Evaluation of Friesian Holstein frozen semen storage time for 33, 30, 27, and 24 years based on sperm morphologic and related variables

Abstract

This study aimed to evaluate Friesian Holstein's frozen semen storage time for 33, 30, 27, and 24 years based on sperm morphologic and related variables. The sample used in this research was frozen semen from Frisian Holstein which were stored in the Sperm Bank of the Singosari Artificial Insemination Center, Malang. The level of MDA was measured using the Enzyme-linked immunosorbent assay method. Examination of the intact acrosomal cap using formol saline was then examined under a microscope. The chromatin maturity examination used a drop of semen immersed in a 10% solution of formaldehyde at room temperature for 30 minutes and rinsed twice with a PBS solution for five and then examined under the microscope. The chromatin integrity examination used a smear of one drop of semen prepared on a glass object, air-dried, and subsequently fixed in a solution of 96% ethanol and acetone at 4°C for 30 minutes, and then examined under the microscope. The morphology and motility examination used a drop of semen to the object glass and then examined under the microscope. The study found that storing sperm for longer periods resulted in higher levels of MDA and lower levels of chromatin maturity and integrity. The levels of MDA were significantly different among the groups stored for 33, 30, 27, and 24 years. The levels of chromatin maturity and integrity were significantly different among the groups stored for 24, 27, 30, and 33 years. However, sperm motility and morphology were not significantly different among the 24, 27, 30, and 33-year storage groups (p>0.05), but the 27-year storage group had the highest normal sperm morphology and motility percentage. It could be concluded that the impact of long-term storage on frozen semen causes changes in variables that have a direct or indirect influence on spermatozoa morphology.