Effects of inbreeding and other systematic effects on fertility of Black Forest Draught horses in Germany

Background

The Black Forest Draught horse (BFDH) is an endangered German coldblood breed with its origin in the area of the Black Forest in South Germany. In this retrospective study, the influence of the inbreeding coefficient on foaling rates was investigated using records from ten breeding seasons. Due to the small population size of BFDH, the level of inbreeding is increasing and may have an effect on foaling rates.The data of the present study included all coverings reported for 1024 BFDH mares in the years 2001–2009. These mares were covered by 32 BFDH stallions from the State Stud Marbach. Data from 4534 estrus cycles was used to calculate per cycle foaling rate (CFR). Pedigree data contained all studbook data up to the foundation of the breed as early as 1836. The level of inbreeding of the mare, stallion and expected foal along with other systematic effects on CFR were analysed using a generalized linear mixed model approach. Stallion was employed as a random effect. Systematic fixed effects were month of mating, mating type, age of the mare and stallion, reproductive status of the mare and stallion line of the mare. Inbreeding coefficients of the stallion, mare and expected foal were modelled as linear covariates.

Results

The average CFR was 40.9%. The mean inbreeding coefficients of the mares, stallions and expected foals were 7.46, 7.70 and 9.66%. Mating type, age of the mare, reproductive status of the mare and stallion line of the mare had a significant effect.

Conclusions

The results showed that the mating type, stallion line of the mare, sire, age and reproductive status of the mare exerted the largest influences on CFR in BFDH. Inbreeding coefficients of the stallion, mare and expected foal were not significantly related with CFR.

     

Management and fertility of mares bred with frozen semen.

Semen quality, mare status and mare management during estrus will have the greatest impact on pregnancy rates when breeding mares with frozen semen. If semen quality is not optimal, mare selection and reproductive management are crucial in determining the outcome. In addition to mare selection, client communication is a key factor in a frozen semen program. Old maiden mares and problem mares should be monitored for normal cyclicity and all, except young maidens, should have at least a uterine culture and cytology performed. Mares with positive bacterial cultures and cytologies should be treated at least three consecutive days when in estrus with the proper antibiotic. With frozen semen, timing the ovulation is highly desirable in order to reduce the interval between breeding and ovulation. The use of ovulation inducing agents such as human chorionic gonadotropin (hCG) or the GnRH analogue, deslorelin, are critical components to accurately time the insemination with frozen semen. Most hCG treated mares ovulate 48h post-treatment (12-72h) while most deslorelin (Ovuplant) treated mares ovulate 36-42h post-treatment. However, mares bred more than once during the breeding cycle appear to have a slight but consistent increase in pregnancy rate compared to mares bred only once pre- or post-ovulation. In addition, the "capacitation-like" changes inflicted on the sperm during the process of freezing and thawing appear to be responsible for the shorter longevity of cryopreserved sperm. Therefore, breeding closer to ovulation should increase the fertility for most stallions with frozen semen. Recent evidence would suggest that breeding close to the uterotubal junction increases the sperm numbers in the oviduct increasing the chances of pregnancy. Post-breeding examinations aid in determining ovulation and uterine fluid accumulations so that post-breeding therapies can be instituted if needed. Average pregnancy rates per cycle of mares bred with frozen semen are between 30 and 40% with a wide range between sires. Stallion and mare status are major factors in determining the success of frozen semen inseminations. Pregnancy rates are lower for barren and old maiden mares as well as those mares treated for uterine infections during the same cycle of the insemination. To maximize fertility with frozen semen, a careful selection of the stallions and mares, with proper client communication is critical. Dedication and commitment of mare owner and inseminator will have the most significant impact on the pregnancy rates.     

Capacitation, acrosome function and chromatin structure in stallion sperm

In general, fertility in breeding stallions is lower and more variable than in the other farm animal species, primarily because selection is based on pedigree, looks and/or athletic performance, with little consideration of fertility or fertility potential. Moreover, because the average stallion breeds only a limited number of mares per year and in-field fertility is influenced significantly by non-stallion factors such as management and mare fertility, meaningful fertility data are hard to come-by. Unfortunately, generating usable figures would involve impractically high costs, time and numbers of mares. Instead, a breeding soundness examination (BSE), based on assessments of sperm number, motility and morphological normality and of mating ability, is often carried out with the ostensible aim of identifying animals with the "potential for good fertility". In fact, the BSE generally succeeds only in ruling out those stallions with a very clear reason for sub-fertility, and still fails to identify some seriously sub-fertile animals. Thus, the routine BSE has very limited use as a predictor of subsequent fertility. This paper reviews assays developed for identifying capacitated, acrosome-reacted and DNA-damaged sperm, and assesses their utility for improving our ability to predict a stallion's fertility prior to the onset of his breeding career.     

Pregnancy rates and sexual behavior under pasture breeding conditions in mares

Pony mares (n=480) and 16 stallions were assigned to four herds of 60 mares and one stallion (large herds) and to 12 herds of 20 mares and one stallion (small herds). The stallions remained with the herds continuously for all of the large herds and seven of the small herds. In the five remaining small herds the stallion was put into a herd for three hours every two days for 12 observation periods. Pregnancy rates and day of ovulation were estimated by size of embryonal enlargements. Mean pregnancy rates of 51% and 54% were obtained in the small herds and 42% in the large herds during a 48-day period (equivalent to two estrous cycles). Pregnancy rates for herds with the stallion present continuously were higher (P<0.01) for the small herds than for the large herds for days 1-24 (42% versus 19%). There was no effect of herd size on number of mares becoming pregnant per herd on days 1-24, but more mares (P<0.01) became pregnant during days 25-48 in the large herds (13.2 mares per herd versus 1.8). In the herds in which the stallion was present intermittently, the number of times that the stallion rebred the same mare when more than one mare was in estrus was greater (P<0.01) than what would be expected to occur by chance (observed, 21%; expected, 11%). Repeated breeding of the same mare seemed related to the availability or activity of the mare, since such mares more frequently followed and positioned themselves in the vicinity of the stallion. Most of the interferences by a mare which involved keeping the stallion and another mare apart were directed at the mare, whereas most of the interferences during mounting were directed at the stallion (P<0.01). Mares were more likely (P<0.01) to interfere when in estrus than when in nonestrus. When interfering mares were in nonestrus, their hostility was usually directed at the stallion (92%), whereas when in estrus their interference was more frequently directed at a mare (73%, P<0.01).     

The equine frozen semen industry.

Recent acceptance of frozen semen as a method to produce registered foals by two of the worlds largest breed associations, the American Quarter Horse and American Paint Horse, has stimulated new interest in frozen semen technology. This review will: (a) attempt to identify the major impediments to the development of the frozen semen industry, (b) suggest alternative methods for marketing and application of frozen semen, and (c) present the results of a recent study in our laboratory. The objective of which was to compare pregnancy rates of insemination with cooled and frozen semen. Major impediments to the development of the frozen semen industry include 1. Lower fertility with frozen semen as compared to cooled semen for many stallions. 2. Increased costs associated with management of mares for AI with frozen semen using current insemination protocols. 3. Unfavorable marketing practices for frozen semen. Reports of fertility with cooled transported semen in commercial breeding programs indicate seasonal pregnancy rates ranging from 60 to 90%. We compiled data from three commercial transported cooled semen programs in which semen from 16 stallions was used for insemination of 850 mares throughout North America by local veterinarians. During the 1999 and 2000 breeding seasons, first cycle and seasonal pregnancy rates of 59.4 and 74.7% were obtained. During that same period, first cycle and seasonal pregnancy rates of 51.3 and 75.6% were obtained following insemination of 876 mares with frozen semen from 106 different stallions processed by our laboratory and distributed through our commercial distribution program. First cycle and seasonal pregnancy rates were higher for mares bred outside of North America than for mares bred within North America (53.5 and 81.9 versus 49.4 and 65.6%, respectively). Seasonal pregnancy rates were higher presumably because of the better mare management employed for mares bred with exported semen and the fact that some of the domestic mares were switched to cooled semen insemination after a failed first cycle attempt with frozen semen. These data support the position that comparable seasonal pregnancy rates may be obtained using frozen and liquid cooled semen in a commercial setting.     

Tests for cooperative behaviour between stallions

Breeding groups with multiple stallions occur sympatrically with single-stallion breeding groups in feral horse, Equus caballus, populations. Mutualism and reciprocal altruism between stallions have been proposed to explain the origin and functioning of multistallion bands. However, empirical support for these hypotheses is contradictory and incomplete. Furthermore, there are no explicit tests of the predictions that each hypothesis makes about stallion behaviour and social structure. We compared nine multistallion and 18 single-stallion bands in the Kaimanawa Ranges, New Zealand. Compared with agonistic behaviours, affiliative behaviours were relatively unimportant in the relationships between stallions within bands. The number of stallions in the band did not have a positive influence on mare group size, stability, home range quality or reproductive success in bands. Furthermore, there was a positive relationship between aggression ('intolerance') by the dominant towards subordinate stallions and the subordinates' effort in mare group defence ('helping') but a negative relationship between helping effort by subordinates and their proximity to, and mating with, the bands' mares. Therefore, the predictions of the mutualism and reciprocal altruism hypotheses were not supported. Indeed, for some of the predictions we found the opposite outcomes to be true. Multistallion bands had significantly poorer reproductive success, and dominant stallions were less tolerant of subordinates that helped most and reduced their access to mares. Nevertheless, in all other respects Kaimanawa stallions in multistallion bands behaved like those described elsewhere. Thus, we reject cooperative hypotheses for multimale breeding groups in horses and discuss the mate parasitism and consort hypotheses as better alternatives. Copyright 2000 The Association for the Study of Animal Behaviour.     

Equine frozen semen: freezability and fertility field results

The freezability of stallion semen defined as the number of selected ejaculates/total number of ejaculates frozen from 161 different stallions was analyzed. Of the stallions, 19, 30, 27 and 24% had a freezability of 0%, 0 to 33%, 33 to 66%, over 66%, respectively In 85 different stallions, the correlation of freezability between first and second year was 0.60 (P < 0.001). The relationship between fertility with fresh and frozen semen and freezability was analyzed in 40 stallions whose freezability and fertility information was recorded during 5 years. There was a strong relationship between fertility of fresh semen and semen freezability (P < 0.001). However, the relationship between fertility of frozen semen and freezability was not as marked (P < 0.05). Analysis of the field fertility per cycle results when mares were bred with 300 or 150 x 10(6) total spermatozoa at different frequencies until ovulation indicated that mares that were inseminated 2 times or more per estrus show an improved fertility in comparison with mares inseminated only once (34%, n = 1576 vs 26%, n = 626; P < 0.001). Foaling rate when mares were inseminated with frozen semen (1858 mares during 8 breeding seasons) was mainly influenced by mare age (< 16 years: 54% vs >/= 16 years 42% p < 0.001). Date of first insemination (before May 15: 58% vs after May 15: 37%) also had a significant effect on foaling rate (P < 0.001).     

Age-specific fertility rates of the Jeju pony (<Emphasis Type="Italic"> Equus caballus</Emphasis>)

Age-specific fertility is an essential parameter of life history. Here we report age-specific fertility rates, measured as the number of foals per mare per year, for Jeju ponies aged 2–28 years. The total sample consisted of 545 foals produced by 178 mares from 1988 to 2002. The mean fertility rate across all ages of the mares was 0.65 foals/mare per year (±0.24 SD). The fertility rates were above average for the 7- to 8- and 19- to 20-year-old mares, whereas they were relatively low for mares under 4 years old. The fertility rates tended to increase with the age of the stallion, but the relationship was not significant (n =15, P =0.09). The incidence of inter-birth interval was not associated with the age that the mare first reproduced (n =64 mares, P =0.99). However, mares that reproduced later in life tended to have a reduced fertility rate due to an increase in the duration of inter-birth intervals relative to mares that reproduced earlier (n =4 years of first reproduction, P =0.068). The fertility rates of Jeju ponies were lower than for other horses, perhaps because only one stallion was introduced to a relatively large herd every year. We suggest that the introduction of more stallions to the herd each year would increase fertility rates.     

Genetic study of gestation length in Andalusian and Arabian mares

The length of gestation in Andalusian, or Spanish Purebred (SPB) and Arabian (AB) mares reared in Spain was analysed, based on 766 spontaneous full-term deliveries appertaining to 141 mares of SPB breed and 72 mares of AB breed in 31 breeding seasons. The data were obtained from the Yeguada Militar de Jerez de la Frontera stud farm in Cádiz, Spain. The mean length of gestation was of 336.8+/-0.48 days in the SPB mares and 340.3+/-0.63 days in AB mares. To assess the accurate prediction of time of birth the potential effect of a number of factors was investigated. The influences of the breed, mare, month and year of mating, age of the mother, number of births and sex of the foal were statistically significant. The factor have the greatest influence over the gestation length was the mare itself, with a correlation among consecutive births of around 0.4. The effect of inbreeding, both of the mare and foal, was negligible. Gestation length shortened as the breeding season progressed: in both breeds, a delay of 1 month in mating corresponded to a decrease of 3 days in the gestation length. According to our results, gestation length decrease as the mare gets older, with the shortest gestation periods when the mare is 10-12 years old, and from this point on, it slowly increases. The gestation period shortens as the 4th or 5th birth approaches, and then gets progressively longer. The range of variation in gestation length due to the number of births to the mare is of 2.9 days for the AB mares, and 2.2 days for SPB mares. The heritability for the gestation length for AB and the SPB breeds was 0.2, with a repeatability of 0.36 and 0.37, for SPB and AB breeds, respectively. With the data from both breeds, and using a classical approach, the response to selection was estimated if mares with extreme gestation lengths were culled, i.e. lengths which are under 310 days, or over 360 days. According to our results, in the case of SPB, a decrease of 14-45% would occur in the number of extreme gestation lengths, while in the AB breed, this value would decrease from 2 to 39%.     

Exposure to stallion accelerates the onset of mares' cyclicity

Horses (Equus caballus) belong to the group of seasonally polyestrous mammals. Estrous cycles typically start with increasing daylight length after winter, but mares can differ greatly in the timing of onset of regular estrus cycles. Here, we test whether spatial proximity to a stallion also plays a role. Twenty-two anestrous mares were either exposed to one of two stallions (without direct physical contact) or not exposed (controls) under experimental conditions during two consecutive springs (February to April). Ovarian activity was monitored via transrectal ultrasound and stallion's direct contact time with each mare was determined three times per week for one hour each. We found that mares exposed to a stallion ovulated earlier and more often during the observational period than mares that were not exposed to stallions. Neither stallion identity nor direct contact time, mare age, body condition, size of her largest follicle at the onset of the experiment, or parasite burden significantly affected the onset of cyclicity. In conclusion, the timing of estrous cycles and cycle frequency, i.e., crucial aspects of female reproductive strategy, strongly depend on how the mares perceive their social environment. Exposing mares to the proximity of a stallion can therefore be an alternative to, for example, light programs or elaborated hormonal therapies to start the breeding season earlier and increase the number of estrous cycles in horses.     

Repeatability of preovulatory follicular diameter and uterine edema pattern in two consecutive cycles in the mare and how they are influenced by ovulation inductors

Follicular diameter is used as a guiding tool to predict ovulation in the mare. However, the great range in preovulatory follicular diameter makes prediction of optimal breeding time based on follicular diameter unreliable. Uterine edema pattern is also useful to determine the best time to breed, since intensity of edema tends to dissipate as ovulation approaches, however, not every mare follows this pattern. The aims of this study were to assess the repeatability of preovulatory follicular diameter and uterine edema pattern in two consecutive spontaneous cycles and to determine how induction treatments (hCG, PGF(2)alpha and GnRH analogues) influence them. Fifty-three mares were followed during two consecutive cycles and scanned three times a day from 2 to 3 days before ovulation. During the first cycle, mares had a spontaneous ovulation and in the consecutive cycle mares received either: (a) no hormonal treatment; (b) 1500 IU hCG; (c) 125-250 microg Cloprostenol or (d) 2.1 mg Deslorelin implant. Mares ovulated consistently from similar follicular diameters in two consecutive spontaneous cycles (r=0.89; P<0.000). All three induction treatments had a significant effect on reducing the preovulatory follicular diameter (P<0.005). Mares showed fair correlation in uterine edema patterns in both consecutive non-induced cycles (r=0.71; P<0.005). In conclusion mares in consecutive cycles ovulated from consistent follicular diameters. Follicular diameters recorded from previous ovulations can be relied on to predict the optimal breeding time in successive cycles especially in mares that ovulate from unusually small follicles.     

Factors influencing conception rates of Arab mares in Tunisia

In order to investigate the factors affecting the reproduction efficiency of Arab breeding mares in Tunisia, breeding data corresponding to 2340 mated cycles, over 5 years (from 2000 to 2004), from 555 mares offspring of 50 sires and bred with 17 stallions were analysed using a multivariate logistic regression. We chose logit link function and binomial distribution and we used log-likelihood-ratio (LL) and Wald tests (X² Wald) to test the mean values. The factors of variation included in the model were the year, the stallion, the age of the mare at mating, the sire and the category of the mare. Logistic regression results showed that age and sire affected both the overall conception rate and the first-cycle conception rate of the mares whilst category and stallion influenced only the overall conception rate. To our knowledge, this is the first study showing that the sire of the mare can affect both the first-cycle conception rate and the overall conception rate of the mares.     

Sexual behavior following introduction of a stallion into a group of mares

A rested stallion was introduced daily for 30 days into each of three herds of 20 mares. Observations of sexual and mating behavior were made for one hour. The stallion remained with a herd until another stallion was introduced the following day. Other mares were isolated from stallions and were bred by artificial insemination. The diameter or growth rate of the preovulatory follicle for the six days preceding ovulation and the length of the interovulatory interval for mares which did not become pregnant were not affected significantly by the presence of a stallion. The number of breedings per hour of observation (2.4 ±0.2) and the length of the interval from introduction of a stallion into the herd of mares to first breeding (12 ±1 min) were significantly different among stallions, but the length of the interval between breedings (17 ±2 min) was not. The mean number of breedings per stallion per hour was not affected significantly by the number of posturing (estrous) mares. The number of times that the stallion rebred the same mare when more than one mare postured during the observation hour (49%) was greater (P<0.01) than what would be expected to occur by chance (30%). The hypothesis that breeding occurs preferentially in those estrous mares that are closest to ovulation was not supported, except for significantly lower breeding activity in posturing mares on days 8 and 7 before ovulation and on days 0 (day of ovulation) and ?1 (26% bred) than on days 6 to 1 (52%).     

Retained placenta in Friesian mares: incidence, and potential risk factors with special emphasis on gestational length

During the foaling seasons of 1999 and 2000, the incidence of retained placenta in 495 normal parturitions of 436 Friesian brood mares was studied. Retained placenta was defined as a failure to expel all fetal membranes within 3 h of the delivery of the foal. Furthermore, the sex of the foal, month of breeding, sire and dam's sire, age of the mare, and time of day of foaling, were studied as factors that might be associated with retained placenta in Friesian mares after normal foalings, and with gestational length. The analysis was carried out using marginal logistic regression, and mixed linear regression, respectively. The incidence of retained placenta was 54%. Mean length of gestation was 331.6 days. Colts were carried 1.5 days longer than fillies. Mares bred in July-September had a 4-day shorter gestation period (329 days) than mares bred earlier in the year. There was a mare, sire, and dam's sire effect on gestational length, and a mare effect on the occurrence of retained placenta. Mares foaling at 4 and >17 years of age, tended to have a lower incidence of retained placenta than mares foaling at 5-17 years of age. No association was found between the occurrence of retained placenta, and gestational length, sex of the foal, month of breeding, dam's sire, and time of day of foaling. It was concluded that the observed high incidence of retained placenta indicates that the Friesian breed of horses has a higher risk for retained placenta than other breeds of horses.     

Equine lymphocyte antigens and reproduction in the Standardbred mare

Equine lymphocyte antigen (ELA) gene frequencies were estimated for pacing and trotting Standardbred mares residing on a breeding farm in central Ohio. The ELA gene frequencies for Ohio Standardbreds did not differ significantly from the ELA gene frequencies of Kentucky Standardbreds, determined by Bailey (1983). No significant differences were found in the distribution of ELA class I antigens in horses with lower overall fertility or a history of abortion on the investigated breeding farm. Likewise, no significant association was observed when the ELA types of both the mare and the stallion to which she was mated were compared with the reproductive efficiency of the mare.     

Equine lymphocyte antigens and reproduction in the Standardbred mare

Summary. Equine lymphocyte antigen (ELA) gene frequencies were estimated for pacing and trotting Standardbred mares residing on a breeding farm in central Ohio. The ELA gene frequencies for Ohio Standardbreds did not differ significantly from the ELA gene frequencies of Kentucky Standardbreds, determined by Bailey (1983). No significant differences were found in the distribution of ELA class I antigens in horses with lower overall fertility or a history of abortion on the investigated breeding farm. Likewise, no significant association was observed when the ELA types of both the mare and the stallion to which she was mated were compared with the reproductive efficiency of the mare.     

When should a mare go for assisted reproduction?

The use of assisted reproductive techniques (ART) has helped owners to produce offspring from valuable mares that were considered infertile using standard breeding techniques. Before referring a mare for an ART, the practitioner should be able to identify the underlying cause of subfertility of the mare. The objective of this review is to provide information regarding embryo transfer, oocyte transfer and intracytoplasmic sperm injection, the three most common ART used in equine practice. Knowing the complexity as well as the risks of these techniques, enables practitioners to refer a subfertile mare to the least complex and most appropriate and successful ART that can overcome specific causes of infertility.     

Herding and snaking by the harem stallion in domestic herds

Four herds of pony mares, each consisting of a stallion and six mares, were used to characterize the nature of herding by the stallion and the factors that induced the herding behavior. Herding behaviors were compared among four successive treatments (six mares alone, stallion added, two new mares added, and entire herd moved to a new pasture). A new treatment was initiated every 7 days and behavior was studied for 5 consecutive days (Days 1-5) for each treatment. Observations were made every 10 min during a 2-h period for each day. The extent of herding was quantitated by the mean distances between mares. The extent of snaking (herding with the head and neck extended and ears held back) was scored 0, 1, 2, or 3 (nil, minimal, intermediate, and maximal, respectively). The mean distance among the original mares on Day 1 when the mares were alone was 5.0 mare lengths and was reduced (P < 0.05) to 1.9 mare lengths when the stallion was added. The mean distance among the original mares of an established stallion/mare herd (3.8 mare lengths) was reduced (P < 0.05) on the day the herd was moved to a new pasture (1.9 mare lengths), similar to the effect of the introduction of the stallion. Scores for the extent of snaking, as well as the extent of herding, were highest (P < 0.05) on Day 1 when the stallion was added or the stallion/mare herd was moved to a new pasture. The extent of herding and snaking decreased (P < 0.05) by Day 2 and was seen only occasionally on Days 3-5. The addition of new mares to the herd did not induce herding of the original mares. However, the new mares maintained mean distances of 8-12 mare lengths from the original mares, resulting primarily from chasing by the stallion. By Day 4, the distances between the new and original mares were not different (P > 0.05) from the distances among the original mares.     

Subclinical infection and periodic shedding of equid herpesvirus 3

The temporary disruption of reproductive activities due to equine coital exanthema (ECE), caused by equid herpesvirus 3 (EHV-3), at thoroughbred breeding facilities and embryo transfer centres, has an appreciable economic impact. The aim of the present study was to estimate the prevalence of excretion of EHV-3 in mares without clinical symptoms under field conditions and the re-excretion patterns of the virus in two seropositive (presumably latently infected) mares maintained in isolation for 11 mo. The EHV-3 virus was detected in perineal-vaginal swabs by real time PCR in 14 (6%) of 220 thoroughbred mares without clinical symptoms at the time of breeding. In the two isolated mares, re-excretion of EHV-3 was demonstrated on two occasions, 3 mo apart (each for a 3 d interval) in one mare, and on only 1 d in the other mare. Antibodies against EHV-3 were identified by seroneutralization in 105 (48%) of the thoroughbred mares, and during the entire period in the two isolated mares. Therefore, the present study provided evidence of EHV-3 shedders in a healthy mare population under both field and isolation conditions. Furthermore, at least two periods of spontaneous EHV-3 reactivation and re-excretion in the presence of serum antibodies occurred in one mare in an 11 mo interval. These findings could assist in the design and implementation of measures to minimize the spread of EHV-3 and control ECE outbreaks.     

The effects of age, season and fertility status on plasma and intratesticular insulin-like growth factor I concentration in stallions

The purposes of this study were to establish the basal plasma and testicular insulin-like growth factor-I (IGF-I) values for stallions ranging in age from 6 months to 23 years and to determine if IGF-I could be used as a marker for declining fertility. Blood and testes were obtained from 28 light horse stallions and colts. Of the 28 stallions, 22 were considered fertile and were categorized by age (<2 y, 5 to 10 y, 11 to 15 y, and 16 to 23 y); 12 age-matched stallions were grouped as to fertility status (fertile, subfertile, infertile); and all 28 stallions were grouped as to season of castration (breeding season vs. non-breeding season). In colts less than 2 years of age, IGF-I concentrations in plasma and testicular extracts were higher (P < 0.01) than in the other age groups and were higher in the breeding season than in the non-breeding season (P < 0.01). No significant differences in plasma or testicular extract concentrations of IGF-I were found among fertility groups. The results of this study demonstrate that plasma and testicular IGF-I levels are high in stallions younger than 2 years of age and then decline and plateau in stallions older than 5 years of age, suggesting that IGF-I may be involved in testicular development. The results allude to a possible seasonal effect on IGF-I production. However, it is difficult to separate true seasonality and the effect of age as only those stallions less than 2 years old exhibited variation between seasons. The IGF-I does not appear to have a direct relationship with declined fertility in the stallions tested, suggesting that IGF-I may not be a reliable biomarker for the diagnosis of subfertility and infertility.