Leptin secretion in horses: effects of dexamethasone, gender, and testosterone

Five experiments were performed to evaluate the effects of dexamethasone (DEX), gender, and testosterone on plasma leptin concentrations in horses. In experiment 1, plasma leptin, insulin, glucose, and IGF-1 concentrations were increased (P < 0.01) in stallions following five daily injections of DEX (125 microg/kg BW). In experiment 2, leptin concentrations increased (P < 0.01) in mares, geldings, and stallions following a single injection of DEX, and the response was greater (P < 0.01) in mares and geldings than in stallions. The gender effect was confounded by differences in body condition scores and diet; however, based on stepwise regression analysis, both BCS and gender were significant sources of variation in the best fit model for pre-DEX leptin concentrations (R(2) = 0.65) and for maximum leptin response to DEX (R(2) = 0.75). In experiment 3, in which mares and stallions were pair-matched based on age and body condition and fed similar diets, mares again had higher (P < 0.01) leptin concentrations than stallions after DEX treatment as used in experiment 2. In experiment 4, there was no difference (P > 0.1) in plasma leptin response in mares following four single-injection doses of DEX from 15.6 to 125 microg/kg BW. In experiment 5, treatment of mares with testosterone propionate every other day for 5 days did not alter (P > 0.1) plasma leptin concentrations or the leptin response to DEX. In conclusion, multiple injections of DEX increase leptin concentrations in stallions, as does a single injection in mares (as low as 15.6 microg/kg BW), geldings and stallions. The greater leptin levels observed in mares and geldings relative to stallions were due partially to their greater body condition and partially to the presence of hyperleptinemic individuals; however, even after accounting for body condition and diet, mares still had greater leptin concentrations than stallions after DEX administration. Elevation of testosterone levels in mares for approximately 10 days did not alter leptin concentrations in mares.     

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.     

Evaluation of the sexual dimorphism in Mangalarga Marchador horses using discriminant analysis

This study used discriminant analysis to evaluate body measures and sexual dimorphism in Mangalarga Marchador horses. Discriminant analysis is a multivariate method that generates functions to classify animals using a prior criterion. In this study a prior criterion is male or female. The study analyzed 25 linear and 11 angular measures obtained from 25 stallions and 56 mares registered with the Mangalarga Marchador Brazilian Breeders Association. Knee girth, cannon girth, chest width, chest girth, back-loin length, hip width, distance from elbow to knee, hind limb hoof length, head width, shoulder length, and body length were found to be linear measures that provided evidence of sexual dimorphism. Angular measures with evidence of sexual dimorphism were shoulder–humerus, metacarpal–phalanx, coxae–femur, and femur–tibial angles. Stallions were larger than mares for almost all linear measures, except for back-loin length, hip width, and distance from elbow to knee. Shoulder–humerus and coxal–femur angles were larger in mares, while metacarpal–phalanx and femur–tibial angles were larger in stallions. Using linear measures, two mares were classified as stallions and three stallions were classified as mares. For angular measures, two other mares were classified as stallions and three other stallions were classified as mares. The discriminant functions can be useful for the selection of horses and to avoid registration of animals not meeting the phenotypic standards of the breed association.     

Epizotiology and phylogeny of equine arteritis virus in hucul horses

The aim of the study was to determine the situation of equine arteritis virus (EAV) infections in hucul horses. A total of 176 horses (154 mares and 22 stallions) from the biggest hucul horse stud in Poland were tested. Antibodies against EAV were detected in 97 (55.1%) horses. The EAV seroprevalence among mares was 53.2% while in stallions - 68.2%. The percentage of positive mares increased with their age, thus amongst the mares of less than 2 years of age the percentage was 32.5%, while in the group of 3-5 years old increased to 59.4% and in the mares in the age of 6-10 years and older than 10 years 89.5% and 95% were seropositive, respectively. Among 11 seropositive stallions five were supposed to be shedders of EAV with their semen. It is likely that those persistently infected stallions were the reservoirs of the virus in the stud. Genetic studies using of ORF5 gene showed high homology between the viruses detected in the semen of those stallions what suggested lateral transmission between the stallions sharing the same stable. Persistent infection in an immature stallion, which has not yet been used for breeding, was established as a result of infection via respiratory route. Phylogenetic analysis confirmed that all hucul viruses shared the same ancestor and as most of EAV strains dominating in Polish horse population belonged to the European origin EAV subgroup (EU-1).     

The variations of white blood cell count in Lipizzan horses

Total and differential leucocyte counts (lymphocytes, neutrophils, monocytes, eosinophils and basophils) were measured in 140 stallions, 101 mares and 25 foals of Lipizzan breed. The values fell in the normal ranges for warm-blooded horses. Differences between mares and stallions were not significant with the exception of foals, having higher white blood cell, neutrophil, lymphocyte, monocyte and basophil values in females than in males. Foals exhibited an age-related increase of total leucocyte count during the first 4 months of life, accompanied by a decrease in neutrophil and increase in lymphocyte and eosinophil counts. In mares and stallions, the total number of leucocytes, lymphocytes, monocytes and basophils significantly decreased but the number of neutrophils and eosinophils remained almost unchanged with age gain.     

Effect of age on immunocytochemical staining characteristics of adenohypophyseal cells in Mongolian pony mares and stallions

OBJECTIVE: To determine the effect of age on immunocytochemical staining characteristics of adenohypophyseal cells of Mongolian pony mares and stallions. ANIMALS: 35 Mongolian ponies. PROCEDURE: Pituitary glands from 1- to 22-year-old horses of both sexes were collected at a commercial slaughterhouse in China and allocated into 7 groups according to age and sex: prepubertal stallions (n = 5; 1 to 2 years old), young stallions (6; 3 to 7 years old), middle-aged stallions (4; 10 to 12 years old), old stallions (5; 15 to 22 years old), young mares (3; 3 to 7 years old), middle-aged mares (5; 10 to 12 years old), and old mares (7; 15 to 22 years old). Pituitary glands were processed for microscopy, and sections were immunocytochemically stained for various pituitary hormones. The percentages and cell areas of 6 types of adenohypophyseal cells were determined by use of morphometry. RESULTS: Age-related alterations in adenohypophyseal cells were detected in horses > 15 years old. Percentage of somatotrophs decreased with age regardless of sex, and percentage of lactotrophs increased with age in stallions. In old mares, percentage of follicle-stimulating hormone cells increased, whereas cell area of follicle-stimulating hormone and luteinizing hormone cells decreased. Differences between sexes for percentage of somatotrophs and lactotrophs, and cell area of follicle-stimulating hormone and luteinizing hormone cells were not evident in old horses. CONCLUSIONS AND CLINICAL RELEVANCE: Differences among percentages and cell areas of pituitary cell types in old horses may be associated with degeneration of the sex organs.     

The influence of mare numbers, ejaculation frequency and month on the fertility of Thoroughbred stallions

Reason for performing the study: Although considerable variation in per cycle pregnancy rates exists between Thoroughbred (TB) stallions, there is little information on factors that may influence this figure. Objective: To assess the influence of month, mare numbers and mating frequency on the fertility of TB stallions standing on studfarms in East Anglia, England. Methods: The daily breeding records of 31 TB stallions mating 3034 mares on 4851 occasions during the 2010 season were surveyed and related to first scan pregnancy rates. The influences of mare book size, month, number of matings per day and mating frequency or abstinence on per mating pregnancy rates were analysed. Results: The overall per mating pregnancy rate for all the stallions was 59.6%, but for individual stallions the figures ranged from 19.0% to 80.1%. The first mating occurred on 9 February and the last on 24 July and the per mating pregnancy rate per month was significantly reduced in June and July. The number of mares mated by individual stallions ranged from 15 to 161, giving a mean overall workload of 160 matings per 100 mares. The per mating pregnancy rate was not related to book size, the number of matings in the season or the mating frequency per day. However, some stallions showed differences in per mating pregnancy rate related to month or the number of ejaculations in the preceding 3 days. Conclusions: The majority of TB stallions are able to maintain good fertility despite large books of mares. However, 5 of the 31 stallions surveyed showed a per mating pregnancy rate of ≤50%. Potential relevance: This survey has identified wide differences between the per mating pregnancy rate in TB stallions. Identification of the factors involved through more comprehensive surveys would provide useful information for mare and stallion owners.     

Effect of gender and exercise on haematological and biochemical parameters in Holsteiner horses

The objective of this study was to assess the influence of exercise of average intensity in the haematological and biochemical values, as well as acidic resistance of erythrocytes in mares and stallions of Holsteiner breed. A total of seventeen horses of Holstein breed (seven mares and 10 stallions aged 6 years) were used in this study. The blood samples were assessed for haematocrit (HCT) value, haemoglobin concentration (HGB), the amount of red blood cells (RBC), white blood cells (WBC), platelets (PLT), leucogram, mean corpuscular haemoglobin concentration (MCHC), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), red cell distribution width (RDW) and platelet distribution width (PDW). Serum concentrations of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) as well as oxidative stress biomarkers were analysed. Stallions showed a significant increase in leucocytes and granulocytes amount, as well as erythrocytes, haemoglobin and haematocrit levels after exercise test. Pre‐exercise level of mean corpuscular haemoglobin concentration was higher in stallions. At the same time, mares showed significant decrease in platelet volume after exercise test. Physical effort in stallions leads to significant increase in aspartate aminotransferase activity. Trained mares and stallions showed a decrease in lipid peroxidation after exercise. Exercise also caused increase in oxidative modified protein of erythrocytes in stallions indicating by exercise‐induced oxidative stress. The resistance of erythrocytes in 0.1 m HCl was similar between females and males. No statistically significant differences in the percentage of haemolysed erythrocytes before and after exercise were observed.     

Occurrence of equine coital exanthema in pastured draft horses and isolation of equine herpesvirus 3 from progenital lesions

During the period from 2001 to the following year, progenital diseases had been epidemic among the draft stallions and mares pastured together in Iwate Prefecture, the northeastern district of Japan. A stallion and 8 of 31 mares were affected in 2001, and 1 of 2 stallions and 10 of 36 mares in 2002. The clinical symptoms consisted of the formation of papules, pustules, ulcers and scabs on the progenital skin and mucosa in stallions and mares. In 2002, Equine herpesvirus 3 (EHV3) was isolated from 2 mares and the glycoprotein G gene of the virus detected from a stallion and 4 mares by polymerase chain reaction. Serum neutralizing tests showed that 12 of 38 horses, 10 clinically and 2 subclinically affected, changed to be positive for the EHV3 antibody. The results suggest that the horses were affected with equine coital exanthema (ECE) through coitus. Five mares with the antibody at the pre-pastured period may have been the possible origins of EHV3 infection in 2002, although the exact origin in 2001 remains unknown. The artificial insemination was performed for the prevention of ECE spreading through coitus on the pasture in 2003. There was no epidemic of the disease in 31 mares, although 3 mares with the antibody at the pre-pastured period showed the significant increase in the titers during the pastured period.     

Capsule types of Klebsiella pneumoniae isolated from the genital tract of mares with metritis, extra-genital sites of healthy mares and the genital tract of stallions

A survey of K. pneumoniae was performed on cervical swabs, feces and nasal swabs of mares and on samples from the genital tract of stallions from 1980 to 1986 in south-western Hokkaido, Japan. K1 was the predominant type (79 of 88, 89.8%) in the metritis cases due to K. pneumoniae in mares of racing breeds. The same type was isolated from semen and swabs of the fossa glandis of 6 of 20 (30.0%) of the stallions of racing breeds. Heavily encapsulated and less heavily encapsulated K1 strains were isolated from the stallions. Mares bred to stallions carrying heavily encapsulated strains developed metritis, while those bred to stallions carrying less heavily encapsulated strains did not. K39 was isolated from cervical swabs solely from metritis-infected mares of draft breeds and not from any mares of the racing breeds examined. Untypable strains were isolated from cervical swabs in 7 of 88 (8.0%) metritis cases of mares of racing breeds and from semen in 7 of 19 (36.8%) stallions of racing breeds and they were predominant in feces (19 of 21, 90.5%) and nasal swabs (3 of 4, 75.0%) of healthy mares of racing breeds.     

Genetic and Environmental Factors Influencing Gestation Length in Lusitano Horses

Breeding and parturition records collected over a period of 35 years in the Alter Real stud of Lusitano horses were used to calculate gestation length (GL). The 1027 gestations by 209 mares mated to 60 stallions had a mean GL of 338.1 ± 9.26 days. The mixed model analysis of variance indicated that the sex of the foal and inbreeding of the dam and foal had no significant effect on GL (P > .05). On the other hand, GL increased linearly with mare age, with an estimated regression coefficient of 0.155 ± 0.069 days/year (P < .05). Year and conception month affected GL (P < .05), with longer gestations observed when breeding occurred between January and April, followed by a decline of about 5 and 10 days in GL of mares bred in May and in June–July, respectively. Important differences were observed between stallions (P < .05), with most stallions resulting in a distribution of GL in an interval of ±5 days. The inclusion of the mare as a random effect in the mixed model resulted in an estimated repeatability of GL of 0.427, indicating that mares tend to be regular in having long or short gestations across their lifetime. Variance components estimated in an Animal Model resulted in heritability estimates of 0.39 for maternal genetic effects and 0.19 for direct genetic effects, with no association between the two components. Overall, the mare seems to have the major genetic influence on GL in Lusitano horses, but environmental factors, such as month of conception and also the age of the mare, have a noticeable effect.     

Pregnancies attained after collection and transfer of oocytes from ovaries of five euthanatized mares

After euthanasia, ovaries were removed from 5 horses and shipped to a laboratory where 46 oocytes were collected. The oocytes were cultured for 24 to 30 hours, and 36 oocytes were transferred to 10 recipient mares via flank laparotomies. Recipient mares were inseminated with semen from various stallions. Sixteen days after transfer, 4 of the recipients were pregnant with at least 1 embryonic vesicle. Embryonic death occurred in 3 recipients, whereas a healthy live foal was born from 1 recipient. Ovaries from valuable mares can be a source of viable oocytes after death of the mare. For shipping to a laboratory, fluctuations in temperature should be minimized and the ovaries should not be chilled.     

Serological surveillance of equine viral arteritis in the United Kingdom since the outbreak in 1993

Serological analysis of blood samples submitted to the Animal Health Trust showed that during 1995, 185 of 9203 unvaccinated horses (2.0 per cent) tested positive for antibodies to equine arteritis virus (EAV), and that during 1996, 46 of 8851 unvaccinated horses (0.52 per cent) tested positive. During both years thoroughbreds were the predominant breed tested and only a small proportion of these (<0.3 per cent), consisting predominantly of imported mares, were seropositive. In contrast, among standardbred horses, from which samples were actively solicited in 1995, 84 of 454 (18.5 per cent) were seropositive. Among standardbreds there was a difference in prevalence between types of horses, with 3.7 per cent of racing horses, 25 per cent of non-racing horses and 41 per cent of stallions testing seropositive. Investigations of seropositive stallions identified during 1994 and 1995 demonstrated that clinically inapparent equine viral arteritis (EVA) had occurred previously in the UK. Of 50 seropositive stallions, nearly half were standardbreds and nearly all had been imported from either North America or the European Union. Whether 34 seropositive stallions were shedding virus in their semen was established either by test mating, by the serology of the covered mares, or by investigation by MAFF following the introduction of the Equine Viral Arteritis Order 1995. Nine of the stallions (26.5 per cent) were identified as presumptive shedders of EAV in semen and among specific breeds, viral shedding was identified in six of 15 (40 per cent) standardbreds and three of nine (33 per cent) warmbloods. In contrast with the outbreak of EVA in the UK in 1993, no signs of disease typical of EAV infection were reported during these investigations, even in mares test mated to stallions shedding the virus.     

Concentrations of prolactin, luteinizing hormone and follicle stimulating hormone in pituitary and serum of horses: effect of sex, season and reproductive state

Pituitary and serum from 86 male or female horses of various reproductive states were collected in the normal breeding season (summer) and in the nonbreeding season (winter) at a commercial slaughterhouse. Concentrations of prolactin (PRL), luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured by radioimmunoassay. Concentrations of pregnant mare serum gonadotropin and reproductive steroids in serum and gross appearance of the reproductive tract and gonads were used to catagorize reproductive state. Concentrations of PRL were higher (P less than .01) in summer than in winter in pituitary and serum of mares, stallions and geldings. In summer, mares had higher (P less than .01) concentrations of PRL in serum than stallions. In mares, concentrations of LH in pituitary were higher (P less than .05) in summer than in winter. Concentrations of LH in serum were higher (P less than .01) in summer than in winter in mares and geldings, higher (P less than .01) in mares than in stallions in summer, higher (P less than .01) in geldings than in stallions in summer and higher (P less than .01) in mares with low serum progesterone (P) concentrations than in mares with high P concentrations in summer. Concentrations of FSH in pituitary and serum did not differ between summer and winter for any type of horse.(ABSTRACT TRUNCATED AT 250 WORDS)     

Superovulation in cycling mares using equine follicle stimulating hormone (eFSH)

Superovulation would potentially increase the efficiency and decrease the cost of embryo transfer by increasing embryo collection rates. Other potential clinical applications include improving pregnancy rates from frozen semen, treatment of subfertility in stallions and mares, and induction of ovulation in transitional mares. The objective of this study was to evaluate the efficacy of purified equine follicle stimulating hormone (eFSH; Bioniche Animal Health USA, Inc., Athens, GA) in inducing superovulation in cycling mares. In the first experiment, 49 normal, cycling mares were used in a study at Colorado State University. Mares were assigned to 1 of 3 groups: group 1, controls (n = 29) and groups 2 and 3, eFSH-treated (n = 10/group). Treated mares were administered 25 mg of eFSH twice daily beginning 5 or 6 days after ovulation (group 2). Mares received 250 (of cloprostenol on the second day of eFSH treatment. Administration of eFSH continued until the majority of follicles reached a diameter of 35 mm, at which time a deslorelin implant was administered. Group 3 mares (n = 10) received 12 mg of eFSH twice daily starting on day 5 or 6. The treatment regimen was identical to that of group 2. Mares in all 3 groups were bred with semen from 1 of 4 stallions. Pregnancy status was determined at 14 to 16 days after ovulation.In experiment 2, 16 light-horse mares were used during the physiologic breeding season in Brazil. On the first cycle, mares served as controls, and on the second cycle, mares were administered 12 mg of eFSH twice daily until a majority of follicles were 35 mm in diameter, at which time human chorionic gonadotropin (hCG) was administered. Mares were inseminated on both cycles, and embryo collection attempts were performed 7 or 8 days after ovulation.Mares treated with 25 mg of eFSH developed a greater number of follicles (35 mm) and ovulated a greater number of follicles than control mares. However, the number of pregnancies obtained per mare was not different between control mares and those receiving 25 mg of eFSH twice daily. Mares treated with 12 mg of eFSH and administered either hCG or deslorelin also developed more follicles than untreated controls. Mares receiving eFSH followed by hCG ovulated a greater number of follicles than control mares, whereas the number of ovulations from mares receiving eFSH followed by deslorelin was similar to that of control mares. Pregnancy rate for mares induced to ovulate with hCG was higher than that of control mares, whereas the pregnancy rate for eFSH-treated mares induced to ovulate with deslorelin did not differ from that of the controls. Overall, 80% of mares administered eFSH had multiple ovulations compared with 10.3% of the control mares.In experiment 2, the number of large follicles was greater in the eFSH-treated cycle than the previous untreated cycle. In addition, the number of ovulations during the cycle in which mares were treated with eFSH was greater (3.6) than for the control cycle (1.0). The average number of embryos recovered per mare for the eFSH cycle (1.9 ± 0.3) was greater than the embryo recovery rate for the control cycle (0.5 ± 0.3).In summary, the highest ovulation and the highest pregnancy and embryo recovery rates were obtained after administration of 12 mg of eFSH twice daily followed by 2500 IU of hCG. Superovulation with eFSH increased pregnancy rate and embryo recovery rate and, thus, the efficiency of the embryo transfer program.

Introduction

Induction of multiple ovulations or superovulation has been an elusive goal in the mare. Superovulation would potentially increase the efficiency and decrease the cost of embryo transfer by increasing embryo collection rates.[1 and 2] Superovulation also has been suggested as a critical requirement for other types of assisted reproductive technology in the horse, including oocyte transfer and gamete intrafallopian transfer. [2 and 3] Unfortunately, techniques used successfully to superovulate ruminants, such as administration of porcine follicle stimulating hormone and equine chorionic gonadotropin have little effect in the mare. [4 and 5]The most consistent therapy used to induce multiple ovulations in mares has been administration of purified equine pituitary gonadotropins. Equine pituitary extract (EPE) is a purified gonadotropin preparation containing approximately 6% to 10% LH and 2% to 4% FSH.[6] EPE has been used for many years to induce multiple ovulations in mares [7, 8 and 9] and increase the embryo recovery rate from embryo transfer donor mares. [10] Recently, a highly purified equine FSH product has become available commercially.The objectives of this study were to evaluate the efficacy of purified eFSH in inducing superovulation in cycling mares and to determine the relationship between ovulation rate and pregnancy rate or embryo collection rate in superovulated mares.

Materials and methods

Experiment 1

Forty-nine normally cycling mares, ranging in age from 3 to 12 years, were used in a study at Colorado State University. Group 1 (control) mares (n = 29) were examined daily when in estrus by transrectal ultrasonography. Mares were administered an implant containing 2.1 mg deslorelin (Ovuplant, Ft. Dodge Animal Health, Ft. Dodge, IA) subcutaneously in the vulva when a follicle 35 mm in diameter was detected. Mares were bred with frozen semen (800 million spermatozoa; minimum of 30% progressive motility) from 1 of 4 stallions 33 and 48 hours after deslorelin administration. The deslorelin implants were removed after detection of ovulation.[11] Pregnancy status was determined at 14 and 16 days after ovulation.Group 2 mares (n = 10) were administered 25 mg of eFSH (Bioniche Animal Health USA, Inc., Athens, GA) intramuscularly twice daily beginning 5 or 6 days after ovulation was detected. Mares received 250 g cloprostenol (Estrumate, Schering-Plough Animal Health, Omaha, NE) intramuscularly on the second day of eFSH treatment. Administration of eFSH continued until a majority of follicles reached a diameter of 35 mm, at which time a deslorelin implant was administered. Mares were subsequently bred with the same frozen semen used for control mares, and pregnancy examinations were performed as described above.Group 3 mares (n = 10) received 12 mg of eFSH twice daily starting 5 or 6 days after ovulation and were administered 250 μg cloprostenol on the second day of treatment. Mares were randomly selected to receive either a deslorelin implant (n = 5) or 2500 IU of human chorionic gonadotropin (hCG) intravenously (n = 5) to induce ovulation when a majority of follicles reached a diameter of 35 mm. Mares were bred with frozen semen and examined for pregnancy as described above.

Experiment 2

Sixteen cycling light-horse mares were used during the physiologic breeding season in Brazil. Reproductive activity was monitored by transrectal palpation and ultrasonography every 3 days during diestrus and daily during estrus. On the first cycle, mares were administered 2500 IU hCG intravenously once a follicle 35 mm was detected. Mares were subsequently inseminated with pooled fresh semen from 2 stallions (1 billion motile sperm) daily until ovulation was detected. An embryo collection procedure was performed 7 days after ovulation. Mares were subsequently administered cloprostenol, and eFSH treatment was initiated. Mares received 12 mg eFSH twice daily until a majority of follicles were 35 mm in diameter, at which time hCG was administered. Mares were inseminated and embryo collection attempts were performed as described previously.

Statistical analysis

In experiment 1, 1-way analysis of variance with F protected LSD was used to analyze quantitative data. Pregnancies per ovulation were analyzed by x² analysis. In experiment 2, number of large follicles, ovulation rate, and embryo recovery rate were compared by Student,'s t-test. Data are presented as the mean S.E.M. Differences were considered to be statistically significant at p < .05, unless otherwise indicated.

Results

In experiment 1, mares treated with 25 mg eFSH twice daily developed a greater number of follicles 35 mm in diameter (p = .001) and ovulated a greater number of follicles (p = .003) than control mares (Table 1). However, the number of pregnancies obtained per mare was not significantly different between the control group and the group receiving 25 mg eFSH (p = .9518). Mares treated with 12 mg eFSH and administered either hCG or deslorelin to induce ovulation also developed more follicles 35 mm (p = .0016 and .0003, respectively) than untreated controls. Mares receiving eFSH followed by hCG ovulated a greater number of follicles (p = .003) than control mares, whereas the number of ovulations for mares receiving eFSH followed by deslorelin was similar to that of control mares (p = .3463). Pregnancy rate for mares induced to ovulate with hCG was higher (p = .0119) than that of control mares, whereas the pregnancy rate for eFSH-treated mares induced to ovulate with deslorelin did not differ from that of controls (p = .692). Pregnancy rate per ovulation was not significantly different between control mares (54.5%) and mares treated with eFSH followed by hCG (52.9%). The lowest pregnancy rate per ovulation was for mares stimulated with 25 mg eFSH and induced to ovulate with deslorelin. The mean number of days mares were treated with 25 mg or 12 mg of eFSH was 7.8 ± 0.4 and 7.5 ± 0.5 days, respectively. Overall, 80.0% of mares administered eFSH had multiple ovulations compared with 10.3% of control mares.     

Genetic variability in Hanoverian warmblood horses using pedigree analysis

A data set constituting a total of 310,109 Hanoverian warmblood horses was analyzed to ascertain the genetic variability, coefficients of inbreeding, and gene contributions of foreign populations. The reference population contained all Hanoverian horses born from 1980 to 2000. In addition, Hanoverian stallions born from 1980 to 1995 and Hanoverian breeding mares from the birth years 1980 to 1995 with registered foals were analyzed for the same genetic parameters. The average complete generation equivalent was approximately 8.43 for the reference population. The mean coefficient of inbreeding was 1.33, 1.19, and 1.29% for the reference population, stallions, and breeding mares, respectively. The effective number of founders was largest in stallions (364.3) and smallest in the reference population (244.9). The ratio between the effective number of founders and the effective number of ancestors was 3.15 for the reference population, 3.25 for the stallions, and 3.06 for the breeding mares. The effective population size in the Hanoverian warmblood reference population was 372.34. English Thoroughbreds contributed nearly 35% of the genes to the Hanoverian reference population and even slightly greater contributions (39%) to the stallions. Trakehner and Arab horses contributed approximately 8 and 2.7%, respectively, to the Hanoverian gene pool. The most important male ancestors were Aldermann I from the A/E line, Fling from the F/W line, and Absatz from the Trakehner line, whereas the breeding mare Costane had the greatest contribution to the reference population, stallions, and breeding mares. From 1996 onward, the stallions Weltmeyer and Donnerhall had the largest genetic impact on the Hanoverian horse population.     

CASE STUDY: Use of Modified Phosphate-Buffered Saline as a Component of Semen Extenders Allows the Use of Transported Semen from Two Stallions

Conception rates for mares bred with transported-cooled and fresh stallion semen were collected over a 4-yr period (1998–2002) for two stallions. Both stallions stood at a commercial breeding farm. Semen from both stallions was used immediately after collection on the farm and after 24 to 48 h of cold storage when transported to locations in the U.S. and Canada. Semen for insemination of mares located on the farm was extended with a commercially available skim milk glucose extender (SKMG). Spermatozoal motility following cold storage for spermatozoa diluted in SKMG extender was unacceptable. Thus, semen from both stallions was centrifuged, and spermatozoa were resuspended in SKMG supplemented with modified PBS. In a previous study, the percentage of motile spermatozoa increased following centrifugation and reconstitution of the sperm pellet in SKMG-PBS as compared with semen dilution in SKMG (Stallion A: 15% vs 47%; Stallion B: 18% vs 43%). In the current study, 22 of 25 (88%) and 3 of 4 (75%) mares conceived with transported-cooled semen from Stallions A and B, respectively. Conception rates for mares inseminated with transported semen did not differ (P>0.05) from those inseminated on the farm with fresh semen. These data illustrate that stallion owners can modify standard cooled semen processing procedures and semen extender composition to improve post-storage spermatozoa motility and to obtain acceptable fertility.     

Reproductive Performance of Thoroughbred Mares in Sweden

During 1997–2001, a total of 430 Thoroughbred mares were mated by one of the two Thoroughbred stallions on a well‐managed stud farm in the central part of Sweden. On arrival, a thorough gynaecological examination of all mares was performed. An early pregnancy examination was performed on days 14 and 15 after ovulation. The overall conception rates for the two stallions were 92.2 and 88.8%, and the corresponding live foal rates were 82.6 and 75.2%, respectively. The mean number of served oestrous cycles was 1.60 per mare per conception and 1.86 per live foal. The first and second cycle conception rates are reported for different age groups, mare categories and month of mating. The age of the mares had a significant influence on the live foal rate, being lower for mares >13 years of age. Resorption and abortion occurred in higher percentages among mares >8 years of age. The highest embryonic death occurred among mares with a history of being barren, or with having resorbed or aborted foetuses during the previous season. The month of mating had no significant influence on the first cycle conception rate or the second cycle conception rate. The frequency of twinning was 10.5% and the results of manual crushing of one of the conceptuses was successful in 88.9% of all cases, according to the ultrasound scanning 2 days later.     

Demographic analysis of the Canadian Standardbred broodmare herd

This study derives demographic parameters for the Canadian Standardbred broodmare herd by analysis of industry mating reports for the years 1986–1989 inclusive. The reference population for the study was all mares bred to Canadian-registered stallions. Number of matings evaluated ranged from 9169 to 9837 in each year. In establishing the size of the national herd, data from registration files for 1986 also were used to estimate the number of Canadian mares bred to US stallions. The national broodmare herd was estimated to consist of 12 237 mares, of which 7.7% annually were bred to US-registered stallions. An estimated 16.7% of the national herd were inactive (not presented for breeding) in any year. Probability of temporary absence from the breeding herd was influenced by age, decreasing from 2 to 6 years, then increasing thereafter. Age-specific culling rates revealed that mares less than 6 years of age were subject to more intense selection pressure than was evident for mares more than 10 years of age. Inconsistent presentation for breeding appeared to be a feature of the management of young broodmares. Overall herd culling rate was estimated to be 10.5%, equivalent to an average breeding life of 9.5 years. It was determined that the average broodmare breeds for 7.9, or 83%, of available years. The results clearly indicate that the reproductive advantage enjoyed by mares 6 years of age results, in part, from more consistent presentation for breeding.     

Reproductive efficiency of Flatrace and National Hunt Thoroughbred mares and stallions in England

REASONS FOR PERFORMING STUDY: Previous surveys of reproductive efficiency in British Thoroughbreds included only mares and stallions standing on studfarms in and around Newmarket. The present study was widened to compare Flatrace (FR) (Group A) and National Hunt (NH) (Group B) mares and stallions on studfarms throughout England. OBJECTIVES: To assess the influences of mare type, status and age, and veterinary manipulations on reproductive efficiency parameters. To compare the inherent fertility of stallions, based on singleton and twin pregnancy rates and pregnancy loss rates, in Groups A and B Thoroughbred breeding stock. METHODS: Managers of 24 FR and 9 NH public studfarms were asked to complete a questionnaire for each mated oestrous cycle shown by 2321 Group A and 1052 Group B mares throughout the 2002 mating season. Parameters such as per cycle singleton and twin pregnancy rates, and pregnancy loss rates were noted, and the success of hormone treatments to induce oestrus and ovulation assessed. The number of matings per oestrus and per pregnancy were recorded, together with the incidence and effectiveness of uterine and other veterinary treatments. The inherent fertility of 84 Group A and 43 Group B stallions in the study, as measured by the singleton and twin early pregnancy rates and the pregnancy loss rates recorded in the mares they mated, was also estimated. RESULTS: Per cycle early pregnancy (Days 13-16) was 63.2% for Group A and 65.3% for Group B mares; and 10.3% and 13.1%, respectively, of those pregnancies were twins or triplets. Early, middle and late pregnancy loss rates were 7.2% vs. 8.0% (Days 15-42), 3.6% vs. 6.1% (Days 42-1st October) and 2.7% vs. 2.1% (October-foaling), respectively. Matings per oestrus and per early pregnancy were significantly higher in Group B vs. Group A mares. For stallions that mated > or = 30 mares, overall early pregnancy rates per cycle in mares mated ranged from 30-89% across the 2 groups. CONCLUSIONS: No major differences in reproductive efficiency were identified between FR and NH mares and stallions. Increasing mare age was the single biggest limiting factor to an otherwise high rate of fertility in well-managed English Thoroughbreds. POTENTIAL RELEVANCE: This study identified factors that influence reproductive efficiency in the Thoroughbred.