Application of the sperm mobility assay to primary broiler breeder stock

Journal of Applied Poultry Research

Volumn 15, Issue 2, 2006, Pages 280-286

  Froman, D P ^a  

^a OREGON STATE UNIVERSITY   (United States)

Author keywords

Broiler breeder; Chicken; Fertility; Semen quality; Sperm

Indexed keywords

GALLUS GALLUS;

EID: 33745660524     PISSN: 10566171     EISSN: None     Source Type: Journal    
DOI: 10.1093/japr/15.2.280     Document Type: Article

References (28)

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14. Animal Reproduction Systems, Chino, CA.
15. The Animal Reproduction Systems (ARS) sperm mobility assay was performed as follows. Unless noted, reagents and equipment were provided by ARS. First, 2 anodized metal cuvette warming blocks were set upon a slide warmer (ARS Model 581A MOD 1) set at 41°C. Sealed, standard polystyrene cuvettes containing 6% (wt/vol) Accudenz were prewarmed within the warming blocks. Each cuvette contained 3 mL. In addition, 2 standard polystyrene cuvettes were set in a warming block. Each was filled with approximately 3 mL of mobility buffer and covered with a plastic cap. Reagent temperature was confirmed with a Type-K beaded thermocouple connected to a digital hand-held thermometer (VWR International. http://www.vwrsp.com). A 100-μL subsample of an ejaculate was placed in a 12 × 75 mm polystyrene test tube (VWR International). An 11.4-μL volume of semen was removed with a positive displacement pipet and added to 3.42 mL of 3% (wt/vol) NaCl dispensed into a standard polystyrene cuvette. The cuvette was covered with a plastic cap. and sperm was mixed by cyclic inversion of the cuvette. Sperm concentration was determined with an ARS 596A Mobility Analyzer, which measures sperm concentration, calculates buffer volume required for the assay, and measures sperm mobility as an index. After the foil tab was removed from a cuvette, the calculated volume of prewarmed buffer (e.g., 0.385 mL corresponding to an observed sperm concentration of 4.35 × 109/mL) was pipetted into an empty 12 × 75 polystyrene test tube. A 50-μL volume of semen was added with a positive-displacement pipet, and the contents were mixed by gentle vortexing. Thus, a sperm suspension was produced that contained 5 × 108 sperm/mL. A 300-μL volume of sperm suspension was immediately overlaid on the Accudenz solution. The cuvette was gently removed from the warming block after 5 min of incubation and then transferred to the well of the ARS 596A Mobility Analyzer to measure sperm mobility.
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17. 2 (TES-buffered saline). The TES-buffered saline was used for preparing sperm suspensions and a 6% (wt/vol) Accudenz solution.
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23. Bowling, E. R., D. P. Froman, A. J. Davis, and J. L. Wilson. 2003. Attributes of broiler breeder males characterized by low and high sperm mobility. Poult. Sci. 82:1796-1801.
24. Froman, D. P., 2005, unpublished data.
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28. The sperm mobility assay entailed overlaying a sperm cell suspension on a solution of 6% (wt/vol) Accudenz in a cuvette, incubating the cuvette for 5 min at body temperature, and then measuring the extent to which sperm have penetrated the underlying Accudenz layer. Rendering sperm immotile by heat denaturation prior to overlay resulted in a negligible sperm mobility index because immotile sperm are immobile. In contrast, mobile sperm readily penetrated the Accudenz solution.