Biological activity of LH during the peripartum period and the estrous cycle of the ewe

Biological activity of luteinizing hormone (LH) is related to the degree of glycosylation of the glycoprotein hormone. The objectives of this study were to determine changes in biologically (BLH) and immunologically (ILH) active LH concentrations in plasma (in vitro bioassay and radioimmunoassay, respectively) and in the ratio of BLH to ILH (B:I) during the peripartum period and during the estrous cycle of the ewe. Blood samples were collected daily 4 days before through 4 days after parturition and during one estrous cycle. Also, samples were collected at 15-min intervals for 6 hr on Days 3 and 12 of the estrous cycle to quantify the influence of an elevated plasma concentration of progesterone (P) on the episodic secretion profiles of BLH. Progesterone concentration was determined on the 4th days pre- and post-partum, on each day of the estrous cycle and at hourly intervals on Days 3 and 12 of the cycle to investigate the hypothesis of an inverse relationship between P and BLH. The BLH and ILH concentrations were low during the peripartum period, and the B:I ratio did not increase by the 4th day postpartum. Mean ILH concentration was greater (P less than .05) in the postpartum than during the prepartum period. During the estrous cycle, mean daily B:I ratio was consistently above unity except for the day of estrus. The pre-ovulatory LH surge (BLH and ILH) was associated with a decrease (P less than .05) in the mean B:I ratio to 0.0065. Mean concentrations of BLH and ILH in plasma samples collected every 15 min on Day 12 were similar to Day 3 of the cycle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Sexual Activity Influences the Secretion of Reproductive Hormones in the Stallion

Contents The aim of this study was to investigate the effect of sexual activity on concentrations of reproductive hormones in plasma of stallions. In the first experiment, two groups of stallions were monitored for secretion of luteinizing hormone (LH), testosterone and oestradiol from the beginning until shortly after the end of the breeding season. One group of animals were reserve stallions not used for breeding (group 1, n = 10), the other group consisted of active breeding sires (group 2, n = 8). Blood samples were withdrawn from March to August at 14-day intervals. In sexually nonactive stallions (group 1), seasonal variations in LH, testosterone and oestradiol occurred and concentrations of these hormones reached a maximum in May (p < 0.05). In the breeding stallions (group 2), no significant changes in the concentrations of these hormones were found between March and August. Concentrations of LH and testosterone were significantly lower in breeding stallions than in reserve stallions at most blood sampling times (p < 0.05). In the reserve stallions, oestradiol concentrations were significantly higher than in the breeding stallions in April and in June (p < 0.05). In a second experiment, the effect of regular sexual activity (semen collection three times per week) on the concentration of LH, testosterone and oestradiol was tested in a group of breeding stallions after a period of sexual rest for several weeks. Blood samples were taken once daily starting the day before the first semen collection was performed. Testosterone concentration significantly decreased in the first days after semen collection started (p < 0.05), while LH secretion was only transiently decreased and no effects on oestradiol concentration were found. In both experiments, semen parameters were within the normal range of fertile stallions. No correlations between the sexual drive of the stallions and concentration of reproductive hormones occurred. It can be concluded that in the stallion the secretion of reproductive hormones is influenced by sexual activity. Regular semen collection seems to inhibit testosterone release by unknown mechanisms while the effects on LH and oestradiol secretion are less pronounced.     

Patterns of secretion of luteinizing hormone, follicle stimulating hormone and testosterone in stallions during the summer and winter

Samples of jugular blood were drawn from each of five stallions every 15 min for 12 h during the summer and winter to determine the short-term fluctuations in plasma concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH) and testosterone. Concentrations of LH and FSH were generally not pulsatile, although one stallion exhibited three distinct pulses in these hormones during the winter. In general, patterns of secretion of all three hormones were similar in both seasons and the number of significant rises in hormonal concentrations did not differ between seasons. Concentrations of LH and FSH were positively correlated (P less than .05) for eight of the ten sampling periods, indicating a close relationship between the secretion rates of these two gonadotropins. Testosterone concentrations varied in an episodic manner during the 12-h period, and all stallions exhibited at least one episode of high testosterone secretion regardless of the pattern of LH concentrations. The response in testosterone concentrations to the three LH pulses exhibited by the one stallion in winter was not the same for each pulse. The correlations between a single random sample and mean concentrations over the 12-h period were high (r between .88 and .99) for all three hormones, indicating that a single sample of blood would be representative of overall concentrations. It appears that the stallion differs from males of other domestic species in that concentrations of gonadotropins and testosterone vary in a much less pulsatile manner.     

Effects on testosterone, LH and cortisol concentrations, and on testicular ultrasonographic appearance of induced testicular degeneration in bulls.

It is well known that heat stress has a detrimental effect on testicular functions. In addition to the alteration of semen quality and testicular damage, reproductive hormone secretion can be altered. The objective of this study was to describe changes in plasma concentrations of testosterone, LH and cortisol, as well as in testicular ultrasonographic appearance after induced testicular degeneration. Four Swedish Red and White bulls, aged 3 years, were used. They were fed according to Swedish standards. The scrotum was covered with an insulation device during 96 h. Semen was collected weekly 3 times before and up to 4 months after insulation. Testicular ultrasonography and clinical genital examination were performed with the same intervals. Heparinized blood samples were taken from the jugular vein at 2 h interval during 24 h every 2 weeks during the study. Blood samples were tested for the content of testosterone, LH and cortisol. Data were analysed, using one way analysis of variance of seminal data, clinical examination data as well as 24 h hormonal output data as percentage of mean individual pretreatment values. The use of a 5 MHz B-mode ultrasound unit did not contribute with an objective estimation of the degree of testicular degeneration. In 3 of the bulls testosterone levels had a tendency to decrease and LH to increase during the time of severe degeneration, whereas an opposite trend was seen during the regenerative phase, changes becoming significant 15 weeks after scrotal insulation. Variation between animals was big. Cortisol levels had a decreasing trend, changes being significant only in individual bulls at 10 and 15 weeks after scrotal insulation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Semen parameters and hormone concentrations in stallions subjected to long-term estrogen administration

Eight pony stallions were paired by estimated daily sperm output (DSO) and randomly assigned to one of two treatments in a randomized block experiment. Stallions received 44 μg/kg BW estradiol cypionate (ECP) or an equivalent volume of physiological saline solution on alternate days during the breeding season. Blood samples collected immediately preceding each injection were assayed for luteinizing hormone (LH), estradiol-17β (E₂) and testosterone (T). Semen was collected twice weekly, 3.5 days apart, to evaluate sperm motility and total number of sperm per ejaculate. Prior to and after 4, 8 and 12 weeks on treatment, semen was collected once daily for 7 days to determine DSO. Data were separated into 9 periods (10 days each) for statistical analysis and subjected to analysis of variance for a randomized block design to determine treatment effects.There were no differences (p>.05) between groups for DSO or LH prior to initiation of treatment. Testosterone was higher (p<.05) in ECP stallions compared with C stallions prior to treatment and at all time points measured. As expected, E₂ was higher (p<.05) in the ECP stallions compared to C stallions after 20 days (period 2) of treatment and for the remainder of the experiment. However, E₂ was higher (p<.05) in the C group prior to treatment, but there was no difference between the groups after 10 d of treatment (period 1). ECP stallions had higher (p<.05) DSO than C stallions after 30 d on treatment. After 40 and 50 d (periods 4 and 5), ECP stallions demonstrated higher (p<.05) total sperm per period than C stallions. This was preceded by higher (p<.05) LH values for ECP stallions than for C stallions after 10 and 20 d (periods 1 and 2). No differences were found between the ECP and C groups for LH between 30 and 60 d. Although numerically higher, no significant differences (p>.05) were seen after 60 days for DSO or after 60, 70 or 80 days for total sperm per period. ECP stallions had higher (p<.05) DSO and total sperm per period after 90 d than C stallions. Additionally, LH remained significantly higher (p<.05) in the ECP group after 60 days (periods 7, 8 and 9). Elevated LH concentrations in ECP stallions demonstrated that estrogen treatment did not inhibit LH secretion in this study.     

Sperm morphology in Estonian and Tori breed stallions

The standard procedure for assessing the breeding potential of a stallion includes the parameter total number of spermatozoa classified as morphologically normal. This study investigated sperm morphology of fresh semen in randomly chosen Estonian (E, n = 8) and Tori (T, n = 7) breed stallions with proven fertility. Two ejaculates were examined from each stallion. An aliquot from each ejaculate was fixed in 1 mL formol-saline immediately after collection and examined with phase-contrast microscope at a magnification 1000x for all types of morphological abnormalities. Furthermore smears were prepared and stained according to Williams (carbolfuchsin-eosin) for a more detailed examination of the sperm heads with light microscope at a magnification 1000x. Analysis of variance was applied to the data, and results are presented as LSmeans (+/- SE). One T stallion that had a disturbance in the spermatogenesis and one 22-year-old E stallion were not included in the analyses. The T stallions had on average 57.5 +/- 4.1% and the E-stallions 74.4 +/- 3.8% morphologically normal spermatozoa (p = 0.012). In 4 of 7 T stallions and 7 of 8 E stallions both ejaculates had > 50% morphologically normal spermatozoa. There was a significant difference between breeds in mean percentage of proximal droplets (17.3 +/- 2.7% and 2.9 +/- 2.5% for T and E stallions, respectively; p = 0.003).     

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.     

Seminal characteristics of two- and three-year-old quarter horse stallions

Seminal and testicular characteristics were compared in 19 three-year-old and 23 two-year-old Quarter Horse stallions. Semen was collected, and testicular evaluations made on all stallions once in April or May and again 60 days later. Semen was collected from stallions twice, one hour apart, at each evaluation. Average testicular tone and scrotal width were greater in three-year-olds than in two-year-olds. Ejaculates of three-year-olds had higher motility scores (66% vs 47%) sperm movement (3.2 vs 2.4), normality (77 vs 71), and total number of normal, motile spermatozoa per ejaculate (4.3 × 10⁹ vs 2.2 × 10⁹). Two-year-olds had twice as many cytoplasmic droplets (11% vs 5%) as three-year-olds. Average sperm concentrations per ml of gel-free semen and gel-free volume of ejaculates were not different. Volume of ejaculate and motility of spermatozoa were positively correlated with pregnancy rate, whereas percent cytoplasmic droplets was negatively correlated with pregnancy rate in both groups of stallions. Pre- and post-ejaculatory urethral, seminal, and preputial cultures were examined for pathologic organisms on all stallions. Eighteen of 42 had positive cultures for Klebsiella spp., Pseudomonas spp., β-Strep spp., or E. coli. Nine two-year-olds and 3 three-year-olds had positive cultures for Klebsiella spp. All but one of these stallions were housed on wood shavings and allowed limited exercise. Three-year-old stallions had superior seminal quality as compared with two-year-olds.     

The effects of equine somatotropin on pituitary and testicular function in the stallion during the nonbreeding season

An experiment was conducted to determine the effects of equine somatotropin on the reproductive axis of the stallion during the nonbreeding season. Adult stallions were treated with equine somatotropin (20 μg/kg body weight [BW]; n = 5) or saline (n = 4) daily for 21 days starting in January. During the last week of treatment, stallions were subjected to low- and high-dose injections of luteinizing hormone (LH), as well as low- and high-dose injections of gonadotropin-releasing hormone (GnRH) and thyrotropin-releasing hormone (TRH). Two months after the onset of somatotropin treatment, semen was collected from all stallions every other day for 14 days. Treatment with equine somatotropin increased (P < .001) daily IGF-1 concentrations but had no effect (P > .1) on concentrations of LH, follicle-stimulating hormone (FSH), or testosterone. The testosterone responses to injections of LH were similar (P > .1) between treatments. Likewise, the LH, FSH, prolactin, and testosterone responses to the injections of GnRH/TRH were similar (P > .1) between groups. At seminal collections, stallions treated with somatotropin exhibited greater volumes of gel-free semen (P < .01) and gel (P < .05) and had decreased time until ejaculation (P < .05). In conclusion, somatotropin treatment for 21 days may alter the long-term accessory gland contribution to seminal volume but does not appear to alter pituitary gonadotrope function or testicular testosterone secretion.     

Biological and immunological luteinizing hormone activity and blood metabolites in postpartum Brahman cows

Multiparous Brahman cows were assigned by order of calving and sex of calf to groups to be fed to maintain body condition score (BCS) of 6 or greater (M; n = 10) or to lose BCS (L; n = 10). Blood samples were collected weekly for progesterone analysis and at 15, 30 and 45 d after parturition at 15-min intervals for 6 h for determination of immunological (ILH) and biological (BLH) luteinizing hormone. Serum concentrations of ILH were determined using a double antibody RIA procedure, whereas BLH was determined using a rat interstitial cell-testosterone bioassay (RICT). By 45 d after parturition 7 of 10 M cows had returned to estrus. Therefore, comparisons between groups were made on d 15 and 30 postpartum. Cows in the M group had a shorter (P less than .001) interval to first estrus (46.7 d) than did L cows (91.2 d). The concentrations of bioactive and immunoactive LH were parallel between d 15 and 30 postcalving. However, a day x treatment interaction (P less than .05) showed that episodic BLH concentrations (ng/ml) decreased with day postpartum in L, but increased in M cows from d 15 to 30 postcalving. Likewise, relative biological activity, as measured by B:I ratios, decreased between d 15 and 30 in L cows, whereas it increased in M cows during the same period (B:I x day interaction; P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)     

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 propionate treatment of stallions: Effects on secretion of luteinizing hormone and follicle stimulating hormone in daily samples and after administration of gonadotropin releasing hormone

Ten stallions were used to determine if the stallion responds to administration of testosterone propionate (TP) with an increase in follicle stimulating hormone (FSH) secretion after administration of gonadotropin releasing hormone (GnRH) as has been previously observed for geldings and intact and ovariectomized mares. Five stallions were treated with TP (350 μg/kg of body weight) in safflower oil every other day for 11 days; control stallions received injections of safflower oil. The response to GnRH (1.0 μg/kg of body weight) was determined for all stallions before the onset of treatment (GnRH I) and at the end of treatment (GnRH II). Blood samples were also withdrawn daily from 3 days prior to treatment through GnRH II. Treatment with TP decreased (P<.10) concentrations of FSH in daily blood samples. However, treatment with TP did not affect (P>.10) the GnRH-induced secretion of FSH. Concentrations of luteinizing hormone (LH) decreased (P<.05) in daily blood samples averaged over both groups of stallions and were lower (P<.10) in TP-treated stallions than in controls during the latter days of treatment. We conclude that TP administration to stallions does not alter the FSH response to GnRH as has been observed for geldings and for mares of several reproductive states.     

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.     

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.     

Pregnancies following artificial insemination with spermatozoa from problem stallion ejaculates processed by single layer centrifugation with Androcoll-E

Some stallions produce ejaculates of low quality and/or low fertility when used for artificial insemination (AI). The purpose of these five case studies was to use Single Layer Centrifugation (SLC) to select the best spermatozoa from 'problem' ejaculates for subsequent use in AI. Sperm quality, in terms of motility, morphology and chromatin integrity, was improved in the SLC-selected samples compared to the corresponding uncentrifuged samples, with the exception of one stallion thought to have ampullary stasis. In this stallion, neither the incidence of spermatozoa with detached heads nor the proportion of damaged chromatin was decreased by SLC, in contrast to previous results. Pregnancies were obtained after using SLC-selected spermatozoa from the five stallions for AI, indicating that the spermatozoa were functional after SLC. Overall, the results suggest that SLC may be useful when preparing AI doses from some 'problem' ejaculates.     

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.     

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.     

Sperm Production and Sperm Morphology of Swedish Warmblood Stallions

The present study investigated daily sperm output and sperm morphology of fresh semen in eight Swedish Warmblood stallions aged 5–8 years. They were used for artificial insemination, and their fertility during the breeding season of semen collection exceeded 60% per cycle. One ejaculate of semen was collected daily for 10 consecutive days from each stallion. The gel-free volume was measured, and the sperm concentration was assessed with a Bürker chamber. The volume of gel-free fraction was multiplied by the sperm concentration to give the total number of spermatozoa (TSN). Sperm morphology was examined in ejaculates collected on days 2, 5 and 10. An aliquot from each ejaculate was fixed in 1 ml formol–saline immediately after collection and examined under a phase-contrast microscope (magnification 1000×) to assess morphological abnormalities. Furthermore smears were prepared and stained according to Williams (carbolfuchsin–eosin) for a more detailed examination of the sperm heads under a light microscope (magnification 1000×). Analysis of variance was applied to data. Total spermatozoa number decreased progressively during the first 8 days of collection, and daily sperm output (DSO) was calculated as mean TSN of collections on days 8–10, being 6.4 × 10⁹ spermatozoa. The overall percentages of morphologically normal spermatozoa in ejaculates collected on days 2, 5 and 10 were above 70%, being significantly lower in ejaculate 2 (68.6%) compared with ejaculates 5 and 10 (72.9% respectively 75.3%).     

Effects of sexual stimulation, with and without ejaculation, on serum concentrations of LH, FSH, testosterone, cortisol and prolactin in stallions

Six lighthorse stallions with previous sexual experience were used to determine the short-term effects of sexual stimulation (SS; 5 min exposure to an estrous mare), SS plus ejaculation (SSE), and no stimulation (control) on serum concentrations of LH, FSH, testosterone, cortisol and prolactin. Stallions received one treatment per day on d 1, 4 and 7. Treatments were assigned such that each stallion 1) received each treatment once and 2) experienced a unique sequence of treatments. Neither SS nor SSE had any consistent effects on LH or FSH concentrations. Testosterone concentrations during control bleedings increased (P less than .05) with time. This increase was suppressed (P less than .05) by both SS and SSE. Cortisol concentrations increased (P less than .05) immediately after SS and SSE. Cortisol concentrations also tended to increase during the control bleedings, but only in stallions that previously had been exposed to SS or SSE. Prolactin concentrations increased (P less than .05) immediately after SS and SSE and tended to rise during control bleedings in stallions previously exposed to SS or SSE. We conclude that 1) prolactin and cortisol were secreted rapidly in response to SS and SSE, 2) the rise in cortisol concentrations likely suppressed testosterone secretion within the next hour, and 3) stallions appeared to associate the distant sounds of other stallions with their own previous exposure to SS and SSE, resulting in a cortisol response (and perhaps a prolactin response) even in the absence of direct stimulation.     

Morphology and Chromatin Integrity of Stallion Spermatozoa Prepared by Density Gradient and Single Layer Centrifugation Through Silica Colloids

The objective was to investigate whether it is possible to improve the quality of stallion semen, with respect to sperm morphology and chromatin integrity, both of which have been linked to fertility, using either density gradient centrifugation (DGC) or a new method, hereby named single layer centrifugation (SLC). The two methods of colloidal centrifugation were evaluated using 38 ejaculates from 10 stallions. Sperm morphology, subjective motility and sperm chromatin integrity were compared in uncentrifuged samples and in centrifuged sperm preparations. The proportion of morphologically normal spermatozoa varied between stallions (p < 0.001) and was increased by both methods of colloidal centrifugation (median value before centrifugation 67.5%; after SLC 78%; after DGC 77%; p < 0.001). The incidence of certain abnormalities was reduced, e.g. proximal cytoplasmic droplets were reduced from 12.9% to 8.8% (p < 0.001), and mid-piece defects from 5.3% to 1.4% (p < 0.05). Similarly, sperm motility and chromatin integrity were significantly improved (p < 0.001), with no difference between the two centrifugation methods. Centrifugation through colloids can enrich the proportions of stallion spermatozoa with normal morphology and normal chromatin structure in sperm preparations. The new method, SLC, was as effective as DGC in selecting motile stallion spermatozoa with normal morphology and intact chromatin. SLC, being simpler to use than DGC, would be appropriate for routine use by stud personnel to improve stallion sperm quality in insemination doses.