Factors affecting sperm recovery rates and survival after centrifugation of equine semen

Conventional centrifugation protocols result in important sperm losses during removal of the supernatant. In this study, the effect of centrifugation force (400 or 900 × g), duration (5 or 10 min), and column height (20 or 40 mL; Experiment 1); sperm concentration (25, 50, and 100 × 10[superscript 6]/mL; Experiment 2), and centrifugation medium (EZ-Mixin CST [Animal Reproduction Systems, Chino, CA, USA], INRA96 [IMV Technologies, Maple Grove, MN, USA], or VMDZ [Partnar Animal Health, Port Huron, MI, USA]; Experiment 3) on sperm recovery and survival after centrifugation and cooling and storage were evaluated. Overall, sperm survival was not affected by the combination of centrifugation protocol and cooling. Total sperm yield was highest after centrifugation for 10 min at 400 × g in 20-mL columns (95.6 ± 5%, mean ± SD) or 900 × g in 20-mL (99.2 ± 0.8%) or 40-mL (91.4 ± 4.5%) columns, and at 900 × g for 5 min in 20-mL columns (93.8 ± 8.9%; P < 0.0001). Total (TMY) and progressively motile sperm yield followed a similar pattern (P < 0.0001). Sperm yields were not significantly different among samples centrifuged at various sperm concentrations. However, centrifugation at 100 × 10[superscript 6]/mL resulted in significantly lower total sperm yield (83.8 ± 10.7%) and TMY (81.7 ± 6.8%) compared with noncentrifuged semen. Centrifugation in VMDZ resulted in significantly lower TMY (69.3 ± 22.6%), progressively motile sperm yield (63.5 ± 18.2%), viable yield (60.9 ± 36.5%), and survival of progressively motile sperm after cooling (21 ± 10.8%) compared with noncentrifuged semen. In conclusion, centrifuging volumes of ≤ 20 mL minimized sperm losses with conventional protocols. With 40-mL columns, it may be recommended to increase the centrifugal force to 900 × g for 10 min and dilute the semen to a sperm concentration of 25 to 50 × 10[superscript 6]/mL in a milk- or fractionated milk-based medium. The semen extender VMDZ did not seem well suited for centrifugation of equine semen

Analysis of airborne Doppler lidar, Doppler radar and tall tower measurements of atmospheric flows in quiescent and stormy weather

The first experiment to combine airborne Doppler Lidar and ground-based dual Doppler Radar measurements of wind to detail the lower tropospheric flows in quiescent and stormy weather was conducted in central Oklahoma during four days in June-July 1981. Data from these unique remote sensing instruments, coupled with data from conventional in-situ facilities, i.e., 500-m meteorological tower, rawinsonde, and surface based sensors, were analyzed to enhance understanding of wind, waves and turbulence. The purposes of the study were to: (1) compare winds mapped by ground-based dual Doppler radars, airborne Doppler lidar, and anemometers on a tower; (2) compare measured atmospheric boundary layer flow with flows predicted by theoretical models; (3) investigate the kinematic structure of air mass boundaries that precede the development of severe storms; and (4) study the kinematic structure of thunderstorm phenomena (downdrafts, gust fronts, etc.) that produce wind shear and turbulence hazardous to aircraft operations. The report consists of three parts: Part 1, Intercomparison of Wind Data from Airborne Lidar, Ground-Based Radars and Instrumented 444 m Tower; Part 2, The Structure of the Convective Atmospheric Boundary Layer as Revealed by Lidar and Doppler Radars; and Part 3, Doppler Lidar Observations in Thunderstorm Environments

Effects of follicular aspiration and flushing, and the genotype of the fetus on circulating progesterone levels during pregnancy in the mare.

When aspirating ovarian follicles in pregnant mares to obtain oocytes for in vitro fertilisation (IVF), the effect of the manipulation on circulating concentrations of progesterone may be an important consideration in terms of the maintenance of pregnancy. The object of this study was to compare the effects of 3 different forms of transvaginal ultrasound-guided follicle aspiration (Treatment 1, no aspiration, n = 4; Treatment 2, aspirate only follicles \u3e or =20 mm in diameter, n = 7; Treatment 3, aspirate all visible follicles, n = 7) on peripheral plasma progesterone concentrations between Days 21 and 150 of gestation in 9 mares carrying intraspecies horse and 9 mares carrying interspecies mule conceptuses. The 3 follicle aspiration treatments were applied at the peak of each follicular wave as determined by follicular mapping by means of transrectal ultrasonography on alternate days. The plasma progesterone profile in mares undergoing Treatment 1 was in close agreement with those reported previously in pregnant mares. A decline in plasma progesterone levels occurred after Day 53 of gestation in Treatments 2 and 3 mares, indicating that the follicular aspiration procedures did interfere with the formation of secondary corpora lutea. However, the levels in individual mares never dropped low enough to endanger the pregnancy. Mares carrying mule pregnancies exhibited higher mean plasma progesterone concentrations between Days 39 and 45 of gestation than mares carrying horse pregnancies, equivalent levels between Days 46 and 66 despite the lower circulating concentrations of chorionic gonadotrophin (mule CG) in their blood during this period and lower progesterone levels between Days 67 and 150 of gestation. The results indicate that the primary corpus luteum in the pregnant mare may be more sensitive to mule CG than horse CG. Furthermore, the earlier disappearance of CG from the circulation in mares carrying mule fetuses is reflected by an earlier decline in plasma progesterone concentrations in this type of equine pregnancy

Concentrations of nitric oxide in equine preovulatory follicles before and after administration of human chorionic gonadotropin

In the present study, follicular fluids of estrous mares treated with saline solution (Control) or nitric oxide synthase (NOS) inhibitors were analyzed for nitric oxide (NO), estradiol-17β (E2) and progesterone (P4) concentrations before and 36 h after administration of human chorionic gonadotropin (hCG). Follicular fluids obtained before (0 h) hCG administration from control mares had lower concentrations of NO than those obtained 36 h after administration ofhCG (58.3 ± 17.8 μmol versus 340.4 ± 57.7 μmol; P \u3c 0.05). A similar pattern was also noted for intrafollicular P4 in control mares, which had lower concentrations of intrafollicular P4 before hCG than 36 h post-hCG administration (P \u3c 0.05). As expected, E2 concentrations of control follicles sampled before hCG administration were higher than those sampled 36 h post-hCG administration (P \u3c 0.05). However, the E2 concentrations in follicles of mares treated with the NOS inhibitors Nω-nitro-L-arginine methyl ester (L-NAME) or aminoguanidine (AG) did not decrease after hCG administration, unlike those in control mares (P \u3e 0.10). In addition, mares treated with NOS inhibitors had lower intrafollicular concentrations of NO and P4 than control mares, both before and after hCG administration (P \u3c 0.05). Increased intrafollicular concentrations of NO in control, hCG-stimulated mares provide evidence for the presence of an NO-generating system in the equine preovulatory follicle that is likely upregulated following administration of hCG. © 2003 Elsevier Science Inc. All rights reserved