Qualitative indicators of sperm from producing goats before and after cryopreservation

E. Korochkina, A. V. Trifonova, E. Finageev, D. Glavatskaya, V. Pushkina
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Abstract

Currently, artificial insemination, as one of the types of assisted reproductive technologies, is widely used in dairy and beef cattle breeding.  The  same  cannot  be  said  about such a promising direction in the agricultural industry as goat breeding. One of the limiting  factors  is  the  negative  impact  of  low temperature on the morphofunctional charateristics  of  sperm  of  breeding  goats.  The purpose of this research was to test a protocol of sperm cryopreservation of stud goats with  modification  of  prepreparation  and subsequent assessment of the quality indicators of sperm before and after its deep freezing. A comprehensive assessment of sperm quality (volume, concentration, morphology, motility)  of  goats  (n=10)  was  carried  out using generally accepted methods and protocols. The assessment of sperm quality indicators included five stages: after sperm collection, two hours after cooling, after thawing: 0 hours, 1 and 2 hours. According on the obtained results, the sperm of breeding goats ha low cryoresistance. After cryopreservation  (0,  1  and  2  hours  after  thawing), there is an increase in the number of sperm with tail damage by 7.5% (p≤0.05), 15.5% and 21.8% (p≤0.01), and also a decrease in the number of progressively moving sperm by 1.4; 1.6 and 2.5 times (p≤0.01) compared with  the  results  of  the  assessment  0  hours after collection. The use of a deep two-phase sperm freezing protocol allows maintaining the viability of sperm with a progression of movements equal to 54.2±5.1% and a number of morphologically normal sperm equal to  64.1±1.9%.  In  this  case,  the  prepreparation  of  sperm  for  the  cryopreservation  process  (current  protocol)  includes sperm  centrifugation  (mode:  7000  rpm  for 15  minutes),  removal  of  seminal  plasma, dilution  1:4  (OptiXcell  diluent),  cooling  (4 hours at 4℃); sperm cryopreservation protocol: 1. immersion of goblets with paillettes 4 cm above liquid nitrogen for 7 minutes; 2. complete immersion in liquid nitrogen.
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冷冻前后生产山羊精子的质量指标
目前,人工授精作为辅助生殖技术的一种,已广泛应用于奶牛和肉牛的繁殖。 而在山羊育种这一农业领域前景广阔的方向上,情况却并非如此。限制因素之一是低温对种羊精子形态功能特征的负面影响。 本研究的目的是对种用山羊的精子冷冻保存方案进行测试,并对精子冷冻前和冷冻后的预处理和质量指标评估进行修改。采用公认的方法和规程对山羊(n=10)的精子质量(数量、浓度、形态、活力)进行了全面评估。精子质量指标评估包括五个阶段:采精后、冷却后两小时、解冻后 0 小时、1 小时和 2 小时:0小时、1小时和2小时。结果表明,育种山羊的精子具有较低的抗冷冻性。冷冻保存后(解冻后 0、1 和 2 小时),与采集后 0 小时的评估结果相比,尾部受损的精子数量分别增加了 7.5%(p≤0.05)、15.5% 和 21.8%(p≤0.01),逐渐移动的精子数量分别减少了 1.4、1.6 和 2.5 倍(p≤0.01)。使用两相精子深度冷冻方案可以保持精子的存活率,精子运动速度为(54.2±5.1%),形态正常的精子数量为(64.1±1.9%)。 在这种情况下,精子冷冻保存前的准备工作(现行方案)包括精子离心(模式:7000 转/分,15 分钟)、去除精浆、1:4 稀释(OptiXcell 稀释液)、冷却(4℃ 4 小时);精子冷冻保存方案:1. 在液氮上方 4 厘米处浸泡高脚杯 7 分钟; 2. 完全浸泡在液氮中。
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