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Öğe 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, SenaThe 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.Öğe A novel polymerase chain reaction to detect brucella canis in dogs(KAFKAS UNIV, VETERINER FAKULTESI DERGISI, 2015) Aras, Zeki; Taspinar, Mehmet; Aydin, IbrahimIn this study, the specific polymerase chain reaction has been standardized and evaluated for the direct diagnosis of Brucella canis in vaginal swab samples from dogs. The specific primer sets are directed to the 16S-23S rRNA inter-space region of Brucella spp. and the deletion of 351 bp in BMEI1426-BMEI1427 in B. canis. A total of 21 references and field strains and 35 vaginal swab samples were used for the evaluation of the polymerase chain reaction. It found that polymerase chain reaction is positive for B. canis DNA indicated by only amplification of 214 bp product. It detected at least 2.7 x 10(1) CFU/g of bacteria diluted in vaginal swab samples indicates that the polymerase chain reaction can be used as a practical alternative for bacterial isolation. The novel polymerase chain reaction provides a simple and rapid for the detection of B. canis in clinical and field samples in one step and in short time about 24 h.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.