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Öğe Can linoleic acid improve the quality of frozen thawed bull sperm?(Cryo-Letters, 2014) Büyükleblebici, Serhat; Taşdemir, Umut; Tuncer, Pürhan Barbaros; Durmaz, Emre; Özgürtaş, Taner; Büyükleblebici, Olga; Coşkun, Erdem; Gürcan, İsmail SafaBACKGROUND: Cryopreservation is known to have a detrimental effect on the motility, viability and membrane integrity of sperm cells. OBJECTIVE: The aim of this study was to investigate the effect of various amount of linoleic acid supplementation to the Tris extender, on bull sperm parameters, DNA integrity and oxidative stress after freeze-thawing. METHODS: Ejaculates were split into five aliquots and extended to a final concentration of 18x10 6 spermatozoa/ml with the base extender containing different doses of linoleic acid 0.125 ml, (L125); 0.250 ml (L250); 0.5 ml (L500), 1 ml (L1000) and no additive (control; L0). The extended samples were equilibrated slowly to 4°C for 4 h and then froze using a digital freezing machine. Frozen straws were thawed individually in water bath at 37°C for 30 s to analyse progressive motility and sperm motion characteristics as well as membrane integrity. Biochemical assays were performed in a spectrophotometer using commercial kits. DNA damage was evaluated by Comet Assay. RESULT: The addition of various linoleic acid did not improve the sperm subjective, CASA and progressive motilities, sperm motility characteristics and DNA integrity (P>0.05). L500 exhibited the greatest values for membrane integrity than that of the other groups (P<0.001). All supplementation groups led to lower percentages of tail abnormalities in comparison to the control (P<0.001). L500 and L1000 significantly decreased total abnormalities. In conclusion, our findings showed that L500 linoleic acid supplementation in semen extender was of great beneficial effect on frozen-thawed bull semen in terms of morphology and plasma membrane integrity. © CryoLetters.Öğe Comparing ethylene glycol with glycerol and with or without dithiothreitol and sucrose for cryopreservation of bull semen in egg-yolk containing extenders(Academic Press, 2014) Büyükleblebici, Serhat; Tuncer, Pürhan Barbaros; Bucak, Mustafa Numan; Taşdemir, Umut; Eken, Ayşe; Büyükleblebici, Olga; Durmaz, Emre; Sarıözkan, Serpil; Endirlik, Burcu ÜnlüThere are few studies performed for investigating the roles of different ratio and cryoprotectants with dithiothreitol or sucrose on sperm motility characteristics and antioxidant capacities of post-thawed bull spermatozoa. The objectives of this study were to compare glycerol (G) and ethylene glycol (EG) at different concentrations as cryoprotectants and dithiothreitol (D) or sucrose (S) (with/without) as antioxidants in Tris extender for cryopreservation of bull semen. Twenty-four ejaculates obtained from three bulls were included in the study. Each ejaculate was split into four equal aliquots and diluted using both of the Tris extenders with glycerol (5% or 7%) or ethylene glycol (3% or 5%). After that, each extenders were split into three equal aliquots and diluted using both of the dithiothreitol 5 mM or sucrose 25 mM, and control (without additives) was cooled to 4 degrees C and frozen in 0.25-ml French straws. when compared to control, different doses cryoprotectants and antioxidants addition no significantly increased the percentages of post-thaw sperm progressive and motitilities, acrosome abnormality and plasma membrane integrity (P > 0.05). However, EG3 + S yielded the greatest percentages of the total abnormality (P < 0.05). As regard to antioxidant activities G7 and EG5 led to lowest MDA activity with or without D or S but, these results were not supported to the GPx activity (P < 0.01). The sperm motion characteristics such as VAP, VCL, ALH and BCF gave significantly different results (P < 0.05). When compared the DNA integrity, different doses cryoprotectants without antioxidants addition significantly increased the percentages the tail intensity and tail moment (P < 0.05). There were no significant differences observed in non-return rates among all treatment groups (P > 0.05). (C) 2014 Elsevier Inc. All rights reserved.Öğe Comparison of cryoprotective effects of lycopene and cysteamine in different cryoprotectants on bull semen and fertility results(Wiley-Blackwell, 2014) Tuncer, Pürhan Barbaros; Büyükleblebici, Serhat; Eken, Ayşe; Taşdemir, Umut; Durmaz, Emre; Büyükleblebici, Olga; Çoşkun, ErdemContents The objectives of this study were to compare glycerol and ethylene glycol at different concentrations as cryoprotectants and lycopene or cysteamine (with/without) as antioxidants in Tris extender for bull semen. Twenty-four ejaculates were obtained from three bulls. Each ejaculate was split into four equal aliquots and diluted using both of the Tris extenders with glycerol (5% or 7%) or ethylene glycol (3% or 5%). After that, each extenders were split into three equal aliquots and added using both of the cysteamine 5mm or lycopene 500g/ml, and control (without additives). The addition of 7% glycerol with cysteamine, 5% ethylene glycol with cysteamine and 3% ethylene glycol with cysteamine groups gave the lowest CASA motility than the other groups. However, 7% glycerol and 7% glycerol with lycopene resulted in a better rate of CASA progressive motility compared with that of other groups. Generally, all the lycopene groups signed better protective effects on acrosome and total morphology than the other groups. Glycerol 7% and 3% ethylene glycol with lycopene groups yielded to slight higher percentages of membrane integrity assessed by HOST than that of the other groups, but 7% glycerol with cysteamine and 3% ethylene glycol with cysteamine showed the worst percentages of membrane integrity. Glycerol 7% and 5% glycerol with lycopene gave rise to a higher value of VAP, VSL and VCL compared with that of the other groups. On the contrary, adding to 5% glycerol with cysteamine showed negative effect for VAP, VSL, VCL and ALH values. All cryoprotectant groups with lycopene decreased chromatin damage than the other groups. Ethylene glycol 3% led to lower non-return rates of inseminated cows. However, this result was not considered to be statistically important.Öğe Effects of various antioxidants on cryopreserved bull sperm quality(Kafkas Üniversitesi, 2014) Taşdemir, Umut; Tuncer, Pürhan Barbaros; Büyükleblebici, Serhat; Özgürtaş, Taner; Durmaz, Emre; Büyükleblebici, OlgaThe objective of this study was to assess the effects of antioxidant supplement (A), fetuin (F), aminoacid (AS) and cysteine (CY) on the sperm parameters, plasma membrane integrity, chromatin damage and antioxidant activities after freeze-thawing. Ejaculates were split into five aliquots and extended to a final concentration of 15x106 spermatozoa/ml with the Tris base extender containing 0.5 ml A, 2 mg/ml F, 13% AS, 5 mM CY andadditive (C). The extended samples were equlibrated slowly to 4°C during 4 h and then frozen using a digital freezing machine.. Frozen straws were thawed individually in water bath at 37°C for 30 s to analyse progressive motility and sperm motion characteristics as well as membrane integrityBiochemical assays were performed in a spectrophotometer using commercial kits. Chromatin damage was evaluated by Comet Assay. A, F, AS anddid not show better result on the percentages of post-thaw sperm motilities. CY exhibited the greatest value of plasma membrane integrity (P<0.05)Total abnormalities were greater in C and F (17.5±0.57%; 15.5±1.98%, respectively; P<0.05). F had greater chromatin damage results (P<0.05).activity was affected by type of antioxidant, notably CY yielded the lowest results when compared to the other groups (P<0.05). In conclusion, although using antioxidants does not have any influence on the sperm motility after thawing, A, AS and CY cause reduction at abnormal spermatozoa;exhibits the greatest cryoprotective activity on plasma membrane integrity and F caused an increase at chromatin damage.Öğe The comparison of three different cryoprotectants in cryopreservation of Angora goat semen(Kafkas Üniversitesi, 2014) Büyükleblebici, Serhat; Tuncer, Pürhan Barbaros; Taşdemir, Umut; Özgürtaş, Taner; Durmaz, Emre; Büyükleblebici, OlgaThe objective of this study was to evaluate glycerol (G), ethylene glycol (EG) and dimethylsulfoxide (DMSO) which were used two different doses on in vitro semen parameters, antioxidant enzymes activities and DNA damage after the freeze-thaw process in Angora goat semen. Semen samples from 5 mature Angora goats were used in this study. A total number of 40 ejaculates were collected twice a week from the goats using an artificial vagina and the semen pooled to minimize individual variation. Each pooled ejaculate was split into 6 equal aliquots and diluted with tris base extenders supplemented with two different doses of cryoprotectants (G 3%, 6%; EG 3%, 6%; DMSO 3%, 6%). G 3% and 6% was added as a cryoprotectant had better CASA motility (P<0.01) and progressive motility (P<0.001) values when compared to EG and DMSO groups. On the other hand, EG 6% showed the best values of preserved membrane integrity (P<0.01). The evaluation of CASA sperm motions parameters, adverse effects were procured in the groups with DMSO groups when compared to the other groups (P<0.05; P<0.001). G 6% group was the greatest VAP, VSL and VCL values than the other groups (P<0.05; P<0.001). DNA damage was not affected by supplemented different doses of cryoprotectants as well as antioxidant activity (P>0.05). In conclusion, no advantages were found in using EG or DMSO to replace G for freezing of Angora goat sperm
