The effect of addition selected amino acids in extender semen on quality and DNA stability of frozen-thawed Sumba Ongole bull spermatozoa

Abstract

<p class="abstrak1"><span lang="IN">The objective of the current study was to asses the optimal concentration of glutamine, glycine and cysteine amino acids in tris-citric-acid-fructose egg yolks (TCFY) extender on quality of SO bull spermatozoa during freezing and thawing. </span><span lang="EN-US">In t</span><span lang="IN">his study the DNA stability of frozen-thawed Sperm</span><span lang="EN-US"> was </span><span lang="IN">also indentified. Three mature bulls maintained at PT. Karya Anugerah Rumpin, private cattle breeding company, West Java, Indonesia were used as semen donors. Semen was collected using artificial vagina and were evaluated </span><span lang="EN-US">prior</span><span lang="IN"> to freezing. Semen was diluted with TCFY supplemented with different concentrations of amino acids (5, 15 and 25 mM glycine and glutamine, and 3, 5 and 7 mM cysteine) then processed for colling and freezing. Semen quality parameters (subjective motility, viability and membrane and DNA integrity). </span><span lang="EN-US">D</span><span lang="IN">ata showed that in general the effect of addition of selected amino acids (glycine, glutamine and cysteine) </span><span lang="EN-US">in</span><span lang="IN">to TCFY extenders on motility, viability and membrane integrity of SO spermatozoa after cooling were significantly different (p<0.05) higher than</span><span lang="EN-US"> that of</span><span lang="IN"> control. Addition of 15 mM glycine, 15 mM glutamine and 5 mM cysteine resulted in significant (p<0.05) increase post-thawing sperm motility and sperm viability as compared to th</span><span lang="EN-US">at of</span><span lang="IN"> control. Furthermore, when spermatozoa were stained with acridine orange after fixation with acetic alcohol, the DNA integrity of post-thawing spermatozoa showed that all spermatozoa were remain intact. In conclusion </span><span lang="EN-US">,</span><span lang="IN">addition of 15 mM glycine, glutamine and 5 mM cysteine increase the cryoprotecting efficacy of bovine bull cryopreservation extender, and furthermore all DNA spermatozoa were remain intact. </span></p>

Effect of addition an amino acid or its combination with EDTA on DNA integrity and morphometry sperm heads of freeze-dried bovine spermatozoa

“…The addition of antioxidants has important benefits on the quality and fertility of semen after liquid storage at different temperatures (Mata-CXampuzanoa, 2014). Said et al (2019) reported that supplemented amino acids (glutamine, cysteine, glycine) to TCFY extenders improved the spermatozoa quality with concerning the motility, viability and membrane integrity both after the equilibration and post-thaw. However, to date, almost no information studies focus on determining the antioxidant effect on freeze-dried spermatozoa.…”

“…Semen quality consist of volume, concentration, motility, and abnormality of the sperm was evaluated before freeze-drying. Ejaculates fulfilling the minimum standard of sperm motility (70%) and sperm morphologically normal (80%) were used for this study (Said et al, 2019). Semen collection was carried out once a week and conducted six times as replications.…”

“…Based on the data of macroscopic and microscopic evaluation of fresh semen samples used in this study were appeared normal category and feasible to be cryopreserved. Ejaculates fulfilling the minimum standard of sperm motility (70%) and sperm morphologically normal (80%) (Said et al, 2019).…”

“…Several studies have shown the beneficial effect of antioxidant therapy on oxidative stress in mammalian spermatozoa (Moltlagh et al, 2014). There is some evidence that proves that the addition of antioxidants to freezing extenders decreases the detrimental effects of ROS (Luno et al, 2014;Said et al, 2019). Furthermore, the addition of a chelating agent, such as EDTA or EGTA to the FD solution inactivates sperm DNase and protects against disruption of sperm DNA during FD and subsequent storage (Kaneko and Nakagawa, 2006).…”

Computer aided semen analysis (CASA) to determine the quality and fertility of frozen thawed sumba ongole sperm supplemented with amino acids

“…Glutathione production is increased in rabbit spermatozoa when glutamine is added to extender solutions by increasing GSH production, glutamine protects the spermatozoa from ROS-induced cryodamage [6]. Previous research discovered that adding 15 mM glycine, 15 mM glutamine, and 5 mM cysteine to bovine bull cryopreservation extender increased its cryoprotecting efficacy and ensured that all DNA spermatozoa were preserved [7].…”