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Abstract：
The advances in Assisted Reproductive Technologies (ARTs) applied in South American camelid species are still scarce. The aim of this study was to compare the effects of three semen extenders, before and after the cryopreservation of spermatozoa obtained from the vas deferens, on sperm quality parameters and in vitro fertilization rates of llama (Lama glama) oocytes. Mature fertile llama males (Lama glama; n = 6; age: 48-60 mo.; BCS: ~2.7) were included in the study. Sperm samples were collected from each male using the surgical technique of the vas deferens deviation. Then, the sperm samples were pooled and diluted with the Tris-EY, Andromed®, or BioxCell® extender in order to subsequently carry out the sperm cryopreservation process. The sperm quality assessment related to each extender was performed before and after cryopreservation with regard to sperm morphological abnormalities, acrosome integrity, sperm viability, membrane permeability, and sperm motility traits. Moreover, in vitro fertilization (IVF) procedures were carried out to evaluate the in vitro fertility of the cryopreserved sperm samples using each extender. Overall, significant differences were observed before and after cryopreservation regarding acrosome integrity, sperm viability, membrane permeability, and sperm motility traits among the extenders used, where Tris-EY and Andromed® were better than BioxCell® (p < 0.05); however, no differences were observed regarding the sperm morphological abnormalities among extenders (p > 0.05). Moreover, multiple differences were observed with regard to the velocity and linearity kinematic parameters obtained by computerized analysis before and after the cryopreservation process, irrespective of the extender used (p < 0.05). Finally, differences were observed regarding the in vitro fertilization rates among the different extender-derived samples (p < 0.05). In conclusion, the sperm quality using Tris-EY and Andromed® was better before and after cryopreservation compared to that using BioxCell®. Although the number of fertilized oocytes obtained after the IVF process between Tris-EY and Andromed® was similar, Andromed®-derived samples showed the best sperm quality results before and after cryopreservation. This indicates that the cryopreservation extender is a determining factor in significantly improving in vitro fertilization rates when using sperm samples obtained from vas deferens in llama (Lama glama) males.
摘要：
在南美骆驼科物种中应用的辅助生殖技术（ART）的进展仍然很少。这项研究的目的是比较三种精液补充剂的效果，在从输精管获得的精子冷冻保存之前和之后，美洲驼（Lamaglama）卵母细胞的精子质量参数和体外受精率。成熟的可育美洲驼雄性（Lamaglama；n=6；年龄：48-60个月。；BCS：~2.7）纳入研究。使用输精管偏离的手术技术从每个男性收集精子样本。然后，收集精子样本并用Tris-EY稀释,Andromed®,或BioxCell®扩展器，以便随后进行精子冷冻保存过程。在冷冻保存之前和之后对精子形态异常进行与每个延长体相关的精子质量评估，顶体完整性,精子活力，膜渗透性，和精子运动特性。此外，进行体外受精（IVF）程序，以评估使用每种补充剂冷冻保存的精子样品的体外生育力。总的来说,在冷冻保存前后观察到有关顶体完整性的显着差异，精子活力，膜渗透性，和使用的延伸剂中的精子运动特性，其中Tris-EY和Andromed®优于BioxCell®（p<0.05）；然而，在精子形态异常方面没有观察到差异（p>0.05）。此外，在冷冻保存过程之前和之后，通过计算机分析获得的速度和线性运动学参数观察到多个差异，与使用的扩展器无关(p<0.05)。最后,在不同的延伸剂来源的样品中观察到体外受精率的差异（p<0.05）。总之,与使用BioxCell®相比，使用Tris-EY和Andromed®冷冻保存前后的精子质量更好。尽管Tris-EY和Andromed®在IVF过程后获得的受精卵母细胞数量相似，Andromed®衍生的样品在冷冻保存之前和之后显示出最佳的精子质量结果。这表明，当使用从美洲驼（Lamaglama）雄性输精管获得的精子样品时，冷冻保存扩展器是显着提高体外受精率的决定因素。
