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用于捷克金斑母鸡遗传资源项目冷冻保存程序的商业家禽精液稀释剂比较

Comparison of Commercial Poultry Semen Extenders Modified for Cryopreservation Procedure in the Genetic Resource Program of Czech Golden Spotted Hen.

作者信息

Petričáková Kristýna, Janošíková Martina, Ptáček Martin, Zita Lukáš, Savvulidi Filipp Georgijevič, Partyka Agnieszka

机构信息

Department of Animal Sciences, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kamýcká 129, 165 00 Praha, Czech Republic.

Faculty of Veterinary Medicine, Department of Reproduction and Clinic of Farm Animals, Wroclaw University of Environmental and Life Sciences, pl. Grunwaldzki 49, 50-366 Wroclaw, Poland.

出版信息

Animals (Basel). 2022 Oct 21;12(20):2886. doi: 10.3390/ani12202886.

Abstract

Spermatozoa cryoconservation represents an important strategy for partial in vitro or rescue programs designed for threatened livestock populations. The procedure for the semen cryopreservation of the Czech Golden Spotted Hen was proposed due to the lower fertilization rate of poultry semen compared to mammalian species. The aim of this study was to compare commercial extenders designed for liquid storage preservation with the use of a predefined cryoprotectant, and, thus, to propose an important tool for the procedure of the semen cryopreservation of the Czech Golden Spotted Hen. Ejaculates were sampled from four roosters during five semen collection days. The samples were frozen in Poultry media, Raptac and NeXcell extenders supplemented with a 9% N-methylacetamide (NMA) cryoprotectant. Sperm parameters of the total motility (MOT; %), plasma membrane and acrosome intactness (PAI; %), plasma membrane damage (%), acrosome damage (%) and cells with plasma membrane and acrosome damage (%) were assessed using a mobile mCASA analyzer and flow cytometer after the cryopreservation of the insemination doses (IDs). For Poultry media (PAI = 51.11%; MOT = 23.58%) and Raptac (PAI = 52.04%; MOT = 23.13%) extenders with the addition of an NMA cryoprotectant, the comparable results were detected after thawing. For NexCell media, the results were poor (PAI = 7.07%; MOT = 3.83%). Our results indicated two extenders suitable for the cryopreservation procedure, with the applied modification.

摘要

精子冷冻保存是为濒危家畜种群设计的部分体外或拯救计划的一项重要策略。由于家禽精液的受精率低于哺乳动物,因此提出了捷克金斑母鸡精液冷冻保存的程序。本研究的目的是比较为液体储存保存设计的商业稀释剂与使用预定义的冷冻保护剂,从而为捷克金斑母鸡精液冷冻保存程序提出一种重要工具。在五个采精日期间从四只公鸡采集精液样本。样本在添加9% N - 甲基乙酰胺(NMA)冷冻保护剂的家禽培养基、Raptac和NeXcell稀释剂中冷冻。在输精剂量(IDs)冷冻保存后,使用便携式mCASA分析仪和流式细胞仪评估总活力(MOT;%)、质膜和顶体完整性(PAI;%)、质膜损伤(%)、顶体损伤(%)以及质膜和顶体均受损的细胞(%)等精子参数。对于添加了NMA冷冻保护剂的家禽培养基(PAI = 51.11%;MOT = 23.58%)和Raptac(PAI = 52.04%;MOT = 23.13%)稀释剂,解冻后检测到了可比的结果。对于NexCell培养基,结果较差(PAI = 7.07%;MOT = 3.83%)。我们的结果表明,经过应用的改良,有两种稀释剂适用于冷冻保存程序。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db96/9598354/2e11a9044d0c/animals-12-02886-g001.jpg

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