Annual Changes in Day-length,Temperature, and Circulating Reproductive Hormones in Thoroughbred Stallions and Geldings

Changes in follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin, immunoreactive(ir)-inhibin, testosterone, estradiol-17β, and insulin-like growth factor (IGF)-I in Thoroughbred stallions along with changes in prolactin secretion in geldings were studied. The correlations of day-length with changes in the concentrations of these hormones were also studied. Five stallions and thirteen geldings were employed to draw blood samples in monthly basis and radioimmunoassay was performed to measure these hormones. All hormones showed a seasonal pattern, the levels being highest during the breeding season and lowest during the winter months. Most of the hormones were at their highest concentration during the month of April, the mid of spring in northern hemisphere. The concentration of circulating IGF-I also demonstrated seasonality, the peak lying on the month of April. The plasma concentration of prolactin also increased during the breeding season. This phenomenon was similar both in stallions and geldings although geldings had lower concentration than that of stallions. The changes in concentration of prolactin in stallions and geldings correlated more towards the day-length than towards the temperature. These results clearly indicate the seasonality of pituitary and gonadal hormones of Thoroughbred stallions, the activity being highest during the month of April and May of the breeding season.     

Effects of Hemicastration on Testes and Testosterone Concentration in Stallions

The endocrine system is critical to the maintenance of testicular function. The homeostasis of sex hormone levels is orchestrated by positive and negative feedback systems controlled by the hypothalamic-pituitary-gonadal axis. This study investigated the long-term effects of hemicastration on testicular size and function in stallions. Four Thoroughbred stallions, 4–6 years of age, were included in this study. Several parameters, including testicular weight and volume, plasma testosterone concentrations, VASA-positive germ cell populations and cross-sectional areas of the seminiferous tubules were compared in stallions that underwent two hemicastrations, approximately 11 months apart. The weights and volumes of testes harvested at the second hemicastration were significantly higher than those of testes collected at the first hemicastration. However, VASA-positive germ cell populations and the cross-sectional areas of seminiferous tubules were not significantly different between testes harvested at the first and second hemicastrations. Similarly, plasma testosterone concentrations measured weekly for 3 weeks before the first hemicastration, 3 weeks after the first hemicastration, and 3 weeks before the second hemicastration were not significantly different. Our results suggest that hemicastration results in compensatory enlargement of the remaining testis and compensatory steroidogenesis to maintain normal reproductive function in stallions.     

Influences of season and artificial photoperiod on stallions: pituitary and testicular responses to exogenous GnRH

Effects of season and photoperiod on the anterior pituitary gland and testes were studied by responses to exogenous GnRH. Stallions were assigned to one of three treatments: 1) control, exposed to natural day length; 2) S-L, 8 h of light and 16 h dark (8:16) for 20 wk beginning July 16, 1982 then 16:8 from December 2, 1982 until March 5, 1984; or 3) S-S, 8:16 from July 16, 1982 until March 5, 1984. Approximately every 8 wk, stallions were administered GnRH (2 micrograms/kg BW) and blood was sampled at 20-min intervals for 2 h before and 8 h after GnRH administration. Concentrations of LH, FSH and testosterone were determined. Baseline concentrations (mean of pre-GnRH samples) of all hormones fluctuated seasonally (P less than .05), but only LH and testosterone displayed seasonal changes (P less than .05) in maximum response to GnRH (highest concentration above baseline after GnRH). The FSH response to GnRH was not affected (P greater than .05) by season, photoperiod or the season X treatment interaction. Exposure of S-L stallions to 16:8 in December resulted in early recrudescence of baseline concentrations of LH, FSH and testosterone. Maximum concentration of testosterone in response to GnRH was stimulated by 16:8, but the increase in baseline LH concentrations in S-L stallions was not associated with an increase in maximum LH response to GnRH. Seasonal patterns of baseline concentrations of FSH and testosterone and maximum LH response to GnRH in S-S stallions were similar to those for control stallions.(ABSTRACT TRUNCATED AT 250 WORDS)     

Influences of season and artificial photoperiod on stallions: luteinizing hormone follicle-stimulating hormone and testosterone

Influence of day length on seasonal endocrine responses were studied using stallions (seven per group). Treatments included 1) control, with natural day length; 2) 8 h light and 16 h dark (8:16) for 20 wk beginning July 16, 1982 then 16:8 from December 2, 1982 until March 5, 1984 (S-L); or 3) 8:16 from July 16, 1982 until March 5, 1984 (S-S). Blood was sampled hourly for 5 h every 4 wk; sera were pooled within horse, and luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone were quantified. Blood was collected every 20 min for 24 h every 8 wk and 2 wk before and after the December light shift. Samples were assayed for LH. Stallions in all groups underwent seasonal changes (P less than .05) in concentrations of LH, FSH, testosterone and basal concentrations of LH and amplitude of LH pulses. Season X treatment (P less than .05) reflected on early recrudescence of LH, FSH and testosterone concentrations in S-L stallions followed by earlier regression. Except for FSH hormone concentrations were depressed in S-S stallions. Number of LH pulses per 24 h was unaffected by season, treatment or their interaction. Mean amplitude of LH pulses was affected (P less than .05) by season X treatment; maximal values occurred in April vs February for control and S-L stallions, and minimal values occurred in December vs April. The season X treatment interaction (P less than .05) similarly affected basal concentrations of LH. Thus, seasonal changes in concentrations of LH, FSH and testosterone can be driven by photoperiod. Increased peripheral concentrations of LH during seasonal recrudescence of reproductive function apparently results from more LH secreted per discharge without an increased frequency of LH discharges.     

Effects of age,season, and fertility status on plasma and intratesticular immunoreactive (IR) inhibin concentrations in stallions

The nature of the relationship between inhibin and reproductive function in the stallion is yet to be elucidated. Blood and testes from 51 light horse stallions ranging in age from 2 mo to 25 years were collected during the breeding and nonbreeding seasons to study the effects of testicular maturation, aging, season, and fertility status on peripheral and intratesticular concentrations of Ir inhibin and other reproductive hormones. Of the 51 stallions, 12 age-matched stallions (6 fertile, 3 subfertile, and 3 infertile) were used in the fertility study. Blood samples were taken before castration and plasma stored at −20°C for analysis of Ir inhibin, luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone (T), estradiol (E₂), and estrogen conjugates (EC) by radioimmunoassay (RIA). Testes were homogenized and testicular extracts prepared and frozen at −70°C for analysis of Ir inhibin, T, E₂, and EC by RIA. Plasma concentrations of Ir inhibin, LH, FSH, T, E₂, and EC and intratesticular concentrations of Ir inhibin, T, E₂, and EC increased with age (P < 0.01). The most dramatic effect appeared to be during testicular maturation. An aging effect was not observed in adult stallions. A seasonal effect was not detected for any of the plasma hormones, whereas for the intratesticular hormones the only change noted was an increase in T in the nonbreeding season (P < 0.05). Plasma Ir inhibin, E₂, and EC were lower (P < 0.01) and gonadotropins higher (P < 0.05) in infertile stallions. Plasma T levels did not change. Intratesticular Ir inhibin concentrations tended to be lower (P < 0.1) in subfertile stallions and significantly lower (P < 0.01) in infertile stallions, whereas intratesticular steroid levels were not different among the three groups. In conclusion, plasma and intratesticular Ir inhibin concentrations seem to be affected by testicular maturation and fertility status.     

Endocrine and testicular changes associated with season, artificial photoperiod, and the peri-pubertal period in stallions.

The seasonal reproductive cycle of stallions is characterized by an annual regression and recrudescence in testicular function and concentrations of LH, FSH, and testosterone in serum. Maximum reproductive capacity occurs during the increasing day lengths of spring and summer. The annual cycle in LH secretion may reflect a seasonally associated and photosensitive reduction and replenishment in pituitary content of LH. Similar to other seasonal breeders, it appears that stallions may possess an endogenous circannual rhythm in reproductive function that is subject to manipulation by altering the light:dark ratio, i.e., photoperiod. The application of a long-day photoperiod (16 hours light:8 hours dark) in December, following 20 weeks of short days (8 hours light:16 hours dark), was effective in hastening the seasonal sexual recrudescence of stallions but was not effective in prolonging the interval of heightened reproductive capacity. The infantile period in colts lasts approximately 32 weeks and is characterized by low gonadotropin concentrations and little gonadal activity. The start of the pre-pubertal period is marked by changes in the hypothalamic-pituitary axis which result in increased amounts of LH and FSH secretion between 32 and 40 weeks of age. Testosterone concentrations in serum exhibit a dramatic increase at 75 to 80 weeks of age, with puberty (defined as the age when the first ejaculate was obtained containing a minimum of 50 x 10(6) sperm with greater than or equal to 10% progressive motility) occurring at 83 weeks of age.     

Seasonal and photoperiodic regulation of secretion of hormones associated with reproduction in Magang goose ganders

This study examined the reproductive endocrine profile under natural and artificial photoperiods in Magang goose ganders. Group 1 ganders (n=8) served as non-treated controls and were exposed to natural photoperiod throughout the experiment from 13th January to 17th December 2004. Group 2 ganders (n=8) were exposed to 18 h long daily photoperiod for 60 days from 13 January till 15 March 2004 and again to 16 h photoperiod for 75 days till 10th October 2004, and the 11h short photoperiod in the remainder periods of the experiment. In control ganders, plasma LH concentrations were high in normal breeding seasons (August-March) and decreased to low levels in non-breeding season from April to July. Testosterone concentrations changed similarly to that of LH throughout the seasons. Seasonal pattern of PRL concentrations was opposite to those of LH and testosterone, with low values in breeding season and high values in non-breeding season. In artificial photoperiod treated ganders, increasing photoperiod increased PRL and decreased LH and testosterone concentrations, while decreasing photoperiod reversed these changes. There were no seasonal or photoperiod caused changes in plasma T3 concentrations in both control ganders and artificial photoperiod treated ganders. These results demonstrated that in Magang goose ganders that long photoperiod stimulates PRL secretion and decreases LH secretion, which terminates reproductive season in spring and early summer, and short photoperiod stimulates LH secretion and inhibits PRL secretion rendering ganders enter into reproductive season.     

Reproductive characteristics of stallions during the breeding and non-breeding season in a tropical region

The objective of this study was to investigate reproductive characteristics of stallions at a tropical zone in the breeding and non-breeding seasons. The following parameters were assessed: testicular volume; semen quality; and serum concentrations of LH, FSH, and testosterone; in addition to the percentages of germ cells and proportions of germ cells/Sertoli cells by testicular cytology in stallions. Semen was collected from eight adult stallions twice a week during a 12-week period in both seasons (6?weeks before and 6?weeks after the summer and winter solstices). Jugular blood samples were collected periodically for hormone analysis by radioimmunoassay during the same periods. Testicular measures and cytological samples were taken at the end of each period. Mean concentration of testosterone was significantly higher (P?=?0.04) during the breeding season and the proportion of Sertoli cells/100 germ cells in cytological smears was significantly lower during the breeding season (P?=?0.0001). Effects of season were not significant either for testicular volume or for any semen parameter (P?>?0.05). Seasonal changes in the mean concentrations of LH and FSH were not observed (P?>?0.05). There were also no significant differences in the mean percentages of germ cell types between both seasons (P?>?0.05). Lack of seasonal differences in the testicular volume and semen parameters of tropical stallions are probably due to the small variation in duration of natural light between the observed periods, slightly under 3?h.     

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.     

光照对马岗鹅季节性繁殖活动和内分泌的调控

研究了光照对马岗鹅季节性繁殖活动和内分泌的调控。在整个试验期（2004.01.13-12.18）对照组接受自然光照,处理组接受人工控制的长短光照处理。在非繁殖期（4-7月份）,对照组公母鹅血浆PRL浓度升高,LH浓度和公鹅睾酮水平降低,鹅换羽;在繁殖期（8-3月份）,公母鹅血浆PRL水平下降,LH浓度和公鹅睾酮水平则上升。在处理组,延长光照均使公母鹅PRL浓度升高,LH浓度和公鹅睾酮水平降低,鹅群进入休产期并换羽;缩短光照则降低PRL水平,促进LH分泌和公鹅睾酮水平上升,鹅群进入繁殖期。整个试验阶段,两组中公鹅甲状腺素T3水平均无明显季节性变化,处理组和对照组母鹅平均产蛋48.8和26.3枚,前者比后者高85.6%。试验结果表明,长光照抑制,短光照促进马岗鹅繁殖活动,光照通过调节PRL和LH的分泌调控马岗鹅繁殖活动的季节性变化。     

The effect of artificial photoperiod at the end of the breeding season on plasma testosterone concentrations in stallions

Testosterone concentrations in stallions showed a seasonal trend with peak concentrations in the spring (April and May in Britain) and lowest concentrations in the period from December to February. The effect on this pattern of changing the length of the photoperiod at the end of the normal breeding season (mid-summer's day) was studied in 2 experiments. In the first experiment artificial illumination was organised from 21 June to mimic the effect of transfer to a southern hemisphere spring and summer, that is short days becoming longer. The stallions had low concentrations of testosterone in February and high concentrations in April. Concentrations in July, August and September were extremely low with a return to high values in late November/early December. In the second experiment, illumination was maintained at the equivalent of a 16 1/2-hour day from 21 June to mid December. These stallions had high testosterone concentrations in April, after which they fell until August, later rising to a maximum in October. These results are discussed in relation to transfer of stallions between the northern and southern hemispheres.     

Inhibin secretion in the golden hamster (Mesocricetus auratus) testis during active and inactive states of spermatogenesis induced by the restriction of photoperiod

Gonadal function in the male golden hamster (Mesocricetus auratus) was investigated during exposure to a short photoperiod condition. Within 3 weeks of exposure to the short photoperiod condition, FSH and testosterone in the plasma significantly decreased, and subsequently immunoreactive (ir)-inhibin significantly decreased. Testicular contents of ir-inhibin and testosterone, and pituitary contents of LH and FSH also significantly decreased by 3 weeks with regression of weight of testes, epididymis and seminal vesicles and sperm head count. Circulating LH varied but not significantly. Thereafter, all reproductive parameters and secretion of LH, FSH, ir-inhibin and testosterone gradually recovered after 17 weeks of exposure even though animals continued to be subjected to the short photoperiod condition. Plasma concentrations of inhibin B and inhibin pro-alphaC were detectable and were significantly decreased after 15 weeks of exposure to the short photoperiod, but their levels were still detectable. Immunopositive reaction of inhibin alpha and betaB subunits was found in Sertoli cells and Leydig cells in the regressed testes of animals subjected to short photoperiod as was also seen in animals before exposure to the short photoperiod. Although the spermatogenic cycle was suppressed like prepubertal animals, the present study showed that the testicular recovery, so-called refractoriness, is functionally different from the developing stage of immature animals, especially with regard to inhibin secretion. The present results showed that changes in FSH preceded changes in inhibin during the regression and recovery phases, indicating that FSH is a major regulatory factor of inhibin secretion in male golden hamsters. The present study also demonstrated that regressed testes still secrete a small amount of bioactive inhibin during exposure to a short-photoperiod condition.     

Influence of the duration of photoperiod on growth,testicular characteristics and endocrine function of boars

Crossbred boars were used to evaluate the influence of exposure to 8 or 16 hr of light daily from 75 to 175 days of age on growth rate, testicular characteristics and endocrine function. At 160 days of age, concentrations of testosterone in serum (P<.10), the areas under plotted 12 hr testosterone profiles (P<.10) and the number (P<.05) and magnitude (P<.10) of testosterone secretory spikes were increased in boars exposed to 16 hr of light compared to boars in 8 hr light, but concentrations of LH in serum were similar in boars exposed to both treatments. Treatment with GnRH resulted in similar concentrations of LH in serum for both groups of boars. Testosterone in serum after GnRH-mediated LH release was greater at .5 (P<.05) and 1.0 (P<.10) hr following GnRH in boars exposed to 16 hr of light compared to boars at 8 hr, but concentrations of testosterone were similar for both treatments from 1.5 to 4.0 hr after GnRH. Growth rate and testicular and epididymal weights and sperm reserves at 175 days of age were not significantly altered by duration of photoperiod. Boars exposed to 8 hr of light had more hair per unit area than boars exposed to 16 hr of light. We conclude that exposure of prepubertal boars to longer daily photoperiods results in increased concentrations of testosterone in serum at 160 days of age.     

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.     

Testosterone serum profile,semen characteristics and testicular biometry of Mangalarga Marchador stallions in a tropical environment

This study was conducted to characterize the daily profile of testosterone secretion and its mean concentrations in the four seasons as well as to evaluate the semen characteristics and testicular biometry of Mangalarga Marchador stallions throughout the year in a tropical region. Three stallions were submitted to semen collections and evaluation of testicular biometry every 14 days along a year. Blood samples were collected once at the middle of each season, in a 20‐min interval during 24 hr in order to evaluate the testosterone secretion profiles among seasons. Testosterone concentrations along the day were higher at the beginning of the afternoon (from 12:00 to 15:00 hr), but a circadian secretion was not clearly observed. Mean testosterone concentrations did not differ among seasons (p > .05), but a pattern of secretion along the day showed variations with higher concentrations in the afternoon during the winter. Ejaculate volume was higher during summer; however, sperm motility decreased in summer and spring. Total sperm in ejaculate, sperm morphology and testicular biometry kept constant along the year showing no differences among the seasons. The results demonstrated that in a tropical region, reproductive aspects of stallions did not show a clearly defined seasonal variation, and months of autumn and winter were not unsuitable for reproduction of the males.     

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.     

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.     

Clinical observations on changes in concentrations of hormones in plasma of two stallions with thermally-induced testicular degeneration

To investigate effects of thermally-induced testicular degeneration on hormonal and seminal parameters in stallions, the scrotum was insulated for 36 hours in two mature (5-year-old mixed breed and 11-year-old Throughbred) stallions. Semen was collected daily for 10 days (DSO) prior to, and at intervals after, scrotal insulation. When DSO determinations were not being made, semen was collected 3 times weekly. Jugular blood samples were collected at 15-minute intervals for 6 hours from each stallion prior to, and at intervals after, scrotal insulation. A mouse interstitial cell testosterone assay was modified to quantify biologic activity of equine luteinizing hormone (BLH) in plasma samples. Immunoactive luteinizing hormone (ILH) and testosterone (T) concentrations were determined in plasma samples by routine RIA procedures. Percentages of progressively motile and morphologically normal spermatozoa began to decrease by 1 to 2 weeks postinsulation, reached nadir values at 3 to 3-1/2 weeks postinsulation, and returned to preinsulation values by 7 weeks postinsulation. Total number of spermatozoa and total number of progressively motile, morphologically normal spermatozoa in ejaculates at DSO returned to normal by 8 weeks postinsulation in stallion 2 and 12 weeks postinsulation in stallion 1. Concentrations of BLH and ILH increased, and while T concentrations decreased, immediately postinsulation. The increase in ILH concentrations was greater than the increase in BLH concentrations, resulting in a decrease in the BLH:ILH (B:I) ratio. Following the peak in LH secretion immediately postinsulation, LH concentrations gradually decreased while T concentrations increased. The B:I ratio was elevated from 1 to 13 weeks postinsulation compared to immediately postinsulation. In addition to changes in spermatozoal quality in ejaculates, stallion response to scrotal insulation included increased secretion of luteinizing hormone and impaired Leydig cell function (as determined by reduced testosterone concentration in circulating plasma). The proportion of biologically active LH secreted in response to thermal testicular injury increased during the recovery phase.     

The relationship of plasma leptin concentration and puberty in Holstein bull calves (Bos taurus)

The objective of this experiment was to study the changes of plasma leptin concentration during puberty and its relationship with testosterone level and testis dimensions in Holstein bull calves. Six Iranian Holstein bull calves with approximately 6 months of age were used. Semen evaluation was conducted at 1‐month interval to determine the puberty state. To detect the plasma leptin and testosterone changes, blood samples were collected from the jugular vein during pre‐puberty (6–7 months of age), puberty (8–9 months of age) and post‐puberty (10–11 months of age). In addition, body weight (BW), body condition score (BCS) and testicular width and length were measured at 3‐week intervals. The effects of time (age) on total sperm number and percentage of progressive motility of sperm, plasma concentration of leptin and testosterone, amplitude and frequencies of testosterone, BW, BCS, testicular dimensions were significant. Sperm number and progressive motility during post‐puberty were higher than those during puberty and pre‐puberty. Plasma concentration of leptin during the pre‐puberty was higher than those during puberty and post‐puberty (p < 0.01). Mean plasma testosterone concentrations during puberty were higher than those during pre‐puberty (p < 0.05). BW, BCS and testicular dimensions consistently increased throughout the trial. Results indicated that in growing bull calves, plasma concentrations of leptin decreased during puberty, while circulating testosterone increased.     

Recommendations for clinical GnRH challenge testing of stallions

Eight mature light-breed stallions with normal testes size, sperm output and semen quality were used to evaluate response to 3 GnRH challenge regimens in the summer in southeast Texas. Gonadotropin releasing hormone (50 μg) was administered intravenously once to each of eight stallions after three days of sexual rest (50 μg GnRH-1X). The same stallions were administered either 5μg GnRH intravenously once hourly for three injections (5 μg GnRH-3X) and 15μg GnRH intravenously once (15μg GnRH-1X) one and two weeks later. Blood samples were collected prior to and at intervals after GnRH administration. Plasma was immediately separated from blood samples and was frozen until assayed for LH, FSH, estradiol and testosterone concentrations. Percentage changes in hormone concentrations from pre-treatment values (baseline) were analyzed by paired studient'st-test to detect significant rises in hormone concentrations. Group mean percentage changes in hormone concentrations were analyzed by analysis of variance to compare responses among treatments. A computerized peak-detection algorithm (PC Pulsar) was used to detect peaks in LH and testosterone concentrations following 5 μg GnRH-3X and 15 μg GnRH-1X treatment.No differences (P>0.10) were detected in percentage change from baseline concentration for LH, FSH, or testosterone at one or two hours after administration of any of the three regimens of GnRH. When more frequent sampling intervals were analyzed for 5 μg GnRH-3X or 15 μg GnRH-1X treatments, no differences were detected in percentage change from baseline concentration for any hormone at 15, 30 or 60 minutes. Thereafter, percentage changes in concentrations of LH and FSH remained increased for 5μg GnRH-3X compared to 15 μg GnRH-1X treated stallions (P<0.05). Percentage changes in concentrations of testosterone were increased for 5μg GnRH-3X compared to 15 μg GnRH-1X treated stallions from 180–300 min (P<0.05), while no differences (P>0.10) were detected between 5 μg GnRH-3X and 15 μg GnRH-1X treated stallions for changes in concentrations of estradiol throughout the experiment.For 15 μg GnRH-1X treated stallions, maximum concentrations of LH in PC Pulsar-detected peaks occurred most commonly at 15 to 30 minutes (7/8 treatment periods) after GnRH injection. Maximum concentrations of testosterone in PC Pulsar-detected peaks occurred most commonly at 60–120 min (7/8 treatment periods) after GnRH injection.A protocol of blood sampling prior to, and 15, 30, 60 and 120 minutes after, intravenous administration of small doses of GnRH would be practical for challenge testing of stallions during the breeding season. In order to reduce cost of hormone assays, we suggest assay of the pre-challenge blood sample (baseline) could include LH, FSH, testosterone and estradiol concentrations (to assess overall hypothalamic-pituitary-testicularfunction), while only LH and testosterone concentrations need be determined after GnRH administration (to assess pituitary and testicular responsiveness). Assay for LH could be done on only the 15 and 30 minute post-GnRH samples, and assay for testosterone could be done on only the 60 and 120 minute post-GnRH samples. Failure to achieve approximately a 50% increase in LH concentration by 30 minutes after GnRH administration, and/or failure to achieve approximately a 100% increase in testosterone concentration by two hours after GnRH administration, could be further pursued either by treatment with increasing dosages of GnRH, or repeated administration of GnRH at hourly intervals, as has been suggested by other workers.