Freezing/thawing procedures induce enhanced reactive oxygen species (ROS) formation in mammalian sperm and these ROS may be a cause for the decrease in sperm function following cryopreservation. In the present study, we used a chemiluminescence method to detect ROS-induced damage in goat spermatozoa. Iron-induced luminescence of fresh and frozen/thawed sperm cells was assessed using a luminometer. It was shown that the freezing/thawing procedure had a significant effect on some luminescence parameters. Semen freezing significantly increased the values of integral, peak max, T.half (rise) and T.max (peak) parameters. A significant correlation was observed between the percentage of motile spermatozoa and integral, peak max and T.half (rise) parameters. In conclusion, the results of the present study indicate that measurement of induced luminescence can be an alternative, sensitive and relatively simple method for assessing the effect of cryopreservation on oxidative damage to spermatozoa.
The aim of this study was to examine the effect of thymosin beta 4 (Tβ4) during in vitro maturation (IVM) of oocytes and subsequent embryonic development after in vitro fertilization as well as to assess the quality of obtained blastocysts. COCs were matured in vitro in 4 different media: 1. control medium; 2. control media supplemented with 50 ng/mL Tβ4; 3. control media supplemented with 0.5 mg/mL Tβ4; and 4. control media supplemented with 1 mg/mL Tβ4. The quality of the developed blastocysts was analysed by the TUNEL method. The number of cleaved eggs was significantly higher (P < 0.05) when gametes were matured in the presence of 50 ng/mL Tβ4 than it was using the other types of media. Additionally, the largest number of blastocysts was observed when 0.5 mg Tβ4 was added to the medium (P < 0.05). No significant difference was noted in the mean number of apoptotic nuclei per blastocyst or in the mean number of nuclei per blastocyst in any of the analysed groups. In conclusion, Tβ4 supplementation (50 ng/mL) in maturation medium increased the number of cleaved oocytes, and the number of blastocysts obtained increased when 0.5 mg/mL Tβ4 was used. This positive effect was not observed when higher a higher concentration of Tβ4 (1 mg/mL) was used.
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