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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 INC ELSEVIER SCIENCE, 2014) Buyukleblebici, Serhat; Tuncer, Purhan Barbaros; Bucak, Mustafa N.; Tasdemir, Umut; Eken, Ayse; Buyukleblebici, Olga; Durmaz, EmreThere 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 Cryopreservation of bull sperm: Effects of extender supplemented with different cryoprotectants and antioxidants on sperm motility, antioxidant capacity and fertility results(ELSEVIER, 2014) Buyukleblebici, Serhat; Tuncer, Purhan Barbaros; Bucak, Mustafa Numan; Eken, Ayse; Sariozkan, Serpil; Tasdemir, Umut; Endirlik, Burcu UnluThe objectives of this study were to compare glycerol (G) and ethylene glycol (EG) at different concentrations and trehalose (T) or cysteine (C; with/without) in Tris extender for cryopreservation of bull semen. Twenty-four ejaculates obtained from three bulls were included in the study. Each ejaculate was divided into four equal aliquots and diluted using both of the Tris extenders with G (5% or 7%) or EG (3% or 5%). After that, each extenders were divided into three equal aliquots and diluted using both of the 5 mM C or 25 mM T, and control (without additives) was cooled to 4 degrees C and frozen in 0.25 ml French straws. The addition of 3% and 5% EG without antioxidants resulted in the least Computer-Assisted Sperm motility Analysis (CASA) motility as compared with the other groups. Treatment with 25 mM Tin 3% EG beneficially effected acrosome morphology as compared with the other groups. Also, treatment with 3% EG with 25 mM T and 5% EG resulted in a greater rate of total abnormalities. Treatment with 3% G yielded a slightly greater percentage of membrane integrity by Hypo-Osmotic Swelling Test (HOST) assessment than that of the other groups. Treatment with 3% EG with 5 mM C resulted in the greatest concentration of malondialdehyde (MDA). The glutathione peroxidase (GPx) antioxidant activity was increased in the C-treatment groups when compared to the other groups. Treatment with 5% EG and 5 mM C resulted in less chromatin damage and detrimental impacts on tail moment. Treatment with 5% EG led to greater non-return rates of inseminated cows. However, this result was not considered to be statistically important. (C) 2014 Elsevier B.V. All rights reserved.Öğe Effect of glutamine and sugars after bull spermatozoa cryopreservation(ELSEVIER SCIENCE INC, 2011) Tuncer, Purhan Barbaros; Sarıözkan, Serpil; Bucak, Mustafa Numan; Ulutaş, Pınar Alkım; Akalın, Pınar Peker; Büyükleblebici, Serhat; Cantürk, FazileThe objective of this study was to evaluate the effects of the addition of different sugars (raffinose, sucrose, and trehalose) on bull spermatozoa cryopreserved in a commercial extender (Optidyl) supplemented with glutamine on semen parameters, fertilizing ability and superoxide dismutase (SOD) activity. Nine ejaculates for each bull were used in the study. Semen was frozen in five different extenders: raffinose 25 mM plus glutamine 3 mM (RGO), sucrose 25 mM plus glutamine 3 mM (SGO), trehalose 25 mM plus glutamine 3 mM (TGO), glutamine 3 mM (GO) and control (0). Insemination doses were processed so that each 0.25 mL straw contained 15 x 10(6)sperm. Groups of GO and RGO resulted in the higher rates of subjective (54.0 +/- 1.7% and 64.0 +/- 1.1%; P < 0.01) and CASA motilities (53.0 +/- 2.7% and 61.0 +/- 4.4%; P < 0.001), respectively compared to the other groups. The supplementation of additives did not provide an effect on the level of post-thaw sperm CASA progressive motilities, the sperm motion characteristics and pregnancy rates. GO and RGO provided the better protective effect for sperm acrosome (4.0 +/- 0.5% and 12.0 +/- 0.6%) and total abnormalities (5.0 +/- 0.3% and 13.0 +/- 0.7%; P < 0.001), respectively. At the HOST values, the additives did not give to result the protective effect in comparison to Optydil extender without additives (P > 0.05). For pregnancy rates, there were no significant differences among the groups. The supplementation of additives did not provide any significant difference on the level of SOD activity (P > 0.05). It can be also thought that these sugars might have worked with glutamine in a synergy. Thereby, sugars such as raffinose and sucrose with glutamine in freezing extender may be recommended to facilitate bull semen freezability. (C) 2011 Elsevier Inc. All rights reserved.Öğe Effects of Antioxidants on Post-thawed Bovine Sperm and Oxidative Stress Parameters: Antioxidants Protect DNA Integrity Against Cryodamage(Academic Press Inc Elsevier Science, 2010) Bucak, Mustafa Numan; Tuncer, Purhan Barbaros; Sarıözkan, Serpil; Başpınar, Nuri; Taşpınar, Mehmet; Çoyan, Kenan; Bilgili, Ali; Akalın, Pınar Peker; Büyükleblebici, Serhat; Aydos, Sena; Ilgaz, Seda; Sunguroğlu, Asuman; Öztuna, DeryaThis study was conducted to determine the effects of methionine, inositol and carnitine on sperm (motility, abnormality, DNA integrity and in vivo fertility) and oxidative stress parameters (lipid peroxidation, total glutathione and antioxidant potential levels) of bovine semen after the freeze-thawing process. Nine ejaculates, collected with the aid of an artificial vagina twice a week from each Simmental bovine, were included in the study. Each ejaculate, splitted into seven equal groups and diluted in Tris-based extender containing methionine (2.5 and 7.5 mM), carnitine (2.5 and 7.5 mM), inositol (2.5 and 7.5 mM) and no additive (control), was cooled to 5 degrees C and then frozen in 0.25 ml straws. Frozen straws were then thawed individually at 37 degrees C for 20 s in a water bath for the evaluation. The extender supplemented with 7.5 mM doses of carnitine and inositol led to higher subjective motility percentages (61.9 +/- 1.3% and 51.3 +/- 1.6%) compared to the other groups. The addition of methionine and carnitine at doses of 2.5 and 7.5 mM and inositol at doses of 7.5 mM provided a greater protective effect in the percentages of total abnormality in comparison to the control and inositol 2.5 mM (P < 0.001). As regards CASA motility, 7.5 mM carnitine (41.6 +/- 2.9% and 54.2 +/- 4.9%) and inositol (34.9 +/- 2.0% and 47.3 +/- 2.2%) caused insignificant increases in CASA and total motility in comparison to the other groups. All of the antioxidants at 2.5 and 7.5 mM resulted in lower sperm with damaged DNA than that of control, thus reducing the DNA damage (P < 0.05). No significant differences were observed in CASA progressive motility and sperm motion characteristics among the groups. In fertility results based on 59-day non-returns, no significant differences were observed in non-return rates among groups. As regards biochemical parameters, supplementation with antioxidants did not significantly affect LPO and total GSH levels in comparison to the control group (P > 0.05). The maintenance of AOP level in methionine 2.5 mM was demonstrated to be higher (5.06 +/- 0.38 mM) than that of control (0.96 +/- 0.29 mM) following the freeze-thawing (P < 0.001). Supplementation with these antioxidants prior to the cryopreservation process protected the DNA integrity against the cryodamage. Furthermore, future research should focus on the molecular mechanisms of the antioxidative effects of the antioxidants methionine, carnitine and inositol during cryopreservation.Öğe The effects of different egg yolk concentrations used with soy bean lecithin-based extender on semen quality to freeze bull semen(Selçuk Üniversitesi Veterinerlik Fakültesi, 2010) Sariozkan, Serpil; Tuncer, Purhan Barbaros; Bucak, Mustafa Numan; Buyukleblebici, Serhat; Kinet, Huseyin; Bilgen, AliAmaç: Boğa spermasını dondurmak için soya lesitin temelli sulandırıcıya (Bioxcell®) %5 ve 10 konsantrasyonlarında santrifüj edilmiş yumurta sarısı (SYS) katıldı ve bunların çözüm sonu sperm motilite, morfolojik anormallikler ve membran bütünlüğü üzerindeki sinerjistik etkileri değerlendirildi. Gereç ve Yöntem: Her bir Simental boğadan alınan ejakulatlar (n=12) üç eşit miktara ayrıldı ve sırasıyla %5 (B5), %10 (B10) SYS eklenmiş ve hiç SYS (B0) içermeyen soya lesitin temelli sulandırıcı ile sulandırıldı. Ardından standart protokollere göre donduruldu ve çözdürüldü. Spermatozoa kryocanlılığı, in vitro çözüm sonu motilite (CASA), akrozomal ve diğer anormallikler ve plasma membran bütünlüğü (HOST) yönünden değerlendirildi. Bulgular: Simental boğalarda, dondurma ve çözdürme sonrası B5 ile sulandırılan grupta, diğer sulandırıcılarla sulandırılan gruplardan önemli derecede daha yüksek CASA motilite ve CASA progresif motilite oranı elde edilmiştir (p<0.001). Gruplar arasında, VAP, VCL ve ALH yönünden önemli bir farklılık bulunmamıştır (p>0.05). En yüksek VSL (p<0.01) ve LIN değeri (p<0.001) B10 grubundan elde edilmiştir. Membran bütünlüğü oranı, B5 grubunda, diğer gruplara göre önemli derecede yüksek bulunmuştur (p<0.001). Öneri: Soya lesitinle kombine olarak sulandırıcıya %5 SYS eklenmesi boğa spermasının dondurulabilirliğini önemli derecede artırmıştır.Öğe The effects of different sugars on motility, morphology and DNA damage during the liquid storage of rat epididymal sperm at 4 degrees C(ACADEMIC PRESS INC ELSEVIER SCIENCE, 2012) Sariozkan, Serpil; Bucak, Mustafa Numan; Canturk, Fazile; Ozdamar, Sairn; Yay, Arzu; Tuncer, Purhan Barbaros; Ozcan, ServetThis study evaluated the protective effects of supplementation with three different sugars on the motility, morphology and DNA integrity of rat epididymal sperm chilled and stored at 4 degrees C Epididymides were obtained from each donor. Rat epididymal sperm was diluted in Ham's F10 plus raffinose, Ham's F10 plus trehalose, Ham's F10 plus fructose, and Ham's F10 medium for control purposes. Thereafter, the extended sperm were chilled and stored in liquid form at 4 degrees C. Sperm motility, morphological abnormalities and DNA damage were determined at 0 and 12 h after chilling. No significant difference was observed in any of the parameters evaluated at 0 h, before storage (P > 0.05). After 12 h of storage, all sugar additives led to statistically higher motility, normal sperm morphology and DNA integrity in comparison to the control group. Raffinose gave the best motility percentages (32.86 +/- 1.84%) after 12 h of storage at 4 degrees C, compared to the other groups (P < 0.001). In conclusion, Raffinose, trehalose and fructose provided a better protection of sperm functional parameters against chilling injury, in comparison to the control group. Crown Copyright (C) 2012 Published by Elsevier Inc. All rights reserved.Öğe Effects of dithioerythritol on ram semen after the freeze-thawing process(ACADEMIC PRESS INC ELSEVIER SCIENCE, 2011) Baspinar, Nuri; Coyan, Kenan; Bucak, Mustafa Numan; Tuncer, Purhan BarbarosThe aim of this study was to evaluate the effects of dithioerythritol added to cryopreservation extender on the post-thawed sperm parameters, lipid peroxidation and antioxidant activities of Merino ram sperm. Semen samples from 5 mature Merino rams (1 and 2 years of age) were used in the study. Semen samples, which were diluted with a Tris-based extender containing 0.5, 1, and 2 mM dithiothreitol and no antioxidant (control), were cooled to 5 degrees C and frozen in 0.25 ml French straws. Frozen straws were then thawed individually at 37 degrees C for 20s in a water bath for evaluation. The addition of dithioerythritol at 0.5 and 2 mM doses led to higher percentages of subjective motility (62.9 +/- 4.2% and 63.6 +/- 1.8%) compared to control (52.0 +/- 4.9%, P < 0.05). As regards CASA motility, dithioerythritol 0.25 and 2 mM (60.2 +/- 4.5% and 59.6 +/- 1.2%) groups were higher from that of control (44.2 +/- 8.7%, P < 0.05). For the CASA progressive motility, 0.25, 0.5 and 2 mM doses of dithioerythritol (22.0 +/- 2.1%, 21.7 +/- 2.5% and 24.0 +/- 1.2%) had increasing effect in comparison to control (15.0 +/- 2.5%). Dithioerythritol at 1 and 2 mM doses for ALH provided higher values compared to the control (P < 0.001) following the freeze-thawing process. Supplementation with dithiothreitol did not significantly affect the integrities of sperm membrane and acrosome, and mitochondrial activities. No significant differences were observed in biochemical parameters among the groups (P > 0.05). Findings of this study showed that dithioerythritol supplementation in semen extenders, was of greater benefit to sperm motility of frozen-thawed ram sperm. (C) 2011 Elsevier Inc. All rights reserved.Öğe Evaluation of post-thaw quality of Brown-Swiss and Holstein bull semen diluted with different extenders(Selçuk Üniversitesi Veterinerlik Fakültesi, 2010) Sariozkan, Serpil; Tuncer, Purhan Barbaros; Bucak, Mustafa Numan; Buyukleblebici, Serhat; Kinet, Huseyin; Bilgen, AliAmaç: Tris + yumurta sarısı, Bioxcell® ve Optidyl® sulandırıcıları ile sulandırılan İsviçre-Esmeri ve Holştayn boğa spermasının çözdürme sonrası kalitesini değerlendirmektir. Gereç ve Yöntem: Holştayn (n=36) ve Esmer (n=36) boğalardan alınan ejakulatlar üç kısma ayrıldı ve sırasıyla Tris, Optidyl® ve Bioxcell® sulandırıcılarıyla sulandırıldı. Standart protokollere göre donduruldu ve çözdürüldü. Sulandırıcıların etkinliği, çözüm sonu sperm motilitesi, akrozomal ve toplam anormallikler ve plazma membran bütünlüğü değerlendirilerek belirlendi. Bulgular: Holştayn ırkı boğa spermasında, dondurmaçözdürme sonrası en yüksek subjektif (p<0.01), ilerleyen (p<0.001) ve CASA motilite (p<0.001) oranları Optidyl® ile sulandırılan grupta saptanmıştır. Optidyl® sulandırıcısının spermatozoa akrozom ve membran bütünlüğünü diğer sulandırıcılara kıyasla en iyi şekilde koruduğu saptanmıştır (p<0.001). Motilite karakterlerinden VAP, VSL ve LIN yönünden en yüksek değerler Optidyl ve Tris sulandırıcılarından elde edilmiştir (p<0.05). Esmer ırk boğa spermasında ise, sulandırıcılar arasında en düşük çözüm sonu subjektif (p<0.01), CASA (p<0.001) motiliteleri ve membran bütünlüğü (p<0.001) oranı Bioxcell® ile sulandırılmış grupta elde edilmiştir. Optidyl® sulandırıcısı kullanılan grupta, Bioxcell® sulandırıcısına kıyasla daha yüksek ilerleyen motilite oranı elde edilmiştir (p<0.01). Bioxcell® ve Tris sulandırıcısı kullanıldığında daha yüksek oranda akrozomal ve toplam anormallikler saptanmıştır. ALH yönünden, en yüksek değer diğer gruplarla karşılaştırıldığında Optidyl sulandırıcısından elde edilmiştir (p<0.05). Öneri: Holştayn ve İsviçre-Esmeri boğa spermasının dondurulması amacıyla Optidyl® sulandırıcısı Tris + yumurta sarısı ile Bioxcell® sulandırıcılarına tercih edilebilir.Öğe Influence of fetuin and hyaluronan on the post-thaw quality and fertilizing ability of Holstein bull semen(ACADEMIC PRESS INC ELSEVIER SCIENCE, 2015) Sariozkan, Serpil; Bucak, Mustafa Numan; Tuncer, Purhan Barbaros; Buyukleblebici, Serhat; Eken, Ayse; Akay, CemalIt was determined that fetuin and hyaluronan supplementation did not provide any significant effect on the post-thaw subjective and CASA motility percentages and sperm motion characteristics, in comparison to the controls (P > 0.05). Sperm acrosome and total abnormalities were similar in all groups (P > 0.05). Groups M (hyaluronan + fetuin) and H (hyaluronan) displayed a higher rate of sperm membrane integrity, compared to that of Group C (control) (P < 0.01). According to the results of the comet assay, the lowest percentage of sperm with damaged DNA was achieved in Group H, when compared to all of the experimental groups (P < 0.01). Furthermore, all of the additives resulted in a lower rate of sperm with damaged DNA than that of the controls, and thus, reduced DNA damage (P < 0.01). For pregnancy rates, there were no significant differences between the extender groups (P > 0.05). MDA formation was found to be lower in Groups M and F (P < 0.01). In Group M, SOD activity was determined to have significantly increased (23.61 +/- 5.62 U/ml) compared to the other groups (P < 0.01). All experimental groups had a GSH-Px activity higher than that of the control group (P < 0.01). Crown Copyright (C) 2015 Published by Elsevier Inc. All rights reserved.
