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Basal medium eagle solution may improve the post-thaw parameters of kangal dog semen
(2016) Gungor, Sukru; Bucak, Mustafa Numan
Amaç: Bu çalışmada dondurma-çözdürme sonrası Kangal köpeği spermatolojik parametreleri üzerine BME'nin etkisi-nin ortaya konulması amaçlandı.Gereç ve Yöntem: Ejakülatlar hafta iki kez digital maniplas-yon yardımıyla alındı. Alınan ejakülatlar 6 eşit hacme bölü-nerek temel Tris sulandırıcısı içeren (T), %5 gliserol (TG), %5 etilen glikol (TE), %5 gliserol (G) %2.5 BME (TGB2.5), TG %10 BME (TGB10), %5 etilen glikol (E) %2.5 BME (TEB2.5) ve TE %10 BME (TEB10) sulandırıcılar ile su-landırıldı. Sulandırılan spermalar 4C'ta 1.5 saat ekilibras-yon sonrası sıvı azot buharında dondurularak sıvı azotta (-196C) saklandı. Bulgular: Çalışmada progresif motilite oranı %10 BME içe-ren gliserollü sulandırıcı grubunda (%23.19), %5 etilen gli-kol içeren (%14.08) grubuna göre istatistiksel olarak yüksek bulundu. Akrozom bütünlüğü %2.5 BME içeren gliserollü sulandırıcı (%44.99) grubunda, %5 gliserol içeren (%33.63) grubuna göre istatiksel olarak üstünlük gösterdi (P0.05).Öneri: Kangal köpeği spermasının dondurulmasında sulan-dırıcıya eklenen BME çözüm sonu spermatolojik parametre-leri iyileştirebilir.
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 Unlu
The 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.
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, Fazile
The 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.
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, Derya
This 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.
EFFECTS OF ARGININE AND TREHALOSE ON POST-THAWED BOVINE SPERM QUALITY
(AKADEMIAI KIADO ZRT, 2017) Ozturk, Caner; Gungor, Sukru; Ataman, Mehmet Bozkurt; Bucak, Mustafa Numan; Baspinar, Nuri; Ili, Pinar; Inanc, Muhammed Enes
The present study was conducted to examine the protective role of arginine and trehalose on post-thaw bull sperm and oxidative stress parameters. Five ejaculates for each bull were used in the study. Each ejaculate, split into three equal aliquots and diluted at 37 degrees C with base extenders containing 2 mM arginine, 25 mM trehalose and no antioxidant (control) was cooled to 5 degrees C and then frozen. Frozen straws were thawed in a water bath for evaluation. Supplementation of the semen extender with arginine decreased the percentages of post-thawed subjective motility (29 +/- 8.21%), CASA motility (12.2 +/- 5.69%) and progressive motility (3.52 +/- 2.13%), compared with the controls (43 +/- 2.73%, 55.4 +/- 6.78% and 33.48 +/- 4.14%, respectively, P < 0.05). Supplementation of the semen extender with trehalose produced a higher mitochondrial activity and sperm viability (36.3 +/- 3.99% and 44.1 +/- 2.18%) compared with the control (13 +/- 8.15 and 31.7 +/- 3.94%, respectively, P < 0.05). It was established that trehalose (95.1%) and arginine (92.8%) protect DNA integrity compared to the control (90.4%) (P < 0.05). Trehalose supplementation in semen extenders provided great benefit in terms of viability, mitochondrial activity, and intact sperm DNA on frozen-thawed bull sperm.
Effects of cysteine and ergothioneine on post-thawed Merino ram sperm and biochemical parameters
(ACADEMIC PRESS INC ELSEVIER SCIENCE, 2011) Coyan, Kenan; Başpınar, Nuri; Bucak, Mustafa Numan; Akalın, Pınar Peker
The aim of the current study was to evaluate the effects of cysteine and ergothioneine on the post-thawed sperm parameters, lipid peroxidation and antioxidant activities. Semen samples from 5 mature Merino rams were used in the study. Semen samples, which were diluted with a Tris-based extender containing L-Cysteine and L-(+)-Ergothioneine 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 20 s in a water bath for evaluation. Ergothioneine at doses of 2 and 4 mM increased percentages of subjective motility, VSL and VCL, compared to controls following the freeze-thawing (P < 0.001). Ergothioneine at three different doses led to higher rates of progressive motility and VAP, compared to control groups (P < 0.001). Cysteine and ergothioneine at three doses provided the higher rates of ALH, in comparison to no antioxidant group (P < 0.001). As regards CASA motility, supplementation with antioxidants did not provide any significant difference on the percentage of post-thaw sperm CASA motilities, in comparison to the control. In regards of sperm membrane integrity, only cysteine 1 mM provided a greater protective effect, compared to control (P < 0.001). Percentages of sperm with high mitochondrial activity were dramatically increased with cysteine at doses of 1 and 2 mM, compared to control (P < 0.05). No significant differences were observed in sperm acrosome integrities among groups. CAT activity was increased significantly only in cysteine1 mM compared to control group (P < 0.001). Cysteine at doses of 2 and 4 mM showed a tendency of increased activities of CAT when compared to control. But these increases were not statistically significant. Supplementation with antioxidants did not significantly affect activities of SOD and GPx. Findings of this study showed that ergothioneine supplementation in semen extenders, was of greater benefit to motility and motion characteristics of frozen-thawed ram sperm. Crown Copyright (C) 2011 Published by Elsevier Inc. All rights reserved.
Effects of different concentrations of BHT on microscopic and oxidative parameters of Mahabadi goat semen following the freeze-thaw process
(ACADEMIC PRESS INC ELSEVIER SCIENCE, 2013) Naijian, Hamid Reza; Kohram, Hamid; Shahneh, Ahmad Zare; Sharafi, Mohsen; Bucak, Mustafa Numan
Oxidative damage to sperm is one of the main causes for decline in motility and fertility of frozen thawed sperm. Thus, it is crucial to use cryoprotectant agents in extender in order to prevent lethal intracellular ice crystal formation. The present study aims to evaluate the effects of different concentrations of the antioxidant butylated hyroxytoluene (BHT) on sperm parameters post-thaw. Semen was diluted into five equal aliquots of extender containing different concentrations of BHT (0, 0.5, 1, 2 and 4 mM), aspirated into 0.25 mL straws, and equilibrated at 5 degrees C for 2 h. After equilibration, straws were frozen in liquid nitrogen vapor and plunged into liquid nitrogen for storage. Sperm parameters, including motility and progressive motility, viability, membrane integrity, acrosome integrity and capacitation status, were assessed. Malondialdehiyde (MDA) and glutathione peroxidase (GSH-PX) activity were also evaluated after freezing-thawing. Results of this experiment show that addition of 1 mM of BHT to the extender for freezing of goat semen can improve motility, progressive motility and viability (P < 0.05) and reduce the MDA level (P < 0.01). HOST (hypo-osmotic swelling test), acrosome integrity, capacitation status and GSH-PX were not affected by the concentrations of BHT (P > 0.05). Therefore, we conclude that the optimum concentration of BHT for cryopreservation of goat semen is 1 mM. (C) 2013 Elsevier Inc. All rights reserved.
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, Ali
Amaç: 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.
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, Servet
This 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.
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 Barbaros
The 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.
Effects of Hypotaurine, Cysteamine and Aminoacids Solution on Post-Thaw Microscopic and Oxidative Stress Parameters of Angora Goat Semen
(Soc Study Reproduction, 2010) Çoyan, Kenan; Tuncer, Pürhan Barbaros; Sarıözkan, Serpil; Ulutaş, Pınar Alkım; Bucak, Mustafa Numan; Başpınar, Nuri; Özkalp, Birol
This study was conducted to determine the effects of cysteamine, hypotaurine and aminoacids solution (BME) on standard semen parameters, lipid peroxidation and antioxidant activities of Angora goat semen after the freeze–thawing process. Ejaculates collected from four Angora goats were evaluated and pooled at 37 C. Semen samples, which were diluted with a Tris-based extender containing the antioxidants hypotaurine (5 mM) and cysteamine (5 mM), and an aminoacid solution (13%), and an extender containing no antioxidants (control), were cooled to 5 C and frozen in 0.25-ml French straws in liquid nitrogen. Frozen straws were thawed individually at 37 C for 20 s in a water bath for evaluation. Supplementation with cysteamine, hypotaurine and BME caused significant (P < 0.05) increases in sperm motility, and significant (P < 0.05) decreases in total abnormality rates in comparison to the control group. While all in vitro treatments did not affect the acrosomal abnormality rates, hypotaurine and BME but not cysteamine significantly (P < 0.05) increased the HOST results as compared to the control group. Supplementation with antioxidants and BME did not significantly affect MDA levels and CAT activity in comparison to the control group (P > 0.05). The antioxidants hypotaurine and cysteamine decreased SOD activity when compared to the BME group and controls (P < 0.001).
Effects of hypotaurine, cysteamine and aminoacids solution on post-thaw microscopic and oxidative stress parameters of Angora goat semen
(ELSEVIER SCI LTD, 2009) Bucak, Mustafa Numan; Tuncer, Puerhan Barbaros; Sariozkan, Serpil; Ulutas, Pinar Alkim; Coyan, Kenan; Baspinar, Nuri; Ozkalp, Birol
This study was conducted to determine the effects of cysteamine, hypotaurine and aminoacids solution (BME) on standard semen parameters, lipid peroxidation and antioxidant activities of Angora goat semen after the freeze-thawing process. Ejaculates collected from four Angora goats were evaluated and pooled at 37 degrees C. Semen samples, which were diluted with a Tris-based extender containing the antioxidants hypotaurine (5 mM) and cysteamine (5 mM), and an aminoacid solution (13%), and an extender containing no antioxidants (control), were cooled to 5 degrees C and frozen in 0.25-ml French straws in liquid nitrogen. Frozen straws were thawed individually at 37 degrees C for 20 s in a water bath for evaluation. Supplementation with cysteamine, hypotaurine and BME caused significant (P < 0.05) increases in sperm motility, and significant (P < 0.05) decreases in total abnormality rates in comparison to the control group. While all in vitro treatments did not affect the acrosomal abnormality rates, hypotaurine and BME but not cysteamine significantly (P < 0.05) increased the HOST results as compared to the control group. Supplementation with antioxidants and BME did not significantly affect MDA levels and CAT activity in comparison to the control group (P > 0.05). The antioxidants hypotaurine and cysteamine decreased SOD activity when compared to the BME group and controls (P < 0.001). (c) 2009 Elsevier Ltd. All rights reserved.
Effects of ultrasonication on damaged spermatozoa and mitochondrial activity rate
(SCIENTIFIC TECHNICAL RESEARCH COUNCIL TURKEY-TUBITAK, 2016) Peker Akalin, Pinar; Baspinar, Nuri; Coyan, Kenan; Bucak, Mustafa Numan; Gungor, Sukru; Ozturk, Caner
The aim of this study was to describe an optimal sonication procedure for sperm cells. Therefore, we used several parameters such as damaged spermatozoa rate (%), mitochondrial activity rate (%), levels of lipid peroxidation, and total antioxidant potential. Ejaculates were collected from rams (n = 3) and were divided into aliquots and 3-, 6-, and 10-s duration times; 1, 3, 5, and 8 repetitive application groups were established. In the groups with 3-, 6- and 10-s duration times, with the increasing number of repeated applications, damaged spermatozoa rates increased (P < 0.05) while mitochondrial activity rates decreased (P < 0.05). In relation with sonication duration time, total antioxidant potential levels increased (P < 0.05) in single-application groups compared to those in control groups and gradually decreased as the repetitions increased. The most effective results were obtained in the group with 8 repetitions and 10-s duration based on damaged spermatozoa rate and mitochondrial activity rate.
Ergothioneine attenuates the DNA damage of post-thawed Merino ram sperm
(ELSEVIER SCIENCE BV, 2012) Coyan, Kenan; Bucak, Mustafa Numan; Baspinar, Nuri; Taspinar, Mehmet; Aydos, Sena
The objective of the current study was to evaluate the effects of antioxidant ergothioneine added to cryopreservation extender on the DNA integrity 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 ergothioneine at different concentrations and no antioxidant (control), were cooled to 5 degrees C and frozen in 0.25 ml French straws. Frozen straws were then thawed at 37 degrees C for 20s in a water bath for evaluation of sperm DNA damage using the Comet test. The addition of ergothioneine at concentrations of 1, 2 and 4 mM resulted in lower sperm with damaged DNA (5.4, 4.7 and 3.2%, respectively) than that of control (7.9%), thus reducing the DNA damage (P<0.01). Findings of this study showed that the increasing concentrations of ergothioneine in semen extenders, were of greater benefit to DNA integrity of frozen-thawed ram sperm. Crown Copyright (C) 2012 Published by Elsevier B.V. All rights reserved.
Esterified glucomannan improves aflatoxin-induced damage of sperm parameters during liquid storage of ram semen at 5 degrees C
(ACADEMIC PRESS INC ELSEVIER SCIENCE, 2014) Ataman, Mehmet Bozkurt; Bucak, Mustafa Numan; Coyan, Kenan
The aim of the present work was to study the effects of aflatoxin (AF) on sperm parameters in rams, and to determine the protective efficiency of esterified glucomannan (EG) co-administered with AF up to 96 h of the liquid storage of ram semen at 5 degrees C. Thirty-two Merino rams (12-14 months old) were used. The animals were examined for their general health status. To ensure their adaptation to the environment and the new feeding regimen, a 15-day acclimatization programme was applied to the animals, prior to the start of the study. Experimental feeding was continued for ninety-two days. The experimental design consisted of four dietary treatments. The control group (C) was fed with commercial feed. The AF group was fed with commercial feed plus 250 mu g/day of total AF. The EG group received commercial feed plus 2 g/day of EG. The AF + EG group was given commercial feed plus 250 mu g/day of total AF and 2 g/day of EG. In the study, ejaculates were obtained from rams twice a week for 12 weeks, using an electro-ejaculator. After collected, the ejaculates were diluted with a skimmed milk extender, and stored at 5 degrees C. Sperm motility and rates of abnormal and nonviable spermatozoa were determined for the different treatment groups at 5 degrees C at 0, 24, 48, 72 and 96 h of liquid storage. During the first two weeks of the trial, the groups did not statistically differ from each other for sperm motility or rates of abnormal and nonviable spermatozoa at 0, 24, 48 and 96 h of storage. As from the third week, the short-term storage of semen produced statistically significant differences between the AF group and the other treatment groups for sperm parameters (p < 0.05). The administration of aflatoxin was observed to have reduced sperm motility and to have increased the rates of abnormal and nonviable spermatozoa in comparison to the control group (p < 0.05), while EG co-administered with AF was determined to have ameliorated the adverse effects of AF on sperm parameters, and this ameliorative effect continued throughout the short-term storage of semen. On the other hand, aflatoxin administration resulted in the deterioration of the sperm parameters in the following weeks, and the combined administration of EG + AF reversed this adverse effect, thus, bringing the sperm parameters closer to the values of the control group. This study demonstrated that, in rams, AF adversely affected sperm, biochemical and testis parameters, and that the combined administration of EG and AF reversibly improved these adverse effects. (C) 2014 Elsevier Inc. All rights reserved.
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, Ali
Amaç: 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.
Histopathological evaluation of the effects of thymoquinone and resveratrol on the liver in rats administered doxorubicin
(Selçuk Üniversitesi, 2022) Ateş, Mehmet Burak; Özdemir, Özgür; Öztürk, Ali Erdem; Bucak, Mustafa Numan; Bulut, Ayşegül
Aim: The purpose was to investigate the effects of various dosages of thymoquinone and resveratrol (5 and 20 mg/kg) on doxorubicin-induced hepatotoxicity in rats from a pathological standpoint. Materials and Methods: Eighty male Wistar Albino rats were used in this study. Animals were divided in to 10 groups: Control (physiological saline, PO); Doxorubicin (physiological saline, PO and Dox,15mg/kg Dox in 10th days, IP); Thymoquinone -5 (TQ-5, 5 mg/kg TQ, PO); TQ-20 (20 mg/ kg TQ, PO); Resveratrol-5 (Res-5, 5 mg/kg Res, PO); Res-20 (20 mg/kg Res, PO); Dox+TQ-5 ; Dox+TQ-20; Dox+Res-5; Dox+Res-20. After the 21-day experiment, 6 replicates were randomly selected from the groups. After weighing the body weight, the livers of the euthanized rats were dissected and weighed. Routine tissue processing processes were applied to liver samples. Hepatocyte degeneration, necrosis/apoptosis, bile duct hyperplasia, dissociation, congestion, karyomegaly, mononuclear cell infiltration, binuclear hepatocytes, and mitosis were all examined microscopically, and a liver total lesion score was calculated Results: Dox treatment increased relative liver weight, but the TQ and Res groups prevented this increase (p<0.05). The liver total lesion score, which increased with Dox, was shown to be lower in the TQ and Res groups (p<0.05). However, the Dox+TQ-5, Dox+TQ-20, and Dox+Res-20 groups, showed no amelioration in necrosis/apoptosis. Conclusion: TQ and Res (5 and 20 mg/kg) decreased the total liver lesion score induced by Dox. Although TQ and RES diminish degeneration and inflammation, their poor protective effects on necrosis/apoptosis, one of the key criteria, were considered as a limiting cause for their uncontrolled usage.
Influence of ellagic acid and ebselen on sperm and oxidative stress parameters during liquid preservation of ram semen
(ROYAN INST, 2019) Bucak, Mustafa Numan; Bodu, Mustafa; Baspinar, Nuri; Gungor, Sukru; Ili, Pinar; Acibaeva, Begimay; Topraggaleh, Tohid Rezaei; Dursun, Şükrü
Objective: The purpose of the present study was to assess the effects of ellagic acid and ebselen on sperm and oxidative stress parameters during liquid preservation of ram semen. Materials and Methods: In this experimental study, sixty ejaculates from six mature Merino rams were used. In experiment 1, the ejaculates were diluted in base extender contained ellagic acid at 0 (control), 0.5, 1, and 2 mM. In experiment 2, ebselen at 0 (control), 10, 20, and 40 mu M were added to the extender. Sperm motility, viability, mitochondrial membrane potential, DNA integrity, lipid peroxidation (LPO), the antioxidant potential (AOP), and total glutathione (tGSH) were evaluated at 0, 24, 48, and 72 hours of preservation. Results: Supplementation of ellagic acid at 1 and 2 mM resulted in higher sperm motility and viability at 0 hours of storage. Ellagic acid at 2 mM led to higher motility and viability compared to controls after 0, 24, and 48 hours of preservation and increased AOP after 24 and 72 hours. Higher tGSH was at 1 mM ellagic acid, compared to control after 72 hours. Addition of ebselen at a concentration of 40 mu M increased motility at 24 and 48 hours and 10 mu M produced the same effect after 48 and 72 hours of storage as well as higher viability, compared to the controls after 0 hours of storage. Sperm DNA integrity was significantly improved after 24, 48, and 72 hours with the addition of ebselen at 10 mu M, and after 72 hours at 40 mu M. Addition of 40 mM ebselen also reduced the LPO levels after 24 hours of storage compared to the controls. Conclusion: The results showed that supplementation of ellagic acid and ebselen in semen extender has a potential effect on sperm and oxidative stress parameters during liquid preservation of ram semen.
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, Cemal
It 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.
Influence of lycopene and cysteamine on sperm and oxidative stress parameters during liquid storage of ram semen at 5 degrees C
(ELSEVIER SCIENCE BV, 2016) Peker Akalin, Pinar; Bucak, Mustafa Numan; Gungor, Sukru; Baspinar, Nuri; Coyan, Kenan; Dursun, Sukru; Ili, Pinar
Ejaculates were collected from six Merino rams with the aid of an artificial vagina twice a week. The ejaculates containing spermatozoa with >80% forward progressive motility and concentrations higher than 2 x 10(6) spermatozoa/ml were pooled. The present study included two experiments. In experiment 1, each pooled ejaculate was divided into four equal aliquots and diluted (37 degrees C) with the Tris based extender, containing 0 (control), 0.5, 1 and 2 mM lycopene, at a final concentration of approximately 400 x 10(6) sperms/ml (single step dilution), In experiment 2, cysteamine at concentrations of 0 (control), 0.5,1 and 2 mM, was used as an additive in the extender, and the procedure explained above was applied for the division of aliquots and the dilution of semen. Diluted semen samples were kept in glass tubes and cooled from 37 degrees to 5 degrees C in a cold cabinet, and maintained at 5 degrees C. Sperm and oxidative stress parameters were evaluated after 0, 24, 48 and 72 h of storage at 5 degrees C. The extender supplemented with 0.5 mM lycopene resulted in higher mitochondrial activity rate (p<0.05) in comparison to the control group at 72 h of storage. Lycopene at 0.5 mM dose led to higher sperm motility rate (p<0.05) when compared to 2 mM lycopene group at 72 h of liquid storage. As regards oxidative stress parameters, only 2 mM lycopene increased total glutathione levels (p<0.05) at 0 h of storage. The extender supplemented with 1 mM cysteamine gave higher motility (p<0.05) at 48 h compared to control. As regards oxidative stress parameters, 1 and 2 mM cysteamine at 48 h and 1 mM cysteamine at 72 h increased total glutathione levels (p<0.05) compared to control groups. Cysteamine at 1 and 2 mM doses decreased lipid peroxidation (p<0.05) at 0 h of liquid storage compared to control. Our data suggest that lycopene at 0.5 and 2 mM and cysteamine at 1 and 2 mM doses can be added to Tris based extender for improving the ram sperm motility, viability, mitochondrial activity and oxidative stress parameters during the liquid storage. (C) 2016 Elsevier B.V. All rights reserved.

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