Boğa Spermasının Dondurulmasında Yumurta Sarısı İçeren Tris Sulandırıcısına Katalaz, Melatonin ve Kolesterol İlavesinin Sperma Kalitesi ve Fertilite Üzerine Etkisi
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Dosyalar
Tarih
2022
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Selçuk Üniversitesi Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Boğa spermasının dondurulmasında yumurta sarısı içeren tris sulandırıcısına katalaz,
melatonin ve kolesterol ilavesinin sperma kalitesi ve fertilite üzerine etkisinin belirlenmesi
amaçlanmıştır.
Bu amaçla sunulan bu tez çalışmasında normospermi gösteren üç boğadan suni vajen yoluyla
elde edilen ejakülatlar %20 yumurta sarısı ve %6 gliserol içeren tris sulandırıcısı ile sulandırıldı. İlk
bölümde sulandırılan ejakülatlar on eşit parçaya ayrıldı. Gruplar kontrol, melatonin 0,1 mM/ml,
melatonin 1 mM/ml, melatonin 3 mM/ml, katalaz 50 IU/ml, katalaz 150 IU/ml, katalaz 300 IU/ml,
kolesterol-siklodekstrin (CLC) 1,25 µg/ml, CLC 2,5 µg/ml ve CLC 5 µg/ml olacak şekilde oluşturuldu.
Her gruptan 45 adet payet donduruldu. Payetlerin çözdürme sonrası motilite değerleri CASA ile
belirlendi. En yüksek motilite değerlerine sahip gruplar çalışmanın II. bölümünde kullanıldı. Bu
bölümdeki gruplar kontrol, melatonin 0,1 mM/ml, katalaz 150 IU/ml, CLC 2,5 µg/ml, melatonin 1
mM/ml + katalaz 150 IU/ml, melatonin 1 mM/ml + CLC 2,5 µg/ml, katalaz 150 IU/ml + CLC 2,5 µg/ml
ve melatonin 1 mM/ml + katalaz 150 IU/ml + CLC 2,5 µg/ml olacak şekilde oluşturuldu. Çalışmanın
bu bölümünde her gruptan 51 adet payet donduruldu. Çözdürme sonrası motilite değerleri CASA ile
belirlendi. Çalışmanın II. bölümünde A ve C boğalarına ait örneklerde motilite değerleri açısından
gruplar arasında istatistiki farklılık gözlenmedi (p> 0,05). B boğasına ait örneklerde motilite değerleri
açısından gruplar arasında istatistiki farklılık önemli bulundu (p<0,05). Üç boğada en yüksek motilite
değerini sağlayan katalaz 150 IU/ml ve CLC 2,5 µg/ml grupların sperma örnekleri flow sitometride
değerlendirildi. A boğasına ait sperma örneklerinde canlı spermaztozoon (SYBR+) analizleri açısından
gruplar arasında istatistiki farklılık önemli bulundu (p<0,05). A boğasına ait sperma örneklerinde
plazma membran ve akrozom bütünlüğü (FITCH-PI-), mitokondriyal oksidatif stres (MITOSOX+) ve
ROS değerleri (BODIPY+) açısından gruplar arasında istatistiki farklılık gözlenmedi (p>0,05). B ve C
boğalarına ait sperma örneklerinde plazma membran ve akrozom bütünlüğü (FITCH-PI-), canlı
spermatozoon muayenesi (SYBR+), mitokondriyal oksidatif stres (MITOSOX+) ve ROS değerleri
(BODIPY+) açısından gruplar arasında istatistiki farklılık gözlenmedi (p>0,05). A, B ve C boğalarına
ait sperma örneklerinde çözdürme sonrası 0. saate motilite değerleri açısından katalaz (150 IU/ml) grubu
ile kontrol grubu arasındaki istatistiki farklılık önemli bulundu (p<0,05). A, B ve C boğalarına ait
sperma örneklerinde çözdürme sonrası 2. saat motilite değerleri açısından katalaz (150 IU/ml) grubu ile
kontrol grubu ve kolesterol (2,5 μg/ml) grubu ile kontrol grubu arasında istatistiki farklılık önemli
bulundu (p<0,05). A, B ve C boğalarına ait spermalarla yapılan tohumlamaları takiben elde edilen
gebelik oranları açısından gruplar (kontrol, katalaz (150 IU/ml), kolesterol (2,5 μg/ml)) arasında
istatistiki farklılık gözlenmedi (p>0,05).
Sonuç olarak melatonin, katalaz ve CLC’in boğa spermatozoonlarına eklenmesi, dondurma
çözdürme sonrası spermalarda motiliteyi koruduğu, ayrıca katalazın boğa spermatozoonlarına
eklenmesi dondurma çözdürme sonrası plazma membran ve akrozomal membran bütünlüğünü
koruduğu sonucuna varıldı.
It was aimed to determine the effect of catalase, melatonin and cholesterol addition to tris diluent containing egg yolk on the semen quality and fertility in the freezing of bull semen. For this purpose, in the present thesis study, ejaculates obtained from three bulls with normospermia through artificial vagina were diluted with tris diluent containing 20% egg yolk and 6% glycerol. In the first part, diluted ejaculates were split into ten equal parts. The groups were comprised of control, melatonin 0,1 mM/ml, melatonin 1 mM/ml, melatonin 3 mM/ml, catalase 50 IU/ml, catalase 150 IU/ml, catalase 300 IU/ml, cholesterol-cyclodextrin (CLC) 1,25 µg/ml, CLC 2,5 µg/ml and CLC 5 µg/ml. 45 straws were frozen from each group. Motility values of straws after thawing were determined by CASA. The groups with the highest motility values were included in the second part of the study. Groups at this part were made up as control, melatonin 0.1 mM/ml, catalase 150 IU/ml, CLC 2.5 µg/ml, melatonin 1 mM/ml + catalase 150 IU/ml, melatonin 1 mM/ml + CLC 2.5 µg /ml, catalase 150 IU/ml + CLC 2.5 µg/ml and melatonin 1 mM/ml + catalase 150 IU/ml + CLC 2.5 µg/ml. In this part of study, 51 straws were frozen from each group. Post-thaw motility values were determined by CASA. In the second part of the study, no statistically significant difference was observed among the groups in tems of motility values in the samples belonging to A and C bulls (p> 0.05). A statistically significant difference was found among the groups in terms of motility values in the samples of B bulls (p<0.05). The semen samples of catalase 150 IU/ml and CLC 2.5 µg/ml groups, which provided the highest motility value in three bulls, were evaluated in flow cytometry. Statistical difference was found to be significant among the groups in terms of live sperm (SYBR+) analysis of semen samples of A bull (p<0.05). No statistical difference was observed among the groups in terms of plasma membrane and acrosome integrity (FITCH-PI-), mitochondrial oxidative stress (MitoSOX+) and ROS values (BODIPY+) in semen samples of bull A (p>0,05). No statistical difference was observed among the groups in terms of plasma membrane and acrosome integrity (FITCH-PI-), live sperm examination (SYBR+), mitochondrial oxidative stress (MitoSOX+) and ROS values (BODIPY+) in semen samples belonging to bulls B and C (p> 0.05). The statistical difference between the catalase (150 IU/ml) group and the control group was found to be significant in terms of motility values at the zeroth hour after thawing in the semen samples belonging to A, B and C bulls (p<0.05). Statistical difference was found between catalase (150 IU/ml) group and control group and cholesterol (2.5 μg/ml) group and control group in terms of motility values in the second hour after thawing in the semen samples belonging to A, B and C bulls (p<0.05). No statistical difference was observed between the groups (control, catalase (150 IU/ml), cholesterol (2.5 μg/ml)) in terms of pregnancy rates obtained after insemination with the semen belonging to A, B and C bulls (p>0.05). As a result, it was concluded that adding melatonin, catalase and CLC to the bull spermatozoa preserves motility in semen after freezing and thawing, also adding catalase to the bull spermatozoa preserves the integrity of the plasma membrane and acrosomal membrane after freezing and thawing.
It was aimed to determine the effect of catalase, melatonin and cholesterol addition to tris diluent containing egg yolk on the semen quality and fertility in the freezing of bull semen. For this purpose, in the present thesis study, ejaculates obtained from three bulls with normospermia through artificial vagina were diluted with tris diluent containing 20% egg yolk and 6% glycerol. In the first part, diluted ejaculates were split into ten equal parts. The groups were comprised of control, melatonin 0,1 mM/ml, melatonin 1 mM/ml, melatonin 3 mM/ml, catalase 50 IU/ml, catalase 150 IU/ml, catalase 300 IU/ml, cholesterol-cyclodextrin (CLC) 1,25 µg/ml, CLC 2,5 µg/ml and CLC 5 µg/ml. 45 straws were frozen from each group. Motility values of straws after thawing were determined by CASA. The groups with the highest motility values were included in the second part of the study. Groups at this part were made up as control, melatonin 0.1 mM/ml, catalase 150 IU/ml, CLC 2.5 µg/ml, melatonin 1 mM/ml + catalase 150 IU/ml, melatonin 1 mM/ml + CLC 2.5 µg /ml, catalase 150 IU/ml + CLC 2.5 µg/ml and melatonin 1 mM/ml + catalase 150 IU/ml + CLC 2.5 µg/ml. In this part of study, 51 straws were frozen from each group. Post-thaw motility values were determined by CASA. In the second part of the study, no statistically significant difference was observed among the groups in tems of motility values in the samples belonging to A and C bulls (p> 0.05). A statistically significant difference was found among the groups in terms of motility values in the samples of B bulls (p<0.05). The semen samples of catalase 150 IU/ml and CLC 2.5 µg/ml groups, which provided the highest motility value in three bulls, were evaluated in flow cytometry. Statistical difference was found to be significant among the groups in terms of live sperm (SYBR+) analysis of semen samples of A bull (p<0.05). No statistical difference was observed among the groups in terms of plasma membrane and acrosome integrity (FITCH-PI-), mitochondrial oxidative stress (MitoSOX+) and ROS values (BODIPY+) in semen samples of bull A (p>0,05). No statistical difference was observed among the groups in terms of plasma membrane and acrosome integrity (FITCH-PI-), live sperm examination (SYBR+), mitochondrial oxidative stress (MitoSOX+) and ROS values (BODIPY+) in semen samples belonging to bulls B and C (p> 0.05). The statistical difference between the catalase (150 IU/ml) group and the control group was found to be significant in terms of motility values at the zeroth hour after thawing in the semen samples belonging to A, B and C bulls (p<0.05). Statistical difference was found between catalase (150 IU/ml) group and control group and cholesterol (2.5 μg/ml) group and control group in terms of motility values in the second hour after thawing in the semen samples belonging to A, B and C bulls (p<0.05). No statistical difference was observed between the groups (control, catalase (150 IU/ml), cholesterol (2.5 μg/ml)) in terms of pregnancy rates obtained after insemination with the semen belonging to A, B and C bulls (p>0.05). As a result, it was concluded that adding melatonin, catalase and CLC to the bull spermatozoa preserves motility in semen after freezing and thawing, also adding catalase to the bull spermatozoa preserves the integrity of the plasma membrane and acrosomal membrane after freezing and thawing.
Açıklama
Anahtar Kelimeler
Katalaz, Kolesterol, Melatonin, Spermatozoon, Yumurta Sarısı, Catalase, Cholesterol, Egg Yolk
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Yılmaz, M. B., (2022). Boğa Spermasının Dondurulmasında Yumurta Sarısı İçeren Tris Sulandırıcısına Katalaz, Melatonin ve Kolesterol İlavesinin Sperma Kalitesi ve Fertilite Üzerine Etkisi. (Doktora Tezi). Selçuk Üniversitesi, Sağlık Bilimler Enstitüsü, Konya.
