Anti-oxidative status and semen quality during cooled storage in stallions

Activity of the anti-oxidative enzymes glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT), content of thiobarbituric acid reactive substances (TBARS) and SH-groups were determined in native stallion semen (n = 8 stallions). Semen was then diluted in Kenney extender, EquiPro((R)) extender either with or without addition of N-acetyl cysteine or phosphate-buffered saline (PBS) and stored for 72 h at 5 degrees C. Correlations between initial activity of enzymes and development of semen motility and membrane integrity were calculated. Activities of GSH-Px, SOD and CAT immediately after semen collections were 10.0 +/- 0.6 picokatals, 0.40 +/- 0.03 SOD units and 0.70 +/- 0.05 nanokatals/10(6) spermatozoa respectively. TBARS content was 0.06 +/- 0.01 nmol and SH-group content 1.7 +/- 0.5 mmol/10(6) spermatozoa. The loss of motile spermatozoa during storage did not differ between extenders. N-acetyl cysteine had no effect on semen motility and membrane integrity. The loss in membrane-intact spermatozoa was highest (P < 0.05) in semen diluted in PBS. Motility and membrane integrity after addition of extender were positively correlated with GSH-Px and CAT, indicating that anti-oxidative mechanisms contribute to the initial high percentage of motile and membrane-intact spermatozoa. However, in these samples the decrease in semen quality was most pronounced. No correlations existed between initial activity of anti-oxidative enzymes, peroxidation products and semen quality during storage. This indicates that once extender has been added, peroxidative damage to sperm membranes is not the predominant cause of losses in semen quality.     

Prevalence of mycoplasmas in the semen and vaginal swabs of Danish stallions and mares

The reproduction rate of horses is one of the lowest within domestic livestock despite advances the veterinary medicine. Infertility in horses may be due mainly to the lack of suitable selection criteria in the breeding of horses. However, acquired infertility due to genital, bacterial infections may occur. Mycoplasmas have been implicated in genital disorders and infertility of many species including humans and horses. However, their role as commensals or pathogens of the genital tract of horses is still not determined. Bacteriological examinations made on the fossa glandis, urethra, penis and semen of stallions, showed the presence of different Mycoplasma species. Therefore our study aimed to find the prevalence of Mycoplasma species and a possible association with fertility problems in Danish riding horses. Eighty semen samples from stallions and 19 vaginal swab samples from mares were tested by PCR for presence of mycoplasmal DNA. The vaginal swab samples were also cultured in the Mycoplasma specific medium. None of the samples were positive for presence of genital mycoplasmas during the screen. The lack of genital mycoplasmas observed in this study may be due to a very extensive use of artificial insemination of modern sport horses.     

Thermoregulation of the testicle in response to exercise and subsequent effects on semen characteristics of stallions

Stallions (n = 8) were implanted with a thermal sensory device in the muscle of the neck and the subcutaneous tissue of the scrotum and then assigned to either a nonexercise (Non-EX; n = 4) or exercise (EX; n = 4) group. A motorized equine exerciser was used to work EX stallions 30 min/d for 4 d/wk during a 12-wk period from July through October 2010. Temperatures (subcutaneous scrotal, intramuscular neck, and rectal) were recorded at 0, 22, and 30 min after the start of exercise, as well as 60 and 120 min post-exercise. Hourly ambient temperature and relative humidity data were also obtained. Semen was collected at 0, 4, 8, and 12 wk and analyzed for volume, sperm concentration, total sperm numbers, percentages of total and progressively motile sperm, sperm morphologic characteristics, and sperm DNA quality. No effect (P > 0.05) of exercise was observed on any of the measured semen variables. Implantation of thermal sensory devices had no demonstrable acute or chronic effects on the scrotal or neck tissue, indicating that the thermal sensory devices are a safe and effective way to measure subcutaneous scrotal and neck temperatures. At 22 and 30 min of exercise, rectal and neck temperatures increased (P < 0.0001) approximately 1.9 and 2.4°C, respectively, and scrotal temperatures simultaneously increased, although not significantly (P = 0.33), approximately 0.8°C. Correlations existed between scrotal, neck, rectal, and ambient temperatures, with the correlation between scrotal and rectal temperatures being greatest (r(s) = 0.76; P < 0.0001). Although moderate exercise for a short duration in extreme heat and humidity did significantly increase core body temperatures in stallions, scrotal temperatures did not significantly increase, and sperm parameters were unaffected.     

Phantoms for the collection of genital secretions in stallions]

Practical experiences of the phantom method for collection of genital secretions from stallions are reported. Taking a phantom used in the Richard-G?tze-Haus Tier?rztliche Hochschule Hannover as a prototype two further models slightly modified have been constructed, baring a flat hollow in the right side of the caudal phantom body for manual inserting of the Artificial Vagina. These three models fulfill four important conditions for routine use: (1) sufficient sexual attractivity for the stallions; 80-85% successful collections of presecretions out of a total of 1050 using the dummy and 70% successful semen collections from more than 240 in total; (2) solid and resistant construction; (3) easy cleaning and desinfection of the surface of the phantom to get representative samples; (4) firm installation on a hygienic floor.     

Seasonal changes in spermatozoas and fertility of stallions in the tropics]

An attempt was made to express the relationship of spermiological characteristics with the fertility on the basis of seasonal variability of the fertility of stallions and of the ejaculate quality. A positive correlation between the activity of spermatozoa and the fertility ensues from the values of correlation coefficients. There is a negative correlation between the fertility and the concentration of citric acid. In the other characteristics, i.e. in the production of spermatozoa, volume of gel substance, pH, ergothioneine concentration, polarographic and hemolytic activity, the correlation coefficients were very low. It is assumed that these characteristics may influence the livability and preservation of the sperm, but they do not affect its fertilizing ability.     

Increased germ cell loss rates and poor semen quality in stallions with idiopathic testicular degeneration

Five mature stallions with poor semen quality were tentatively diagnosed as having testicular degeration of unknown cause. Testis samples from these five stallions, and from three mature stallions with normal semen quality, were obtained by castration and prepared for histomorphometry. Increased germ cell loss rates during late meiosis and spermiogenesis occurred in the stallions with idiopathic testicular degeneration. Poor semen quality, represented by a low percentage of morphologically normal, progressively motile sperm in ejaculates, appeared to be a good predictor of testicular degeneration in these stallions.     

Contribution of various factors to genetic evaluations of stallions

Genetic evaluations (BLUP) of 262 stallions were obtained for the following subjectively scored traits of the Swedish Riding Horse Quality Events: conformation, gaits, temperament in the gaiting test, jumping ability and temperament in the jumping test. The genetic evaluations were partitioned into the various factors included in the statistical model applied to the data. The partitioned sire solutions were compared with genetic evaluations obtained from application of an animal model. The differences between the evaluations indicated the influence of the genetic merit of mates. The following correlations were found between the estimated breeding values (from the animal model) of sires and their mates for the five traits in the same order as above: 0.101, 0.046, 0.096, 0.004, 0.020. Evaluations of genetic values within birth years (1973–1979) indicated a slightly negative genetic trend in conformation score and slightly positive genetic trend in the other four traits.     

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.     

Effect of GnRH immunisation on hormonal levels, sexual behaviour, semen quality and testicular morphology in mature stallions

The aim of this study was to investigate the effect of gonadotrophin-releasing hormone (GnRH) immunisation on mature stallions that had been used for breeding. Four Standardbred stallions were used in the study: 3 experimental animals and 1 control animal. Semen was collected regularly, i.e. twice/week, during the 4 months prior to the experimental period. The stallions were immunised against GnRH with a GnRH-BSA conjugate. Equimune was used as the adjuvant. The stallions were immunised on 5 occasions, 4 at 2 week intervals, and the fifth 4 weeks after the fourth. Blood samples were taken once a week for analysis of GnRH antibody titre and every third week for testosterone and oestrone sulphate analyses. Semen was collected once a week, and libido and sexual behaviour were observed. Ejaculate volume, sperm concentration, total number of sperm in the ejaculate, sperm motility and sperm morphology were evaluated. Testicular size was measured once a week. At the end of the study, the stallions were castrated, and a histological examination of the testes performed. All immunised stallions produced antibodies against GnRH, and plasma testosterone concentration decreased. However, the effect of immunisation varied between stallions. In 2 of the stallions, high levels of antibodies were found, while in the third, the level was moderate. Four weeks after the first immunisation, a decrease in libido was observed. Two months after the first immunisation, marked changes in semen quality were observed in the 2 stallions with high antibody titres. Fourteen weeks after the first immunisation, the total number of sperm/ejaculate had decreased from >8.6 x 10(9) to <2.7 x 10(9), sperm motility from >59 to <10% and the frequency of morphological normal spermatozoa had decreased from >60 to <14%. The dominating abnormalities were abnormal head shapes, proximal cytoplasmic droplets and detached heads. In the third stallion, only slight changes in semen quality were found. No changes were observed in the control stallion. Decreases in testicular size were noted in all of the experimental stallions. Pronounced histological alterations in the testes were observed in 2 of the stallions. It is concluded that the vaccine was effective in stimulating production of GnRH antibodies and in suppressing testicular function and androgen secretion. However, there was an individual variation in the responses among the stallions and, further, libido was not totally suppressed.     

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.