Unusual ovarian activity in a mare preceding the development of an ovarian granulosa cell tumour.

An 8-year-old mare, with a foal at foot, was inseminated on foal heat with frozen semen, with the resultant pregnancy lost between days 34 and 41. The right ovary developed a large anovulatory follicle that was non-responsive to multiple doses of ovulating agents. The follicle eventually appeared to luteinise, although plasma progesterone concentrations did not reflect this. Another follicle developed, responded to GnRH and resulted in a pregnancy from frozen semen that went to term with a healthy foal. When the mare was examined after foaling, the structure on the right ovary appeared to be a granulosa cell tumour; the left ovary was smaller than normal and non-functional. Surgical removal of the right ovary before increasing photoperiod resulted in a return to function of the left ovary and a pregnancy to frozen semen on the second cycle following removal. Figures showing concentrations of inhibin, progesterone, androstenedione, oestradiol and testosterone are presented for this entire period. Unusual ovarian activity in the mare might be a prelude to the development of a granulosa cell tumour.

Preliminary study of ovarian activity in fillies treated with a GnRH vaccine.

OBJECTIVE: To investigate the effects of two doses (200 and 400 mg) of a water-soluble gonadotrophin-releasing hormone vaccine on the ovarian activity of 2-year-old fillies. DESIGN: A controlled vaccination dose rate experiment. ANIMALS: Six 2-year-old Australian Stock Horse fillies were randomly allocated to three treatment groups; unvaccinated controls, those receiving 200 mg of the vaccine and those receiving 400 mg of the vaccine. RESULTS: Ovarian activity of the treated fillies was suppressed at the peak of breeding season while that of untreated controls continued normally. The control fillies displayed oestrous activity and behaviour. Suppression of ovarian activity occurred for 25 and 30 weeks in the 200 and 400 mg groups, respectively. These differences were not significant. Ovarian activity ceased 2 to 3 weeks after primary vaccination. Antibody titres were low (330) until after the booster immunisation when they rapidly peaked at 22,000 and 28,000 in the 200 mg and 400 mg groups, respectively. Plasma progesterone concentrations of the treated fillies remained below 3.18 nmol/L while GnRH was suppressed. The vaccine had no significant effect on plasma androstenedione concentrations. Recovery from the effect of the vaccine was associated with development of ovarian follicles, normal oestrous behaviour and ovulation. Three of the four treated fillies and one of the controls conceived during the next breeding season and foaled normally. All the treated fillies conceived and produced normal foals in the following two breeding seasons. CONCLUSION: Both dose rates suppressed ovarian function and prevented oestrous behaviour. These effects were reversible and the subsequent fertility of the vaccinated fillies was normal.

Suppression of testicular function using two dose rates of a reversible water soluble gonadotrophin releasing hormone (GnRH) vaccine in colts.

OBJECTIVE: To investigate the effect of two dose rates (200 and 400 ng) of a gonadotrophin releasing hormone (GnRH) vaccine on testicular function. DESIGN: A vaccination dose rate experiment. PROCEDURE: Two injections were administered 4 weeks apart to six colts in each treatment group. To maintain immunosuppression until the end of the breeding season, a third injection was given if antibody titres fell below 1000. RESULTS: Effective antibody titres were present for 12 to 27 weeks. Testosterone concentrations decreased from 2.22 to 0.31 nmol/L 6 weeks after primary vaccination. Androstenedione concentrations decreased from 1.78 to 0.28 nmol/L 5 weeks after vaccination. Testosterone and androstenedione concentrations above 0.69 and 0.87 nmol/L were attained 31 to 43 weeks after vaccination. Mean scrotal widths and lengths decreased over 29 weeks from 9.2 cm and 9.7 cm to 6.7 cm and 7.6 cm. At surgical castration these dimensions were 10.1 cm and 11.0 cm. Mean semen characteristics before vaccination and after recovery were: gel-free volume 16.5 and 13.5 mL, sperm concentration 295.5 x 10(6) and 315.6 x 10(6)/mL, total sperm per ejaculate 4041 x 10(6) and 4657 x 10(6) and live normal spermatozoa 32% and 60%. Histologically, the testes showed active spermatogenesis. The mean testicular parenchyma weights for the 200 and 400 mg groups were 129.0 g and 109.8 g. Daily sperm production per testis and per gram of testis for the 200 and 400 mg groups were 3.7 x 10(8) and 2.8 x 10(6), and 2.3 x 10(8) and 2.0 x 10(6). CONCLUSIONS: Both dose rates suppressed testicular function. Data showed that the vaccine effects were reversible. Individual immune response was less varied in the 200 mg group. Further work is necessary to achieve a less variable response in the immunosuppression of testicular function.

The influence of age and breed on stallion semen.

This study reports on the variation in semen quality and in spermatozoal and behavioral characteristics of 168 stallions representing 9 breeds and ranging in age from 2 to 26 yr. Semen samples were collected into an artificial vagina and the number of mounts and urethral pulsations per semen sample were recorded. Semen characteristics were examined for total volume, gel-free volume, gel volume, color score, mass activity, nonmotile spermatozoa, dead spermatozoa, semen density, spermatozoa concentration, total number of spermatozoa and semen pH. Morphological characteristics of the spermatozoa included abnormal heads, abnormal mid-pieces, abaxial mid-pieces, protoplasmic droplets and abnormal tails. Sources of variation were evaluated and the overall means calculated by least-squares analyses of variance for nonorthogonal data. The significance of breed effects and between stallion variability were estimated using mixed-model procedures. All semen characteristics with the exception of color and urethral pulsations had significant variation due to age. Semen quality (gel-free volume, sperm concentration, total sperm numbers and sperm abnormalities) was poorest in stallions under 3 yr of age and over 11 yr. Significant breed variation was apparent in most characteristics except for pH, semen color, abnormal midpieces and urethral pulsations. It is recommended that both the age and breed of stallion be taken into consideration when evaluating stallion semen.

Immunocastration of colts and immunospeying of fillies.

A series of experiments using an ovalbumin conjugated gonadotrophin releasing hormone was used to stimulate antibody production, suppress testosterone secretion and depress testicular function in yearling and 2 year old colts and fillies. In the preliminary experiment, an injectable oil-based formulation was administered to yearling colts. Testicular development and testosterone secretion were retarded for a period of approximately 28-32 weeks while antibody titres were greater than 1:1000. An implant and water-soluble vaccine (200 and 400 mg) is presently being tested in 2 year old colts. Testicular dimensions and testosterone concentrations decreased following vaccination. Preliminary results indicate that the effects of the implant are reversible and last for approximately 30 weeks. The water-soluble vaccine produced effective antibodies for 10-19 weeks. Plasma testosterone of all colts was reduced to gelding levels (0.2 ng/mL) while antibody titres were greater than 1:1000. Semen evaluation, morphometric analysis and daily sperm production data indicate that the testes recovered normal function. These experiments are continuing and more definitive data on the vaccine and its reversibility will be available at their conclusion. Ovarian activity of the treated fillies (200 and 400 mg) ceased soon after vaccination. Their progesterone concentrations have remained below 1 ng/mL for 20 weeks. The untreated control fillies have continued to cycle normally as determined by ovarian ultrasonography and progesterone concentrations.

Seasonal variation in the estrous cycle of mares in the subtropics.

Data on the estrous cycles and sexual receptivity scores of 7 maiden Australian Stock Horse mares were used to study seasonal variation from December until the following November. Mares were grazed in paddocks containing both native and improved tropical pasture species. The study was conducted in South-East Queensland (27 degrees 27' South latitude). There were no differences between either the sexual receptivity or the lengths of natural (Mean=7.5 days, SEM=0.4) and PGF-induced estruses (Mean=7.6 days, SEM=0.4). Plasma estrogens were not related to differences in sexual receptivity scores between mares. The lowest incidence of estrus occurred at the time of the winter solstice (June 22) in the Southern Hemisphere. Winter estruses (Mean=9.3 days, SEM=0.8) were longer than those of summer (Mean=6.6 days, SEM=0.5) and autumn (Mean=6.6 days, SEM=0.9). Approximately 30% of estruses were anovulatory. Most of these occurred in autumn and winter (P<0.05). There was considerable variation in the occurrence of anovulatory estrus between mares. There were no differences between the lengths of ovulatory and anovulatory estruses. Ovulatory estruses were significantly longer in spring than in summer (P<0.05). There were no seasonal differences between the lengths of anovulatory estruses. The length of diestrus (Mean=16.6 days, SEM=0.3) was not affected by mare or season. Only 3 of the 7 mares did not cycle during winter (anestrus), as confirmed by progesterone concentrations of less than 1 ng/ml. Finally, there was no evidence of mares having 2 breeding seasons per year in this study.

A preliminary study of immunological castration in colts.

This study tested the effectiveness of a conjugated GnRH vaccine for stimulating antibody production, suppressing testosterone secretion and depressing testicular development in yearling colts. Two colts were allocated to each of three groups, (1) control, (2) subcutaneous and (3) intramuscular vaccinations. Two injections of the vaccine were given 11 weeks apart. Liveweight gain was not affected by vaccination but plasma testosterone concentrations in the treated colts were suppressed and their antibody titres to GnRH were greater than 1:1000. Testicular development in the treated colts also was retarded at this time, which was approximately 28 weeks after the first injection. Semen samples, containing spermatozoa, were collected from all the colts prior to their castration at the end of the investigation. Antibody titres and testosterone concentrations in the treated colts had returned to levels similar to those of the controls by the end of the experiment. The testes of the vaccinated colts were still smaller than those of the controls on castration but were, nevertheless, increasing in size. Morphometric analysis of testicular histology and daily sperm production data indicated that the testes of the treated colts were recovering and would probably have regained normal function had they been left in situ.