The objective of this study was to design a protocol to separate spermatozoa from seminal plasma of raw llama semen without prior enzymatic treatment using a single‐layer centrifugation with Androcoll‐E^™ (AE). Two experiments were performed: (a) samples were divided into three aliquots (1 ml) that were deposited on the top of 4, 5 or 6 ml of AE and were centrifuged at 800g for 20 min and (b) samples were divided into two aliquots (1 ml) that were deposited on the top of 4 ml of AE and were centrifuged at 600g or 1,000g for 20 min. Columns of 5 and 6 ml of AE showed a total sperm motility (TM) significantly lower, while in the 4 ml column, this parameter was not different from the TM of samples before the AE treatment. The percentage of spermatozoa with intact and functional membranes, normal morphology and intact acrosomes, as well as the percentages of sperm with highly condensed chromatin, was conserved (p ˃ .05) in the three column heights and in the two centrifugation speeds evaluated. In conclusion, the different column heights of AE (4, 5 and 6 ml) and the different centrifugation speeds used (600, 800 and 1,000g) allow separating spermatozoa of raw llama semen without enzymatic treatment, preserving the evaluated sperm characteristics. However, of all the studied treatments, centrifugation in the 4 ml column of AE at 800g would be the method of choice to process raw llama semen samples destined for reproductive biotechnologies.

中文翻译：

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使用Androcoll-E™无需酶处理即可分离美洲驼精子的新方案。

这项研究的目的是设计一种方案，通过使用单层离心Androcoll-E ^™（AE）进行酶处理，无需事先进行酶处理即可从生的美洲驼精液的精浆中分离精子。进行了两个实验：（a）将样品分成三等份（1 ml），分别置于4、5或6 ml AE的顶部，并以800 g离心20分钟，（b）将样品分成将两个等分试样（1 ml）分别置于4 ml AE的顶部并以600 g或1,000 g离心20分钟 在5 ml和6 ml的AE柱中，总精子活力（TM）显着降低，而在4 ml柱中，该参数与AE处理之前的样品的TM值没有差异。精子与完整且功能性的膜，形态正常和顶体完整率的百分比，以及精子与高度浓缩的染色质的百分比，是保守（p 在三柱高度和在评估的两次离心速度˃0.05）。总之，AE的柱高不同（4、5和6 ml），离心速度也不同（600、800和1,000 g）无需经过酶处理即可分离生驼羊精液的精子，从而保留评估的精子特征。但是，在所有研究的处理方法中，在4毫升AE柱中以800克离心是处理用于生殖生物技术的生美洲驼精液样品的首选方法。