Does using microfluidic sperm sorting chips in bovine IVEP affect blastocyst development?

This study aimed to determine the effect of microfluidic sperm sorting chip on embryo development and quality in the sperm treatment step in in vitro embryo production in cattle. Only A-quality oocytes obtained from the ovaries of Holstein cattle were included in the study. These oocytes were first placed in in vitro maturation medium, and matured oocytes were randomly divided into two groups at the 24th hour of maturation. Oocytes in the first group with the Microfluidic Sperm Sorting Chip (MFSC, n = 154) were put into a fertilization medium with spermatozoa prepared with microfluidic sperm sorting device. Oocytes in the second group (Con, n = 169) were fertilized with spermatozoa prepared by the routine sperm treatment step of the commercial company. The rate of cleavage (85.71% vs. 76.33%, respectively) and of reaching the blastocyst (44.15% vs. 32.54%, respectively) in the MFSC group were higher than the control group. In addition, it was determined that the numbers of ICM (45.8 ± 2.04 vs. 39.2 ± 1.85, respectively), TE (122.13 ± 2.19 vs. 115.0 ± 2.61, respectively), TC (167.93 ± 2.89 vs. 154.2 ± 2.62, respectively) increased in the MFSC group compared to the control group. A statistically significant difference was found in the number of cells with apoptosis per embryo (5.14 ± 0.77 vs. 11.91 ± 0.79) and apoptotic index rates (3.06 ± 0.47 vs. 7.72 ± 0.55%) of the MFSC and Con groups. As a result, we concluded that using microfluidic sperm sorting chips during sperm treatment in bovine IVEP increases the rate of reaching the blastocyst, embryo development, and quality and reduces the possibility of apoptosis in developing blastocysts. For this reason, it is also thought that the use of microfluidic sperm sorting devices during sperm treatment in bovine IVEP may be a new alternative in this field.

The evaluation of xenotransplantation of feline ovarian tissue vitrified by needle immersed vitrification technique into male immunodeficient mice.

In this study, the efficiency of the "Needle Immersed Vitrification" technique was tested on cryopreserved feline ovarian tissue. For vitrification, ovarian fragments (0.5-1.5 mm2) from each ovary were collected; the grafts were exposed to 7.5-15% ethylene glycol and 7.5-15% dimethyl sulfoxide at room temperature and stored in liquid nitrogen at least 1 week. Morphologic examinations, expression of genes such as B cell lymphoma 2, B-cell lymphoma-2-associated X protein, Bone morphogenetic protein 15, zone of polarizing activity, zona pellucida C protein and DNA (cytosine-5)-methyltransferase 1, ultrastructural analysis and viability tests were carried out from collected grafts. Light microscopy examinations revealed the percentage of morphologically normal primordial follicles in a fresh group which was significantly higher than the treatment groups (p < 0.001). Terminal deoxynucleotidyl transferase dUTP nick end labeling and anti-caspase-3 staining observed in oocytes, follicle cells, interstitial tissue showed higher rates of apoptosis for post-vitrification and -transplantation groups than freshly grafted ovarian tissues. Furthermore, we observed significant downregulation of zone of polarizing activity and zona pellucida C protein gene expression in vitrified ovarian tissue grafts than in the fresh grafts (p < 0.05). In conclusion, we suggest that the needle immersed vitrification method is a convenient, cheap, and feasible vitrification method for cat ovarian tissues. However, further studies need to be performed to determine more optimal vitrification solutions and equilibration times for the needle immersed vitrification method in order to adapt it for cat ovaries.

Effect of strategies to increase progesterone levels on fertility of bovine embryo transfer recipients - A meta-analysis.

The pregnancy rate following embryo transfer (ET) is a very important factor in the success of embryo production programs. Different strategies were therefore developed to increase pregnancy rates. The aim of this meta-analysis was to investigate the effects of hormone treatments used to increase the success of embryo transfer programs on pregnancy rates. A meta-analysis was performed of 46 trials from 39 publications involving treated (n = 7856) and control (n = 6663) cattle. The meta-analysis explained the effect size with its 95 % confidence interval (CI) for pregnancy per embryo transfer (P/ET) after hormonal treatment under different moderators. Hormonal support was found to increase P/ET compared to the control group (P < 0.05). However, GnRH treatment was found to increase P/ET by approximately 4.3 % and hCG treatment by 8.0 %. Progesterone supplementation was not found to have a statistically significant effect on P/ET. In addition, GnRH treatment significantly increased P/ET when used to transfer in vitro or frozen-thawed embryos or in studies using cows as recipients. It was observed that hCG treatment had a positive effect on P/ET according to all moderators. Progesterone supplementation significantly increased P/ET when frozen embryos were transferred and reduced P/ET, especially in publications where fresh or in vitro produced embryos were transferred or cows were used as recipients. The results of this meta-analysis showed that the use of GnRH, and hCG, in bovine embryo transfer programs increased P/ET, whereas the use of progesterone had no effect on P/ET. However, it was found that P/ET could increase/decrease depending on the moderator.

Multiple ovulation and embryo transfer during the breeding season in Angora goats: A comparison of fresh and vitrified-thawed embryo transfer.

This study aimed to assess the superovulation response and pregnancy rates of fresh and vitrified-thawed embryos after transfer in Angora goats with comparing transfer at the beginning (BS) and end (ES) of the breeding season. Nine Angora goats were used as donors in both periods. Donor goats were synchronized and superovulated with the FSH and mated with five fertile bucks. At 156 hr following mating, embryos were collected surgically. Recipient Angora goats were divided into two groups at the beginning (fresh, n = 15; vitrified-thawed, n = 15) and end (fresh, n = 8; vitrified-thawed, n = 8) of the breeding season. Fresh or vitrified-thawed grade I embryos (early blastocyst/blastocyst) were transferred surgically to synchronized recipients. On the 30th, 60th and 90th day of transfer, goats were examined by ultrasonography. The number of corpora lutea (CL), total oocyte/embryo and transferable embryos obtained in BS was found to be statistically higher than ES. On the 30th day of transfer, pregnancy rates were 73.30% and 75.00% in the fresh transfer groups in both BS and ES periods; while, rates of 20.00% and 37.50% were found in the vitrified-thawed group, respectively. The embryo survival rates of fresh transfers were 55.55% and 31.25% at BS and ES, respectively. The number of CL, total oocyte/embryo and transferable embryos in the BS was higher than ES following superovulation. Also, fresh embryo transfer can be successfully carried out during the breeding season in Angora goats. Moreover, although pregnancies were obtained following the transfer of vitrified-thawed embryos, they did not sustain on the 60th and 90th days. So, further studies are needed for the vitrified-thawed embryos.

The evaluation of clinicopathological and immunohistochemical findings of cervical clear cell carcinoma in a bitch

Background: Clear cell carcinoma (CCC) of the cervix is a rarely seen malignant tumor which is classified in adenocarcinomas of the endometrium. It is responsible for 2.15-3.0% of endometrial tumors. The diagnosis is difficult and is still controversial in uterine carcinomas. Macroscopically, masses were protruding to endometrial surface. However, the definitive diagnosis was carried by microscopical evaluation. The cells contain clear cytoplasm. The cells are seen in this pattern because there is either glycogenation or secretoric activity in the cells.Case: In the present case, clinicopathologic and immunohistochemical findings of clear cell carcinoma of the cervix were defined in a 3 year-old spayed Golden retriever bitch. Previously spayed bitch, which had vaginal discharge for the last month-long, was submitted to the Department of Obstetrics and Gynecology, Ankara University, Faculty of Veterinary Medicine. In the general clinical examination, vaginoscopy was performed and vaginal cytology was obtained from smear. The source of discharge was detected as cervix uteri. In vaginal cytologic examination, erythrocyte, neutrophile and superficial cells were observed. In abdominal ultrasonography, a hypoechoic mass was detected at cranial part of the urinary bladder. To evaluate the general health condition of the dog, total blood counting and serum biochemistry were analyzed in addition to assessing its hormone prophlye. Estrogen and progesterone levels were evaluated. Estradiol (E2) level was measured as 23 pg/mL and progesterone level was measured as 1.96 ng/mL from collected serum. The cervical mass in diameters of 3x4x2.5 cm was removed in operation. In macroscopical examination, it had spherical and regularly shape. After the macroscopical examination, tissue samples were fixed in 10% buffered formalin. Then, the samples stained with Haematoxylin-Eosin (H&E), Massons trichrome and PAS stainings, respectively.[...](AU)

The evaluation of clinicopathological and immunohistochemical findings of cervical clear cell carcinoma in a bitch

Background: Clear cell carcinoma (CCC) of the cervix is a rarely seen malignant tumor which is classified in adenocarcinomas of the endometrium. It is responsible for 2.15-3.0% of endometrial tumors. The diagnosis is difficult and is still controversial in uterine carcinomas. Macroscopically, masses were protruding to endometrial surface. However, the definitive diagnosis was carried by microscopical evaluation. The cells contain clear cytoplasm. The cells are seen in this pattern because there is either glycogenation or secretoric activity in the cells.Case: In the present case, clinicopathologic and immunohistochemical findings of clear cell carcinoma of the cervix were defined in a 3 year-old spayed Golden retriever bitch. Previously spayed bitch, which had vaginal discharge for the last month-long, was submitted to the Department of Obstetrics and Gynecology, Ankara University, Faculty of Veterinary Medicine. In the general clinical examination, vaginoscopy was performed and vaginal cytology was obtained from smear. The source of discharge was detected as cervix uteri. In vaginal cytologic examination, erythrocyte, neutrophile and superficial cells were observed. In abdominal ultrasonography, a hypoechoic mass was detected at cranial part of the urinary bladder. To evaluate the general health condition of the dog, total blood counting and serum biochemistry were analyzed in addition to assessing its hormone prophlye. Estrogen and progesterone levels were evaluated. Estradiol (E2) level was measured as 23 pg/mL and progesterone level was measured as 1.96 ng/mL from collected serum. The cervical mass in diameters of 3x4x2.5 cm was removed in operation. In macroscopical examination, it had spherical and regularly shape. After the macroscopical examination, tissue samples were fixed in 10% buffered formalin. Then, the samples stained with Haematoxylin-Eosin (H&E), Masson’s trichrome and PAS stainings, respectively.[...]