T. Susilawati
Fakultas Peternakan Universitas Brawijaya Malang

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Sperm Quality, Kinematics, Membrane Integrity, and DNA Fragmentation of Frozen Sexed Semen in Holstein-Friesian Bulls Safa, F.; Yekti, A. P. A.; Utami, P.; Syah, H. A.; Febrianto, N.; Rachmawati, A.; Yuswati, A.; Amaliya, A.; Sulistyowati, D.; Susilawati, T.
Tropical Animal Science Journal Vol. 48 No. 3 (2025): Tropical Animal Science Journal
Publisher : Faculty of Animal Science, IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.5398/tasj.2025.48.3.199

Abstract

Percoll density gradient centrifugation (PDGC) is a widely used method for sperm separation in Indonesia. This study evaluated sperm quality, membrane integrity, acrosome damage, and DNA fragmentation after PDGC-based sperm sexing and cryopreservation. This research was conducted as a laboratory experiment with six treatment groups: fresh semen (T1), post-sexing X-chromosome-bearing sperm (T2), post-sexing Y-chromosome-bearing sperm (T3), post-thawing unsexed semen (T4), post-thawing X-chromosome-bearing sperm (T5), and post-thawing Y-chromosome-bearing sperm (T6). The observed variables included individual motility (IM), viability, concentration, abnormality, total motile sperm (TMS), intact acrosome cap (IAC), membrane integrity (MI), DNA fragmentation, and sperm kinetic variables assessed using computer-assisted sperm analysis (CASA). The results showed that PDGC sexing significantly reduced IM, concentration, TMS, viability, MI, and IAC. Furthermore, the freezing of both unsexed and sexed semen further decreased IM, concentration, TMS, viability, MI, and IAC while increasing sperm abnormalities. DNA fragmentation increased post-sexing and freezing in unsexed semen, indicating potential DNA damage from these processes. Kinetic variable assessment using CASA showed that PDGC maintained sperm motility characteristics, whereas freezing led to a significant decline in sperm kinetics. In conclusion, PDGC is a viable method for sperm sexing in cattle, as it preserves sperm quality, membrane integrity, acrosome integrity, DNA integrity, and motility characteristics while being compatible with semen cryopreservation. These findings confirm that PDGC-processed semen is suitable for artificial insemination (AI).
The Quality of Cryopreserved Semen of Local Chickens Treated with Ringer’s Lactate-Egg Yolk Extender Supplemented with Glycine and Glucose Junaedi; Isnaini, N.; Natsir, M. H.; Susilawati, T.
Tropical Animal Science Journal Vol. 47 No. 2 (2024): Tropical Animal Science Journal
Publisher : Faculty of Animal Science, IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.5398/tasj.2024.47.2.161

Abstract

The aim of this study is to investigate the potential of glucose, glycine, and a combination of glucose and glycine supplemented into Ringer’s lactate extender on the quality of pre- and post-cryopreservation of semen of local chickens. Semen samples were collected from KUB roosters. The research procedures involved formulation of the extender, semen collection, assessment of fresh semen, dilution, packaging, cryopreservation, storage, and subsequent assessment of cryopreserved semen. Parameters assessed in fresh, pre-cryopreservation, and post-thawed semen using computer-assisted semen analysis (CASA), included motility (total motility, progressive, static, and slow motilities), kinematics, spermatozoa morphology, malondialdehyde (MDA) concentration, and scanning electron microscopy (SEM). There were four treatments: T1= control, T2= 50 mM glucose, T3= 60 mM glycine, and T4= a combination of 50 mM glucose and 60 mM glycine. Each treatment was repeated 10 times. The results showed that the addition of 60 mM glycine and the combination of 50 mM glucose and 60 mM glycine effectively preserved total motility, maintained progressive motility, and reduced damage to the bent tail during the equilibration process (prior to cryopreservation). In addition, total motility, progressive motility, DAP, DSL, DCL, VAP, VSL, VCL, STR, and LIN were significantly increased after thawing. This approach also minimized the formation of MDA. Furthermore, observations using SEM showed comparatively more intact acrosomes compared to the other treatments. In conclusion, the addition of 60 mM glycine and a combination of 60 mM glycine and 50 mM glucose to Ringer’s lactate-egg yolk extender proved to be effective in preserving the quality of domestic chicken spermatozoa during semen cryopreservation.