Sieme H, Martinsson G, Rauterberg H, Walter K, Aurich C, Petzoldt R, Klug E
State Stud of Lower Saxony, Celle, Germany.
Reprod Domest Anim. 2003 Apr;38(2):134-40. doi: 10.1046/j.1439-0531.2003.00416.x.
The objective of this research was to improve the techniques in processing chilled and frozen-thawed horse semen. In a preliminary experiment (Exp. I), different techniques for sperm selection and preparation [Swim-up, Glass wool (GW) filtration, Glass wool Sephadex (GWS) filtration; Percoll] were tested for their suitability for equine spermatozoa and results were compared with the routine procedure by dilution (Exp. I). In the main experiment (Exp. II), two sperm preparation techniques (GWS, Leucosorb) refering to the results of Exp. I and a previous study of our group (Pferdcheilkunde 1996 12, 773) were selected for processing complete ejaculates either for cooled-storage or cryopreservation. In a third experiment (Exp. III), pregnancy rates from inseminations with semen processed according to the techniques tested in Exp. II were compared with those obtained with semen processed according to routine procedures. In Exp. I (six stallions, six ejaculates/stallion), between 48 and 92% of spermatozoa were lost following the different sperm selection procedures (p < 0.05). Preparation of sperm increased percentage of progressively motile spermatozoa (pms) [Swim-up, GW, GWS vs dilution, Percoll (p < 0.05)] and decreased percentage of sperm head abnormalities [Swim-up, GW, GWS vs dilution, Percoll (p < 0.05)] probably by not improving the quality of individual cells, but by elimination of spermatozoa of inferior quality. In Exp. II (eight stallions, three ejaculates/stallion) Leucosorb and GWS procedures allowed the filtration of large volumes (extended ejaculates) for routine laboratory practice. GWS and Leucosorb filtration resulted in increased motility, membrane integrity and sperm viability after storage of spermatozoa until 48 h at +5 degrees C when compared with control (diluted) and centrifuged semen (p < 0.05). Significantly more spermatozoa were recovered after centrifugation (87.8 +/- 15.4%) compared with GWS (63.5 +/- 18.6%) and Leucosorb filtration (53.6 +/- 22.3%). GWS or Leucosorb procedure resulted in successful cryopreservation of stallion semen without centrifugation for removal of seminal plasma. The per cycle conception rate of inseminated mares using 200 x 106 pms transferred within 8 h after collection of semen was not affected by GWS filtration or Leucosorb separation when compared with centrifugation (n.s.; Exp. III). In conclusion, GWS and Leucosorb filtration results in the improvement of semen quality and should be considered as a method for stallion semen processing. Additional studies are needed for the evaluation of potentially higher fertilizing ability of stallion spermatozoa separated by techniques for sperm selection.
本研究的目的是改进马冷冻精液和冻融精液的处理技术。在初步实验（实验I）中，测试了不同的精子筛选和制备技术[上浮法、玻璃棉（GW）过滤、玻璃棉葡聚糖凝胶（GWS）过滤； Percoll法]对马精子的适用性，并将结果与常规稀释程序进行比较（实验I）。在主要实验（实验II）中，根据实验I的结果以及本研究小组之前的一项研究（《马口腔学》1996年12期，773页），选择了两种精子制备技术（GWS、Leucosorb）来处理完整的射精精液，用于冷藏或冷冻保存。在第三个实验（实验III）中，将按照实验II中测试的技术处理的精液进行授精后的妊娠率与按照常规程序处理的精液所获得的妊娠率进行比较。在实验I（6匹种马，每匹种马6次射精）中，不同的精子筛选程序后，48%至92%的精子丢失（p<0.05）。精子制备提高了渐进性活动精子（pms）的百分比[上浮法、GW、GWS法与稀释法、Percoll法相比（p<0.05）]，并降低了精子头部异常的百分比[上浮法、GW、GWS法与稀释法、Percoll法相比（p<0.
