ABSTRACT
Unsuccessful rooster fertility following cryopreservation may be linked to specific changes in spermatozoa quality, which can be determined using various methods. These determinations also facilitate the design of improved freezing and thawing processes. Here, we update the current state of methodologies available for the assessment of rooster semen quality after cryopreservation. Computer-assisted sperm analyses (CASA) is one of the main systems used to analyse motion parameters of spermatozoa (total motility, progressive motility and motion parameters). Moreover, fluorescent techniques and flow cytometry can improve the assessment of various aspects of semen quality (viability, acrosome status, mitochondrial potential, lipid peroxidation, DNA damage, lipid peroxidation and cell debris removal) using specific fluorescent markers such as ethidium bromide, Yo-Pro-1, Annexin V, propidium iodide, SYBR-14, PNA, JC-1, BODIPY, acridine orange and DRAQ5. Transmission electron microscopy also yields valuable information on spermatozoa ultrastructure. The application of these techniques to rooster spermatozoa is reviewed in relation to specific freezing techniques, the effects of cryoprotective agents (CPAs) and extenders, and the determination of spermatozoa quality after cryopreservation.
Subject(s)
Chickens , Cryopreservation , Semen Preservation , Animals , Cryopreservation/veterinary , Cryoprotective Agents/pharmacology , Male , Semen Analysis , Semen Preservation/veterinary , Sperm Motility , SpermatozoaABSTRACT
BACKGROUND: Preservation of genetic resources in gene bank is necessary for conservation of endangered poultry species. OBJECTIVE: This study is to characterize Oravka rooster semen quality. MATERIALS AND METHODS: Heterospermic pool was diluted (1:1 by volume) in a freeze medium composed of a commercial diluent (Kobidil+, K), 8% dimethyl sulphoxide (DMSO) or 8% ethylene glycol (EG) or 8% glycerol (GL), and then frozen in liquid nitrogen vapour. RESULTS: Spermatozoa in the GL/K+ had significantly higher number of motile and progressively moving spermatozoa (p < 0.05) than in DMSO/K+ and EG/K+ groups. The percentage of apoptotic and necrotic spermatozoa were significantly higher in the DMSO/K+ and EG/K+ groups compared with the GL/K+ group. Based on the total motility, progressive movement parameters and viability, our study showed that 8% GL diluted in Kobidil+ provided the highest cryoprotective effect on the Oravka rooster spermatozoa.
