Fatih AVDATEK, Mustafa GÜNDOĞAN, Deniz YENİ

Ramlıç Irkı Koç Spermasına Katılan Bor’un Dondurma ve Çözdürme Sonrası Spermatolojik Parametreler, Oksidatif Stres ve DNA Hasarı Üzerine Etkileri

Çalışmada koç ejakülatlarının dondurulması ve çözdürülmesi sürecinde sperma sulandırıcısına katılan farklı dozlarda Borun spermatolojik değerler, oksidatif stress ve DNA hasarı üzerine etkilerinin araştırılması amaçlanmıştır. Araştırmada toplam 5 Ramlıç koçtan sun’i vagina ile alınan ejekülatlar toplandı. Başlıca spermatolojik özellikleri belirlenen normospermi kalitesindeki ejekülatlar birleştirilerek kullanıldı. Araştırma ve kontrol gruplarında spermalar mix ejekülat biçiminde değerlendirildi. Spermaların sulandırılması ve dozlanması farklı Boron dozları (1, 2 ve 4 mM) ve % 5 glyserol içeren Tris ana sulandırıcısı ile yapıldı. Ayrıca Boron içermeyen kontrol grubu da oluşturuldu. Sulandırma ve dozlama işlemleri sonrasında 0.25 ml lik payetlere çekilen spermalar sıvı azot buharında donduruldu. Dondurulan spermalar su banyosunda 37 ⁰ C de 20 saniye tutularak çözdürüldü. Çözdürme sonrası koçlarda spermatozoon motilitesi, morfolojik muayene, membran bütünlüğü (HE testi), DNA hasarı ve oksidatif stres parametreleri (TAS ve TOS) belirlendi. Çözdürülen spermalarda subjektif motilite (% 57.00 ± 1.528) açısından 1 mM bor katılan grupta en yüksek (p

The Effect of Boron Addition on Spermatological Parameters, Oxidative Stress and DNA Damage After Frozen-Thawed Process in Ramlic Ram Semen

The aim of the present study was to investigate the effects of different doses of Boron added toextender on some spermatological parameters, oxidative stress, and DNA damage during postthawedof ram semen. The ejaculate samples were collected by artificial vagina from five Ramlıçrams. After determining the primary spermatological properties, those having normospermie qualitywere pooled. The samples were evaluated as a mix ejaculate in the experiment and control groups.The samples were extended with Tris extender containing 5% glycerol and different Boron doses(1, 2 and 4 mM). Control group was formed with no Boron. After dilution and dosing of the samples,withdrawn in 0.25 mL straws were frozen at vapour of liquoid nitrogen. The frozen semen wasthawed in a water bath at 37 ºC for 25 seconds. After thawing; motility, morphology, DNA integrity(COMET assay) and oxidative stress parameters (TAS and TOS) of frozen thawed ram spermwere determined. In the thawed semen according to subjective motility (57.00±1.528 %), 1 mMboron dose group indicated the highest value (P

PDF

___

O’Hara L, Hanrahan JP, Richardson L, et al. Effect of storage duration, storage temperature, and diluent on the viability and fertility of fresh ram sperm. Theriogenology 2010; 73: 541-549.
Kaymakçı M, Taşkın T. Türkiye koyunculuğunda melezleme çalışmaları. Hayvansal Üretim 2008; 49: 43- 51.
Duydu Y, Başaran N, Üstündağ A, et al. Reproductive toxicity parameters and biological monitoring in occupationally and environmentally boron-exposed persons in Bandirma, Turkey. Arch Toxicol 2011; 85: 589- 600.
Başaran N, Duydu Y, Bolt MH. Reproductive toxicity in boron exposed workers in Bandirma, Turkey. J Trace Elem Med Biol 2012; 26: 165-167.
Saylı BS, Tuccar E, Elhan AH. An assessment of fertility in Boron-exposed Turkish subpopulations. Reprod Toxicol 1998; 12: 297-304.
Korkmaz M, Sayli U, Sayli BS, et al. Estimation of human daily boron exposure in a boron-rich area. Br J Nutr 2007; 98: 571-575.
Korkmaz M, Yenigün M, Bakırdere S, et al. Effects of chronic boron exposure on semen profile. Biol Trace Elem Res 2011; 143: 738-750.
Treinen KA, Chapin RE. Development of testicular lesions in F344 rats after treatment with boric acid. Toxicol Appl Pharmacol 1991; 107: 325-335.
Ku WW, Chapin RE, Wine RN, Gladen BC. Testicular toxicity of boric acid: Relationship of dose to lesion development and lack of recovery in the F344 rat. Reprod Toxicol 1993; 7: 305-319.
Watson PF. Use of giemsa stain to detect the changes in the acrosome of frozen ram spermatozoa. Vet Rec 1975; 97:12-15.
Garner DL, Johnson LA. Viability assessment of mammalian sperm using SYBER-14 and propidium Iodide. Biol Reprod 1995; 53: 276-284.
Nagy S, Jansen J, Topper EK, Gadella BM. A triple-stain flow cytometric method to 96 assess plasma-and acrosome-membrane integrity of cryopreserved bovine sperm immediately after thawing in presence of egg-yolk particles. Biol Reprod 2003; 68: 1828-1835.
Bucak MN, Ataman MB, Baspinar N, et al. Lycopene and resveratrol improve post-thaw bull sperm parameters: Sperm motility, mitochondrial activity and DNA integrity. Andrologia 2015; 47: 545-552.
Garner DL, Thomas CA, Joerg HW, DeJarnette JM, Marshall CE. Fluorometric assessments of mitochondrial function and viability in cryopreserved bovine spermatozoa. Biol Reprod 1997; 57: 1401-1406.
Erel Ö. A novel automated method to measure total antioxidant response against potent free radical reactions. Clinical Biochemistry 2004; 37: 112-119.
Erel Ö. A new automated colorimetric method for measuring total oxidant status. Clinical Biochemistry 2005; 38: 1103-1111.
Hughes CM, Lewis SE, McKelvey-Martin VJ, Thompson W. Reproducibility of human sperm DNA measurements using the alkaline single cell gel electrophoresis assay. Mutat Res 1997: 374: 261-268.
Collins AR. The comet assay for DNA damage and repair: Principles, applications, and limitations. Mol Biotechnol 2004; 26: 249-261.
Tırpan MB, Tekin N. Effects of boron added instead of Tris components on freezing and post thaw quality of Angora buck semen. Ankara Üniv Vet Fak Derg 2015; 62: 295-302.
Elkomy AE, Abd El-hady AM, Elghalid OA. Dietary boron supplementation and its impact on semen characteristics and physiological status of adult male rabbits. Asian J Poult Sci 2015; 9: 85-96.
Hunt CD. Regulation of enzymatic activity. One possible role of dietary boron in higher animals and humans. Biol Trace Elem Res 1998; 66: 205-225.
Brown TF, McCormick ME, Morris DR, Zeringue LK. Effects of dietary boron on mineral balance in sheep. Nutr Res 1989; 9: 503-512.
Hegsted M, Keenan MJ, Siver F, Wozniak P. Effect of boron on vitamin D deficient rats. Biol. Trace Elem Res 1991; 28: 243-255.
Hunt CD, Herbel JL, Nielsen FH. Metabolic responses of postmenopausal women to supplemental dietary boron and aluminum during usual and low magnesium intake: Boron, calcium and magnesium absorption and retention and blood mineral concentrations. Am J Cli Nutr 1997; 65: 803-813.
Hunt CD, Idso JP. Dietary boron as a physiological regulator of the normal inflammatory response: A review and current research progress. J. Trace Elem Exp Med 1999; 12: 221-233.
Scialli AR, Bonde JP, Brüske-Hohlfeld I, et al. An overview of male reproductive workers. Reprod Toxicol 2010; 29: 10-24.
Kayar A, Dokuzeylul B, Kandemir FM, et al. Total oxidant and antioxidant capacities, nitric oxide and malondialdehyde levels in cats seropositive for the feline coronavirus. Veterinarni Medicina 2015; 60: 274-281.
Ghiselli A, Serafini M, Natella F, Scaccini C. Total antioxidant capacity as a tool to assess redox status: Critical view and experimental data. Free Radic Biol Med 2000; 29: 1106-1114
Ince S, Kucukkurt I, Cigerci IH, Fidan AF, Eryavuz A. The effects of dietary Boric Acid and Borax supplementation on lipid peroxidation, antioxidant activity, and DNA damage in rats. Journal of Trace Elements in Medicine and Biology 2010; 24: 161-164.