The acrosome cap allows sperm to penetrate the egg membrane and produce male pronuclei within female chicken eggs, facilitating successful fertilization. Given this, it is important to establish practical methods for evaluating the integrity of the acrosome cap and thus the quality of the rooster’s sperm. There are several established methods for evaluating the acrosomes of mammalian sperm, but none of these methods are suitable for evaluating the acrosome status of rooster spermatozoa. Therefore, a simplified method for evaluating the rooster acrosome is needed. Here we evaluated the usefulness of CBB (coomassie brilliant blue) staining of the acrosome at concentrations of 0.04%, 0.08%, and 0.3% CBB solutions. Our data revealed a clear staining pattern for intact acrosome caps at 0.04% and 0.08% CBB but not at 0.3% CBB. This protocol revealed differences in acrosome integrity between fresh and frozen rooster sperm smears suggesting that CBB staining may facilitate easier semen evaluation in roosters. This protocol allows for the accurate differential staining of acrosome cap in rooster spermatozoa.

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
CONCLUSION
REFERENCES

• Sung Woo Kim(Animal Genetic Resources Research Center, National Institute of Animal Science, RDA, Hamyang 50000, Korea) Corresponding author
• Jae-Yeong Lee(Animal Genetic Resources Research Center, National Institute of Animal Science, RDA, Hamyang 50000, Korea)
• Chan-Lan Kim(Animal Genetic Resources Research Center, National Institute of Animal Science, RDA, Hamyang 50000, Korea)
• Yeong Gyu Ko(Animal Genetic Resources Research Center, National Institute of Animal Science, RDA, Hamyang 50000, Korea)
• Bongki Kim(Department of Animal Resources Science, Kongju National University, Yesan 23439, Korea) Corresponding author