Effects of age, parity, and pregnancy abnormalities on foal birth weight and uterine blood flow in the mare.

Color Doppler sonography has become routine for the evaluation of high-risk pregnancies in human medicine. Previous studies documenting uterine blood flow parameters in the pregnant mare have found a decrease in peripheral blood flow resistance in the first pregnancy weeks and an increase in uterine blood flow, especially in the last trimester of pregnancy. However, these studies involved only a small number of mares. No naturally occurring pregnancy abnormalities occurred that would allow blood flow changes to be retrospectively examined and analyzed. The objective of the present study was to monitor the diameter of the uterine artery, uterine blood flow, and the combined thickness of the uterus and placenta (CTUP) throughout gestation in a large number of pregnant mares of different age and parity. In the present study, 51 warmblood mares were examined by ultrasonography on Days 16 and 30, at monthly intervals until Day 300, and then every 10 days from Day 300 until parturition. After localization of the uterine artery ipsilateral and contralateral to the conceptus, the diameter of each artery, the uterine blood flow (pulsatility index [PI], blood flow volume [BFV], and the presence of early diastolic notch), and the CTUP were measured and correlated to placental and foal birth weight after delivery. Furthermore, the effect of age (3-7 years [n = 16], 8-11 years [n = 17], 12-16 years [n = 18]) and parity (0-2 foals [n = 22], 3-4 foals [n = 15], 5-8 foals [n = 14]) on these parameters were analyzed. The diameter of the uterine artery increased more than threefold in the ipsilateral artery (0.40 ± 0.07-1.33 ± 0.08 cm) and 2.7-fold in the contralateral artery (0.39 ± 0.07-1.07 ± 0.08 cm [P < 0.0001]). The early diastolic notch disappeared in the pulse waves in 98% of the ipsilateral arteries and 85.7% in the contralateral arteries on Day 150 when placentation is complete. Blood flow volume increased 50-fold in the ipsilateral artery during pregnancy and increased dramatically in the last trimester. The median foal weight was 52.6 kg. Mares with heavier foals (>52.6 kg) had a 1.38-fold higher BFV in the last 2 months (P < 0.05) compared with lighter foals. Pulsatility index decreased 2-fold until completion of placentation at around Day 150 and continued to decline until Day 240 where it then stayed constant and at a low level until delivery. Age predominantly influenced PI, whereas the diameter of the uterine arteries, which is correlated to BFV (r ipsilateral = 0.919, P < 0.0001 and r contralateral = 0.909, P < 0.000), was strongly affected by parity. Four mares spontaneously aborted (Days 200, 208, 213, and 246) and four mares spontaneously developed placentitis that was diagnosed by the presence of an increased CTUP and/or placental pathology after delivery. Although not statistically relevant, the aborting mares showed a slightly increased total BFV, but no differences in PI were seen compared with mares without abnormalities of pregnancy. Mares that developed placentitis had a late (Days 150-210) disappearance of the early diastolic notch and an increased PI in the first half of pregnancy. In conclusion, the study documented differences in uterine artery diameter and blood flow in a large number of pregnant mares. Furthermore, this is the first known report to document uteroplacental blood flow changes associated with naturally occurring placentitis and abortion in mares.

Influence of transrectal and transabdominal ultrasound examination on salivary cortisol, heart rate, and heart rate variability in mares.

Pregnancy diagnostics in equine reproduction are routinely performed using transrectal ultrasonography, although it is also possible to visualize the fetus by transabdominal ultrasound examinations from the 90th day of gestation onward. We hypothesized that ultrasound examinations may stress the mare and that the gestational stage status and lactation may influence the mare's stress reaction. To investigate the stress reaction, 25 thoroughbred mares of different age, pregnancy and lactational status underwent a transrectal examination. In pregnant mares, an additional transabdominal examination was performed. Salivary cortisol concentration, mean heart rate, and heart rate variability of mares were assessed to evaluate the reactions of hypothalamic-pituitary-adrenal (HPA) axis and of the autonomic nervous system. Significant differences were observed between lactating and nonlactating mares; with a lower responsiveness to stress in lactating mares. The transrectal ultrasound examination in nonlactating mares induced a significant increase in salivary cortisol (P < 0.05), and in the heart rate variability parameter, ratio of low to high frequencies (P < 0.05). This reflects an activation of the HPA axis and a shift to more sympathetic dominance. In contrast, a transabdominally performed pregnancy check did not induce an activation of the HPA axis over basal level but increased the mean heart rate and low to high frequency ratio. The results of this study indicate that checks of advanced pregnancies can be easily performed by transabdominal ultrasonography. With regard to animal welfare, this technique should be preferred during midgestation in nonlactating mares.

Evaluation of pregnancy and foaling rates after reduction of twin pregnancy via transvaginal ultrasound-guided aspiration in mares.

OBJECTIVE: To assess pregnancy and live foaling rates after reduction of twin pregnancy via transvaginal ultrasound-guided aspiration (TUA) in mares and evaluate effects of gestational period, localization of conceptuses, fluid aspiration volume, and combination of TUA with embryonic or fetal puncture on these outcomes. DESIGN: Clinical trial. ANIMALS: 44 mares pregnant with twins (25 to 62 days of gestation). PROCEDURES: TUA was performed in all mares and combined with embryonic or fetal puncture in 13. Follow-up ultrasonographic examinations were performed by referring veterinarians. Effects of gestational period and TUA-related variables on pregnancy and foaling rates were assessed. RESULTS: Singleton pregnancy (32/44 [73%]), persistent twin pregnancy (3/44 [7%]), or loss of both conceptuses (9/44 [20%]) was confirmed 5 to 7 days after TUA. Two mares with persistent twin pregnancy underwent another TUA, and 1 underwent prostaglandin F(2α)-induced abortion; these were excluded from subsequent analyses. Eighteen of 24 mares reevaluated 3 to 4 weeks after TUA were pregnant with 1 conceptus. Twenty of 41 (49%) mares delivered live singleton foals. Gestational period of TUA did not affect early pregnancy rates. Foaling rate for mares that underwent TUA after day 42 (0/5) was lowest of all groups and was significantly lower than that of mares that underwent TUA between days 31 and 35 (9/14). Three of 13 mares that underwent TUA with embryonic or fetal puncture delivered live foals, compared with 17 of 28 that underwent TUA alone. Effects of other variables were nonsignificant. CONCLUSIONS AND CLINICAL RELEVANCE: TUA was effective for reduction of twin pregnancy in mares, with best results achieved before gestational day 43.

Iodixanol density gradient centrifugation for selecting stallion sperm for cold storage and cryopreservation.

Density gradient centrifugation can be used for selection of sperm of superior quality and removal of seminal plasma for use in artificial insemination. In this study, the use of two-layer iodixanol density gradient centrifugation was evaluated for processing of stallion semen. The protocol includes centrifugation through a 16% iodixanol top layer of 1.090 g mL(-1) and collection of motile and intact sperm on a 30% iodixanol bottom layer of 1.165 g mL(-1). Sperm recovery and effects on sperm quality were determined during cold storage as well as after cryopreservation and compared with ordinary dilution and centrifugation. Two-layer iodixanol density gradient centrifugation allows for selection of greater percentages of morphologically normal and progressively motile sperm compared to ordinary centrifugation. This likely results from collecting sperm on the bottom layer that functions as cushion fluid, which prevents mechanical forces as occur when sperm are packed in a pellet. In addition, percentages of membrane and chromatin integrity are increased when cells are selected based on their density via centrifugation through the top and bottom layers. Removal of seminal plasma and increased initial percentages of motile and membrane intact sperm after iodixanol density gradient centrifugation also result in greater percentages of progressively motile and membrane intact sperm during cold storage as well as after freezing and thawing. In conclusion, the two-layer iodixanol density gradient centrifugation protocol described in this manuscript allows for selection of stallion sperm with greater survival rates for cold storage and cryopreservation.