THE EFFECT OF DIFFERENT INCUBATION TIMES IN QUALITY OF SPERM AFTER SEXING USING THE TWO-GRADIENT EGG WHITE SEDIMENTATION METHOD IN HOLSTEIN FRIESIAN BULLS Hikma Nur Azizah1), Trinil Susilawati1), Anny Amalia2), Deny Sulistyowati2), Andi Rezda Sri Mulyani1), Aulia Puspita Anugra Yekti 1*)
1) Faculty of Animal Science, Universitas Brawijaya, Malang, 65145, Indonesia
2) Singosari National Artificial Insemination Center, Malang, 65153, Indonesia
Abstract
Indonesia persistent dependence on imports to fulfill domestic milk requirements necessitates strategic invention. Consequently, one key initiative to address this deficiency is the application of biotechnology, particulary through artificial insemination and the innovation of spermatozoa sexing techniques. This study aimed to analyze the effect of different incubation times on the quality of sperm after sexing using two gradient egg white sedimentation method in holstein friesian bulls. The evaluated semen quality parameters included motility, viability, concentration, abnormality, and total spermatozoa motil (TSM). This researched was conducted in August to October 2025 at the laboratory of Singosari Artificial Insemination Center. Fresh semen was collected from Holstein friesian bulls with an average motility above 70% and was processed using trico buffer and trico egg yolk diluents. Sexing treatments were applied at three incubation times (20, 30, 40 minutes) with ten replications. Data were analyzed using one way ANOVA in a randomized block design. The 40 minutes incubation time in upper layer showed the highest motility and abnormality, whereas the 20 minute incubation time yielded the highest viability, concentration, and total spermatozoa motil (TSM). In conclusion, the semen quality among all treatments did not show any differences with the motility after sexing remained below the expected standard. Therefore, improvements are needed to help maintain optimal sperm performance after the sexing process.
Keywords: Frisian Holstein, Sexing Spermatozoa, Egg White Sedimentation, Different Times of Incubation, Semen Quality
