Vitrification is becoming a preferred method for pre-implantation embryo cryopreservation. The objective of this study was to determine the differentially expressed genes of in vivo- and in vitro-produced bovine embryos after vitrification. In vitro- (IVF) and in vivo-derived (IVV) bovine blastocysts were identified as follows: in vitro-produced fresh (IVF-F), in vitro-produced vitrified (IVF-V), in vivo-derived fresh (IVV-F), in vivo-derived vitrified (IVV-V). The microarray results showed that 53 genes were differentially regulated between IVF and IVV, and 121 genes were differentially regulated between fresh and vitrified blastocysts (P < 0.05). There were 6, 268, 962, and 17 differentially regulated genes between IVF-F × IVV-F, IVF-V × IVV-V, IVF-F × IVF-V, and IVV-F × IVV-V, respectively (P < 0.05). While gene expression was significantly different between fresh and vitrified IVF blastocysts (P < 0.05), it was similar between fresh and vitrified IVV blastocysts. Significantly up-regulated KEGG pathways included ribosome, oxidative phosphorylation, spliceosome, and oocyte meiosis in the fresh IVF blastocyst samples, while sphingolipid and purine metabolisms were up-regulated in the vitrified IVF blastocyst. The results showed that in vitro bovine blastocyst production protocols used in this study caused no major gene expression differences compared to those of in vivo-produced blastocysts. After vitrification, however, in vitro-produced blastocysts showed major gene expression differences compared to in vivo blastocysts. This study suggests that in vitro-produced embryos are of comparable quality to their in vivo counterparts. Vitrification of in vitro blastocysts, on the other hand, causes significant up-regulation of genes that are involved in stress responses.

The most significant focal points of the embryo transfer technology are as follows: the selection of donors, the response of the selected donor to the superovulation protocol and the obtained number of the transferable embryos. For this purpose, it is suggested that donor selection can be done by anti‐Müllerian hormone (AMH) levels, and embryo production is evaluated. AMH is secreted by the granulosa cells of primordial, pre‐antral and antral follicles below 4 mm in the ovary, independent of FSH. Therefore, the aim of this study was to investigate the relationship between serum AMH levels and the number of corpus luteum (CL), total embryos and transferable embryos that were shaped after a uniform superovulation protocol. For this reason, 48 Simmental cows, which were located at General Directory of Agricultural Enterprises (region, province, etc. instead of the general directorate), were used as donors for the embryo transfer. Blood samples were taken at random, regardless of the stage of animal's sexual cycle. AMH levels were measured by enzyme‐linked fluorescent assay (ELFA) method of the miniVIDAS® (bioMérieux SA) using AMH Bovine Test Kit. According to the statistical analyses of the obtained data, AMH levels were positively correlated with CL and total embryos (p < .05). No significant correlations between AMH and transferable embryos were approved (p > .05). It was also determined that each 200 pg/ml increase in serum AMH level resulted in one increase in CL number. Overall, considering the positive correlation between AMH level and the obtained number of CL and total embryos after a superovulation treatment, it was concluded that measuring blood AMH level prior to any further costly implementation may be an effective method in donor selection.

The aim of the present study was to investigate effectiveness of superovulatory response and embryo yield in Anatolian Black heifers, induced with the administration of two different follicle stimulating hormone. Heifers received a progesterone releasing device (Cue-mate containing 1.56 g progesterone). The heifers were randomly assigned to four groups. Group Folltropin (F) were administered with Folltropin (400 mg) as a control, while those included in Groups Ovagen (O) were administered with Ovagen at doses of 8.8 mg (O1), 11.44 mg (O2) and 17.6 mg (O3) respectively. On the 9th day of the application, the Cue-mate was removed and the heifers received 500 μg prostaglandinF2α in all goups. The heifers were artificially inseminated (AI) using semen obtained from Anatolian Black bulls; in the evening of day 11 and in the morning of day 12 with 12 hours intervals. Embryos were collected by uterine flushing 7 days after AI. The mean number of CL determined as 6.33±0.718 in F, was found to be higher than the numbers obtained with the administration of the three different doses of O (3.82±0.502, 3.50±0.513, 3.58±0.448 respectively; p<0.001). The transferable embryo yield did not differ significantly among the treatment groups (p>0.05). In conclusion, findings show that although the administration of F was ascertained increase the number of CL and the total number of ova/embryos recovered, these increased numbers had no reflection on the number of transferable embryos in Anatolian Black heifers. Yerli Kara Düvelerde Süperovulasyon ve Embriyo Verimi Üzerine Farklı FSH Uygulamalarının EtkileriÖZ Bu çalışmanın amacı Yerli Kara düvelerde süperovulasyon oluşturma ve embriyo elde etme yönünde iki farklı follikül uyarıcı hormonunun etkinliğini gözlemlemekti. Düvelere 1.56 g progesteron içeren Cue-mate uygulandı. Düveler dört gruba ayrıldı. Folltropin (F) grubuna control olarak 400 mg folltropin diğer taraftan Ovagen (O) grubuna ise sırasıyla 8.8 mg (O1), 11.4 (O2) ve 17.6 (O3) olmak üzere Ovagen uygulandı. Süperovulasyon protokolünün 9. gününde Cue-mate çıkarıldı ve gruplardaki tüm ineklere 500 µg prostaglandinF2α yapıldı. 11. gün akşam ve 12. gün sabah olmak üzere 12 saat ara ile tüm ineklere Yerli Kara boğalardan alınan spermalar ile suni tohumlama yapıldı. Suni tohumlama uygulamasından 7 gün sonra embriyo elde etme çalışması ile embriyolar toplandı. Ortalama CL sayıları F grubunda 6.33 ± 0.718 belirlenirken, O grubunda uygulanan üç farklı dozdan (sırasıyla 3.82 ± 0.502, 3.50 ± 0.513, 3.58 ± 0.448; p<0.001) elde edilen CL sayılarından daha yüksek bulundu. Transfer edilebilir embriyo oranları uygulama grupları arasında farklı değildi (p>0.05). Sonuç olarak, bulgular F grubunda yapılan uygulamanın CL, elde edilen embriyo ve ovum sayısını artırmasına rağmen transfer edilebilir embriyo sayısına olumlu bir etkisinin olmadığını gösterdi.

Conventional buffalo semen freezing studies are limited in Anatolian buffaloes, which are overly sensitive to exogenous stimulation. The present study's object was to determine the main features of Anatolian Buffalo semen obtained by artificial vagina method for the first time. A total number of 150 ejaculates were collected from three Anatolian Buffalo bulls (app. 4 years of age). The mean pH, volume and concentration of semen were found 6.63±0.15, 1.61±0.5 ml, 1629±222.67 x10 6 spermatozoa/ml, respectively. The sperm motion characteristics were determined by using a computerassisted sperm analysis system (CASA); the total and progressively motile sperm values were 57.

The aim of the study was to show whether there were some differences among 9 Holstein bulls and within their replications on their ability of in vitro fertilization for in vitro embryo production as cleavage and coming into the blastocyst stage. Semen collected and frozen from nine Holstein bulls with satisfactory in vivo fertilization capabilities for artificial insemination was used for in vitro fertilization. Direct washing method by Brackett and Oliphant medium and 5 or 6 h incubation period were used for in vitro fertilization. Charles Rosenkrans medium was used for in vitro embryo culture. An atmosphere with a higher than 95% relative humidity, 39°C, 5% CO 2 and was used for all in vitro embryo production processes. Totally 2519 A and B quality oocytes were treated for in vitro embryo production. As a result statistically significant (P<0.05) variation was found for cleavage and blastocyst development among bulls. However there was no significant difference (P>0.05) between the replications for each bull as cleavage and coming into blastocyst stage. The results showed varied capabilities of bulls for in vitro fertilization and embryo production and male factor can affect success of in vitro embryo production. Keywords: Bull, In vitro, Embryo, Cleavage, Blastocyst In vitro Embriyo Üretimine Boğa Etkisinin AraştırılmasıÖzet Dokuz farklı Holştayn boğaya ait spermanın kullanıldığı bu in vitro embriyo üretim çalışmasının amacı, hem boğalar arasında hem de boğaların kendi tekrarları arasında yarıklanma ve blastosiste ulaşma oranları bakımından fark olup olmadığının gösterilmesi olmuştur. Suni tohumlama boğası olarak kullanılan ve fertilite sorunu olmayan ve tatminkar düzeyde fertilite oranlarına sahip dokuz Holştayn boğadan alınan ve dondurulan spermalar in vitro fertilizasyon amacıyla kullanılmıştır. İn vitro fertilizasyon için Brackett ve Oliphant mediumu ile Direkt yıkama metodu ve 5-6 saat inkubasyon periyodu; in vitro embriyo kültürü için de Charles Rosenkrans mediumu kullanılmıştır. Kültür periyotlarında %95'in üzerinde bağıl nem, 39°C, %5 CO 2 içeren bir kültür ortamı sağlanmıştır. Toplam 2519 A ve B kalite oosit in vitro embriyo elde etme sürecine alınmıştır. Sonuç olarak yarıklanma ve blastosiste ulaşma oranları bakımından boğalar arasında önemli düzeyde istatistiki farklılık bulunmuştur (P<0.05). Fakat her bir boğanın kendi içindeki tekrarları arasında yarıklanma ve blastosiste ulaşma oranları bakımından önemli düzeyde fark görülmemiştir (P>0.05). Bu sonuçlar boğaların değişik düzeyde in vitro fertilizasyon ve embriyo üretim yeteneklerine sahip olduklarını ve in vitro embriyo üretim başarısı için erkek faktörünün belirleyici olduğunu göstermektedir.