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Öğe Effects of triton X-100 pretreatment of lyophilized and frozen-thawed ram sperm on preimplantation embryo developmental competence(Taylor & Francis Inc, 2022) Üstüner, Burcu; Yağcıoğlu, Selin; Nur, Zekariya; Alçay, Selim; Demir, Kamber; Gökçe, Elif; Pabuccuoğlu, SerhatIn this study, it was aimed to determine the effect of destruction of lyophilized and frozen-thawed ram sperm plasma and acrosomal membrane on development of embryos produced by intracytoplasmic sperm injection (ICSI). Semen samples were divided into two groups for lyophilization (L) and freezing (F). For the removal of the plasma membrane, L and F groups were incubated with Triton X-100 (LTX-100 and FTX-100, respectively). Integrities of the plasma membrane, acrosome and chromatin structure were evaluated. Oocytes were injected with these sperm groups. Although no plasma membrane and acrosome integrities of the L (0.0%) group were detected, the plasma membrane integrity of the F group (69.4%) was significantly higher than the FTX-100 group (23.6%) (p < 0.05). The acrosome integrity of the FTX-100 group (3.80%) was significantly lower than the F group (55.6%) (p < 0.05). The chromatin integrities of L and F groups were higher than the Triton X-100 treated groups (p < 0.05). ICSIs with L, LTX-100, F and FTX-100 sperm were produced similar cleavage and blastocyst rates. In conclusion, data presented here confirm that ram spermatozoa can effectively be lyophilized and injected into oocytes for initiation of embryonic development and Triton X-100 pretreatment is not necessary while using lyophilized and frozen semen.Öğe Long Term Incubation Resilience of Post-Thaw Ram Semen Diluted with Lecithin-Based Extender Supplemented with Bovine Serum Albumin(Kafkas Univ, Veteriner Fakultesi Dergisi, 2019) Alcay, Selim; Toker, Mehmed Berk; Gökçe, Elif; Önder, Nail Tekin; Üstüner, Burcu; Nur, ZekariyaThe objective of the study was to determine the optimum concentration of BSA in lecithin-based extender for post-thawing quality and incubation resilience (0 h, 6 h and 10 h) of ram sperm. Ejaculates were collected from five rams via electro ejaculation. Ejaculates were mixed to obtain pooled semen.Then, pooled semen was diluted with soybean lecithin-based extender without BSA (control) or supplemented with different concentrations of BSA (2.5 mg/mL, 5 mg/mL, 7.5 mg/mL and 10 mg/mL), at a final concentration of 150x106 spermatozoon/mL. Sperm motility, plasma membrane functional integrity (HOST), mitochondrial activity (rhodamine123), capacitation status (CTC), and DNA integrity (TUNEL) were evaluated. At the 10 h incubation, motility, plasma membrane functional integrity and mitochondria! function were better preserved in the BSA5 group compared to the control group. It was determined that high doses of BSA (5 mg/mL, 7.5 mg/mL and 10 mg/mL) affected acrosome reaction.The highest acrosome reaction rates were obtained in BSA10 groups in 6 h and 10 h incubation (P<0.05). TUNEL assay demonstrated that there were no differences among groups for DNA fragmentation at post-thaw and during incubation periods. The study shows that BSA supplemented extenders may have beneficial effect on ram semen parameters at 0 h, 6 h and 10 h of incubation. The results of the present study demonstrated a remarkable advantage of using 5 mg/mL of BSA in 1% lecithin-based extender.Öğe Lyophilized Extender Supplemented with Rainbow Trout (Oncorhynchus mykiss) Seminal Plasma Improves Cryopreservation of Ram Sperm(Kafkas Univ, Veteriner Fakultesi Dergisi, 2022) Üstüner, Burcu; Alçay, Selim; Gökçe, Elif; Yılmaz, M. Melih; Aktar, Ahmet; Hüraydın, Oğuzhan; Nur, ZekariyaThis study aimed to investigate the effect of the Rainbow trout seminal plasma (RTSP) supplemented (control, 10% or 15%) lyophilized extender on freezability of ram semen. Collected semen was pooled and split into two aliquots, and each of the ejaculates was diluted with fresh or lyophilized extenders with RTSP (0%, FC and LC; 10% F10 and L10or 15% F15 and L15) using two-step dilution method. Semen was frozen using the programmable freezing machine. Semen samples were examined for sperm motility, defective acrosomes, plasma membrane integrity and DNA fragmentation at native and post-thaw stage. The highest percentages of post-thaw motility and plasma integrity were observed in the F10, F15, and L15 (P 0.05). There was no significant difference in the rate of post-thaw defective acrosomes when the other extender groups were compared with the control (P 0.05). The highest percentage of post-thaw DNA fragmentation values were observed in the FC group, while the lowest DNA fragmentation was obtained in the F15 but only significant different from FC and LC groups. In conclusion, the findings of this study show that the lyophilized extender with 15% RTSP added can be used successfully for freezing ram semen.Öğe Successful cryopreservation of honey bee drone spermatozoa with royal jelly supplemented extenders(Academic Press Inc Elsevier Science, 2019) Alcay, Selim; Çakmak, Selvinar; Çakmak, İbrahim; Mulkpinar, Emine; Gökçe, Elif; Üstüner, Burcu; Nur, ZekariyaThe aim of the present study was to evaluate different concentrations of royal jelly (RJ) supplemented extenders for post-thawing quality of drone sperm. Semen samples were collected from sexually mature drones. Pooled semen was diluted with extender without RJ (control) or supplemented with different concentrations of RJ (1, 2, 4 or 8%). Sperm motility, plasma membrane functional integrity, and acrosomal integrity were evaluated. At post thaw, the highest sperm motility and acrosomal integrity rates were obtained in the RJ1 group. Functional integrity of sperm membrane was better preserved in the RJ1 and RJ2 groups compare to the other groups. The study shows that RJ supplemented extenders have beneficial effects on drone semen parameters. The results of the present study demonstrated advantage of using 1% RJ supplemented extender.
