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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 Raffinose and hypotaurine improve the post-thawed Merino ram sperm parameters(ACADEMIC PRESS INC ELSEVIER SCIENCE, 2013) Bucak, Mustafa Numan; Keskin, Nazan; Taspinar, Mehmet; Coyan, Kenan; Baspinar, Nuri; Cenariu, Mihai C.; Bilgili, AliThe aim of this study was to determine the effects of raffinose and hypotaurine on sperm parameters after the freeze-thawing of Merino ram sperm. Totally 40 ejaculates of five Merino ram were used in the study. Semen samples, which were diluted with a Tris-based extender containing 10 mM raffinose, 5 mM hypotaurine, 5 mM raffinose +2.5 mM hypotaurine (H + R) and no antioxidant (control), were cooled to 5 degrees C and frozen in 0.25 ml French straws and stored in liquid nitrogen. Frozen straws were then thawed individually at 37 degrees C for 25 s in a water bath for evaluation. The addition of raffinose led to higher percentages of subjective and CASA motilities (47.5 +/- 12.2%, 46.3 +/- 13.6%) compared to controls (38.8 +/- 13.8%, 30.5 +/- 11.7%, P < 0.05). For the CASA progressive motility, 5 mM raffinose (20.12 +/- 8.82%) had increasing effect in comparison to control (10 +/- 7.94%, P <0.05) following the freeze-thawing process. Raffinose and hypotaurine led to higher viability (40.8 +/- 4.68%, 40.8 +/- 4.7%), high sperm mitochondrial activity (29.5 +/- 5.4%, 27.3 +/- 4.9%) and acrosome integrity (50.8 +/- 8.1, 50.7 +/- 4.4) percentages, compared to control groups (31.5 +/- 3.5%, 9.5 +/- 8.2%, 42.8 +/- 7.3%, P < 0.05). H + R group only led to high sperm mitochondrial activity when compared to control group. In the comet test, raffinose and hypotaurine resulted in lower sperm with damaged DNA (6.2% and 3.9%) than that of control (9.1%), reducing the DNA damage. For TUNEL assay, The TUNEL-positive cell was distinguished by distinct nuclear staining. Raffinose and H + R groups resulted in lower sperm with TUNEL-positive cell (1.5 +/- 1.2% and 2.1 +/- 0.9%) than that of control (4.9 +/- 2.5%) (P < 0.05). In conclusion, findings of this study showed that raffinose and hypotaurine supplementation in semen extenders provided a better protection of sperm parameters against cryopreservation injury, in comparison to the control groups. Crown Copyright (C) 2013 Published by Elsevier Inc. All rights reserved.Öğe Van Gölünde Avlanan İnci Kefali Örneklerinde Organik Klorlu İnsektisit Düzeyleri Üzerinde Araştırmalar(Selçuk Üniversitesi, 1996) Sağmanlıgil, Hülya; Bilgili, Ali; Yarsan, Ender; Türel, İdrisIn this research, total one hundred and sixty fishes (chalcalburnus tarichi) were used in 1994 and 1995 years. and heptachlore epoksite was only determined as organochlorine insecticides in these experiments. Heptachlore epoksite levels in fish meat were found as 0.0204 ppm. As a result, Van Lake and fish (chalcalburnus tarichi) was not contaminated with organochlorine insecticides, even if there are some low levels of heptachlore epoksite. This would suggest that contamination of Van Lake with organochlorine insecticides is not significant, therefore it is not life threatening.
