Rijsselaere Tom, Van Soom Ann, Maes Dominiek, de Kruif Aart
Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.
Theriogenology. 2003 Nov;60(8):1553-68. doi: 10.1016/s0093-691x(03)00171-7.
Computerized measuring devices are needed to assess canine semen quality objectively both for research and practical purposes. As internal image settings may influence the results considerably, the effect of different technical settings and semen processing on the parameters assessed by the Hamilton-Thorne Ceros 12.1 semen analyzer (HTR Ceros 12.1) was investigated. The frame rate (15, 30 or 60 frames/s) significantly (P<0.05) influenced most of the measured motility characteristics in experiment 1 while no differences in the motility parameters were found using a different sampling duration (0.5 or 1 s, i.e. 30 or 60 frames scanned) in experiment 2. In experiment 3, an increase in sperm velocity (VAP, VSL, VCL), in linearity and in the percentage of motile and rapidly moving spermatozoa was observed with increasing sperm concentrations (25 x 10(6), 50 x 10(6) or 100 x 10(6) ml(-1)). In experiment 4, a clear effect of the diluent used was visible with higher velocity parameters (VAP, VSL, VCL) and higher percentages of motile, progressive and rapid spermatozoa for semen samples diluted in Hepes-TALP or prostatic fluid in comparison with physiological saline or egg-yolk-Tris extender. In experiment 5, significant (P<0.01) and high correlations were found between the conventional dog semen analysis methods and HTR Ceros 12.1 measurements (n=97 semen samples) for the sperm concentration (r=0.91), the motility (r=0.74) and the progressive motility (r=0.84). In experiment 6, the ejaculates from 21 proven, fertile dogs were compared with the ejaculates of a population (N: 11) of young beagles (1.5 years) but no significant differences in HTR Ceros 12.1 measurements were found between the two groups. Based on our results, diluting dog semen samples to 50 x 10(6) ml(-1) with physiological saline solution and scanning 30 frames at a frame rate of 60 frames/s (i.e. a scanning time of 0.5 s), are the set-up parameters proposed to obtain objective and standardized canine semen motility results using the HTR Ceros 12.1.
为了科研和实际应用目的，需要使用计算机化测量设备来客观评估犬精液质量。由于内部图像设置可能会对结果产生相当大的影响，因此研究了不同技术设置和精液处理对汉密尔顿-索恩Ceros 12.1精液分析仪（HTR Ceros 12.1）所评估参数的影响。在实验1中，帧率（15、30或60帧/秒）对大多数测量的活力特征有显著影响（P<0.05），而在实验2中，使用不同的采样持续时间（0.5或1秒，即扫描30或60帧）时，活力参数没有差异。在实验3中，随着精子浓度增加（25×10⁶、50×10⁶或100×10⁶毫升⁻¹），精子速度（VAP、VSL、VCL）、直线性以及活动精子和快速移动精子的百分比均有所增加。在实验4中，与用生理盐水或蛋黄- Tris稀释液稀释的精液样本相比，用Hepes - TALP或前列腺液稀释的精液样本具有更高的速度参数（VAP、VSL、VCL）以及更高比例的活动、前进和快速精子，可见所用稀释剂有明显影响。在实验5中，对于精子浓度（r = 0.91）、活力（r = 0.74）和前进活力（r = 0.84），传统犬精液分析方法与HTR Ceros 12.1测量结果（n = 97个精液样本）之间存在显著（P<0.01）且高度的相关性。在实验6中，将21只经证实有生育能力的犬的射精样本与一群（N：11）年轻比格犬（1.5岁）的射精样本进行比较，但两组在HTR Ceros 12.1测量结果上未发现显著差异。基于我们的结果，建议使用生理盐水溶液将犬精液样本稀释至50×10⁶毫升⁻¹，并以60帧/秒的帧率扫描30帧（即扫描时间为0.5秒），以此作为使用HTR Ceros 12.1获得客观和标准化犬精液活力结果的设置参数。
