Sperm Penetration Assay as an Indicator of Bull Fertility

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  • PARK Yoo-Jin
    Department of Animal Science & Technology and BET Research Institute, School of Bioresource & Bioscience, Chung-Ang University, Gyeonggi-Do 456-756, Korea
  • MOHAMED El-Sayed A.
    Department of Animal Science & Technology and BET Research Institute, School of Bioresource & Bioscience, Chung-Ang University, Gyeonggi-Do 456-756, Korea
  • OH Shin-Ae
    Department of Animal Science & Technology and BET Research Institute, School of Bioresource & Bioscience, Chung-Ang University, Gyeonggi-Do 456-756, Korea
  • YOON Sung-Jae
    Department of Animal Science & Technology and BET Research Institute, School of Bioresource & Bioscience, Chung-Ang University, Gyeonggi-Do 456-756, Korea
  • KWON Woo-Sung
    Department of Animal Science & Technology and BET Research Institute, School of Bioresource & Bioscience, Chung-Ang University, Gyeonggi-Do 456-756, Korea
  • KIM Heung-Ruil
    Dairy Cattle Improvement Center, National Agricultural Co-operative Federation, Gyeonggi-Do 412-030, Korea
  • LEE Myeung-Sik
    National Institute of Animal Science, Rural Development Association, Kangwon-Do 232-950, Korea
  • LEE Kichoon
    Department of Animal Sciences, The Ohio State University, OH 43210, USA
  • PANG Myung-Geol
    Department of Animal Science & Technology and BET Research Institute, School of Bioresource & Bioscience, Chung-Ang University, Gyeonggi-Do 456-756, Korea

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To predict the fertility of frozen-thawed bull spermatozoa, a sperm penetration assay (SPA) using zona-free hamster oocytes was optimized, and the assay results were compared with data from field fertility expressed as the non-return rate (NRR). To increase sperm penetration, the spermatozoa were pre-incubated and coincubated with oocytes in media containing various concentrations of heparin (0 to 50 μg/ml). Coincubation with 10 μg/ml heparin showed the highest sperm penetration (P<0.05); it is considered to be the optimized SPA method. Sperm fertility index values obtained from WSPA were significantly correlated with the historic average NRR of 46 bulls (P<0.01). To determine the normal range for SPA, we established the lower limits of the sperm fertility index and set the cut-off value at 2.55, at which point the NRR was more than 70%, using the receiver operating characteristic curve. The overall accuracy for the 46 bulls was 95.7% (44/46) for both the low and high NRR, with a sensitivity of 95.5% (21/22) and a specificity of 95.8%. This protocol would make it easier to discriminate bulls according to their sperm fertilizing ability.

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