Donoghue A M, Thistlethwaite D, Donoghue D J, Kirby J D
Germplasm and Gamete Physiology Laboratory, USDA, Beltsville, Maryland 20705, USA.
Poult Sci. 1996 Jun;75(6):785-9. doi: 10.3382/ps.0750785.
The routine determination of sperm concentration not only provides a measure of semen quality but also supplies a means of delivering precise sperm numbers in modern artificial insemination (AI) programs. Methods for estimating sperm numbers in laboratory settings (e.g., hemacytometer) are not practical for use in breeding facilities. A microprocessor-controlled semen analyzer (Densimeter, Model 534-B-Mod1, Animal Reproductive Systems) is commercially available for evaluating stallion semen. We calibrated and modified the semen analyzer to determine sperm concentration in turkey semen and to provide information on the required dilution for a constant AI dose. To calibrate the semen analyzer for turkeys, six samples of pooled semen (10 toms per sample) were collected by abdominal massage. A dilution curve of undiluted and 1:0.5, 1:1, 1:1.5, 1:2, 1:3, 1:5, and 1:10 diluted semen was established using Beltsville Poultry Semen Extender. Sperm concentration in diluted samples was determined by hemacytometer counts. The hemacytometer counts were used to develop an equation for estimating sperm concentration from the semen analyzer's absorbance readings and the equation programmed into the instrument. The newly developed program on the semen analyzer was subsequently validated by comparing instrument values for sperm concentrations with concurrent hemacytometer counts, which compared well to concentrations determined by a previously calibrated photoelectric colorimeter (Klett). Correlation coefficients between the semen analyzer and hemacytometer counts and the Klett and hemacytometer counts were r = 0.996 and r = 0.992, respectively (P < 0.001). The semen analyzer method was accurate and precise and could be beneficial to commercial turkey AI programs because it is easy to use, requires limited technological skills, and provides results for determining sperm concentration and AI dose in approximately 1 min.
常规测定精子浓度不仅能衡量精液质量，还能为现代人工授精（AI）程序提供精确的精子数量。在实验室环境中估算精子数量的方法（如血细胞计数器）在育种设施中并不实用。一种微处理器控制的精液分析仪（密度计，型号534 - B - Mod1，动物生殖系统公司）可用于评估种马精液。我们对该精液分析仪进行了校准和改进，以测定火鸡精液中的精子浓度，并提供恒定AI剂量所需稀释度的信息。为校准用于火鸡的精液分析仪，通过腹部按摩收集了六个混合精液样本（每个样本10只公火鸡）。使用贝尔茨维尔家禽精液稀释液建立了未稀释以及1:0.5、1:1、1:1.5、1:2、1:3、1:5和1:10稀释精液的稀释曲线。通过血细胞计数器计数确定稀释样本中的精子浓度。血细胞计数器计数用于推导一个方程，以便根据精液分析仪的吸光度读数估算精子浓度，并将该方程编入仪器程序。随后，通过将精液分析仪测得的精子浓度值与同时进行的血细胞计数器计数结果进行比较，对精液分析仪上新开发的程序进行了验证，该结果与先前校准的光电比色计（Klett）测定的浓度结果比较良好。精液分析仪与血细胞计数器计数之间以及Klett与血细胞计数器计数之间的相关系数分别为r = 0.996和r = 0.992（P < 0.001）。精液分析仪方法准确且精确，对商业火鸡AI程序可能有益，因为它使用方便，所需技术技能有限，并且能在大约1分钟内提供测定精子浓度和AI剂量的结果。
