Batista M, Niño T, Alamo D, Castro N, Santana M, González F, Cabrera F, Gracia A
Obstetrics and Reproduction, Faculty of Veterinary of Las Palmas, Transmontaña s/n, 35413 Arucas, Spain.
Theriogenology. 2009 May;71(8):1307-15. doi: 10.1016/j.theriogenology.2008.12.024. Epub 2009 Feb 25.
The semen of five Majorera breed bucks was collected and processed to reach a final concentration of 200 x 10(6)spermatozoa/straw in the extender containing 4% of glycerol and 12% of egg yolk. Two freezing techniques were assessed: (LN) straws were frozen and stored in liquid nitrogen, and (ULF) straws were frozen and stored in the ultra-low freezer at -152 degrees C. Semen quality (sperm motility, acrosome integrity and abnormal sperm cells percentages) was determined for different storage times (1, 30, 90 and 365 days of cryopreservation). Thereafter, 150 Majorera goats were assigned to four experimental groups: for groups LN-1 (n=40) and LN-6 (n=35), the goats were transcervically inseminated with frozen-thawed semen stored for 1 and 6 months in liquid nitrogen, respectively, while for groups ULF-1 (n=40) and ULF-6 (n=35), the goats were transcervically inseminated with frozen-thawed semen stored for 1 and 6 months in an ultra-low freezer at -152 degrees C, respectively. The pregnancy rate was determined by transabdominal ultrasound scanning; in addition, the kidding rate and prolificacy were recorded at parturition. In vitro results showed that the freezing protocol did not affect sperm quality with similar values for up to 1 year of cryopreservation. The kidding rates were not significantly different between experimental groups (43.6%, 38.5%, 42.8% and 40.0% for groups LN-1, ULF-1, LN-6 and ULF-6, respectively). In all experimental groups, the kidding rate and prolificacy were significantly higher (p<0.01) in multiparous than in nulliparous goats. Therefore, the in vitro results and fertility trials confirmed the efficiency of the ULF technique for freezing and storage of goat semen.
采集了5只马约雷拉品种公羊的精液,并进行处理,使其在含有4%甘油和12%蛋黄的稀释液中的最终浓度达到200×10⁶精子/细管。评估了两种冷冻技术:(液氮法)细管在液氮中冷冻并储存,(超低温法)细管在-152℃的超低温冰箱中冷冻并储存。测定了不同储存时间(冷冻保存1、30、90和365天)的精液质量(精子活力、顶体完整性和异常精子细胞百分比)。此后,将150只马约雷拉山羊分配到四个实验组:对于LN-1组(n=40)和LN-6组(n=35),山羊分别经子宫颈授精冷冻解冻后在液氮中储存1个月和6个月的精液,而对于ULF-1组(n=40)和ULF-6组(n=35),山羊分别经子宫颈授精冷冻解冻后在-152℃的超低温冰箱中储存1个月和6个月的精液。通过经腹超声扫描确定妊娠率;此外,在分娩时记录产羔率和产仔数。体外结果表明,冷冻方案在长达1年的冷冻保存期内对精子质量没有影响,各项指标值相似。各实验组之间的产羔率没有显著差异(LN-1组、ULF-1组、LN-6组和ULF-6组分别为43.6%、38.5%、42.8%和40.0%)。在所有实验组中,经产山羊的产羔率和产仔数均显著高于初产山羊(p<0.01)。因此,体外结果和生育试验证实了超低温技术在山羊精液冷冻和储存方面的有效性。