The Use of Equine Follicle Stimulating Hormone to Increase Equine Chorionic Gonadotropin in the Pregnant Mare

Equine chorionic gonadotropin (eCG), obtained from pregnant mares, is used for assisted reproductive technologies in laboratory rodents and livestock. The objective of the present study was to use equine follicle-stimulating hormone (eFSH) to increase the incidence of twin pregnancies, through multiple ovulations, and increase eCG. Nineteen light horse–type mares were enrolled in the study. The control group (n = 9) was bred with fresh or cooled semen and given human chorionic gonadotropin (hCG) at the time of breeding. The second group (n = 10) was given 12.5 mg of eFSH intramuscularly twice a day beginning 5–7 days after ovulation. Prostaglandin F2α was administered intramuscularly the second day of eFSH treatment. Treatment with eFSH continued until follicles were >35 mm in diameter, and mares were then given no treatment for 36 hours. The mares were then bred with fresh or cooled semen from the same stallion as the control group and given hCG. Blood samples were taken weekly from day 35 to day 105 after ovulation. Serum concentration of eCG was obtained, and data were analyzed with multivariate analysis using the mixed procedure. Significance was set at P < .05. Data were combined for all mares carrying twins and compared with those carrying singletons. The group of mares carrying twins had higher peak concentrations of eCG and higher values for area under the curve compared with mares carrying singletons (P < .05). These results suggest inducing twins could be a method used to increase eCG production.     

Supplemental l-Arginine Shortens Gestation Length and Increases Mare Uterine Blood Flow before and after Parturition

Supplementing diets with l-Arginine (Arg) improves female reproductive performance and reproductive blood flow in other species. The objectives of this study were to investigate uterine artery blood flow changes before and after parturition, and evaluate blood flow in Arg supplemented and control mares by Doppler ultrasonography. Sixteen light-horse mares began Doppler ultrasonography evaluation, 21 days before expected foaling date (EFD) and continued until day 7 postparturition. The mares under treatment (n = 8) were supplemented with 100 g Arg, once daily, beginning with 21 days before EFD. Blood flow measurements were calculated as pulsatility index (PI) and resistance index (RI) for both uterine arteries, either ipsilateral or contralateral; to uterine horn of established pregnancy; defined gravid uterine artery (GUA) and non-gravid uterine artery (NGUA), respectively. The mares under treatment had a shorter gestation length (337 ± 1.7 days) as compared to control (345 ± 2.1 days; P ≤ .05). No differences in gestation length were observed between groups when examined by age, parity, EFD, or sex of foal. Both GUA and NGUA uterine artery diameter decreased from the day before parturition to day 7 after parturition (P ≤ .001). During this time period, both PI and RI increased (P ≤ .01); indicating less blood flow. A treatment effect was observed with Arginine-treated mares having greater blood flow prepartum in the NGUA (P ≤ .001) and postpartum in the GUA (P ≤ .05), for both indices. The data demonstrated that supplementing mares with Arg shortened gestation length and increased uterine arterial blood flow before and after parturition.     

Treating the Mare with Placentitis: A Clinical Approach

Placentitis is a challenging condition in the mare to both diagnose and treat. The identification of clinical signs, use of ultrasonography, and interpretation of laboratory values (serum progestin and mammary secretion electrolytes) will be described. Current therapies for treating placental infections will be discussed. In addition, recommendations for managing the postpartum mare and neonate will be provided.     

自制孕马全血对凉山半细毛羊不育母羊的治疗观察

用自制孕马全血对体况较好的不育、适龄凉山半细毛羊母羊进行皮下注射治疗。试验表明：该自制药品对体况较好的凉山半细毛羊不育适龄母羊具有一定疗效。     

Embryo Transfer Efficiency of Quarter Horse Athletic Mares

This study was designed to compare embryo recovery rates and pregnancy rates of athletic and breeding Quarter Horse mares in a tropical warm climate. Thirty-nine barrel racing mares in training and 135 breeding mares as control donors were included. During the training period, the ambient temperature ranged from 31°C to 36°C and the average humidity from 70% to 90%. After the detection of a 35-mm follicle by ultrasound, ovulation was induced with 1 mg of deslorelin acetate (i.m), and insemination was performed 24 hours later with cooled and fresh semen from different fertile stallions. Embryos were collected on day 8 postovulation. The body temperature (rectal) was evaluated from eight athletic donor mares randomly selected from the same studied group. A total of 138 and 657 embryo collections were carried out on training and breeding mares, respectively, with a total of 105 (76%) and 466 (71%) embryos collected (P > .05). Similarly, no differences (P > .05) were observed for the pregnancy rates on day 15 (82/105, 78% vs. 370/466, 79%), and day 40 (73/105, 69% vs. 328/466, 70%) between the training and breeding donor mares. Just after training, the body temperature increased to an average of 39.4°C and the respiratory rate from 14.5 to 35.3 breaths per minute. The results of the present study showed that embryo production from appropriately trained donor mares in good condition were similar to non-athletic broodmares.     

Absence of Any Specific Diurnal Pattern in the Time of Ovulation in the Mare

This study investigates a long-held belief that mares tend to ovulate during the night. If this were so, a strategic examination regimen might replace multiple daily examinations when the time of ovulation needs to be known to within a few hours. Over 500 mares were examined at least three times daily to determine the time, to within ±4.25 hours, of 896 ovulations. The time of ovulation was determined as midway between the pre-ovulatory and post-ovulatory examinations. Three broad peaks of incidence of ovulation occurred at 5:00 am, 2:00 pm, and 10:00 pm. The interval between peaks was 9, 8, and 7 hours. during which 34.2% (5:00 am to 2:00 pm), 44.3% (2:00 pm to 10:00 pm), and 21.3% (10:00 pm to 5:00 am) of ovulations occurred. The percentage of ovulations occurring in the period 2:00 pm to 10:00 pm was significantly higher (P < .001) than in the period 10:00 pm to 5:00 am and significantly higher (P < .01) than in the period 5:00 am to 2:00 pm. The data showed that mares ovulated at all times of the day and night, with some increased incidence between 2:00 pm and 10:00 pm. Therefore, no strategic examination regimen was possible.     

Corpus Luteum Vascularization and Progesterone Production in Autumn and Winter Cycles of the Mare: Relationship Between Ultrasonographic Characteristics of Corpora Lutea and Plasma Progesterone Concentration in the Last Cycles Before Anestrus

In 20 estrus cycles of 15 mares, Color Doppler ultrasound of corpora lutea and plasma progesterone concentration (P4) was analyzed on days 6, 10, 14, 16, and 18 after ovulation. Progesterone concentration was positively correlated with corpora lutea cross-sectional area (CSA), vascularized area (VA), and index of vascularization (IV = VA/CSA) (P < .0001). Cross-sectional area, VA, and IV in corpora lutea of mares with P4 < 1 ng/mL were significantly lower than in corpora lutea of mares with P4 > 1 ng/mL. Mares with CSA < 3,473 pixels, VA < 25.5 pixels, and IV < 7.6% were prone to express P4 < 1 ng/mL 25.4, 7.9, and 7.6 times more than mares with higher values, respectively. Corpus luteums analyzed parameters differed significantly between last cycles (LCs) of the breeding season and previous cycles until day 14 after ovulation (P < .05). No significant differences were found in P4 between LCs and previous ones.     

Comparison of the Cytology Brush and Cotton Swab in the Cytological Evaluation of the Endometrium in Mares with Regard to Fertility

The aim of this study was to compare the cytology brush (CB) and cotton swab (CS) in the cytological evaluation of the endometrium in mares with regard to fertility. The study was conducted on 26 mares displaying spontaneous estrus. Samples for cytological evaluation were taken from each mare by using commercially available CS and CB. After sample collection, all mares were mated in the same estrus, and pregnancy diagnosis was performed 14-18 days after last mating. No vaginal cells were found in smears, and the CB technique yielded significantly more cells/high-power field (HPF) than the CS technique. Additionally, more cases of endometritis were diagnosed using the CB than the CS. It was also shown that the degree of inflammation is more important in diagnosis of infertility in the mare than the mere presence or absence of inflammation. In conclusion, further studies are needed to establish more precise criteria for the interpretation of inflammation, especially if samples are collected using the CB.     

Effect of Dose of Cloprostenol on the Interval to Ovulation in the Diestrous Mare: A Retrospective Study

Although the ovulatory effects of prostaglandins are well documented in several domestic species including horses, there has been little attention paid to the use of this ovulatory effect for clinical purposes. Mares often grow large follicles during the luteal phase that may or may not ovulate before progesterone levels decline. Clinical observations of administering prostaglandins in diestrous mares with large follicles suggest that there may be a negative correlation between follicular diameter and interval from treatment to ovulation. The objectives of this study were twofold: to investigate the cloprostenol dose rate effect on interval to ovulation and to confirm the negative correlation between follicular diameter and interval to ovulation. The hypothesis tested was that high doses of cloprostenol given in diestrus to mares with larger follicles would induce ovulation more rapidly than in mares given lower doses or with smaller follicles. To test the hypothesis, a total of 1,234 estrous cycles were induced with different doses of cloprostenol (ranging from 8.75 to 625 μg). All mares had at least one follicle of 28 mm or larger. Dominant follicles were followed by transrectal ultrasound examinations every other day until ovulation was detected. There was a significant effect of dose (P < .000) and follicular diameter (P < .000) on the interval from treatment to ovulation. The shortest mean interval (2.4 days) was observed after administration of 625 μg in mares with follicles 36 mm or larger, whereas the longest (4.9 days) occurred after 8.75 μg in follicles of 28 to 31 mm.     

The Effect of Manual Removal of Placenta Immediately after Foaling on Subsequent Fertility Parameters in the Mare

Retained placenta is considered to be a common problem in postpartum mares. The incidence varies from 6% to 54% depending on the breed, with higher incidence in heavy draught mares than in light-weight mares. Retained placenta has been linked to lower postpartum oxytocin concentration, impaired uterine involution, and dystocia. The objective of this study was to assess the effect of early manual removal of placenta immediately postpartum on subsequent fertility parameters (development of free intrauterine fluid, inflammatory status of endometrium, and on pregnancy rates) and to compare them with mares with spontaneous expulsion of placenta. A total of 29 mares, mainly Irish draught, were closely monitored during foaling by closed circuit television and allocated to two groups: (1) mares that expelled the placenta spontaneously within 3 hours of foaling; and (2) mares that were cleansed manually immediately after foal delivery. All mares were examined and scanned 5 and 9 days postpartum, and free intrauterine fluid was recorded; endometrial swabs were taken 9 days postpartum for endometrial cytology and culture. None of the fertility parameters analyzed showed statistical difference between groups 1 and 2. Therefore, it can be concluded that early manual removal of placenta has no detrimental effects on subsequent fertility of mares and, therefore, can be recommended when a veterinarian attends a foaling.     

Effect of Follicle Size and Follicle-Stimulating Hormone on Ovulation Induction and Embryo Recovery in the Mare

The timing of ovulation is an important component to many equine breeding strategies. The action of luteinizing hormone on ovulation induction has been recognized; however, potential effects of follicle-stimulating hormone (FSH) have been less defined. Objectives of this study were to determine whether (1) mares could be induced to ovulate follicles ≤30 mm; (2) equine FSH (eFSH) has a positive effect on ovulation induction, and (3) ovulation of small follicles would affect embryo recovery. Light-horse mares (n = 12) between 4 and 10 years of age were assigned to treatments when they had a dominant growing follicle with a mean diameter of 24, 28, or 35 ± 2 mm and endometrial edema. Treatments were (1) H35, human chorionic gonadotropin (hCG) at 35 ± 2 mm; (2) F35, eFSH at 35 ± 2 mm; (3) H28, hCG at 28 ± 2 mm; (4) FH28, eFSH and hCG at 28 ± 2 mm; (5) D28, deslorelin (gonadotropin-releasing hormone [GnRH] analog) at 28 ± 2 mm; (6) FH24/H24, hCG or eFSH and hCG at 24 ± 2 mm. Mares’ reproductive tracts were scanned at 24 ± 2-hour intervals after treatment to detect ovulation. Mares were inseminated, and embryos were collected. Numbers of mares that ovulated within 48 ± 2 hours after treatment were: H35, 8/8 (100%); F35, 8/14 (57%); H28, 7/12 (58%); FH28, 9/12 (75%); D28, 3/7 (43%) and FH/H24, 4/14 (29%). The number of mares that ovulated in 48 ± 2 hours for H35 was not different from that for FH28 but was higher (P < .05) than all other groups. Embryo recovery rates, diameters, developmental stages, and morphology scores were not different for mares ovulating 48 hours or less versus more than 48 hours after treatment or among treatment groups. Results of this study demonstrate that follicles ≤30 mm can be induced to ovulate with no effect on embryo recovery or quality, as assessed by stereomicroscopy.     

The Effect of Sulpiride Treatment During the Periovulatory Period on Prolactin Concentration and Ovulation in the Mare

Sixteen estrous cycles from 10 cyclic mares were randomly assigned to a control or sulpiride group (n = 8 each). All mares received 1,500 IU of human chorionic gonadotropin (hCG) (hour 0) during estrus with a follicular diameter ≥32 mm. Mares were scanned every 12 hours until ovulation. In the treatment group, beginning at hour 0, each mare received 1.5 mg/kg of sulpiride every 12 hours intra-muscularly until ovulation or formation of a luteinized unruptured follicle (LUF). Concentrations of luteinizing hormone (LH) and prolactin (PRL) were measured by radioimmunoassay. In each group, there were 10 preovulatory follicles for the eight cycles. The ovulation rate (9/10, 90%) was similar in the control and sulpiride groups. Two mares formed an LUF, which was first detected at hours 48 and 72 for the sulpiride and control mares, respectively. The interval from hCG to ovulation was 49.5 ± 11.1 and 43.5 ± 5.8 hours, for the control and sulpiride groups, respectively (P > .5). LH followed the typical preovulatory surge pattern, with no difference between groups (P > .5). Sulpiride administration increased PRL concentration in treated mares at 24 (P < .1), 36, and 48 hours (P < .05) after treatment. In conclusion, sulpiride administration every 12 hours increased PRL concentration in treated mares after 24 hours of the beginning of treatment. However, at this time window and concentration, PRL did not have any effect on ovulation. The control mare that developed an LUF had a PRL concentration similar to other ovulatory control mares (always ≤10 ng/mL).     

Randomized Comparative Trial of Acupuncture and Exercise Versus Uterine Ecbolics in the Treatment of Persistent Postbreeding Endometritis in Mares

Although exercise and acupuncture are frequently used therapies to treat persistent postbreeding endometritis, their efficacy to date is unproven. The objective of this study was to determine if exercise and acupuncture are effective methods to reduce intrauterine fluid and compare the effectiveness of these treatments to the use of uterine ecbolics. Twelve mares susceptible to postbreeding endometritis were enrolled in the study with a randomized cross-over design using both positive and negative controls. During each estrous cycle, mares were randomized into one of six treatment groups, including stall rest (SR), oxytocin, cloprostenol, exercise, electroacupuncture, and oxytocin and exercise. Each mare was challenged with an insemination dose of 500 × 10⁶ dead sperm at time 0 hours. Intrauterine fluid measurements were taken at 0, 4, 24, 48, 72, and 96 hours postbreeding. Associations between treatment efficacy and fluid clearance were investigated using a random-effects logistic regression model that controlled for positive uterine culture. Compared with the SR negative control, exercise was the most effective treatment and had 29.7 times increased odds of fluid clearance. The second most effective treatment was oxytocin alone, with 16.9 times increased odds of fluid clearance. This was followed by cloprostenol that had 10.6 greater odds of fluid clearance, and finally, the treatment that combined exercise with oxytocin had 8.4 times greater odds of fluid clearance. Results from this study confirm that exercise and exercise combined with oxytocin are effective methods to clear intrauterine fluid.     

Laparoscopic Anatomy of the Abdomen and Laparoscopic Ligating Loops,Electrocoagulation, and a Novel Modified Electroligation Ovariectomy in Standing Mare

The purpose of this study was to provide a detailed laparoscopic anatomy of the caudal abdominal region of mare in a standing position and to evaluate and modify a technique for standing laparoscopic ovariectomy using combination between hand-tied ligating loop and electrocoagulation techniques, as the ligating loops, electrocoagulation, and modified electroligation laparoscopic ovariectomy were applied using nine adult mares. Laparoscopy was practical and effective for direct visual examination of internal abdominal organs in the mare. Ventral dislocation of abdominal viscera after pneumoperitoneum was established with the mare in standing position, which provided an excellent inspection of the dorsal and ventral structures in the peritoneal cavity on the right and left sides. Standing laparoscopic ovariectomy using an electroligation modified method was considered a safe and effective method for hemostasis of the mesovarium, technically easy, time saving, and economical. The mean surgical time for bilateral ovariectomy was 40 ± 7.63, 60 ± 5.25, and 85 ± 6.43 minutes for electroligation-modified technique, ligating loops technique, and electrocoagulation technique, respectively.     

Effect of Timing of Postovulatory Insemination Relative to Human Chorionic Gonadotropin/Buserelin Treatment With 1 Straw of Frozen-Thawed Semen on Mare Fertility

The reproductive management of mares for frozen semen artificial insemination (AI) can be costly and labor intensive. Predicting the exact time of ovulation can be challenging even when ovulation-inducing drugs are used. The main objective of this retrospective study was to determine whether there was an effect of interval between examinations to detect ovulation on likelihood of pregnancy and early embryonic loss in mares after postovulatory breeding with a single straw of frozen/thawed semen. The second objective was to determine the efficacy of two different drugs (human chorionic gonadotropin vs. buserelin) for timely induction of ovulation. The length of the interval from penultimate check to ovulation had no significant effect on pregnancy or embryo loss rates (4 hours: 34.1% and 13.3% vs. 8 hours: 26.1% and 0% and 16 hours: 34.5% and 10%, respectively) nor did the ovulation-inducing drug used, number of the cycle, or the stallion. In conclusion, there appears to be no advantage of checking mares for ovulation during the late evening and night hours when using a postovulatory AI protocol and ovulation-inducing drugs.     

Comparison of Cell-Free Fetal DNA Plasma Content Used to Sex Determination Between Three Trimesters of Pregnancy in Torkaman Pregnant Mare

This investigation aimed to compare the cell-free fetal DNA (cffDNA) plasma present in three trimesters of pregnancy in Torkaman pregnant mare. Peripheral blood samples of 32 pregnant mares in three trimesters of pregnancy were collected in tubes containing ethylenediaminetetraacetic acid at three time points. Circulating cffDNA was extracted from 3 mL of maternal plasma. Using outer and inner primers, a conventional polymerase chain reaction was performed for the sex-determining region Y (SRY) gene present in the Y chromosome. Of the total 32 Torkaman pregnant mares, 24 were carrying male fetuses and eight were carrying female fetuses. In total, the accuracy of the test was 48.75%, 68.75%, and 75% in the first, second, and third trimesters of pregnancy, respectively. The sensitivities were 25%, 58.32%, and 66.66%, respectively, whereas their specificities were 100% in all trimesters. In conclusion, the SRY gene can permit the detection of equine fetal sex with good accuracy through cffDNA analysis in maternal plasma just in the third trimester of pregnancy, although specificity in all duration of pregnancy was 100%.     

Plasma Testosterone in Cows Related to Day of Gestation and Sex of the Foetus,and Plasma Testosterone Levels Around Parturition

No significant differences in plasma testosterone level were observed between cows carrying a male foetus and cows carrying a female foetus at any ten-day interval from day 35 of gestation until parturition. Reported higher abortion rates for male than for female foetuses would thus appear not to be due to effects of foetal testosterone on the maternal endocrine balance. In spite of a great individual variation in plasma testosterone values at similar stages of gestation, certain trends are evident. From the 35th to the 80th day of gestation the average concentration was 90–100 pg/ml. Later it rose and reached 200 pg/ml on the 180th day, remaining at this level until after partus. During the first day after parturition plasma testosterone fell significantly and stabilized around 120 pg/ml.     

Proposal and Validation of a Scale of Composite Measure Reactivity Score to Characterize the Reactivity in Horses During Handling

This study aimed to identify the parameters related to the expression of the reactivity in horses during handling and based on that proposed and validated a scale of composite measure reactivity score to characterize horse's reactivity. To this end, the first stage (S1) proposed the scale and the second (S2) validated it. In S1, 364 Lusitano horses were evaluated, 188 were adult breeding mares (4–12 years old), and 176 were foals (males/females, aged from 2 months to 2 years). During hooves trimming, vermifuge application, palpation scores were assigned to behaviors of movement, ears and eyes position, breathing, vocalization, and urination. A response parameter called reactivity was attributed to each animal, ranging from score 1 (nonreactive/calm) to score 4 (very reactive/aggressive). The verification of the possible parameters (age, behavior), which explains the response parameter (reactivity), was taken using ordinal proportional odds model. Movement, breathing, ears and eyes position, vocalization, and age appear to explain the reactivity of horses during handling (P < .01). Therefore, based on these parameters, it was possible to propose two scales of composite measure reactivity score: one to characterize the mares and another the foals. On S2, the proposed scale was validated by the simultaneous application of Forced Human Approach Test, another commonly used test to evaluate the reactivity in horses, with a correlation of 0.97 (P < .05). The assessment of the reactivity of horses during handling by a composite measure reactivity score scale is valid, and easy to apply, without disrupting daily routine and override the impact of individual differences.     

Doppler evaluation of maternal and fetal vessels during normal gestation in the bitch

The aim of this work is to evaluate the haemodynamic characteristics of maternal and fetal vessels during normal pregnancy in bitches, using Colour and Pulsed wave Doppler ultrasonography, in order to obtain more information about maternal and fetal circulation. The blood waveforms of the uteroplacental arteries, aorta, caudal cava vein and umbilical cord of the fetuses were recorded weekly in 16 pregnant bitches. Also, the measurements of Peak Systolic, End Diastolic Velocities, Resistance and Pulsatility Indices were carried out. Uteroplacental blood flow was biphasic while the ones of the umbilical artery and aorta were first systolic and then diastolic. The cava showed a typical waveform of venous vessels. During gestation the EDV and PSV of fetal vessels increased (alpha<0.05) while the PI and RI of all vessels examined decreased (alpha<0.05) except for the IP of the Aorta. The Doppler ultrasonography was used to study the characteristics of maternal and fetal vessel flow and their progressive changes during pregnancy. This study can be considered a further contribution in diagnosing and monitoring high-risk pregnancies in Veterinary Medicine.     

Effect of handling-induced stress on the results of spectral Doppler echocardiography in falcons

In order to evaluate the effects of habituation to handling on the Doppler-derived velocity of diastolic ventricular inflow and systolic aortic outflow, falcons that were accustomed to regular handling were compared with falcons that had been handled very little. The mean heart rate of the frequently handled birds was 111 beats per minute less than the non-habituated birds. All the mean blood flow velocities recorded were much higher in the non-habituated birds than those accustomed to handling. Taking into account the difficulty of objectively quantifying the level of stress suffered by any individual bird, the question is, if meaningful clinical or reference values for Doppler-derived blood flow velocities may be obtained in unsedated or non-anaesthetized birds?