1st d of estrus or received 5 mg of estradiol valerate i.m. at d 11 to 15 after the onset of estrus (pseudopregnant; n = 9). Gilts were treated with prostaglandin F2 alpha (PGF2 alpha, 15 and 10 mg) at 12-h intervals on d 44 of pregnancy or pseudopregnancy. The day of abortion or luteolysis (progesterone less than .2 ng/mL) was considered d 0. Six pregnant and four pseudopregnant gilts received s.c. an E2 capsule (24 mg of E2) on d -20 and additional E2 capsules on d -13 and -6. The E2 capsules were removed on the day after PGF2 alpha administration. Blood samples were collected at 12-h intervals from d -21 to -3, at 6-h intervals from d -2 to 21 or the onset of estrus, and at 15-min intervals for 8 h on d -2, 1, 4, 7, 10, 14, and 18. After each 8-h sampling period, gilts were treated i.v. with GnRH at .5 micrograms/kg of BW and blood samples collected at 10-min intervals for 3 h. A greater (P less than .05) proportion of sham-treated gilts than of E2-treated gilts exhibited a preovulatory-like LH surge after abortion/luteolysis. It was evident that E2 supplementation before luteolysis reduced the ability of pregnant and pseudopregnant gilts to return to estrus.     

Effects of exogenous estradiol-17 beta and progesterone on naloxone-reversible inhibition of the release of luteinizing hormone in ewes

The role of endogenous opioids in controlling luteinizing hormone (LH) secretion was studied by injecting the opioid antagonist naloxone into intact and ovariectomized ewes that were treated with estradiol-17 beta (E2) and progesterone (P4). The existence of a naloxone-reversible inhibition of LH release was examined in five experiments using a total of 52 mature ewes. Naloxone at a dosage of 1 mg/kg disinhibited release of LH and abruptly increased serum concentrations of LH in a variety of experimental models. This naloxone-reversible inhibition of LH secretion was apparent in all experimental models that involved P4-induced inhibition of basal LH secretion but not in one model in which P4 inhibited the LH surge. Specific effects of E2 on naloxone-reversible inhibition of LH varied among experimental models. When prolonged administration of P4 alone appeared to lose its LH-inhibitory potency, E2 restored inhibition of LH as well as the naloxone-reversible state. Whenever E2 acted synergistically to suppress basal LH secretion in models involving brief (5 d) exposure to P4, E2 appeared to antagonize the naloxone-reversible state. In summary, P4-induced suppression of LH secretion appeared to be mediated by endogenous opioids, but the apparent interaction of E2 and opioids in LH suppression varied among experiments.     

Uterine involution in mares treated with progesterone and estradiol-17 beta

Bacteriology, histology, and scanning electron microscopy were used to evaluate uterine involution in 27 mares treated with daily injections of 150 mg of progesterone and 10 mg of estradiol-17 beta, commencing within 18 hours of parturition. These findings were compared with those for 24 untreated mares at postpartum day 10 or 11. The treatment resulted in significantly (P less than 0.05) greater uterine gland proliferation. Gland density was significantly (P less than 0.05) greater in mares treated for 6 to 10 days than in those treated 2 to 5 days. The proportion of ciliated cells to secretory cells lining the endometrial surface was significantly (P less than 0.05) greater in mares during delayed foal estrus than in those at postpartum days 10 to 11. The proportion of ciliated to secretory cells increased with increasing duration of treatment. It was concluded that treatment with progesterone and estradiol-17 beta allowed additional time for uterine involution in the early postpartum period.     

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.     

Activities of ketone body utilising enzymes in tissues of fed and fasted sheep.

The activities of 3-hydroxybutyrate dehydrogenase, 3-ketoacid CoA-transferase and acetyl-CoA acetyltransferase have been measured in the kidney, heart and brain of fed and four-day fasted ewes. The results indicate tha there is a decrease in the capacity of these organs to catabolise ketone bodies in fasting ketosis.     

Concentration of IGF-I and estradiol-17beta in the blood plasma of pregnant cattle

Blood samples were collected from pregnant cows, heifers and also from non pregnant cows serving as controls. The determination of insulin-like-growth-factor-I (IGF-I) and estradiol-17 beta (E2) were performed immunologically. In the non pregnant cows E2 remained unchanged. IGF-I decreased around parturition and increased again commencing the 6th week of lactation. Compared to nonpregnant animals E2 but not IGF-I was slightly elevated during the first 10 weeks of pregnancy. During the 10th up to 20th week of pregnancy the mean values of IGF-I were increased as well as the ones of E2. During the late pregnancy the values of IGF-I in heifers are obviously more elevated as in lactating pregnant or non-pregnant cows; analogous high values were determined in pregnant cows only during the dry period. Before parturition even a negative correlation exists between IGF-I and E2. It is concluded that IGF-I is predominantly regulated by other factors.     

Effects of natural mating stimuli on serum luteinizing hormone, testosterone and estradiol-17 beta in yearling beef bulls

The objective of this study was to determine the effect of natural mating stimuli on serum concentrations of LH, testosterone (T) and estradiol-17 beta (E2) in beef bulls. Twenty sexually experienced, yearling beef bulls were bled every 15 min during a 9-h period, 4 h before and 5 h after exposure to estrual females. For exposure, each bull was placed individually in an isolated pen with two restrained estrual heifers for 10 min or until one service was achieved. Timing and number of all behavioral events, including flehmen responses, abortive mounts and services, were recorded for each bull by two independent observers. Of the 20 bulls, 9 bulls mounted and were removed immediately after achieving a service, 8 bulls mounted without achieving a service and 3 bulls exhibited no interest during exposure. Twelve bulls achieved fewer than three and eight bulls achieved three or more flehmen responses during exposure. Postexposure responses in LH, T and E2 were not consistently correlated with number of mounts or presence or absence of a service. However, postexposure LH and T, but not E2, responses were highly correlated with number of flehmen responses achieved (r = .40 to .66; P = .08 to .001). In bulls that achieved three or more flehmen responses, serum LH increased within 30 min after exposure (P = .02) and serum T was increased dramatically within 1 h after exposure (P less than .01), compared with preexposure hormone concentrations, regardless of the number of mounts and regardless of the presence or absence of a service. Natural mating stimuli had no effect on serum E2, and mounting activity alone and mounting that culminated in a service did not necessarily result in increased LH or T in beef bulls. However, number of flehmen responses achieved during exposure to females dramatically influenced postexposure serum LH and T concentrations in beef bulls.     

Effects of estradiol-17 beta, naloxone and gonadotropin releasing hormone on postpartum secretion of luteinizing hormone in fall-lambing ewes

The interaction among exogenous estradiol-17 beta, naloxone and gonadotropin releasing hormone (GnRH) in the control of luteinizing hormone (LH) secretion was studied in intact postpartum ewes nursing their offspring. One-half of 30 fall-lambing ewes were implanted subcutaneously with an estradiol-17 beta containing Silastic capsule between postpartum d 1 and 12 which doubled their serum concentrations of estradiol (16.0 +/- .1 vs 8.4 +/- .1 pg/ml). Blood samples were collected from implanted and non-implanted ewes at 15-min intervals for 5 h on d 3, 8, 13, 20 and 28 postpartum. Pre-injection samples were collected for 1 h, and ewes were injected with saline, naloxone (NAL;1 mg/kg) or GnRH (100 micrograms/ewe). When averaged across all days and implant groups, serum LH in the three post-NAL samples was higher (P less than .05) than in the three pre-NAL samples (3.6 +/- 1.2 vs .6 +/- .2 ng/ml). Post-GnRH concentrations of serum LH were lower (P less than .05) in estradiol-implanted ewes than in non-implanted ewes on d 8 and 13, but there were no differences in any LH characteristics on d 20 and 28 after implant removal on d 12. In non-implanted ewes, serum LH responses to GnRH increased (P less than .05) eightfold from d 3 (3.8 +/- 1.4 ng/ml) to d 8 (31.6 +/- 1.4 ng/ml), remained elevated through d 20, but declined by d 28 (10.8 +/- 1.4 ng/ml).(ABSTRACT TRUNCATED AT 250 WORDS)     

Ghrelin differentially modulates the GH secretory response to GHRH between the fed and fasted states in sheep

The effect of energy balance on the growth hormone (GH) secretory responsiveness to growth hormone-releasing hormone (GHRH) has not been determined in ruminant animals. Therefore, we examined the effects of intravenous injections of 0, 3.3, and 6.6 μg ghrelin/kg body weight (BW), with and without GHRH at 0.25 μg/kg BW, on GH secretory responsiveness in both the fed and fasted sheep. The injections were carried out at 48 h (Fasting state) and 3 h (Satiety state) after feeding. Blood samples were taken every 10 minutes, from 30 minutes before to 120 minutes after the injection. Low (3.3 μg/kg BW) and high (6.6 μg/kg BW) doses of ghrelin stimulated GH secretion significantly (P < .05) greater in the Satiety state than in the Fasting state. Growth hormone-releasing hormone plus both doses of ghrelin stimulated GH secretion significantly (P < .05) greater in the Satiety state than in the Fasting state. Ghrelin and GHRH exerted a synergistic effect in the Satiety state, but not in the Fasting state. Plasma ghrelin levels were maintained significantly (P < .05) greater in the Fasting state than in the Satiety state except the temporal increases after ghrelin administration. Plasma free fatty acid (FFA) concentrations were significantly (P < .01) greater in the Fasting state than in the Satiety state. In conclusion, the present study has demonstrated for the first time that ghrelin differentially modulates GH secretory response to GHRH according to feeding states in ruminant animals.     

Relationships between LH and estradiol-17 beta after removal of luteal progesterone in the ewe

Three experiments were conducted to examine the relationship between systemic concentrations of luteinizing hormone (LH) and estradiol-17 beta (E2) after withdrawal of progesterone in cycling ewes. In Exp. 1, ewes were assigned randomly to one of three treatments: laparotomy (C), removal of the luteal ovary (ULO), or ULO plus anesthesia with sodium pentobarbital for 6 h beginning 4 h after surgery. Anesthesia was used in an attempt to block the expected increase in tonic secretion of LH. Patterns of LH and E2 in these three groups did not differ during the 24-h experimental period. In Exp. 2, a longer period of anesthesia was utilized. Forty-eight ewes were assigned at random to one of four treatments: C, ULO, lutectomy or an intrafollicular injection of prostaglandin F2 alpha (PGF2 alpha). One-half of the ewes in each group were anesthetized with sodium pentobarbital from initiation of treatment (0 h) until 10 h after surgery. Sodium pentobarbital did not suppress the increases in LH and E2 after progesterone withdrawal. The regression of concentrations of E2 on concentration of LH was not significant. In Exp. 3, ewes were infused with either saline or dopamine after receiving an im injection of PGF2 alpha. Tonic secretion of LH increased after 4 h in ewes infused with saline, but not in ewes infused with dopamine. Despite the suppression of LH, concentrations of E2 increased in dopamine-treated ewes as in control ewes. Therefore, the initial increase in E2 after a decline of progesterone in cycling ewes is independent of increases in LH.     

High plasma estradiol-17beta levels in dogs with benign prostatic hyperplasia and azoospermia

The semen quality of 22 dogs (4 to 7 years old) with benign prostatic hyperplasia (BPH) was examined at the hospital of our university, and 4 of the 22 BPH dogs were diagnosed as azoospermic. The mean peripheral plasma estradiol-17beta (E2) level (17.3 pg/ml) of the 18 BPH dogs with spermatogenic function was higher than that of 5 normal male dogs and their mean T level (1.7 ng/ml) was lower. The mean E2 level (27.3 pg/ml) of the 4 BPH dogs with azoospermia was significantly higher than the value in the BPH dogs with spermatogenic function (P<0.01), and the mean T level (1.1 ng/ml) was significantly lower (P<0.05). Five normal male dogs were given 10 intramuscular injections of estradiol benzoate (E2B) 5 microg/kg, at 3-day intervals to investigate the relationship between high plasma E2 levels and the cause of the BPH and azoospermia. Their testes and prostates were measured and biopsied both before and 30 days after the start of E2B injections. At 30 days after the start of the E2B injections, the mean peripheral plasma T levels had decreased by half, and the mean testicular volume had decreased to 88% of original volume. The numbers of spermatocytes, spermatids, and spermatozoa in the seminiferous tubules of all of the dogs were significantly lower (P<0.05, 0.01). In addition, the mean prostatic volume increased to 130%, the mean height of the glandular epithelium decreased, and the glandular lumen became increased in diameter. These findings indicate that both BPH and serious spermatogenic dysfunction may be simultaneously induced by protracted high plasma E2 levels in dogs.     

Effect of estradiol-17 beta treatment of gilts on blood mononuclear cell functions in vitro.

Porcine blood mononuclear cells (BMC) were exposed to prepartum concentration of estrogen in gilts before acquisition (in vivo), and their subsequent reactivity (in vitro) was explored. In a cross-over experimental designed study, 6 ovariectomized gilts were injected once with 3.75 mg of estradiol-17 beta benzoate in arachidic oil or with arachidic oil only during 2 experiments. The ability of their BMC to proliferate in response to stimulation with phytohemagglutinin, concanavalin A, and pokeweed mitrogen was assayed in cultures of blood and in cultures of purified BMC. After 2 days of mitogen stimulation, activity of accessible interleukin 2 was quantified in supernatants obtained from cultures of purified BMC and supernatants of blood cultures stimulated with pokeweed mitogen. Also, production of immunoglobulins by purified BMC in response to polyclonal stimuli was measured. Three days after treatment with estradiol, the proliferative response was suppressed in blood cultures stimulated with concanavalin A (P less than 0.05) and phytohemagglutinin (P less than 0.07). Effects of estradiol treatment were not found in any of the assays performed with purified BMC. We, therefore, assumed that in vivo exposure to estradiol can affect the function of porcine BMC; however, this was only evident when the in vitro assays were performed on blood cultures.     

Factors affecting retention of estradiol-17 beta silicone rubber implants in ears of steers

A series of trials were conducted to identify the factors causing loss of estradiol-17 beta (E2-beta) silicone rubber implants from the ears of cattle and to evaluate methods of reducing this loss. Surface application of cattle feces to the ears before implanting resulted in an increase in loss of implants compared with the loss from dry, clean ears (30.6 vs 8.6%; P less than .05). Washing ears with a povidone-iodine antiseptic solution before implanting or treating implant sites with an antibiotic after implanting reduced (P less than .05) implant loss when ears were coated with the fecal slurry. Coating silicone rubber implants with .5 to 2 mg of oxytetracycline hydrochloride (OTC) reduced (P less than .0001) implant loss from 39.8 to 13.8% when ears were coated with fecal slurry. When silicone rubber implants with a 1.5-mg coating of OTC were implanted in cattle before submerging in a dipping vat, implant loss was reduced from 6.2 to 2.7%. In studies designed to evaluate mechanical factors affecting implant loss, implants that were placed in the middle of the ear in tight skin moved .79 cm toward the insertion site during a 14-d period after administration compared with 2.82 cm when placed in the base of the ear. When placed in the middle of the ear in tight skin, only 2 of 399 (.5%) implants were lost from steers submerged in a dipping vat immediately following implantation compared with 42 of 394 (10.7%) when placed in the base of the ear (P less than .0001).(ABSTRACT TRUNCATED AT 250 WORDS)     

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.