Effect of administration of adrenocorticotropic hormone on plasma concentrations of testosterone, luteinizing hormone, follicle stimulating hormone and cortisol in stallions

In boars and rabbits, administration of adrenocorticotropic hormone (ACTH) results in a testis-dependent, short-term increase in concentrations of testosterone in peripheral plasma. This experiment was designed to assess the short-term effects of a single ACTH injection on plasma concentrations of testosterone, luteinizing hormone (LH), follicle stimulating hormone (FSH) and cortisol in stallions. Eight light horse and two pony stallions were paired by age and weight and then one of each pair was randomly assigned to the treatment (ACTH, .2 IU/kg of body weight) or control (vehicle) group. Injection of ACTH increased (P<.01) plasma concentrations of cortisol by approximately twofold in the first 60 minutes; control stallions showed no change (P>.10) in concentrations of cortisol during the blood sampling period. Control stallions exhibited a midday increase (P>.05) in concentrations of testosterone similar to that reported previously; ACTH treatment prevented or delayed this increase such that concentrations of testosterone in treated stallions were lower (P<.05) than in controls 4 to 5 hours after injection of ACTH. Treatment with ACTH had no effect (P<.10) on plasma concentrations of LH or FSH up to 12 hours after injection.     

Induced gonadotropin release in adrenocorticotropin-treated bulls and steers

The effect of adrenocorticotropin hormone (ACTH) on plasma cortisol and on gonadotropin releasing hormone (GnRH)-induced release of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone was determined in nine Holstein bulls and 12 Holstein steers. Treatments consisted of animals receiving either GnRH (200 micrograms, Group G), ACTH (.45 IU/kg BW, Group A) or a combination of ACTH followed 2 h later by GnRH (Group AG). Group G steers and bulls had elevated plasma LH and FSH within .5 h after GnRH injection and plasma testosterone was increased by 1 h after GnRH injection in bulls. In Group A, plasma cortisol was elevated by .5 h after ACTH injection in both steers and bulls, but plasma LH and FSH were unaffected. In Group A bulls, testosterone was reduced after ACTH injection. In Group AG, ACTH caused an immediate increase in plasma cortisol in both steers and bulls, but did not affect the increase in either plasma LH or FSH in response to GnRH in steers. In Group AG bulls, ACTH did not prevent an increase in either plasma LH, FSH or testosterone in response to GnRH compared with basal concentrations. However, magnitude of systemic FSH response was reduced compared with response in Group G bulls, but plasma LH and testosterone were not reduced. The results indicate that ACTH caused an increase in plasma cortisol, but did not adversely affect LH or FSH response to GnRH in steers and bulls. Further, while testosterone was decreased after ACTH alone, neither ACTH nor resulting increased plasma cortisol resulted in decreased testosterone production in the bull after GnRH stimulation.     

Effects of human chorionic gonadotrophin and castration on plasma gonadal steroid hormones of the dog

An intramuscular injection of 500 I.U. of human chorionic gonadotrophin resulted in an increase of plasma testosterone and progesterone concentrations in the intact male dog, but had no effect on plasma 17B-estradiol concentration. Castration caused a rapid decrease in concentration of testosterone, progesterone, and 17B-estradiol, indicating that the tests were the major organs producing these hormones in the male dog.     

Prolonged effect of a single injection of human chorionic gonadotrophin on plasma testosterone and oestrone sulphate concentrations in mature stallions

The long term effect of a single injection of 6,000 iu of human chorionic gonadotrophin (hCG) was studied in two pony stallions. Peripheral plasma samples were analysed for testosterone and oestrone sulphate. Testosterone concentrations were markedly elevated for five days after injection in both stallions. No adverse effects of these high concentrations were observed on concentrations later in the experiment. There was an initial increase in oestrone sulphate in one stallion, after which concentrations decreased to below pre-injection levels. The other stallion (whose initial oestrone sulphate concentrations were somewhat higher) showed no rise in response to hCG but did show a significant decline from five days after injection. Whether this suppression is an effect of the high testosterone concentrations remains to be determined.     

Effect of GnRH and hCG administration on plasma LH and testosterone concentrations in normal stallions, aged stallions and stallions with lack of libido

Gonadotrophin-releasing hormone (GnRH) (a single intravenous injection with 0.042 mg busereline acetate) was administered to control stallions (n=5), aged stallions (n=5) and stallions with lack of libido (n=5). Jugular blood samples were taken at -10, 0, 10, 20, 40 and 80 minutes after treatment and measured for luteinizing hormone (LH) and testosterone concentrations. A single intravenous injection of hCG (3000 IE) was given 1 day later. Venous blood samples were taken at -60, 0, 15, 30, 60, 120, and 240 minutes after treatment and measured for the testosterone concentration. The experiment was performed in the breeding season. There was a wide variation between stallions in basal concentrations of LH and testosterone. The treatment groups all showed a significant increase in LH and testosterone concentrations after treatment with GnRH. There was a significant difference (P<0.05) between the control, the lack of libido stallions and the aged stallions in the production of LH before and after stimulation with GnRH. The aged stallions had higher basal LH concentrations. GnRH induced a rise in plasma LH in all groups, but the greatest response was observed in aged stallions. No response to GnRH was seen with respect to plasma testosterone. There was an increase in plasma testosterone following hCG; however, this increase was very small in aged stallions. After stimulation with hCG the control and lack of libido stallions had a significant increase (P<0.05) in testosterone production. In conclusion, stimulation with either GnRH or hCG can be a valuable method to test whether the function of the stallion's reproductive endocrine system is optimal.     

Plasma steroid profiles following follicle-stimulating hormone or equine chorionic gonadotropin injection in cows chronically treated with gonadotropin-releasing hormone agonist

Plasma steroid profiles following follicle-stimulating hormone (FSH) or equine chorionic gonadotropin (eCG) injection were studied in chronically gonadotropin releasing hormone agonist (GnRH-A)-treated cows. Follicular development and irINH secretion were stimulated by FSH or eCG injection. The plasma concentrations of estradiol-17 beta (E(2)) and testosterone (T) were markedly increased following eCG injection. However, significant increases of the plasma E(2) and T concentrations were not detected in FSH-treated cows. Ovulation of developed follicles were depended on the hCG injection in both groups. These results show: 1) Follicular response to an exogenous gonadotropin is still remained, 2) Ovulation of developed follicles is induced by hCG injection and 3) FSH and eCG cause disparate plasma steroid profiles, under the influence of repeated GnRH-A treatment.     

Testosterone effects on gonadotropin response to GNRH: cows and pony mares

Effects of testosterone propionate (TP) treatment on plasma concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) before and after an injection of gonadotropin releasing hormone (GnRH) were studied using ovariectomized cows and pony mares. An initial injection of GnRH (1 microgram/kg of body weight) was followed by either TP treatment or control injections for 10 (cows) or 11 (ponies) d. A second GnRH injection was administered 1 d after the last TP or oil injection. Concentrations of LH and FSH were determined in samples of plasma taken before and after each GnRH injection. Control injections did not alter the response to GnRH (area under curve) nor the pre-GnRH concentrations of LH and FSH in ovariectomized cows or ponies. Testosterone treatment increased (P less than .01) the FSH release in response to GnRH in ovariectomized mares by 4.9-fold; there was no effect in cows, even though average daily testosterone concentrations were 59% higher than in pony mares. Testosterone treatment reduced the LH release in response to GnRH by 26% in ovariectomized mares (P less than .05) and by 17% in ovariectomized cows (P approximately equal to .051). These results are consistent with a model that involves ovarian androgens in the regulation of FSH secretion in the estrous cycle of the mare, but do not support such a model in the cow.     

Using a GnRH agonist to obtain an index of testosterone secretory capacity in the cockatiel (Nymphicus hollandicus) and sulphur-crested cockatoo (Cacatua galerita)

Objective   Validation of a stimulation test for determining the steroidogenic capacity of the parrot testis. The major aim was to characterise testosterone secretion after injection of a gonadotropin-releasing hormone agonist (GnRHa), then use the test to investigate seasonal reproduction in the male cockatiel.
Procedure   A synthetic GnRHa (buserelin; 8.0 µg of peptide/kg bodyweight) was injected IM into male cockatiels (n = 7) and sulphur-crested cockatoos (n = 3) and serial blood samples collected at 0, 30, 60, 90 and 120 min after administration. Once validated, the technique was subsequently used to examine seasonal changes (23 months) in the testosterone profile of a captive cockatiel population.
Results   Injection of buserelin resulted in a significant increase in the testosterone concentration of cockatiel plasma, with maximal concentrations occurring at approximately 60 (1.33 ± 0.08 ng/mL) to 90 min (1.22 ± 0.08 ng/mL) after injection. Although no clear pattern of seasonal variation in testosterone secretion was detected in cockatiel plasma, samples taken 60 and 90 min after administration showed a significant increase in all seasons. Injection of buserelin in the sulphur-crested cockatoo also resulted in increased testosterone secretion, with maximal concentrations obtained after 90 min.
Conclusion   Buserelin can be used to obtain a reliable index of the prevailing testosterone capacity of the cockatiel and cockatoo testis. With further studies, this test may be incorporated into clinical assessment of reproductive status.     

Effect of prostaglandin F2 alpha on adrenal-produced steroid hormones in cows

Ovariectomized, nonlactating cows were treated with IM injections of either physiologic saline solution or prostaglandin F2 alpha. Plasma concentrations of cortisol increased significantly by 30 to 60 minutes after injection of prostaglandin F2 alpha, but there were no significant increases in plasma concentrations of estradiol, progesterone, or testosterone. After saline solution treatment, there were no increases in any of the hormones measured.     

The stimulation of testosterone and LH secretion by synthetic GnRH in the male Cape porcupine

The effects of GnRH stimulation on plasma testosterone and luteinizing hormone (LH) levels in Cape porcupine males were examined by analysing plasma collected before and after an intravenous injection of GnRH. In six mature males and one subadult, which were given an intravenous injection of 0,5 ml saline, levels of plasma testosterone and LH did not increase. Four weeks later an intravenous GnRH challenge (40 μ?) caused plasma testosterone to rise three-fold and LH to rise 10-15-fold within 180 min in five of the mature males. Peaks of plasma testosterone and LH occurred 90 and 120 min, respectively, after stimulation, and baseline and peak levels of both hormones were significantly related.     

Leptin secretion in horses: effects of dexamethasone, gender, and testosterone

Five experiments were performed to evaluate the effects of dexamethasone (DEX), gender, and testosterone on plasma leptin concentrations in horses. In experiment 1, plasma leptin, insulin, glucose, and IGF-1 concentrations were increased (P < 0.01) in stallions following five daily injections of DEX (125 microg/kg BW). In experiment 2, leptin concentrations increased (P < 0.01) in mares, geldings, and stallions following a single injection of DEX, and the response was greater (P < 0.01) in mares and geldings than in stallions. The gender effect was confounded by differences in body condition scores and diet; however, based on stepwise regression analysis, both BCS and gender were significant sources of variation in the best fit model for pre-DEX leptin concentrations (R(2) = 0.65) and for maximum leptin response to DEX (R(2) = 0.75). In experiment 3, in which mares and stallions were pair-matched based on age and body condition and fed similar diets, mares again had higher (P < 0.01) leptin concentrations than stallions after DEX treatment as used in experiment 2. In experiment 4, there was no difference (P > 0.1) in plasma leptin response in mares following four single-injection doses of DEX from 15.6 to 125 microg/kg BW. In experiment 5, treatment of mares with testosterone propionate every other day for 5 days did not alter (P > 0.1) plasma leptin concentrations or the leptin response to DEX. In conclusion, multiple injections of DEX increase leptin concentrations in stallions, as does a single injection in mares (as low as 15.6 microg/kg BW), geldings and stallions. The greater leptin levels observed in mares and geldings relative to stallions were due partially to their greater body condition and partially to the presence of hyperleptinemic individuals; however, even after accounting for body condition and diet, mares still had greater leptin concentrations than stallions after DEX administration. Elevation of testosterone levels in mares for approximately 10 days did not alter leptin concentrations in mares.     

Melatonin improves testicular haemodynamics,echotexture and testosterone production in Ossimi rams during the breeding season

The objective was to determine effects of a single parenteral dose of melatonin on testicular blood flow indices, testicular echogenicity and plasma testosterone concentrations in rams during the physiological breeding season. We hypothesized that melatonin enhances testicular blood flow, echogenicity and plasma testosterone concentrations during the breeding season in rams. During the breeding season, 12 sexually mature Ossimi rams were randomly allocated to either a melatonin group (n = 8) that received 18 mg of melatonin in 1 ml of corn oil (injected SC) or a control group (n = 4) that received 1 ml corn oil only. Blood collection and ultrasonographic assessment of the testes and supratesticular arteries were conducted immediately before treatment (W0) and once weekly for 6 weeks after melatonin injection (W1-W6). Mean plasma testosterone concentrations were greater (p < .05; at least 1 ng/ml) in the melatonin-treated group compared to the control group from W4 to W6 after treatment. A decrease (p < .05) in both resistive index (RI) and pulsatility index (PI) began 1 week after melatonin injection (W1) and persisted until the end of the experiment, with mean RI and PI values in the melatonin group lower (p < .05) than those in the control group on W3 and W4. Furthermore, plasma testosterone concentrations in melatonin-treated rams were inversely correlated to both RI and PI (r = −.7 and −.6, respectively, p < .01). Testicular echogenicity decreased (p < .05) 1 week after melatonin injection (W1) and remained lower (p < .05) in the melatonin-treated group compared to the control group until the end of the study (W6). In conclusion, melatonin administration significantly altered testicular blood flow and echogenicity and increased plasma testosterone concentrations in Ossimi rams during the breeding season.     

Influence of long-term treatment with equine somatotropin (EquiGen) on gonadal function in stallions with poor semen quality

The aim of the present study was to investigate the spermatogenic and Leydig cell activity in stallions with impaired semen quality after treatment with equine somatotropin. Experiments were performed using 18 adult clinically healthy stallions with poor semen quality which did not pass breeding soundness evaluation. The animals were randomly divided into a treatment (n = 9) and a control (n = 9) group. Over a period of 90 days, nine stallions received a daily intramuscular injection of 10 mg recombinant equine somatotropin (EquiGen, BresaGen Limited, Adelaide, Australia) and 9 control animals were injected with the same amount of physiological saline solution. During and until 2 months after treatment, semen characteristics and daily sperm output as well as plasma testosterone concentrations were determined monthly in all stallions. In addition, testosterone concentration measurement after stimulation with hCG was performed in all animals immediately before and at the end of the treatment period as well as 2 months later. Our results demonstrate that equine somatotropin (EquiGen) given daily in a dose of 10 mg per animal during 90 days had no significant effect neither on plasma testosterone concentrations and hCG-induced testosterone release nor on semen quality parameters in adult stallions with poor semen characteristics.     

Androgen and oestrogen response to a single injection of hCG in cryptorchid horses

Androgen (testosterone and androstenedione) and oestrogen (oestradiol -17 beta and oestrone) concentrations were measured by radio-immunoassay in the peripheral plasma of two geldings (five-years-old), three bilateral cryptorchids (two, two and a half, and five-years-old) and three normal intact stallions (four, five and five and a half-years-old) before and after a single injection of 10,000 iu human chorionic gonadotrophin (hCG). In the stallions, hCG administration resulted in an immediate sharp increase of conjugated oestrogens and a more gradual increase of unconjugated androgens. In the cryptorchids, the unconjugated androgens increased following a similar pattern to that observed in the stallions, but reached lower peak values, whereas the conjugated oestrogens showed only a very slight increase. In the stallions and cryptorchids, the maximum oestrogen levels were reached two days after injection, whereas the maximal levels for androgens were reached a day later. In the geldings, hCG injection had no effect on plasma steroid levels. It is suggested that the measurement of unconjugated androgens (testosterone or/and androstenedione) before and three days after intravenous injection of 10,000 iu hCG may prove useful for the diagnosis of cryptorchidism or exploration of testicular function in stallions.     

The function of the pituitary-testicular axis in dogs prior to and following surgical or chemical castration with the GnRH-agonist deslorelin

Chemical castration, that is the reduction of circulating testosterone concentrations to castrate levels by administration of a GnRH-agonist implant, is a popular alternative to surgical castration in male dogs. Detailed information concerning the pituitary-testicular axis following administration of a GnRH-agonist implant is still scarce. Therefore, GnRH-stimulation tests were performed in male dogs, prior to and after surgical and chemical castration. This approach also allowed us to determine plasma concentrations of testosterone and oestradiol in intact male dogs for future reference and to directly compare the effects of surgical and chemical castration on the pituitary-testicular axis. In intact male dogs (n = 42) of different breeds GnRH administration induced increased plasma LH, FSH, oestradiol and testosterone concentrations. After surgical castration basal and GnRH-induced plasma FSH and LH concentrations increased pronouncedly. Additionally, basal and GnRH-induced plasma oestradiol and testosterone concentrations decreased after surgical castration. After chemical castration, with a slow-release implant containing the GnRH-agonist deslorelin, plasma LH and FSH concentrations were lower than prior to castration and lower compared with the same interval after surgical castration. Consequently, plasma oestradiol and testosterone concentrations were lowered to values similar to those after surgical castration. GnRH administration to the chemically castrated male dogs induced a significant increase in the plasma concentrations of LH, but not of FSH. In conclusion, after administration of the deslorelin implant, the plasma concentrations of oestradiol and testosterone did not differ significantly from the surgically castrated animals. After GnRH-stimulation, none of the dogs went to pre-treatment testosterone levels. However, at the moment of assessment at 4,4 months (mean 133 days ± SEM 4 days), the pituitary gonadotrophs were responsive to GnRH in implanted dogs. The increase of LH, but not of FSH, following GnRH administration indicates a differential regulation of the release of these gonadotrophins, which needs to be considered when GnRH-stimulation tests are performed in implanted dogs.     

Clearance rate of gonadotrophin releasing hormone in peripheral plasma of the pig.

Gonadotrophin releasing hormone was administered as an intravenous bolus injection into four boars and four ovariectomized sows. Radioimmunoassay of concentrations of gonadotrophin releasing hormone in blood collected periodically after injection indicated a biexponential decline suggesting a rapid distribution to the extracellular fluid and a slower elimination by metabolism. A mean half-life value of 2.12 +/- 0.95 (SD) minutes was calculated for the first component and of 13.15 +/- 2.55 minutes was calculated for the second component of the decline in gonadotrophin releasing hormone concentrations. No significant difference was detected between boars and sows for half-life value of either component. In the four boars, luteinizing hormone values reached a peak in plasma 20 minutes after injection and that of testosterone at 90 minutes after gonadotrophin releasing hormone treatment.     

The castration of male lambs by immunization against GnRH

The objective of this study was to evaluate the effect of a GnRH-vaccine in the ram lamb. Experiments were performed using 20 male lambs, randomly divided into a test (GnRH-immunization) and control group (physiological NaCl-solution). At a body weight of 20 kg (age 2-3 months) and three weeks later, all animals of the test group received 2 ml of Improvac (CSL Limited, Parkville, Victoria, Australia). The body weight as well as the blood testosterone concentration were measured weekly for 16 weeks. Thereafter, blood samples for testosterone analysis were taken monthly in immunized lams only. After the booster injection testicular growth was suppressed and plasma testosterone remained at low values < 0.1 ng/ml for at least 12 weeks. The mean corresponding testosterone concentrations for the control lambs ranged between 0.1 and 0.9 ng/ml plasma. An increase of testosterone was observed in 8 of 10 immunized animals between 3 to 7 months after the booster dose. The control lambs showed a tendency for better growth rate than vaccinated animals, but the difference was not significant. Our results demonstrate that in prepubertal ram lambs two immunizations with Improvac, three weeks apart, can suppress testosterone secretion and testicular growth at least for three months after the booster injection. For a suppression of reproductive function longer than three months after the second vaccination, a third immunization is needed at this time or when testicular growth is beginning.     

Testicular blood flow and plasma concentrations of testosterone and total estrogen in the stallion after the administration of human chorionic gonadotropin

The goal of this study was to investigate for the first time a possible association between plasma concentrations of testosterone and total estrogen and testicular blood flow in the stallion. Correlations between these variables were calculated before and after administration of human chorionic gonadotropin (hCG). Eight mature warmblood stallions received 5,000 IU hCG intravenously, and four stallions received solvent only. Testicular blood flow in the left and right testicular arteries was assessed using colour Doppler sonography by measuring blood flow volume (BFV) and pulsatility index (PI) immediately before (time 0) and 1, 3, 6, 12, 24, 72, 120 and 168 h after hCG administration. EDTA blood samples were collected after each examination from a jugular vein to measure plasma testosterone and total estrogen concentrations. After treatment, the BFV increased and was elevated at 1 h and between 12 and 24 h. The profile of the PI was contrary to that of the BFV throughout the study period. Following hCG, there was a biphasic increase in testosterone concentration with maxima between 1 and 3 h and between 24 and 72 h, and there was a monophasic increase in the total estrogen concentration with a maximum between 6 and 24 h. At time 0, the total estrogen concentration correlated significantly with BFV (r=0.90; P<0.05) but the testosterone concentration did not (P>0.05). The testosterone and total estrogen concentrations did not correlate with PI (P>0.05). The total estrogen concentration, but not testosterone, correlated well with BFV after injection of hCG (P<0.05). The results of this study indicated that the testicular blood flow volume of the stallion may be regulated by estrogens, but additional studies are necessary to investigate whether there is a causal relationship.     

Testosterone concentration in the seminal plasma of cocks

Testosterone concentrations in the seminal plasma of cocks ranged from 0.46 to 5.05 ng/ml and were substantially lower than in blood plasma. No significant variation was noted in seminal plasma testosterone concentrations during the light phase of the day, whereas the concentration in blood declined over this period. Spermatozoal concentration and seminal testosterone decreased in the third sample of the semen collected sequentially at 3 h intervals. Testosterone concentrations in seminal plasma (1.57 +/- 0.17 ng/ml) and in the semen from the ductus deferens (1.34 +/- 0.24 ng/ml) were not significantly different.     

Plasma testosterone and LH responses to LHRH in double-muscled bulls treated with trenbolone acetate and zeranol

Twenty-four double-muscled Belgian White Blue bulls were assigned, according to body weight, to two groups with and without treatment with anabolic agents. Implants containing 140 mg trenbolone and 36 mg zeranol (Forplix) were inserted sc on the upper face of the ear flap. Plasma concentrations of testosterone and luteinizing hormone (LH) were determined at d 0, 30 and 60 of the experimental period. Mean testosterone levels at d 0 for treated and controls were, respectively, 2.1 and 1.7 ng/ml during the 10-h sampling period. At d 30 and 60, testosterone levels were strongly depressed in implanted bulls (.2 ng/ml) as compared with 2.5 and 1.7 ng/ml in control bulls (P less than .001 at d 30 and P less than .01 at d 60). Average plasma LH concentrations were identical in the two groups at d 0 and 60 (1.1 and 1.5 ng/ml, respectively), but showed a slight decrease at d 30 in the treated group (P less than .10). The pulsatile character of LH and testosterone profiles was abolished by the Forplix treatment. Luteinizing hormone-releasing hormone (LHRH) injection at d 0 was followed in both groups by an immediate and sharp increase in plasma LH concentrations. The LH response reached a maximal value between 20 to 40 min postinjection and then declined rapidly. On the contrary, Forplix treatment strongly reduced LH and testosterone responses to LHRH stimulation in treated animals. Average daily gain and feed to gain ratios were 1.087 +/- .127 and 7.52 +/- .32 kg, respectively, for the control bulls and 1.335 +/- .092 and 6.24 +/- .24 kg for the Forplix-treated bulls, thus clearly showing a beneficial effect of Forplix treatment.