Gaffari TÜRK, Seyfettin GÜR, İzzet KARAHAN

In vitro effects of prostaglandin F2 $alpha$ and metamizol on the motility of diluted bull semen

Bu çalışma, 4 °C'da kısa süreli saklanan sulandırılmış boğa spermasının motilitesi üzerine prostaglandin $F_2$ alfa $(PGF_{2alpha})$ ve metamizolun in vitro etkilerini araştırmak amacıyla yapıldı. Araştırmada, 3 yaşında, sağlıklı 3 adet İsviçre Esmeri boğa kullanıldı. Her bir boğadan 3 hafta boyunca haftada iki kez olmak üzere 6 sperma örneği alındı. Her sperma örneğinin 1 ml'si sperma miktarı, kitle hareketi, başlangıç motilitesi, spermatozoon yoğunluğu ve anormal spermatozoon oranı'nın belirlenmesi için ayrıldı. Her boğanın sperma örneklerinin geri kalanı 1:10 oranında (sperma:sulandırıcı) izotonik sodyum sitrat+yumurta sarısı sulandırıcısıyla sulandırıldı. Daha sonra sulandırılmış sperma örnekleri eşit hacimde (0,5 ml) 12 steril tüpe konuldu. Bu örneklerin 6 tanesine $PGF_{2alpha}$'nın farklı miktarları (0, 125, 250, 500, 1000 and 1500 µg), benzer şekilde metamizol de aynı miktarlarda diğer 6 örneğe 37 °C 'de ilave edildi. $PGF_{2alpha}$ ve metamizol ilavesinden sonra sperma örneklerinin tümü 4 °C 'de saklandı. Bu örneklerde spermatozoon motilitesi 0, 1, 2, 4, 24, 48, 72, 96, 120, 144, 168, 192 ve 216. saatlerde belirlendi. Spermatolojik özellikler yönünden boğalar arasında başlangıçta bir farklılık olmadığı tespit edildi. Sulandırılmış ve 4 °C 'de saklanan sperma örneklerine 125 ve 250 µg PGF2a'nın ilave edilmesi, $PGF_{2alpha}$ bulunmayan sulandırılmış sperma örneklerine göre 24. saatten sonra spermatozoon motilitesinde istatiksel olarak anlamlı (P < 0,05) bir artış oluşturdu. Bununla birlikte, $PGF_{2alpha}$ bulunmayan sulandırılmış sperma örneklerine göre, 500 ve 1000 µg $PGF_{2alpha}$ ilave edilmesi 4. saatten sonra, 1500 µg $PGF_{2alpha}$ ilave edilmesi ise 2. saatten sonra spermatozoon motilitesinde anlamlı (P < 0,05) azalmalara neden oldu. Diğer yandan, metamizol bulunmayan sulandırılmış sperma örnekleriyle karşılaştırıldığında 500 ve 1000 µg metamizol ilave edilmesi sırasıyla 48 ve 24. saatten sonra; metamizol'un 500, 1000 ve 1500 µg'lık miktarları ise 4. saatten sonra 4°C 'de saklanan sperma örneklerinin motilitesinde anlamlı (P < 0,05) azalmalara neden oldu. Sonuç olarak, sulandırılmış boğa sperma örneklerine $PGF_{2alpha}$'nın düşük miktarlarda ilave edilmesinin 4 °C 'de kısa süreli saklanması esnasında motilitenin daha uzun süre korunmasına yardımcı olabileceği kanaatine varıldı.

Sulandırılmış boğa spermalarının motilitesi üzerine prostaglandin F2 $alpha$ ve metamizol'un in vitro etkileri

The purpose of the present study was to investigate in vitro effects of prostaglandin $F_2$ alpha ($PGF_{2alpha}$) and metamizol on the motility of diluted bull semen during short-term storage at 4 °C. In this study, 3 healthy adult Brown-Swiss bulls, each 3 years old, were used. Semen samples were collected with an artificial vagina. Samples were taken from each bull twice a week over the course of 3 weeks (6 samples per bull). One millilitre of semen was removed from each sample for the determination of semen volume, mass activity, initial motility, sperm concentration, and abnormal sperm rate. The remainder of the semen samples of each bull were diluted to a concentration of 1:10 (semen:extender) with isotonic sodium citrate + egg yolk solution. The diluted semen samples were then divided into 12 sterile tubes of equal volume (0.5 ml). Different amounts of PGF2a [0 ($PGF_{2alpha}$ -free), 125, 250, 500, 1000, and 1500 µg] were added to 6 of these samples at 37 °C. Similarly, different amounts of metamizol [0 (metamizolfree), 125, 250, 500, 1000, and 1500 µg] were added to the other 6 samples at 37 °C. After the supplementation of $PGF_{2alpha}$ or metamizol, all semen samples were stored at 4 °C. The sperm motility of the samples was examined at 0, 1, 2, 4, 24, 48, 72, 96, 120, 144, 168, 192, and 216 h.No significant differences in spermatological characteristics were determined between the bulls. Sperm supplementation with 125 µg and 250 µg $PGF_{2alpha}$ caused a statistically significant increase (P < 0.05) in the motility of diluted semen stored at 4 °C after 24 h versus diluted $PGF_{2alpha}$ -free semen. However, the supplementation with 500 µg and 1000 µg $PGF_{2alpha}$ caused significant motility decreases after 4 h, while 1500 µg PGF2a also caused a decrease after 2 h compared to diluted $PGF_{2alpha}$ -free semen (P < 0.05). On the other hand, metamizol supplementation of 125 µg and 250 µg significantly decreased the motility of diluted semen stored at 4 °C after 48 hand 24 h, respectively, while 500 µg, 1000 µg, and 1500 µg of metamizol reduced motility after 4 h (P < 0.05) in comparison to the diluted metamizol-free semen. In conclusion, the addition of small amounts of $PGF_{2alpha}$ to diluted bull semen samples may help to maintain sperm motility during short-term storage at 4 °C.

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