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Öğe Comparision of different cryoprotectants on slow freezing of in vivo derived Saanen goats embryos(ANKARA UNIV PRESS, 2018) Cizmeci, Sakine Ulkum; Guler, Mehmet; Kaymaz, MustafaThis study aimed to determine the effects of different cryoprotectants on the viability of Saanen goats embryos which were frozen and thawed with slow freezing method. The study was conducted on 15 Saanen goats and 3 bucks. Fluorogeston acetate (20 mg) impregnated intravaginal sponges were inserted in goats for 12 days regardless of the sexual cycle. Starting on the 9th day of intravaginal sponge administration, follicle stimulating hormone (FSH) was injected intramuscularly, every 12 hours at decreasing doses (50-50; 30-30; 20-20 mg) for 3 days. Goats were mated naturally 24 hours after removal of the sponges. Embryos were recovered by laparotomic uterine flushing on the 7th day after the first mating. Collected embryos were frozen by using 3 different cryoprotectants [ethylene glycol (EG), glycerol, and dimetil sulfoksit DMSO)] with slow freezing technique. Thawed embryos were incubated at 38.5 degrees C and 5% CO2. The viability of embryos was evaluated at the 24th, the 48th, the 72nd hours after thawing. Superovulation response (>= 4 Cl), embryo recovery rate and transferable embryo rate were found to be 93.3%, 72.3% and 58%, respectively. Viability rates of frozen and thawed embryos at the 24th, the 48th, the 72nd hours were found respectively to be 64.86%; 56.76%; 54.05% in EG group, 54.55%, 45.45%; 36.36% in glycerol group and 46.88%; 40.63%; 28.13% in DMSO group. Viability rates of the frozen embryos with EG were statistically better than embryos frozen with glycerol and DMSO (P<0.05) at 72nd hour. Survival rates of blastocysts frozen were 76%; 64%; 60%; 54.6% in EG group 45.5%; 36.4% in glycerol group, and 42.1%; 36.8%; 21.1% in DMSO group and at 72nd hour the difference between EG and DMSO group was significant (P<0.05). In conclusion, viability of embryos at the 24th, the 48th, the 72nd hours after thawing in EG group was significantly higher than the embryos frozen with other cryoprotectants.Öğe The effect of anti-mullerian hormone and progesterone concentrations on superovulation response and embryo yield in goats(ELSEVIER SCIENCE INC, 2020) Alkan, Kübra Karakaş; Alkan, Hasan; Kaymaz, MustafaWe aimed to evaluate the relationship of anti-Mullerian hormone (AMH) and progesterone concentrations with superovulation response in goats and to determine donors exhibiting better superovulation response by measuring AMH concentrations. For this, blood samples were collected from multiparous Angora goats (n = 24) for measuring the progesterone and AMH concentrations on the day the synchronization protocol was initiated (Day 0), on the day of the first FSH administration (Day 9), on the day the progesterone source was removed (Day 11), and on the day of uterine flushing. Descriptive statistics (mean, standard deviation, median, minimum value, maximum value, and percentile) were given for superovulation response and embryo yield. To compare the differences between the two groups, the Student's t-test was used. The relationship between two continuous variables was assessed by the Pearson Correlation Coefficient. The AMH cutoff values in superovulation responses were evaluated by ROC analysis on the day the synchronization protocol was initiated. A strong positive correlation was found between the AMH concentrations measured on the day the synchronization protocol was initiated (Day 0), on the day of the first FSH administration (Day 9), and on the day of removal of the progesterone source (Day 11) and the count of total corpus luteum (CL), total oocyte/embryo, transferable embryo, and Code I quality embryo (P < 0.05). Furthermore, AMH concentration increased on the day the synchronization protocol was initiated, the donor's superovulation response increased as well. The cutoff value was 4.74 ng/ml, as assessed by the ROC curve analysis conducted for selecting donors exhibiting better superovulation responses. The sensitivity and specificity of the selected cutoff value were found to be quite high (P < 0.01). However, a positive correlation was noted between the progesterone concentrations measured on the day of uterine flushing and total CL count, total oocyte/embryo count, transferable embryo count, and Code I quality embryo count (P < 0.01). In conclusion, it was determined that an increase in AMH concentrations in goats led to an increase in the total CL count, embryo count, and embryo quality and that AMH measurement could be used to identify donors that responded better to superovulation. Additionally, a positive correlation was found between the progesterone concentration measured on the day of uterine flushing and the total CL count, transferable embryo count, and embryo quality. (C) 2019 Elsevier Inc. All rights reserved.Öğe Immunohistochemical and qPCR determination of the expression and serum level of anti-Mullerian hormone in pre-pubertal, intact and ovarian remnant syndrome detected bitches(WILEY, 2019) Alkan, Kübra Karakaş; Ceylan, Ahmet; Alkan, Hasan; Özen, Doğukan; Bayraktaroğlu, Alev Gürol; Kaymaz, MustafaThe aim of this study was to determine the serum concentrations, ovarian presence and expression of anti-Mullerian hormone (AMH) in pre-pubertal, bitches with signs of ovarian remnant syndrome (ORS) and intact bitches. In addition, we aimed to verify the suitability of serum AMH concentrations for diagnostic purposes in sterilized bitches and/or in suspected cases of ORS in the field of veterinary medicine. For this purpose, 36 healthy female dogs divided into six groups: proestrus, oestrus, dioestrus, anoestrus, pre-pubertal and ORS. Serum AMH concentrations were determined by electrochemiluminescence immunoassay, and ovarian presence and distribution of AMH was confirmed by immunohistochemical and qPCR techniques. According to the results of qPCR, while the expression values of AMH were at the highest concentrations in the proestrus and oestrus, there was a statistically significant decrease in these values at the later stages of the cycle (p < 0.05). According to hormone analysis, the serum AMH values of the ORS group had decreased significantly compared with the proestrus and oestrus (p < 0.05). Although serum AMH levels of ORS group were increased compared with anestrus and pre-pubertal groups, this increase was statistically non-significant (p > 0.05). Immunohistochemically, AMH expression was first observed in the granulosa cells of primordial follicles in folliculogenesis. Expression values were the highest in the proestrous and oestrus groups, but values from bitches in later stages of the cycle were statistically significant decrease in comparison with these groups (p < 0.05). As a result, AMH concentration and expression were found to be higher in proestrus and oestrus than in other periods (p < 0.05). In addition, the measurable level of AMH concentration in bitches with ORS is an indication that it can be used in the diagnosis of ORS.Öğe In vivo elde edilen Saanen keçisi embriyolarının yavaş dondurulması üzerine farklı kriyoprotektanların etkisinin karşılaştırılması(2018) Çizmeci, Sakine Ülküm; Güler, Mehmet; Kaymaz, MustafaBu çalışmada farklı kriyoprotektanların, yavaş dondurma yöntemi ile dondurulup çözdürülmüş Saanen keçisi embriyolarının canlılığı üzerine etkilerinin belirlenmesi amaçlanmıştır. Çalışmanın hayvan materyalini 15 baş Saanen ırkı keçi ve 3 baş teke oluşturdu. Keçilere siklusun dönemi gözetilmeksizin 12 gün süreyle fluorogeston asetat (20 mg) emdirilmiş intravaginal sünger yerleştirildi. Sünger uygulamasının 9. gününden itibaren 3 gün süreyle 12 saat arayla azalan dozlarda folikül uyarıcı hormon (FSH) (5050; 30-30; 20-20 mg) kas içi yolla enjekte edildi. Sünger çıkarıldıktan 24 saat sonra doğal aşım yaptırıldı. İlk aşımdan sonraki 7. gün laparatomik uterus yıkaması yapılarak embriyolar elde edildi. Toplanan embriyolar 3 farklı kriyoprotektan [etilen glikol (EG), gliserol ve dimetil sulfoksit (DMSO)] kullanılarak yavaş dondurma yöntemiyle donduruldu. Çözdürülen embriyolar 38.5 ºC'de %5 CO2'de inkübasyona bırakıldı ve 24, 48 ve 72. saatlerde canlılık muayeneleri yapıldı. Çalışmada süperovulasyon cevabı (4 Cl) %93.3, embriyo toplama oranı %72.3, transfer edilebilir embriyo oranı ise %58 olarak belirlendi. Dondurulup çözdürülen embriyoların 24, 48 ve 72. saatlerdeki canlılık oranları EG'de %64.86; %56.76; %54.05, gliserolde %54.55; %45.45; %36.36, DMSO da ise %46.88; %40.63; %28.13 olarak belirlendi. Embriyoların 72. saate ulaşmalarında EG ile dondurulanların gliserol ve DMSO ile dondurulanlardan istatistiki olarak daha iyi olduğu tespit edildi (P0.05). Saatlere göre blastosistlerin yaşama oranları EG'de %76; %64; %60, gliserolde %54.6; %45.5; %36.4 ve DMSO'da %42.1; %36.8; %21.1 olarak belirlendi ve 72. saatte EG ve DMSO arasında farkın önemli olduğu görüldü (P0.05). Yapılan çalışma sonucunda çözdürme sonrası 24, 48 ve 72. saatlerdeki canlılık oranları EG ile dondurulan embriyolarda diğer kriyoprotektanlarla dondurulanlardan daha yüksek olduğu belirlendi
