This study was the first to combine the addition of antioxidants to a skim milk–egg yolk (SM–EY) extender and different equilibration periods to obtain higher quality post-thawed Kacang buck semen. This study aimed to determine the effects of green tea extract (GTE) on the quality of frozen Kacang goat sperm equilibrated for one and two hours. The pool of Kacang buck ejaculate was equally divided into four portions and was diluted in an SM–EY extender that contained four doses of 0, 0.05, 0.10, and 0.15 mg of GTE/100 mL for T0, T1, T2, and T3 groups, respectively. The aliquots were treated for an equilibration period of 1–2 h before further processing as frozen semen. Post-thawed semen quality was evaluated for sperm quality. The Sanger method was used for DNA sequencing, and the amino acid sequence was read using MEGA v.7.0. The post-thawed semen of the T2 group that was equilibrated for one hour had the highest semen quality. Pre-freezing motility had the highest determination coefficient compared to post-thawed sperm motility. This study is the first to report amino acid mutation due to freeze–thawing. The frequency of amino acid mutations revealed that T2 was the least mutated amino acid. Glycine, valine, leucine, serine, and asparagine strongly correlated to post-thawed sperm motility. It can be concluded that a combination of 0.1 mg GTE/100 mL extender as an antioxidant and one-hour equilibration period resulted in the best post-thawed Kacang buck semen quality.

The study aimed to determine the addition of green tea extract (GTE) in extender on the quality and DNA mutation of post-thawed Kacang buck sperm. The sperm DNA mutation was observed on nicotinamide adenine dinucleotide hydride (NADH) dehydrogenase 1 (ND1) of mitochondrial Deoxyribonucleic Acid (mtDNA). A pool of 12 Kacang buck ejaculates was diluted in skim milk-egg yolk extender contained 0, 0.05, 0.10, and 0.15 mg of GTE/100 mL for T0, T1, T2, and T3 group, respectively. Each of the aliquot groups was packaged in 0.25 mL French mini straw contained 60 million alive sperm and froze according to the protocol. The ND1 mtDNA amplification of samples was carried out Polymerase Chain Reaction machine, followed by DNA sequencing using the Sanger method. Meanwhile, the phylogenetic tree was constructed using the neighbor-joining (NJ) method with MEGA 7.0 software. The results showed that the T2 group maintained the highest quality for Kacang buck post-thawed semen. There was the highest percentages of sperms viability, motility, intact plasma membrane (IPM), the lowest of malondialdehyde (MDA) concentration, sperm DNA fragmentation (SDF), the total and types of ND1 mtDNA mutation frequency. The phylogenetic tree analysis revealed that the clade of the T2 group was most closely related to the sequence reference. However, there was no correlation between the semen quality parameters (sperm viability, motility, IPM, MDA concentration, and SDF) with ND1 mtDNA mutation of post-thawed Kacang buck semen. It could be concluded that GTE was useful as an antioxidant for Kacang buck semen extender for frozen sperm.

No abstract

This study aimed to determine the relationship between heat detector reading results and the feature of vaginal swab preparations as a parameter for determining estrus in Peranakan Etawa goats. Five healthy non-pregnant goats weighing 20-30 kg, were synchronized using PGF2α (7.5 mg Dinoprost, Enzaprost) intramuscularly twice with 11 days interval. Vaginal cytology examination began the day after the second PGF2α injection. The heat detector (Draminski®, Dramiński SA, Poland) is an estrus detecting device based on the amount of electric current flowing through the vaginal mucus. Spearman correlation analysis showed a relationship (p <0.05) between heat detector reading and estrus detection based on vaginal cytology. The heat detector readings were very significantly correlated positively (p <0.01) with the number of epithelial cells (coefficient correlation, r= 0.564), but negatively correlated with the number of cornified cells (r = –0.854), and also significantly correlated positively (p <0.05) to the number of leukocytes (r = 0.487). The estrus stages were significantly correlated positively (p <0.05) only to the number of leukocytes (r = 0.918). Likewise, the number of epithelial cells was highly correlated negatively (p <0.01) only to cornified cells (r = –0,643), and the number of cornified cells was significantly correlated negatively (p <0.05) only to the number of leukocytes (r = –0.472). It could be concluded that the features of the proestrus, estrus and metestrus stages based on the goat vaginal cytology was correlated with the reading results of the heat detector.

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.