INFLUENCE OF DIFFERENT BUFFERS ON THE VIABILITY CHARACTERISTICS OF CANINE SPERMATOZOA DURING STORAGE AT 4–6 °C
DOI: http://dx.doi.org/10.30970/sbi.2003.893
Abstract
Background. Maintaining high fertilization capacity of spermatozoa is a key factor for successful artificial insemination in dogs. Given that the use of chilled semen provides higher pregnancy rates and larger litter sizes compared with frozen semen, the demand for this method of gamete preservation is increasing. Preservation of sperm fertilizing ability in an in vitro system directly depends on the storage medium; however, analysis of current literature indicates that most studies focus on the effects of antioxidants, energy substrates, or membrane protectants, whereas the impact of buffer system parameters on gamete homeostasis during cooling remains insufficiently studied. Therefore, the aim of this study was to determine the influence of different buffers on the characteristics of chilled canine semen at 4–6 °C.
Materials and Methods. Extenders differing in their buffer system were added to the second fraction of canine semen: T-BSA (TRIS, pH 6.2), hT-BSA (TRIS, pH 7.3), and HEPES-BSA (HEPES, pH 6.2). Sperm motility, membrane integrity (HOS test), and the level of DNA fragmentation (SCD method, HALOMAX test system) were evaluated every 24 hours over a period of 10 days.
Results and Discussion. It was established that the T-BSA extender (pH 6.2) most effectively ensures long-term preservation of the functional parameters of canine spermatozoa during chilled storage at 4–6 °C. At the same time, DNA integrity remained a stable parameter in all groups, even under conditions of complete loss of sperm motility. Thus, optimization of the buffer parameters of the extender is a key factor in prolonging the shelf life of chilled canine semen in clinical practice.
Conclusion. The T-BSA extender (pH 6.2) was found to be the most effective for preserving the functional parameters of canine spermatozoa during prolonged storage at 4–6 °C. DNA integrity remained a stable parameter in all groups, even in the case of complete loss of motility. Therefore, optimization of the buffer parameters of the extender is a key factor for extending the shelf life of chilled canine semen in clinical practice.
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