Ultrasound-guided fetal thorax compression to reduce post-fixation twins in the mare.

BACKGROUND: Management of twin pregnancy after conceptus vesicle fixation in the horse is challenging because the reduction techniques described are either invasive, difficult to perform or associated with disappointing success rates. OBJECTIVES: To evaluate the success of transrectal ultrasound-guided fetal thorax compression for reducing post-fixation twin pregnancy in mares. STUDY DESIGN: Retrospective clinical study. METHODS: Sixteen mares were presented for twin reduction between 51 and 79 days of gestation. History obtained from the owner and/or referring veterinarian detailed information regarding the mare (age, breed), pregnancy (day of gestation, dizygotic versus monozygotic twins, unilateral versus bilateral fixation), treatment and outcome (one live fetus at discharge; live singleton at foaling) after twin reduction. Transrectal fetal thorax compression was performed under ultrasound guidance by two experienced operators. RESULTS: Overall 9 of 16 twin pregnancies were successfully reduced and the likelihood of success was significantly higher in dizygotic than monozygotic twins. The procedure was successful in 9 of 10 dizygotic twins but unsuccessful in all six cases of monozygotic twins. Among the dizygotic twins, two mares lost the pregnancy after discharge from the clinic, seven mares delivered a healthy foal of normal size. MAIN LIMITATIONS: Small case number. CONCLUSIONS: Transrectal ultrasound-guided fetal thorax compression is a minimally-invasive and successful technique for reducing dizygotic twin pregnancies at approximately 2 months of gestation, but does not lead to any live births in cases of monozygotic twins.

In Vitro-Produced Equine Blastocysts Exhibit Greater Dispersal and Intermingling of Inner Cell Mass Cells than In Vivo Embryos.

In vitro production (IVP) of equine embryos is increasingly popular in clinical practice but suffers from higher incidences of early embryonic loss and monozygotic twin development than transfer of in vivo derived (IVD) embryos. Early embryo development is classically characterized by two cell fate decisions: (1) first, trophectoderm (TE) cells differentiate from inner cell mass (ICM); (2) second, the ICM segregates into epiblast (EPI) and primitive endoderm (PE). This study examined the influence of embryo type (IVD versus IVP), developmental stage or speed, and culture environment (in vitro versus in vivo) on the expression of the cell lineage markers, CDX-2 (TE), SOX-2 (EPI) and GATA-6 (PE). The numbers and distribution of cells expressing the three lineage markers were evaluated in day 7 IVD early blastocysts (n = 3) and blastocysts (n = 3), and in IVP embryos first identified as blastocysts after 7 (fast development, n = 5) or 9 (slow development, n = 9) days. Furthermore, day 7 IVP blastocysts were examined after additional culture for 2 days either in vitro (n = 5) or in vivo (after transfer into recipient mares, n = 3). In IVD early blastocysts, SOX-2 positive cells were encircled by GATA-6 positive cells in the ICM, with SOX-2 co-expression in some presumed PE cells. In IVD blastocysts, SOX-2 expression was exclusive to the compacted presumptive EPI, while GATA-6 and CDX-2 expression were consistent with PE and TE specification, respectively. In IVP blastocysts, SOX-2 and GATA-6 positive cells were intermingled and relatively dispersed, and co-expression of SOX-2 or GATA-6 was evident in some CDX-2 positive TE cells. IVP blastocysts had lower TE and total cell numbers than IVD blastocysts and displayed larger mean inter-EPI cell distances; these features were more pronounced in slower-developing IVP blastocysts. Transferring IVP blastocysts into recipient mares led to the compaction of SOX-2 positive cells into a presumptive EPI, whereas extended in vitro culture did not. In conclusion, IVP equine embryos have a poorly compacted ICM with intermingled EPI and PE cells; features accentuated in slowly developing embryos but remedied by transfer to a recipient mare.

In vitro aging of stallion spermatozoa during prolonged storage at 5°C.

Artificial insemination with chilled stallion semen is hampered by a limited period of maximum fertility maintenance (24-48 h). This study used multiparametric flow cytometry to simultaneously measure reactive oxygen species (ROS) production, mitochondrial function or [Ca2+ ]i and plasma membrane fluidity in viable, acrosome-intact spermatozoa, with the aim of providing insight into changes in sperm function during storage at 5°C. High proportions of viable and acrosome-intact spermatozoa (71 ± 8%) remained after 96 h of storage demonstrating that the basic integrity of the cells was well preserved (n = 17 stallions). In addition, more than 90% of viable, acrosome-intact spermatozoa had active mitochondria and low intra-cellular or mitochondrial ROS levels. By contrast, the percentage of viable, acrosome-intact sperm with low plasma membrane fluidity and low [Ca2+ ]i decreased over time (1 h: 63 ± 16%, 96 h: 29 ± 18%; p < 0.05). The [Ca2+ ]i in viable sperm rose 3.1-fold (p < 0.05) over the 4 days, and fewer spermatozoa responded to bicarbonate stimulation (1 h: 46 ± 17%, 96 h: 19 ± 12%) with an increase in plasma membrane fluidity following prolonged storage. Overall, prolonged storage of stallion semen at 5°C resulted in disturbed calcium homeostasis and increased plasma membrane fluidity. The decline in fertility of stallion semen during cooled-storage may therefore relate to aspects of in vitro aging (changes in plasma membrane fluidity and intracellular calcium) which impairs capacitation-associated cell functions.

A Modified Flotation Density Gradient Centrifugation Technique Improves the Semen Quality of Stallions with a High DNA Fragmentation Index.

Sperm DNA fragmentation compromises fertilization and early embryo development. Since spermatozoa lack the machinery to repair DNA damage, to improve the likelihood of establishing a healthy pregnancy, it is preferable to process ejaculates of stallions with a high sperm DNA fragmentation index (DFI) before artificial insemination or intracytoplasmic sperm injection. The aim of this study was to examine a modified flotation density gradient centrifugation (DGC) technique in which semen was diluted with a colloid solution (Opti-prepTM) to increase its density prior to layering between colloid layers of lower and higher density. The optimal Opti-prepTM solution (20-60%) for use as the bottom/cushion layer was first determined, followed by a comparison between a modified sedimentation DGC and the modified flotation DGC technique, using different Opti-prepTM solutions (20%, 25% and 30%) as the top layer. Finally, the most efficient DGC technique was selected to process ejaculates from Friesian stallions (n = 3) with high sperm DFI (>20%). The optimal Opti-prepTM solution for the cushion layer was 40%. The modified sedimentation technique resulted in two different sperm populations, whereas the modified flotation technique yielded three populations. Among the variants tested, the modified flotation DGC using 20% Opti-prepTM as the top layer yielded the best results; the average sperm recovery was 57%; the DFI decreased significantly (from 12% to 4%) and the other sperm quality parameters, including progressive and total motility, percentages of spermatozoa with normal morphology and viable spermatozoa with an intact acrosome, all increased (p < 0.05). In Friesian stallions with high sperm DFI, the modified flotation DGC markedly decreased the DFI (from 31% to 5%) and significantly improved the other semen quality parameters, although sperm recovery was low (approximately 20%). In conclusion, stallion sperm DFI and other sperm quality parameters can be markedly improved using a modified flotation DGC technique employing a 40% Opti-prepTM cushion and a 20% top layer.

Relationships between blood and follicular fluid urea nitrogen concentrations and between blood urea nitrogen and embryo survival in mares.

High blood urea nitrogen (BUN) concentration is linked to low fertility in cows and ewes; however, this relationship has not been reported in mares. The study characterized the relationship between BUN and follicular fluid urea nitrogen (FUN) during follicle growth (Experiment 1) and the impact of BUN from embryo donors on the pregnancy outcome of recipient mares (Experiment 2). In experiment one, follicular fluid and blood samples were collected from mares during diestrus with growing follicles and during estrus with pre-ovulatory follicles (n = 16 and 10 mares, respectively). In experiment two, BUN concentrations of embryo donors were related to pregnancy outcome after embryo transfer. In experiment one, there was a strong positive correlation between BUN and FUN (R = 0.83; P < 0.0001), with higher BUN in mares with growing follicles than with preovulatory follicles (P = 0.004) and higher FUN in growing follicles than in preovulatory follicles (P = 0.031). In experiment two, BUN was higher in donor mares that produced unsuccessful embryos compared to donor mares that produced embryos resulting in successful pregnancies at D14 (P < 0.03). Additionally, there was an effect of age (P = 0.01) and interaction between age and lactation (P = 0.009) in donor mares for embryo survival after embryo transfer. Donor mares with unsuccessful embryos were older than donor mares with successful embryos. Therefore, these experiments showed that BUN was related to follicular fluid environment as well as to the survival of Day 7-8 embryos after transfer to recipient mares.

A retrospective study on semen quality parameters from four different Dutch horse breeds with different levels of inbreeding.

A high degree of inbreeding has been reported to negatively impact semen quality in Friesian horses and Shetland ponies. Both breeds are characterized by a closed studbook, small population size, and high incidence of inbreeding. The Dutch Warmblood studbook (KWPN: Koninklijk Warmblood Paardenstamboek Nederland) is a much larger studbook with two distinct populations: the KWPN-Riding horses, managed as an 'open' studbook, and the KWPN-Harness horses, representing a much smaller subpopulation within the KWPN breed and managed as an 'almost closed' studbook. It was recently reported that the degree of inbreeding in KWPN-Harness horses has increased in recent decades due to the small gene pool; however, the degree of inbreeding is still lower than that of Friesian horses and Shetland ponies. We hypothesized that a high or rising degree of inbreeding might negatively impact semen quality. In the present study, we retrospectively compared semen quality parameters of stallions from four different breeds or types (Friesian Horses, Shetland Ponies, KWPN-Riding horses, and KWPN-Harness horses), each reported with different degrees of inbreeding. Semen concentration, and percentages of motile, morphologically normal and live spermatozoa, and the total number of morphologically normal, progressive motile spermatozoa per ejaculate (TNM) were analyzed for 2832 semen evaluations performed over a 15-year period. KWPN-Harness horses had a significantly lower sperm concentration, % motile spermatozoa and % live spermatozoa than KWPN-Riding horses but the % motile and % morphologically normal spermatozoa and TNM in both KWPN-Harness and KWPN-Riding horses were significantly higher than in Friesian horses and Shetland ponies. These results suggest a lower semen quality in KWPN-Harness than KWPN-Riding horses, potentially as a result of a higher coefficient of inbreeding. The negative trend observed in the KWPN-Harness horses may be a warning sign, and breeders or stud books should monitor the degree of inbreeding carefully to avoid a further reduction in semen quality, to the levels observed in Friesian horses and Shetland ponies.

A practical guide to estimate the age of the early CL by ultrasonography in mares examined for the first time to be used as recipients in a commercial embryo transfer program.

The echogenicity of the early CL undergoes obvious changes during the first 5 days of luteal development. This phenomenon could potentially be used to estimate the age of the CL in mares which have not been checked frequently for the diagnosis of ovulation. The objective of this retrospective study was to determine the accuracy of estimating the age of the early CL (Day 0 to Day 4; Day 0 = Day of ovulation) in recipient mares examined for the first time in the breeding season to be used in a commercial embryo transfer (ET) program. The post-transfer pregnancy rate and embryo loss of 28 recipient mares whose CL age was estimated at the first scan of the breeding season (at least 2 months gap from the previous examination) were compared to that of 41 recipients in which the Day of ovulation had been determined by daily examinations and that received an embryo on the respective day. There was no difference (P > 0.1) in pregnancy (85.7 and 85.3%) or early embryonic loss (12.5% and 11.4%) between groups, respectively.

In vitro production of horse embryos predisposes to micronucleus formation, whereas time to blastocyst formation affects likelihood of pregnancy.

Invitro embryo production is an increasingly popular means of breeding horses. However, success is limited by a high incidence of early embryo loss. Although there are various possible causes of pregnancy failure, chromosomal abnormalities, including aneuploidy, are important potential contributors. This study evaluated the frequency of micronucleus formation as a proxy for aneuploidy in invitro-produced (IVP) and invivo-derived horse blastocysts. Associations between IVP embryo morphology, frequency of nuclear abnormalities and the likelihood of pregnancy were investigated. IVP blastocysts exhibited a higher frequency of cells with micronuclei than invivo-derived embryos (10% vs 1% respectively; P=0.05). This indication of chromosomal instability may explain the higher incidence of pregnancy failure after transfer of IVP embryos. However, the frequency of micronuclei was not correlated with brightfield microscopic morphological characteristics. Nevertheless, IVP embryos reaching the blastocyst stage after Day 9 of invitro culture were less likely to yield a pregnancy than embryos that developed to blastocysts before Day 9 (27% vs 69%), and embryos that had expanded before transfer were more likely to undergo embryonic death than those that had not expanded (44% vs 10%). These findings indicate that current embryo culture conditions are suboptimal and that the speed of embryo development is correlated with pregnancy survival.

In vitro-produced horse embryos exhibit a very narrow window of acceptable recipient mare uterine synchrony compared with in vivo-derived embryos.

In recent years, the number of equine invitro-produced embryos (IVP) has increased markedly; as yet, there are few reports on what constitutes an 'ideal' recipient for an IVP embryo. This study retrospectively investigated the effects of recipient mare oestrous cycle characteristics on the likelihood of pregnancy after transfer of IVP (n=264) and invivo-derived embryos (n=262). IVP embryos tolerated only a narrow window of recipient mare 'synchrony', with transfer on Day 4 after ovulation resulting in a higher likelihood of ongoing pregnancy (69%) than transfer on Days 3, 5 or 6 (53.2%, 41.3% and 23.1% respectively; P=0.02). In contrast, Day 8 invivo-derived embryos tolerated a wide range of uterine (a)synchrony, with no difference in pregnancy or pregnancy loss for recipients that ovulated between Day 4 and Day 9 before transfer. However, transferring invivo-derived embryos to recipients that had a longer oestrus preceding transfer resulted in higher Day 12 and ongoing pregnancy rate (P<0.01). This effect was not significant in IVP embryos. In conclusion, Day 6-8 IVP blastocysts survive best after transfer to Day 4 recipient mares; Day 8 invivo-derived embryos survive equally well in Day 4-9 recipients, but do better in mares that have a long preceding oestrus.

Mare and stallion effects on blastocyst production in a commercial equine ovum pick-up-intracytoplasmic sperm injection program.

This study retrospectively examined the degree to which success within a commercial ovum pick-up (OPU)-intracytoplasmic sperm injection (ICSI) program varied between individual mares and stallions. Over 2 years, 552 OPU sessions were performed on 323 privately owned warmblood mares. For mares that yielded at least one blastocyst during the first OPU-ICSI cycle, there was a 77% likelihood of success during subsequent attempts; conversely, when the first cycle yielded no blastocyst, the likelihood of failure (no embryo) in subsequent cycles was 62%. In mares subjected to four or more OPU sessions, the mean percentage of blastocysts per injected oocyte was 20.5% (range 1.4-46.7%), whereas the mean number of blastocysts per OPU-ICSI session was 1.67 (0.2-4.2). Age did not differ significantly between mares that yielded good or poor results. The number of recovered oocytes per OPU was positively associated with the likelihood of success (P<0.001). Although there were considerable between-stallion differences, most stallions (14/16) clustered between 15.6% and 26.8% blastocysts per injected oocyte, and the number of blastocysts per OPU (mean 1.4; range 0.2-2.2) was less variable than among mares. In conclusion, although both mare and stallion affect the success of OPU-ICSI, mare identity and the number of oocytes recovered appear to be the most reliable predictors of success.

The recipient's Day after ovulation and the number of corpora lutea influence the likelihood of pregnancy in mares following transfer of ICSI frozen embryos.

Asynchronous embryo transfer is an excellent tool to investigate how subtle differences in the uterine environment affect embryo development and survival. Progesterone secreted from the corpus luteum following ovulation is one of the main factors responsible for establishing endometrial receptivity for the pre-implantation embryo via complex alterations in the expression of genes involved in the secretion of the histotroph. The objective of this retrospective study was to determine whether the recipient's Day after ovulation and the number of CL at ET influence the pregnancy rates of IVP horse embryos. The study included 650 heterologous frozen ICSI horse embryo transfer cycles and evaluated the pregnancy and ongoing pregnancy rate. The ongoing pregnancy was significantly lower in recipient mares with ET performed 5 and 6 days after ovulation (47.4% and 37.5%, respectively) than in recipients with ET 4 days after ovulation (67.3%). Furthermore, Day 5 recipient mares (Day 0 = Day of ovulation) with 2 corpora lutea (CL) at the time of ET had lower ongoing pregnancy rate (36.1%) than Day 5 recipient mares with 1 CL (51.9%). In contrast, the presence of 2 CL was associated with a higher ongoing pregnancy rate (75.8%) in recipient mares with a less advanced uterine stage at the time of ET (Day 3 and 4 after ovulation), compared to recipients with only 1 CL at ET (62.7%). In conclusion, both the number of days after ovulation and the number of CL recorded in the recipient mare at ET influenced the ongoing clinical pregnancy rate. This study highlights the importance of establishing exactly when progesterone rises above a threshold (relative to the Day of ovulation) when trying to determine the optimal window for transferring an IVF/IVP embryo.

The Effect of Different Flushing Media Used to Aspirate Follicles on the Outcome of a Commercial Ovum Pickup-ICSI Program in Mares.

The in vitro production of embryos by ovum pickup (OPU) and intracytoplasmic sperm injection (ICSI) is gaining popularity among horse breeders and veterinarians. Various collection media are available for flushing follicles during OPU. The objective of this study was to determine whether the type of flushing media used to aspirate follicles and collect oocytes influences the outcome of a commercial equine OPU-ICSI program. Two commercial embryo flushing media (EFM1 and EFM2) supplemented with heparin were compared with a flushing media designed specifically for the collection of oocytes (oocyte flushing media [OFM]) on the outcome of OPU-ICSI parameters in 234 Warmblood mares. The OPU-ICSI performed in mares using one of the EFM1 resulted in a lower (P < .05) blastocyst rate and blastocysts per OPU-ICSI session (11.9 ± 13.2%, 0.88 ± 1.3) than the OFM (19.2 ± 15.2%, 1.24 ± 1.2). Unlike the EFM2 solution, the heparin used to prepare the EFM1 contained preservatives including benzyl alcohol, a component known to alter the oocyte membrane, which might have been responsible for the lower developmental competence of oocytes collected with EFM1. In conclusion, exposure of oocytes (<1.5 hours) to one of the flushing medium tested in this study affected negatively the outcome of the OPU-ICSI commercial program when compared with flushing media designed for collection of equine oocytes. Care should be taken when choosing the components of the flushing media used to collect oocytes. Further research should be carried out to confirm the potential negative effect of the preservatives used in multidose heparin vials.

A retrospective comparison of the efficiency of different assisted reproductive techniques in the horse, emphasizing the impact of maternal age.

Advancing maternal age is known to negatively affect fertility in the horse. This age-related decrease in fertility has been linked primarily to reduced oocyte quality rather than to impaired uterine function. In the past decade, the use of ovum pick-up (OPU) and ICSI to produce foals has rapidly gaining popularity amongst sport horse breeders. However, it is not yet known how maternal age influences the efficiency of a commercial OPU-ICSI program and whether the age effect is similar to that observed for other ART in the horse. To answer this question, reproductive records of 289 mares bred by natural mating (NM), 328 mares bred by AI, 205 embryo donor mares (AI-EF-ET), and 473 mares submitted for OPU-ICSI and ET were analyzed retrospectively using a regression model to investigate the effects of maternal age and breeding technique on the likelihood of producing a viable pregnancy. The reproductive efficiency (quantified as the proportion of mares that yielded at least one Day 45 pregnancy) of the different breeding techniques NM, AI, AI-EF-ET and OPU-ICSI-ET was 63.3, 43.9, 45.8 and 37.4%, respectively (P < 0.05). However, the frequent production of multiple embryos per ICSI session (up to 10 embryos in one attempt), makes OPU-ICSI-ET as effective as AI-EF-ET when measured in terms of the mean number of Day 45 pregnant recipients per donor mare. Increasing maternal age was associated with a reduction (P < 0.05) in the reproductive efficiency of all breeding techniques (NM, AI, AI-EF-ET) except OPU-ICSI-ET (P > 0.05). In the OPU-ICSI-ET group, increasing maternal age was associated with a lower number of follicles aspirated and oocytes recovered per mare. Nevertheless, the percentage of blastocysts per injected oocyte, and post-ET likelihoods of pregnancy and pregnancy loss were not influenced by the age of the oocyte donor mare (P > 0.05).

Effect of embryo transfer technique on the likelihood of pregnancy in the mare: a comparison of conventional and Wilsher's forceps-assisted transfer.

The aim of this study was to compare the success of embryo transfer (ET) performed by veterinarians with different degrees of experience using one of two ET techniques. Over three years, 179 embryos were transferred by three operators with moderate to high experience using a 'conventional' manual technique, and 170 embryos were transferred by four operators with little or no previous ET experience using a 'Wilsher' technique (Polansky speculum and Wilsher's cervical forceps). The pregnancy status of recipient mares at the first pregnancy diagnosis and the embryo loss by the last examination were compared between groups and within group among operators. More recipients became pregnant (P<0.005) following ET with the Wilsher technique (157/170; 92.3 per cent) than with the conventional technique (127/179; 70.9 per cent), while the incidence of pregnancy loss did not differ between the groups (9.1 v 7.9 per cent, respectively). For the conventional technique, there was a significant operator effect (P<0.01) on the percentage of pregnant recipients at the first examination (50.9-79.7 per cent); no operator effect was apparent for the Wilsher technique (90.9-93.4 per cent). In conclusion, the Wilsher technique yielded high pregnancy rates (>90 per cent).

The influence of age, antral follicle count and diestrous ovulations on estrous cycle characteristics of mares.

Reproductive aging must be well understood to optimize the reproductive management of older mares and to predict their reproductive life-span. The objectives of this study were to: 1) examine age-related differences in follicular dynamics, endocrine profiles, and primordial follicle counts, 2) evaluate the influence of antral follicle count (AFC) on age-related changes in follicular parameters, and 3) determine the influence of diestrous ovulations on the estrous cycle. Young (3-8yr; n = 10), middle-aged (9-18 yr; n = 16), and old (>18 yr; n = 19) light horse mares were examined with transrectal ultrasonography to monitor follicular growth over two consecutive estrous cycles. Jugular blood samples were taken and plasma progesterone and FSH concentrations were determined by an enzyme immunoassay and radioimmunoassay, respectively. Both interovulatory intervals and follicular phases were longer and the day of follicle deviation occurred later in aged mares. Furthermore, older mares had a tendency to ovulate smaller follicles. Neither follicular growth rate, the number of ovulations nor the length of luteal phase was influenced by mare age. Interestingly, as mare age increased, mares with low AFC had longer interovulatory intervals and follicular phases than mares with medium or high AFC. In addition, the number of primordial follicles declined with an increase in mare age but varied considerably between mares of the same age. Progesterone concentrations were positively influenced by age, whereas FSH concentrations were not, despite that FSH concentrations appeared higher in aged mares during the follicular phase. Estrous cycles with a diestrous ovulation had a longer interovulatory interval as well as a longer follicular and luteal phase while day of deviation occurred later. Progesterone concentrations were significantly higher on day 14 and 16 in estrous cycles with a diestrous ovulation than without a diestrous ovulation. In conclusion, aging in mares is associated with changes in follicular parameters which in turn are closely linked to differences in antral follicle count suggesting a relationship with ovarian reserve. Therefore, determination of antral follicle counts in aged mares can provide valuable information about the reproductive aging process. Finally, diestrous ovulations have a significant influence on different estrous cycle parameters.

Biological Functions and Clinical Applications of Anti-Müllerian Hormone in Stallions and Mares.

Anti-Müllerian hormone (AMH) plays a major role in sexual differentiation, Leydig cell differentiation, and folliculogenesis. In addition, AMH has clinical value in equine practice. In stallions, AMH can serve as an endocrine marker for equine cryptorchidism and as an immunohistochemical marker for Sertoli cell tumors. Considering that AMH is also an ovarian specific product, intact mares can be differentiated from ovariectomized mares. Peripheral AMH concentrations reflect the follicular population in mares, and therefore, are useful in the assessment of ovarian reserve and reproductive life-span of aged mares. Last, AMH is particularly suitable as a diagnostic marker for equine granulosa cell tumors.

Induction of ovulation in seasonally anestrous mares under ambient lights using recombinant equine FSH (reFSH).

Traditionally, mares are put under artificial lights to advance the first ovulation of the year. The aim of the present study was to determine the efficacy of recombinant equine FSH (reFSH) in stimulating follicular development and advancing the first ovulation of the year in seasonally anestrous mares compared with anestrous mares given a placebo. Both groups of mares were housed under ambient light conditions. Sixty deep anestrous mares of light horse breeds (follicular diameters ≤ 20 mm in diameter and progesterone <1 ng/mL) were maintained under a natural photoperiod at three different sites: University of California, Davis, Colorado State University, and University of Kentucky Gluck Centre. Twenty mares at each site were randomly allocated to receive either 0.65 mg of reFSH (group A: treatment; n = 10) or a placebo (group B: control; n = 10) twice daily by im beginning on January 31. Treatment continued until one or more preovulatory follicles developed or up to a maximum of 15 days. Randomized treatments were blinded. Follicular development was closely monitored by transrectal ultrasonography. When the largest follicle reached ≥ 35 mm in diameter, reFSH treatment was discontinued and an injection of 2500 international units of hCG was administered iv 36 hours later to induce ovulation. Jugular blood samples were collected daily from all mares at University of California, Davis, and processed for LH, FSH, progesterone, estradiol-17ß, and immunoreactive-inhibin by RIA. All 30 mares receiving reFSH (group A) developed follicles ≥ 35 mm within 7.4 ± 1.6 days of treatment. Twenty-three of the 30 reFSH-treated mares (group A) ovulated within 72 hours after hCG administration. In contrast, mares in group B (placebo, control) did not exhibit significant follicular development and none ovulated within the 15-day observation period. Mares in group A had significantly higher plasma levels of FSH, estradiol-17ß, and immunoreactive-inhibin during treatment but did not exhibit a preovulatory LH surge. Mares administered reFSH returned to anestrus and spontaneously ovulated at a similar calendar date as control mares. These data indicate that reFSH was effective in stimulating the development of ovarian follicles and advancing the first ovulation of the year in seasonally anestrous mares under ambient lights but was not successful in inducing continued cyclicity.

Serum anti-Müllerian hormone concentrations in stallions: developmental changes, seasonal variation, and differences between intact stallions, cryptorchid stallions, and geldings.

Anti-Müllerian hormone (AMH), a homodimeric glycoprotein, is secreted early in fetal life when it exerts a crucial function in sexual differentiation. The secretion of AMH in male humans persists after birth and is characterized by high prepubertal concentrations followed by a significant decrease at the onset of puberty. The expression of AMH in the normal and cryptorchid equine testis is well characterized but data regarding circulating AMH concentrations are lacking. The objectives of this study were to determine serum AMH concentrations in neonatal colts and fillies, prepubertal colts, and postpubertal stallions, and to evaluate variations in serum AMH related to season and gonadal status of stallions. In addition, we examined the presence and determined concentrations of AMH in seminal plasma of mature stallions. Serum AMH concentrations were significantly higher in neonatal colts than in neonatal fillies. Moreover, concentrations of AMH are high in prepubertal colts whereas significantly lower concentrations were detected after puberty. In intact mature stallions, season influenced AMH concentrations with significantly higher concentrations during spring and summer. Serum AMH concentrations were significantly higher in cryptorchid stallions compared with intact stallions or geldings. Finally, AMH was identified in seminal plasma of intact mature stallions, but there was no significant correlation between serum and seminal plasma AMH concentrations. In conclusion, serum AMH concentration varies with sex in the neonatal period, postnatal sexual development and season, and serum AMH concentration can be used as a biomarker for the presence of testicular tissue.

Evaluation of risk factors, management, and outcome associated with rectal tears in horses: 99 cases (1985-2006).

OBJECTIVE: To identify risk factors for rectal tears in horses; assess the effect of initiating cause on tear location, size, and distance from anus; and determine short-term survival rate among horses with various grades of rectal tears. DESIGN: Retrospective case series. ANIMALS: 99 horses. PROCEDURES: Medical records for horses with a rectal tear were reviewed, and data including age; sex; breed; cause, location, and size of the tear and its distance from the anus; tear grade; treatment; and outcome (short-term survival [ie, survival to discharge from the hospital] vs non-survival) were recorded. Data for age, sex, and breed of horses with rectal tears were compared with data for all horses evaluated at the hospital during the same interval to determine risk factors for rectal tears. RESULTS: Arabians, American Miniature Horses, mares, and horses > 9 years of age were more likely to develop a rectal tear than other breeds, males, or younger horses. Dystocia had a significant influence on rectal tear size. Location of a rectal tear and its distance from the anus were not associated with cause. Applied treatments for grade 1, 2, and 3 rectal tears were effective, unlike treatments for grade 4 rectal tears. Irrespective of treatment, the overall short-term survival rate among horses with grade 1, 2, 3, and 4 rectal tears was 100%, 100%, 38%, and 2%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Accurate identification of risk factors could help practitioners and owners implement adequate measures to prevent the development of rectal tears in horses.