Ahmed Hussain, Andrabi S Murtaza Hassan, Jahan Sarwat
Animal Reproduction Laboratory, Animal Sciences Institute, National Agricultural Research Centre, Islamabad, Pakistan; Reproductive Physiology Laboratory, Department of Animal Sciences, Quaid-i-Azam University, Islamabad, Pakistan.
Animal Reproduction Laboratory, Animal Sciences Institute, National Agricultural Research Centre, Islamabad, Pakistan.
Theriogenology. 2016 Oct 1;86(6):1516-1522. doi: 10.1016/j.theriogenology.2016.05.010. Epub 2016 May 24.
The present study was carried out to assess various postthaw semen quality parameters for the prediction of fertility in buffalo bull during low-breeding season. Semen (30 ejaculates) was collected from five adult buffalo bulls with artificial vagina (42 °C). Sperm motility parameters, velocity distribution, motion kinematics, and subpopulations were analyzed by computer-aided sperm motion analyzer (CASA). Moreover, sperm visual motility, supravital plasma membrane integrity, viability/acrosome integrity, viability/mitochondrial transmembrane potential, DNA fragmentation/integrity, and morphology were analyzed by phase-contrast microscope, supravital hypoosmotic swelling test, Trypan blue/Giemsa staining, propidium iodide/"5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanine iodide" (JC-1) fluorochromes, neutral comet assay/acridine orange assay and wet mount technique, respectively. Outcome of 528 inseminations was analyzed for in vivo fertility. Pearson's correlation coefficients revealed that sperm progressive motility (%), rapid velocity (%), average path velocity (μm/s), straight line velocity (μm/s), subpopulation one (most rapid, and progressive) of motile spermatozoa (%), supravital plasma membrane integrity (%), and viable spermatozoa with intact acrosome (%) were significantly correlated with in vivo fertility (r = 0.64, P < 0.01; r = 0.57, P < 0.01; r = 0.52, P < 0.01; r = 0.56, P < 0.01; r = 0.73, P < 0.001; r = 0.74, P < 0.001; r = 0.88, P < 0.001); whereas nonviable spermatozoa with damaged acrosome or low-mitochondrial transmembrane potential and comet length (μm) of neutral comet assay were negatively associated with in vivo fertility (r = -0.79, r = -0.75, P < 0.001, and r = -0.60, P < 0.05, respectively). Multiple regression analysis reported that combination of semen quality parameters as predictor of fertility were better (R(2) adjusted = 81.30%, P < 0.001) as compared with single parameter (R(2) adjusted = 50.20%, P < 0.007). It is concluded that assessment of CASA parameters and some other sperm structural and functional parameters, that is, integrity of plasma membrane and acrosome, and transmembrane potential of mitochondria were able to predict the in vivo fertility of water buffalo bull during low-breeding season.
本研究旨在评估冻融后精液的各种质量参数,以预测水牛公牛在低繁殖季节的生育能力。从五头成年水牛公牛身上用人工阴道(42°C)采集精液(30次射精)。通过计算机辅助精子运动分析仪(CASA)分析精子活力参数、速度分布、运动动力学和亚群。此外,通过相差显微镜、活体低渗肿胀试验、台盼蓝/吉姆萨染色、碘化丙啶/“5,5',6,6'-四氯-1,1',3,3'-四乙基苯并咪唑基碳菁碘化物”(JC-1)荧光染料、中性彗星试验/吖啶橙试验和湿片技术分别分析精子的视觉活力、活体细胞膜完整性、活力/顶体完整性、活力/线粒体跨膜电位、DNA片段化/完整性和形态。分析了528次授精的体内生育结果。皮尔逊相关系数显示,精子前向运动率(%)、快速速度(%)、平均路径速度(μm/s)、直线速度(μm/s)、活动精子的亚群一(最快且最具前向性)(%)、活体细胞膜完整性(%)和顶体完整的活精子(%)与体内生育能力显著相关(r = 0.64,P < 0.01;r = 0.57,P < 0.01;r = 0.52,P < 0.01;r = 0.56,P < 0.01;r = 0.73,P < 0.001;r = 0.74,P < 0.001;r = 0.88,P < 0.001);而顶体受损或线粒体跨膜电位低的无活力精子以及中性彗星试验的彗星长度(μm)与体内生育能力呈负相关(r = -0.79,r = -0.75,P < 0.001,以及r = -0.60,P < 0.05)。多元回归分析表明,与单个参数相比,将精液质量参数组合作为生育能力预测指标更好(调整后的R(2) = 81.30%,P < 0.001)(调整后的R(2) = 50.20%,P < 0.007)。得出结论,评估CASA参数以及其他一些精子结构和功能参数,即细胞膜和顶体的完整性以及线粒体的跨膜电位,能够预测水牛公牛在低繁殖季节的体内生育能力。