The effect of duration of preservation on the quality, MDA level, and DNA damage of post-thawed Bali cattle bull sperm

Abstract

Frozen semen quality is one of many factors that promote successfully of artificial insemination. Preservation at 5ºC is one of the steps of semen freezing to obtain high fertility sperm after added with the extender. This study was aimed to determined the sperm quality (motility, viability, and membrane integrity), MDA level and DNA damage of post-thawed sperm after being preserved at 5ºC for different duration. Bali cattle bull semen collected by artificial vagina. Macroscopic and microscopic evaluation of ejaculate was conducted first. Qualified semen was diluted in Tris Aminomethan-Egg yolk and then devided into two treatment group: preserved at 5ºC for 4 hours (first group) and 22 hours (second group), and continued to processed into frozen semen, and stored overnight. Pre-freezing and post-thawed of semen analysis was conducted based on SNI: 4869-1:2017 of The National Standardization Agency of Indonesia. Measurement of levels of Malondialdehyde (MDA) was conducted by spectrophotometry at 532 nm wavelength, meanwhile measurement of sperms DNA damage percentage was performed by Toluidine blue staining. The result of showed that quality of post-thawed sperm (motility, viability, and membrane integrity) was higher (P<0.05), and MDA level and DNA damage were lower (P<0.05) in preservation at 5°C for 22 hours compared to those of 4 hours. It could be concluded that preservation at 5°C longer (22 hours) means sperm had longer chance to adapt with the extender, this implies the higher quality and lower lipid peroxidation.

Effect of green tea extract in extender of Simmental bull semen on pregnancy rate of recipients

“…This result also better compared to an earlier study that addition of 0.75% GTE has reached motility, viability, and IPM as 52.64%, 64.32%, and 53.34%, respectively [ 1 ]. The lowest DNA fragmentation of this study (3%) was lower compared to the lowest (11.5%) observed in Bali cattle bulls [ 11 ].…”

“…DNA fragmentation in 100 spermatozoa was examined under the light microscope (Olympus BX-53, Japan) at 400× magnification. Bright blue spermatozoa heads indicated intact chromatin integrity (normal DNA), whereas dark blue heads indicated reduced chromatin integrity (fragmented DNA) [ 11 ].…”

Addition of L-arginine in skim milk extender maintains goat spermatozoa quality in chilled temperature for five days

“…Semen is cooled to 5°C and stored for several days until it is used for insemination [2,3]. In bull semen, preservation at 5°C for 22 h results in a higher quality of postthawed sperm (motility, viability, and membrane integrity), and lower malondialdehyde (MDA) and DNA damage, compared to those of 4 h [4]. The problem is that the life span of the spermatozoa after ejaculation is only 8 h and is mediated by increasing concentration of lactic acid, a metabolic product of the spermatozoa itself, which causes lipid peroxidation [5,6].…”

The enriched Y-bearing sperm combined with delayed fixed-time artificial insemination for obtaining male Simmental crossbred offspring

“…The semen samples were frozen following the Indonesian National Standard (SNI) with the number SNI: 4869-1:2017 (production and analysis of frozen semen) of the National Standardization Agency of Indonesia [15]. The semen was diluted in a tris-egg yolk extender, after which it was equilibrated at 5°C in a cold handling cabinet (Minitüb, Germany) for 22 h [16] after which it was packaged in 0.25 mL French straws containing 50 × 10 6 live spermatozoa and sealed. The straws were exposed to liquid nitrogen vapor (−140°C) for 10 min and stored in liquid nitrogen (−196°C) for a month before they were assessed and used for AI [17].…”