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

Abstract

Objective: The aim of this study was to ascertain the effects of adding green tea extract (GTE) to skim milk-egg yolk (SM-EY) extender on both the quality of post-thawed bull semen and the pregnancy rates of the recipient cows.Methods: Twelve ejaculates from four Simmental bulls, aged 3 to 5 years and weighing 900 to 950 kg, were diluted SM-EY extender, added with 0, 0.05, 0.1, and 0.15 mg GTE/100 mL extender and then frozen. After four weeks storage in liquid nitrogen, the sperm were thawed and evaluated for viability, motility, intact plasma membrane (IPM), and DNA fragmentation. Meanwhile, the estrus cycles of 48 recipient cows were synchronized by intramuscular administration of a single injection of 5 mg prostaglandin F2α. Estrus cows were divided into four equal groups and inseminated artificially 18 to 20 h after the onset of estrus by using semen from each extender group. Pregnancy was diagnosed by measuring serum progesterone levels at 21 days, followed by transrectal palpation 90 days after insemination.Results: The findings revealed that adding 0.1 mg of GTE/100 mL extender produced the highest percentages of sperm viability (70.67%±1.75%), motility (69.17%±1.47%), and IPM (69.23%±1.21%) and the lowest percentage of DNA fragmentation (3.00%±0.50%). The pregnancy diagnosis revealed that all cows (36/36) inseminated using frozen semen in GTE addition extender were pregnant (pregnancy rate 100%), whereas the pregnancy rate of the control group was 83.33% (10/12).Conclusion: It may be concluded that 0.1 mg GTE/100 mL extender yields the best quality of spermatozoa and that all variants doses of GTE in extender produce a higher pregnancy rate among recipient cows.

Green tea extract increases the quality and reduced DNA mutation of post-thawed Kacang buck sperm

“…The T2 group had the best semen quality, as it had the highest post-thawed viability, motility, and IPM and the lowest MDA concentration and SDF. This result was consistent with findings that GTE's addition to an extender maintained the motility, viability, IPM, and DNA integrity of Simmental bull sperm [ 27 ]. In boar semen, GTE supplementation also reduced ROS production during freezing, as evidenced by higher motility and viability of the GTE-supplemented group compared with that of the control group [ 44 ].…”

“…The extract was then evaporated in an acid chamber until a solid was formed and then freeze-dried to obtain ready-to-use dried GTE. The extract was stored at −20 °C [ 27 ].…”

“…The skim milk solution was added to 5 mL of egg yolk derived from laboratory chicken eggs to obtain 100 mL of the skim milk-egg yolk mix. Next, penicillin (1.000 IU/mL) and streptomycin (1 mg/mL) were added to the mixture [ 27 ]. The aliquot was divided into four equal portions, for the control group (T0, without GTE) and the treatment groups T1 (0.05 mg GTE), T2 (0.1 mg GTE), and T3 (0.15 mg GTE) added per 100 mL extender.…”

“…Toluidine blue staining was used for SDF assessment. Bright blue spermatozoa heads indicated intact chromatin integrity (normal DNA), whereas dark blue heads indicated reduced chromatin integrity (fragmented DNA) [ 27 ]. The sperm viability, motility, IPM, and SDF measurement was conducted in 400 spermatozoa under the light microscope (Olympus BX-53, Japan) at 400× magnification.…”

Kualitas post-thawing semen domba Merino dalam bahan pengencer berbasis susu skim-kuning telur yang ditambah isolat crude protein Tirosine Kinase

“…Susu bubuk skim (Merck 115338) 10 gram dilarutkan dalam 100 mL akuades, dipanaskan hingga 92-95 °C selama 10 menit, lalu didinginkan hingga 37 °C. Kuning telur (dari telur ayam untuk penggunaan laboratorium) 5 mL ditambah dengan larutan susu skim 100 mL, ditambah penisilin dan streptomisin masingmasing 1.000 IU/mL dan 0,1 mg/mL (Susilowati et al, 2021). Campuran tersebut dibagi menjadi dua bagian yang sama untuk kelompok kontrol (P0, tanpa penambahan isolat crude PTK), dan kelompok perlakuan T1 yang ditambah isolat crude PTK 1,597 mg/ml pengencer.…”