Campos Lívia Batista, Peixoto Gislayne Christianne Xavier, da Silva Andreia Maria, Souza Ana Liza Paz, de Souza Castelo Thibério, Maia Keilla Moreira, Pereira Alexsandra Fernandes, Silva Alexandre Rodrigues
Laboratory of Animal Germplasm Conservation, Federal Rural University of Semi-Arid (UFERSA), BR 110, Km 47, Mossoro, RN, 59625-900, Brazil.
Laboratory of Animal Germplasm Conservation, Federal Rural University of Semi-Arid (UFERSA), BR 110, Km 47, Mossoro, RN, 59625-900, Brazil.
Theriogenology. 2017 Apr 1;92:57-62. doi: 10.1016/j.theriogenology.2017.01.008. Epub 2017 Jan 5.
In collared peccaries, the development of artificial insemination (AI) is scarce, requiring search for alternative methods for the evaluation of sperm fertilizing ability. Thus, the aims of this study were to estimate the binding capability of collared peccaries sperm, using swine oocytes and the egg perivitelline membrane, and to evaluate the prognostic value of sperm parameters on the in vitro interactions among sperm and heterologous substrates. Eleven ejaculates were collected by eletroejaculation and evaluated for viability and morphology by light microscopy, for functionality by hypo-osmotic swelling test, for plasma membrane integrity by epifluorescence microscopy, and for sperm motility by computerized analysis. Subsequently, for analysis of the in vitro interactions, sperm samples were cultured in an incubation medium with swine oocytes and egg perivitelline membrane for 18 h and 20 min, respectively, at 38.5 °C and humidified atmosphere. The sperm-oocyte interaction rate was 100% with sperm penetrating 19.8+ 5.5% of oocytes. The average values of bound sperm and penetrated sperm per oocyte were 39.4 + 4.6 and 2.5 + 0.7, respectively. Already for perivitelline membrane binding assay, all samples presented sperm bound (100%) with average of 140.6 ± 19.4 bound sperm (range 33.9-308.7). Moreover, positive correlations were observed for the number of sperm bound to swine oocytes and osmotic response (r = 68.5%; P = 0.02), membrane integrity (r = 65.1%; P = 0.03), and straightness (r = 66.5%; (P = 0.03), as weel as for the number of sperm bound to egg perivitelline membrane and sperm viability (r = 74.0%; P = 0.01), total motility (r = 63.6%; P = 0.04), and linearity (r = 70.5%; P = 0.02). Finally, a negative correlation among slow (r = -80.5%; P = 0.01) and static (r = -84.3%; P = 0.01) sperm with the egg perivitelline membrane was observed. In conclusion, swine oocytes and perivitelline membrane can be used as indicators for the functional evaluation of the binding capability of sperm derived from collared peccaries. These tests could be incorporated into the routine of semen technologies.
在领西猯中，人工授精（AI）技术的发展较为匮乏，因此需要寻找评估精子受精能力的替代方法。本研究的目的是利用猪的卵母细胞和卵周膜来评估领西猯精子的结合能力，并评估精子参数对精子与异源底物体外相互作用的预后价值。通过电刺激采精收集了11份精液样本，利用光学显微镜评估精子的活力和形态，通过低渗肿胀试验评估精子功能，利用落射荧光显微镜评估精子质膜完整性，利用计算机分析评估精子活力。随后，为了分析体外相互作用，将精子样本分别与猪卵母细胞和卵周膜在38.5℃、湿润的培养箱中孵育18小时20分钟。精子与卵母细胞的相互作用率为100%，有19.8±5.5%的卵母细胞被精子穿透。每个卵母细胞上结合的精子和穿透的精子的平均值分别为39.4±4.6和2.5±0.7。在卵周膜结合试验中，所有样本均有精子结合（100%），每个样本平均有140.6±19.4个精子结合（范围为33.9 - 308.7）。此外，观察到与猪卵母细胞结合的精子数量与渗透反应（r = 68.5%；P = 0.02）、膜完整性（r = 65.1%；P = 0.03）和直线性（r = 66.5%；P = 0.03）呈正相关，与卵周膜结合的精子数量与精子活力（r = 74.0%；P = 0.01）、总活力（r = 63.6%；P = 0.04）和线性度（r = 70.5%；P = 0.02）也呈正相关。最后，观察到慢速（r = -80.5%；P = 0.01）和静止（r = -84.3%；P = 0.01）精子与卵周膜之间呈负相关。总之，猪卵母细胞和卵周膜可作为评估领西猯精子结合能力功能的指标。这些检测方法可纳入精液技术的常规操作中。
