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.

Utilization of endogenous and dietary urea in the large intestine of the mature horse.

The dynamics of N metabolism in mature horses were investigated when they were fed on a low-N diet or the same diet supplemented with sufficient urea or soybean meal to meet their theoretical N requirements. There were no differences in DM, organic matter or neutral-detergent-fibre digestibilities for the three diets. N digestibilities and digestible-N intakes were similar for the urea- and soyabean-supplemented diets and very low for the low-N diet. For all three diets plasma urea was degraded in the digestive tract to NH3 which was utilized by the bacterial population as a N source. NH3 was absorbed from the large intestine into the blood and converted to urea. NH3 was also incorporated into plasma proteins. The horses fed on the low-N diet degraded a greater proportion of endogenous urea in the digestive tract than did horses fed on the urea-or soyabean-supplemented diets. However, the horses fed on the urea diet had the highest degradation rate of urea. The quantity of urea degraded in the digestive tract of horses fed on the urea-supplemented and the low-N diets could not compensate for a lack of dietary crude protein in these diets. The horses were in a negative N balance when fed on the low-N and urea-supplemented diets and a positive N balance when fed on the soyabean-supplemented diet. Dietary urea supplementation did not benefit the horses.

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.

Milk and water intakes of foals sucking grazing mares.

Intakes of milk and milk nutrients were determined for 8 foals at 11-18 days of age and for 10 foals at 30-44 days and 60-74 days of age while sucking grazing mares. Water intakes (sources other than milk) of the foals were determined at 30-44 days and 60-74 days of age. Five of the 10 mares were fed a protein supplement (24% crude protein) in addition to grazing during the stud season. The protein supplement did not influence foal intakes of milk and milk nutrients, milk composition, weight gains of the mares or the growth rate of the foals. Foal milk intakes increased (P less than 0.05) from 16.9 kg/day at 11-18 days to 18.1 kg/day at 60-74 days of age. The water intakes of the foals increased (P less than 0.01) from 3.9 kg/day at 30-44 days to 5.5 kg/day at 60-74 days of age. Total fluid intakes per kg foal liveweight were 246, 202 and 172 g at 11-18, 30-44 and 60-74 days of age, respectively. For each kg of weight gain, foals consumed 12.8, 15.7 and 16.4 kg milk at 11-18, 30-44 and 60-74 days of age. Stage of lactation had a significant effect on the total solids, lactose and protein content of milk. The fat and gross energy content of milk remained constant.

Effect of lupin (Lupinus angustifolius) supplementation on ovarian and pituitary activity in ewes.

In each of three experiments, thirty seasonally anoestrous Border Leicester ewes were fed on a maintenance ration of oaten chaff. Fifteen of them were given a supplement of 500 g lupin grain per head per day. The ewes were treated with 10 mg follicle stimulating hormone (Expt 1), 600 I.U. pregnant mare serum gonadotrophin (Expt 2) and either 150 or 300 micrograms gonadotrophin releasing hormone (Expt 3) to determine whether the ovaries and/or the anterior pituitary were capable of responding to the nutrient status of the animals and influencing ovulation rate. In each experiment, the number and size of corpora lutea and follicles in the lupin-supplemented and -unsupplemented groups were similar. It was concluded that the mechanism by which lupins increase the ovulation rate is probably neural and not a result of direct effect on either the pituitary or the ovaries.

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.

Ovarian activity in fillies treated with anabolic steroids prior to the onset of puberty.

This study investigated the effects of zero, low (0.55), medium (1.1) and high (2.2 mg/kg LW) boldenone undecylenate on the reproductive physiology of sixteen fillies from the ages of 7 to 11 months until 2 years. Ovarian activity measured by cross-sectional area, total number of ovulations, total number of follicles and the number of large follicles (greater than 30 mm in diameter) was significantly reduced by steroid treatments. Of fillies that ovulated during the first breeding season, the age of first ovulation and the second breeding season was significantly delayed in the high dose group. Final clitoral diameters and the increase in clitoral diameter over the period of the study were significantly greater in the treated groups.

Effects of a protein deficient diet and urea supplementation on lactating mares.

Six lactating mares were fed either a low protein diet or the same diet with added urea ad libitum over 71 days. The quantity consumed by the mares, milk intakes of their foals, milk composition, plasma urea nitrogen (PUN), mare liveweight changes and foal growth rates were measured. The mares were unable to consume sufficient quantities of either diet to meet their nitrogen requirements and all lost weight. Adding urea to the diet significantly increased PUN in mares and foals, raised urea concentrations in the milk, decreased the mares' feed intake and significantly increased their weight loss. Low concentrations of protein were measured in milk from mares on both diets. Milk intakes of all the foals were reduced and they showed poor growth rates.

A study of nonsurgical embryo transfer in the mare.

The success rate of nonsurgical embryo recovery was influenced neither by year nor by season within years. The preferred method of nonsurgical embryo transfer was by Cassou pistolette. From a total of 15 attempts to transfer embryos nonsurgically, 9 (60%) were successful. Of the five attempts during February through April 1982, only one was successful in producing a live foal. The degree of synchrony between the ovulations of the donors and recipients in these five attempts ranged from +3 to -3 d. The recipient of the successful transfer ovulated on the same day as the donor. Eight of the ten attempts during September through December 1982 produced live foals. Synchronization of ovulations between the donors and recipients in these transfers ranged from 0 to -2 d. Repeated attempts to recover embryos had no deleterious effects on fertility of the donors.

Synthesis and absorption of cysteine from the hindgut of the horse.

The extent to which cysteine synthesised by microbes within the hindgut of the horse is incorporated into plasma cysteine was estimated by an isotopic technique in two horses fed four different diets. The results showed that between 1 per cent and 6 per cent of the plasma cysteine was of microbial origin. It is argued that the maximum contribution of microbial cysteine, and presumably other amino acids of microbial origin, to the plasma pool is 12 per cent of the net supply. These data support the hypothesis that microbial amino acid synthesis within the hindgut of the horse does not significantly affect its amino acid status.

Morphological characteristics of stallion spermatozoa.

A study of the morphological characteristics of stallion spermatozoa was conducted at the semen laboratory of the Department of Animal Production during four breeding seasons. A total of 590 ejaculates collected from 216 stallions aged from 2 to 26 years and including 13 breeds or colour types was examined. Overall means for the spermatozoal characteristics of these stallions and the classes of head and tail abnormalities are presented and compared with results of other works. Scanning electron micrographs are included to illustrate recognised abnormalities.

Seasonal and circadian changes of testosterone levels in the peripheral blood plasma of stallions and their relation to semen quality.

Three stallions were bled each hour for 25 h at 28-day intervals throughout 1 year. Testosterone levels were pulsatile. Pulse frequency and pulse amplitude were higher in the summer months than at other times (P less than 0.01). The number of testosterone pulses also varied throughout the day, with the greatest frequency occurring in the afternoon (14.00-17.00 h) and at night (22.00-01.00 h). Mean testosterone levels were highest in the summer (P less than 0.01) but showed a secondary, smaller increase in the autumn. Semen characteristics were assessed from measurements of 222 stallions. Semen volume was greatest in the summer (P less than 0.01) but both the concentration (P less than 0.01) and the total number of spermatozoa per ejaculate (P less than 0.05) were highest in the autumn. These results show that the highest testosterone concentrations in peripheral plasma are not necessarily associated temporally with optimum semen quality.

Characteristics and fertility of stallion semen.

A study of stallion fertility was conducted on commercial studs in eastern Australia over 4 breeding seasons (1974/75-1977/78). Data from 47 stallions (66 stallion seasons) aged 2-26 years and representing 7 breeds and 1664 mares were used to relate seminal characteristics to fertility. Percentage pregnancies per service was the most sensitive measure of fertility with stallions on commercial studs. Breed and age of stallion, breeding season and the referral status of the stallion ('suspect' or 'normal') had significant (P less than 0.01) effects on fertility. The semen characteristics that were clearly associated with percentage pregnancies per service were: total volume, gel-free volume, sperm concentration, total numbers of spermatozoa and total number of live spermatozoa. Approximate threshold levels for these characteristics are presented which could provide a diagnostic basis for the classification of stallions as being suitable or doubtful for breeding purposes.

The repeatability of seminal characteristics of stallions.

Fifteen seminal characteristics were measured in ejaculates from 4 laboratory stallions and from 164 commercial stud stallions. Complete field and laboratory data were available from 536 and 531 ejaculates, respectively. These were obtained over 4 breeding seasons (1974/75-1977/78) and 9 breeds were represented. Stallions at commercial studs produced 1-13 ejaculates at intervals of approximately 4 weeks and ranging from 1 h to 1 year apart. Intra-class correlations or 'repeatability' of each seminal characteristic were calculated. Significant between-stallion variation occurred in all characteristics except for pH of gel-free semen. Low repeatabilities (less than 0.3) were estimated for gel-free volume, pH of gel-free semen and the percentage of abnormal mid-pieces. Moderate repeatabilities (0.3 to 0.6) occurred for total volume, gel volume, motility, percentage of dead spermatozoa, sperm concentration, total sperm number and the percentage of sperm head abnormalities, tail abnormalities, loose heads and cytoplasmic droplets. It is suggested that if a seminal examination shows characteristics below or near recommended minimum levels, an additional 1 or 2 samples should be examined before a stallion is classed as likely to have doubtful fertility.

Collection of semen from stallions at stud.

Semen was collected from 222 stallions of 13 breed or colour types in southern Queensland and northern New South Wales. A total of 648 collection attempts were made, using an artificial vagina, during 4 consecutive breeding seasons (1974/5 to 1977/8). Modifications were made to the techniques used by previous workers because collections were made at commercial studs using minimal animal restraint. Of all collection attempts, 621 (96%) were successful, while at least one semen sample was collected from each of 216 stallions (97%). There were no significant relationships between stallion collection failures and breeding season, time of year or age and breed of stallion. Time of year (seasons and months) was the only factor having a significant relationship with collection failures; highest failure rates occurring in autumn and winter. The techniques described are applicable for breeding soundness examinations of untrained stallions and for collection of semen for artificial insemination.

Haemolytic disease in new born calves.

Neonatal haemolytic disease was diagnosed in a Bos indicus cross bred beef herd in south-east Queensland. The evidence suggested that this was due to B. argentina vaccination for babesiosis while non-pregnant or less than one month pregnant. The cows received 5 or 6 vaccinations during their breeding life, which indicated that antibody levels can persist for long periods. The autopsy, histopathological and haematological findings indicated that the mechanism was disseminated intra-vascular coagulation which occurred in calves dying within 24 hours of birth. This resulted in fibrin deposition in pulmonary capillaries and severe pulmonary oedema.