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使用自然感染的卵母细胞对体外胚胎生产中猪圆环病毒2型感染的评估。

Evaluation of porcine circovirus type 2 infection in in vitro embryo production using naturally infected oocytes.

作者信息

Weng Xiao-Gang, Liu Yan, Zhou Si-Han, Zhang Yu-Ting, Shao Yu-Tong, Xu Qian-Qian, Liu Zhong-Hua

机构信息

Key Laboratory of Animal Cellular and Genetics Engineering of Heilongjiang Province, College of Life Science, Northeast Agricultural University, Harbin, 150030, Heilongjiang, PR China.

Key Laboratory of Animal Cellular and Genetics Engineering of Heilongjiang Province, College of Life Science, Northeast Agricultural University, Harbin, 150030, Heilongjiang, PR China.

出版信息

Theriogenology. 2019 Mar 1;126:75-80. doi: 10.1016/j.theriogenology.2018.12.009. Epub 2018 Dec 4.

Abstract

In vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT) are important breeding techniques for livestock. High-quality MII oocytes produced from in vitro maturation (IVM) are required for the two techniques listed above. The ovaries used for IVM operations are primarily acquired from commercial abattoirs, and the pathogen status of slaughtered animals becomes an unavoidable issue. Our previous monitoring data showed that porcine circovirus type 2 (PCV-2) is the main pathogen present in ovaries from abattoirs. However, the characteristics and effects of PCV-2 infection in oocyte maturation and in vitro production (IVP) of embryos are unclear, and currently there are no relevant studies. Therefore, the aim of this study was to determine the PCV-2 infection pattern and determine whether it affects oocyte in vitro maturation and IVP embryo development. More than five hundred ovaries and five thousand oocytes were utilized in the present study. Polymerase chain reaction (PCR) was used to detect PCV-2 DNA in ovaries, follicular fluid (FF), oocytes, cumulus cells and IVP embryos. The effects of viral infections on the rate of oocyte maturation and IVP embryo development were evaluated. We also analyzed the number of copies of the virus in the IVM and IVP process by absolute quantitative fluorescence PCR. Our study showed that the prevalent virus subgenotype in ovaries was PCV-2a. PCV-2a infection did not significantly affect ovarian/oocyte morphology and maturation. Moreover, virus infection did not have a significant effect on the development of the IVP embryos except for a reduction in IVF blastocyst cell numbers. Further tests showed that the viral copy numbers fluctuated at different stages between the IVP embryos and culture medium. For the first time, this study identified the infection pattern of naturally sourced PCV-2 in the course of oocyte maturation and embryo development.

摘要

体外受精(IVF)和体细胞核移植(SCNT)是家畜重要的繁殖技术。上述两种技术都需要体外成熟(IVM)产生的高质量MII期卵母细胞。用于IVM操作的卵巢主要从商业屠宰场获取,屠宰动物的病原体状况成为一个不可避免的问题。我们之前的监测数据表明,猪圆环病毒2型(PCV - 2)是屠宰场卵巢中的主要病原体。然而,PCV - 2感染在卵母细胞成熟和胚胎体外生产(IVP)中的特征及影响尚不清楚,目前也没有相关研究。因此,本研究的目的是确定PCV - 2的感染模式,并确定其是否影响卵母细胞体外成熟和IVP胚胎发育。本研究使用了五百多个卵巢和五千多个卵母细胞。采用聚合酶链反应(PCR)检测卵巢、卵泡液(FF)、卵母细胞、卵丘细胞和IVP胚胎中的PCV - 2 DNA。评估病毒感染对卵母细胞成熟率和IVP胚胎发育的影响。我们还通过绝对定量荧光PCR分析了IVM和IVP过程中病毒的拷贝数。我们的研究表明,卵巢中流行的病毒亚型是PCV - 2a。PCV - 2a感染对卵巢/卵母细胞形态和成熟没有显著影响。此外,除了体外受精囊胚细胞数量减少外,病毒感染对IVP胚胎的发育没有显著影响。进一步的测试表明,IVP胚胎和培养基中病毒拷贝数在不同阶段有所波动。本研究首次确定了天然来源的PCV - 2在卵母细胞成熟和胚胎发育过程中的感染模式。

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