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The aim of the current study was to evaluate the effect of egg yolk and different soybean lecithin concentrations on the efficiency of ram semen cryopreservation and to test the fertilizing ability of frozen-thawed ram semen. Ejaculates with a thick consistency, rapid wave motion (3-5), and >70% initial motility were pooled. Pooled semen were then divided into four groups and diluted at 1/5 (semen/extender) with 1%, 3%, 6% lecithin (L1, L3 and L6) or 20% egg yolk (EY20) using the two-step dilution method. As expected, the results of the current study showed that both motility and the rates of defective acrosomes in sperm were negatively affected by the cryopreservation procedure (P<0.001). The motility values of at 5°C and post-thawed semen in the EY20 group were significantly higher than those in the L1, L3 and L6 groups (P<0.05). There were no differences in motility rates among the lecithin groups at the dilution, cooling, equilibration or post thawing stages (P>0.05). The results of in vitro fertilization, as assessed by the rate of blastocyst formation, were more successful in the EY20 group than those noted in different lecithin groups. In conclusion, freezing ram semen with an extender containing egg yolk could yield better post-thaw sperm parameters and embryonic development compared to lecithin containing extenders. Keywords: Cryopreservation, Lecithin, Egg yolk, Ram semen, IVF Tris-Bazlı Sulandırıcılarda Yumurta Sarısı ve Soya Lesitininin Eritme Sonrası Koç Spermasının Kalitesi ve in vitro Fertilite Üzerine Etkisi ÖzetBu çalışmanın amacı, yumurta sarısı ve farklı lesitin konsantrasyonlarının koç spermasının dondurulabilirliği üzerine etkisini değerlendirmek ve dondurup çözdürülen koç spermasının fertilizasyon yeteneğini tespit etmektir. Kitle hareketi (3-5) ve >%70 motiliteye sahip ejakülatlar birleştirildi (pooling). Pooling yapılan sperma dört gruba bölündü ve %1, %3, %6 (L1, L3 ve L6) lesitin veya %20 yumurta sarısı (EY20) içeren sulandırıcılar ile 1/5 (sperma/sulandırıcı) oranında iki aşamalı sulandırma yöntemi kullanılarak sulandırıldı. Bu çalışmanın sonucunda; motilite ve akrozomal bozukluk oranlarının dondurma prosedüründen olumsuz yönde etkilendiği tespit edildi (P<0.001). EY20 grubunun 5°C'de ve eritme sonrası motilite değerleri L1, L3 ve L6 gruplarına göre yüksek bulundu (P<0.05). Sulandırma, soğutma, ekilibrasyon ve eritme sonrası aşamalarda motilite oranları bakımından lesitin grupları arasında farklılık saptanmadı (P>0.05). Blastosist oranları bakımından değerlendirilen in vitro fertilizasyon sonuçlarına göre, EY20 grubunun farklı lesitin gruplarından daha başarılı olduğu tespit edildi. Sonuç olarak, yumurta sarısı içeren sulandırıcı ile dondurulan koç spermasının lesitin gruplarına göre eritme sonrası spermatolojik özellikler ve embriyonik gelişim bakımından daha üstün olduğu tespit edildi.

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International audienceThe aim of the present study was to investigate the suitability of the water test and hypo-osmotic swelling (HOS) test for analyzing honeybee semen. In particular, the relationships between these tests of the integrity of the sperm plasma membrane and tests of sperm motility and sperm viability were measured using SYBR-14/PI. To this end, semen was obtained from mature drones (16 days or older) collected from four colonies. The means of the per-drone sperm concentration, sperm motility, and live spermatozoa were 1.47 × 106, 76.0%, and 87.2%, respectively. The percentage of spermatozoa with swollen tails was 92.2% as measured with the water test. With the HOS tests at 50, 100, and 150 mOsm/kg, the percentages of spermatozoa with swollen tails were 94.2%, 90.5%, and 85.6%, respectively, after a 30-min incubation and 92.2%, 90.5%, and 88.0%, respectively, after a 60-min incubation. It was observed that subjecting honeybee spermatozoa to a hypo-osmotic solution resulted in clearly identifiable swollen-tail spermatozoa. The percentages of swollen-tail spermatozoa obtained using the water and HOS tests were higher than the percentages of viable and motile spermatozoa. Similar results were obtained using the water test and the HOS test at 50 mOsm/kg. It was concluded that HOS and water tests are suitable and simple measures of the functional integrity of the plasma membranes of bee spermatozoa, and they may be useful additions to standard methods of semen evaluation in mammals

This study investigates the effect of freezing rates on the spermatological parameters of frozen and thawed Saanen goat semen. Equilibrated semen was frozen at 4 different freezing rates from +5 °C to -150 °C (G10: 10 °C/min, G12: 12 °C/min, G15: 15 °C/min, and G24: 24 °C/min) and stored in liquid nitrogen until use. Semen samples were examined for sperm motility, defective acrosomes (FITC-PSA), and DNA integrity using TUNEL after dilution with extender A at equilibration and postthaw stage. There was no significant difference among the freezing stages in terms of DNA fragmentation (P > 0.05). DNA integrity was partially affected by the freezing rate. The increase of freezing rate from 10 °C/min to 24 °C/min between +5 °C and -150 °C resulted in higher postthaw DNA damage. The study found that the freeze-thawing process is detrimental to postthawed goat semen motility (P < 0.05), acrosome integrity (P < 0.05), and DNA integrity (P > 0.05). Although the freezing rates used in the present study had no effect on postthaw sperm motility and acrosome integrity (P > 0.05), sperm DNA integrity was affected.