Sperm chromatin structure assay of bulls qualified for artificial insemination

Abstract

The goal of our study was to find the relationship between fertility of bulls qualified for Al and the percentage of spermatozoa with abnormal chromatin structure as an independent parameter. We used the frozen semen of 8 mature bulls from one Al center. Each bull was represented by 3 ejaculates collected with at least 2-week intervals. Bull fertility was calculated on the basis of non-return ratio and was expressed as a scale where 100 points represented the average fertility of all the Al center's bulls. Bulls with lower or higher fertility received a lower or higher score respectively. Fertility scores of bulls used in the study ranged from 83 to 104. Semen was processed according to the SCSA (sperm chromatin structure assay) method and was analyzed by flow cytometry. “Artificial” αt (αt=red/green+red fluorescence) and red fluorescence histograms were used for calculation of COMPαt, SDαt, %Red, %PeakR and MeanR parameters. The percentage of spermatozoa with abnormal chromatin ranged from 1.2% to 23.8%. A large variation among ejaculates was found for bulls with lower fertility. Fertility correlated significantly with COMPαt (−0.50, P < 0.05), SDαt (−0.55, P < 0.01), %Red (−0.53, P < 0.01), %PeakR (−0.58, P < 0.01) and MeanR (−0.45, P < 0.05). The SCSA method has a practical application in analyzing spermatogenesis disorders in bulls. If regularly applied, it allows us to identify and eliminate ejaculates with a high level of sperm chromatin abnormalities.