Kangal köpeği spermasının dondurulmasında farklı antioksidanlar ve kriyoprotektanların etkisi
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Dosyalar
Tarih
2015
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Yayıncı
Selçuk Üniversitesi Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Sunulan tez çalışmasında, Kangal köpeklerinden alınan ejakülatlar birleştirilerek 3 farklı deney grubu oluşturuldu. Her deney grubu altı farklı alt gruptan oluştu. Deney grupları temel Tris sulandırıcısıyla (T) sulandırıldı. Birinci deney grubu, %5 gliserol (G)+kuersetin (K) 50 µM (TGK50), %5G + K 200 µM (TGK200), %5 etilen glikol (E)+ K 50 µM (TEK50), %5 E+K200 µM (TEK200), %5G (TG) ve %5E (TE) içeren Tris sulandırıcılarıyla sulandırılan spermalarla oluşturuldu. İkinci deney grubu, TG + Ergotiyonin (E) 0,5 mM (TGE0.5), TG + E 2 mM (TGE2), TE + E 0,5 mM (TEE0.5), TE + E 2 mM (TEE2), %5 TG ve %5 TE içeren Tris sulandırıcılarıyla sulandırılan spermalarla oluşturuldu. Üçüncü deney grubu, TG + BME %2,5 (TGB2.5), TG+ BME %10 (TGB10), TE+ BME %2,5 (TEB2.5), TE+ BME %10 (TEB10), % 5TG ve %5 TE içeren Tris sulandırıcılarıyla sulandırılan spermalarla oluşturuldu. Üç farklı deney gruplarında sulandırılan spermalar 4oC'ta 1,5 saat ekilibrasyon sonrası sıvı azot buharında dondurularak sıvı azotta (~-196ºC) saklandı. Spermalar sulandırma ve dondurma/çözdürme sonrası subjektif spermatozoon motilitesi, çözüm sonu CASA motiliteleri, spermatozoa hareket özellikleri, spermatozoa canlılığı (SYBR-14/PI), mitokondriyal aktivitesi (JC-1/PI), akrozom bütünlüğü (FITC/PI) ve DNA bütünlüğü yönünden değerlendirildi. Birinci deney grubunda çözüm sonu subjektif motilite değerleri TGK50 (%55,70±0,71), TGK200 (%60,00±1,09), TEK200 (%56,40±1,42) gruplarında, TG (%50,70±1,70) ve TE (%51,40±0,92) gruplarına göre yüksek bulundu. Progresif motilite TGK200 (%27,14±1,08) grubunda, TE (%20,56±1,07) grubuna göre istatiksel olarak yüksekti. TGK50 ve TGK200 grupları canlılık, akrozom bütünlüğü ve mitokondriyal aktivite oranları bakımından (sırasıyla, %59,69±1,84, %60,49±2,54; %56,68±3,17, %52,67±3,57; %58,34±6,17, %57,53±6,08) TE grubuna (sırasıyla, %48,68±2,41, %37,95±1,76, %43,25±2,10) göre istatiksel olarak üstünlük gösterdi. DNA bütünlüğü korunmasında TGK200 (%95,75±0,52) grubu, TE (%93,41±0,74) grubuna göre istatiksel olarak daha yüksek koruma sağladı (P<0.05). İkinci deney grubunda çözüm sonu progresif motilite oranı TGE2 (%28,71±1,52) grubunda, TG (%15,81±0,49) ve TE (%15,23±2,39) gruplarına göre yüksek bulundu. Canlılık ve akrozom bütünlüğü TGE2 (sırasıyla, %66,37±3,24; 59,87±1,27) grubunda, TG (%58,42±2,44; 54,69±0,85) ve TE (%49,99±2,52; 47,06±1,48) gruplarına göre istatiksel olarak üstünlük gösterdi (P<0.05). Üçüncü deney grubunda progresif motilite oranı TGB10 (%23,19±1,61) grubunda, TE (%14,08±3,07) grubuna göre istatistiksel olarak yüksek bulundu. Akrozom bütünlüğü TGB2.5 (%44,99±4,97) grubunda, TG (%14,08±3,07) grubuna göre istatiksel olarak üstünlük gösterdi (P<0.05). Sonuç olarak, Kangal köpeği spermasında, farklı kriyoprotektanlarla antioksidan kombinasyonunun çözüm sonrası spermatolojik parametreler üzerinde olumlu koruma sağladığı sonucuna ulaşıldı.
In the presented thesis, ejaculates collected from Kangal dogs were allocated to three different experimental groups. Each experimental group comprised six subgroups, which were established by dividing the collected semen into six equal aliquots. All subgroups were diluted with a Tris-based extender (T). The first experimental group included semen samples diluted with Tris-based extenders containing 5% glycerol (G)+quercetin (K) 50 µM (TGK50), 5% G+K 200 µM (TGK200), 5% ethylene glycol (E)+K 50 µM (TEK50), 5% E+K 200 µM (TEK200), 5% G (TG) and 5% E (TE). The second experimental group included semen samples diluted with Tris-based extenders containing TG+Ergothionine (E) 0.5 mM (TGE0.5), TG+E 2mM (TGE2), TE+E 0.5 mM (TEE0.5), TE+E 2 mM (TEE2), TG and TE. The third experimental group was composed of semen samples diluted with Tris-based extenders containing TG+BME 2.5% (TGB2.5), TG+BME 10% (TGB10), TE+BME 2.5% (TEB2.5), TE+BME 10% (TEB10), TG and TE. The diluted semen samples, belonging to the three different experimental groups, were equilibrated at 4◦C for 1.5 hours and then frozen in liquid nitrogen vapour and stored in liquid nitrogen (~-194ºC). Following dilution and freeze-thawing, the semen samples were assessed for subjective sperm motility, post-thaw CASA motility, sperm motility parameters, sperm viability (SYBR-14/PI), mitochondrial activity (JC-1/PI), acrosome integrity (FITC/PI) and DNA integrity. In the first experimental group, the post-thaw subjective motility values of the TGK50 (55.70±0.71%), TGK200 (60.00±1.09%), and TEK200 (56.40±1.42%) were higher than those of the TG (50.70±1.70%) and TE (51.40±0.92%). The progressive motility rate was significantly higher in TGK200 (27.14±1.08%), when compared to TE (20.56±1.07%). In TGK50 and TGK200, the viability, acrosome integrity and mitochondrial activity rates (59.69±1.84%, 60.49±2.54%; 56.68±3.17%, 52.67±3.57%; 58.34±6.17%, and 57.53±6.08%, respectively) were significantly higher than those of TE (48.68±2.41%, 37.95±1.76%, 43.25±2.10%, respectively). The DNA integrity rate was higher in TGK200 (95.75±0.52%), compared to TE (93.41±0.74%) group (P<0.05). In the second experimental group, the post-thaw progressive motility rate was higher in the TGE2 (28.71±1.52%), compared to TG (15.81±0.49%) and TE (15.23±2.39%). Furthermore, the viability and acrosome integrity rates were significantly higher in TGE2 (66.37±3.24% and 59.87±1.27%, respectively) than in TG (58.42±2.44%; 54.69±0.85%) and TE (49.99±2.52%; 47.06±1.48%) (P<0.05). In the third experimental group, the progressive motility rate of the TGB10 (23.19±1.61%) was significantly higher than that of the TE (14.08±3.07%). The acrosome integrity rate of the subgroup TGB2.5 (44.99±4.97%) was significantly higher than that of TG (14.08±3.07%) (P<0.05). In conclusion, it was determined that different cryoprotectant and antioxidant combinations improved the post-thaw quality of Kangal dog semen.
In the presented thesis, ejaculates collected from Kangal dogs were allocated to three different experimental groups. Each experimental group comprised six subgroups, which were established by dividing the collected semen into six equal aliquots. All subgroups were diluted with a Tris-based extender (T). The first experimental group included semen samples diluted with Tris-based extenders containing 5% glycerol (G)+quercetin (K) 50 µM (TGK50), 5% G+K 200 µM (TGK200), 5% ethylene glycol (E)+K 50 µM (TEK50), 5% E+K 200 µM (TEK200), 5% G (TG) and 5% E (TE). The second experimental group included semen samples diluted with Tris-based extenders containing TG+Ergothionine (E) 0.5 mM (TGE0.5), TG+E 2mM (TGE2), TE+E 0.5 mM (TEE0.5), TE+E 2 mM (TEE2), TG and TE. The third experimental group was composed of semen samples diluted with Tris-based extenders containing TG+BME 2.5% (TGB2.5), TG+BME 10% (TGB10), TE+BME 2.5% (TEB2.5), TE+BME 10% (TEB10), TG and TE. The diluted semen samples, belonging to the three different experimental groups, were equilibrated at 4◦C for 1.5 hours and then frozen in liquid nitrogen vapour and stored in liquid nitrogen (~-194ºC). Following dilution and freeze-thawing, the semen samples were assessed for subjective sperm motility, post-thaw CASA motility, sperm motility parameters, sperm viability (SYBR-14/PI), mitochondrial activity (JC-1/PI), acrosome integrity (FITC/PI) and DNA integrity. In the first experimental group, the post-thaw subjective motility values of the TGK50 (55.70±0.71%), TGK200 (60.00±1.09%), and TEK200 (56.40±1.42%) were higher than those of the TG (50.70±1.70%) and TE (51.40±0.92%). The progressive motility rate was significantly higher in TGK200 (27.14±1.08%), when compared to TE (20.56±1.07%). In TGK50 and TGK200, the viability, acrosome integrity and mitochondrial activity rates (59.69±1.84%, 60.49±2.54%; 56.68±3.17%, 52.67±3.57%; 58.34±6.17%, and 57.53±6.08%, respectively) were significantly higher than those of TE (48.68±2.41%, 37.95±1.76%, 43.25±2.10%, respectively). The DNA integrity rate was higher in TGK200 (95.75±0.52%), compared to TE (93.41±0.74%) group (P<0.05). In the second experimental group, the post-thaw progressive motility rate was higher in the TGE2 (28.71±1.52%), compared to TG (15.81±0.49%) and TE (15.23±2.39%). Furthermore, the viability and acrosome integrity rates were significantly higher in TGE2 (66.37±3.24% and 59.87±1.27%, respectively) than in TG (58.42±2.44%; 54.69±0.85%) and TE (49.99±2.52%; 47.06±1.48%) (P<0.05). In the third experimental group, the progressive motility rate of the TGB10 (23.19±1.61%) was significantly higher than that of the TE (14.08±3.07%). The acrosome integrity rate of the subgroup TGB2.5 (44.99±4.97%) was significantly higher than that of TG (14.08±3.07%) (P<0.05). In conclusion, it was determined that different cryoprotectant and antioxidant combinations improved the post-thaw quality of Kangal dog semen.
Açıklama
Anahtar Kelimeler
Antioksidanlar, Antioxidants, Dondurarak saklama, Cryopreservation, Kriyoprotektif ajanlar, Cryoprotective agents, Köpekler, Dogs, Sivas, Kangal, Spermatozoa
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Güngör, Ş. (2015). Kangal köpeği spermasının dondurulmasında farklı antioksidanlar ve kriyoprotektanların etkisi. Selçuk Üniversitesi, Yayımlanmış doktora tezi, Konya.
