Sperm Quality in Cryopreserved Holstein Friesian Bull Semen and Bacterial Load as Affected by Different Extenders
Anggita Dian Pramudhita1, Putri Utami1, Habib Asshidiq Syah1, Arik Yuswati1, Asri Nurul Huda2, Anny Amaliya3, Deny Sulistyowati3, Lilik Eka Radiati4, Kuswati5, Trinil Susilawati1 and Aulia Puspita Anugra Yekti1*

1Department of Animal Reproduction and Breeding, Faculty of Animal Science, Universitas Brawijaya, Malang 65145, Indonesia
2Department of Animal Nutrition, Faculty of Animal Science, Universitas Brawijaya, Malang 65145, Indonesia
3Singosari National Artificial Insemination Center, Malang 65153, Indonesia
4Departement of Animal Product Technology, Faculty of Animal Science, Universitas Brawijaya, Malang 65145, Indonesia
5Department of Animal Production, Faculty of Animal Science, Universitas Brawijaya, Malang 65145, Indonesia
*auliapay[at]ub.ac.id

Abstract

This study compared the effects of two different extenders, tris aminomethane egg yolk and modified trico egg yolk, on post-thaw sperm quality and bacterial load in cryopreserved semen from a Holstein Friesian bull. Semen samples were divided into two treatment groups, each consisting of five semen straws (0.25 ml), which were cryopreserved using a Tris aminomethane egg yolk or Trico egg yolk extender. Post-thaw evaluations included sperm motility, viability, abnormality, membrane and acrosome integrity, capacitation status, DNA fragmentation, sperm kinematics, and bacterial load. Data were analyzed using paired t-tests and bacterial content was analyzed descriptively. The results showed that the Tris aminomethane extender significantly enhanced several sperm quality parameters post-thawing compared to Trico egg yolk. Specifically, Tris aminomethane significantly improved intact acrosome (70.36%) and increased the percentage of non-capacitated spermatozoa (67.92%) Furthermore, significantly higher values were observed in parameters related to sperm kinematics, including Distance Average Path (DAP, 33.38%), Distance Curvilinear (DCL, 62.22%), Velocity Average Path (VAP, 84.31%), and Curvilinear Velocity (VCL, 155.06%). Meanwhile, trico egg yolk extender caused significantly higher values in sperm straightness (STR, 84.02%), linearity (LIN, 56.22%), and wobble (WOB, 65.40%). Other parameters, including viability, abnormality, total motile sperm, DNA fragmentation, sperm kinematic parameters (motility, progressive motility, straight-line distance, linear velocity, lateral head amplitude, and beat cross frequency) did not differ significantly between the two extenders. Meanwhile, the bacterial of E. coli and Mycoplasma was higher in the trico extender, while Staphylococcus was more prevalent in tris aminomethane extender. These findings indicate that both extenders can maintain the quality of frozen semen. The results of this study show that Trico egg yolk extender is a promising and cost-effective alternative, with comparable performance in several sperm quality, but the effectiveness of bacterial inhibition needs to be re-evaluated in terms of the dosage of antibiotics used.

Keywords: Bacterial contamination, cryopreservation, sperm quality, Tris aminomethane extender, Trico extender.

Topic: Animal reproduction and breeding