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    Effect of Ergothineine and Quercetin Additions in to the In Vitro Embriyo Development
    (2024) Bodu, Mustafa; Ozturk, Ali Erdem; Elcin, Murat; Atay, Yunus Emre; Narlicay, Salih; Ataman, Mehmet Bozkurt; Bucak, Mustafa Numan
    Aim: This study aimed to investigate the effects of ergothioneine and quercetin on the in vitro development of bovine embryos. Materials and Methods: Cumulus-oocyte complexes (COCs) were collected via ovary aspiration from a local abattoir and cultured in vitro for maturation. After maturation, in vitro fertilization was performed. The pronuclear embryos were divided into three groups: control, 10 ?M L-ergothioneine, and 10 ?M quercetin supplemented. After the addition of antioxidants to the CR1aa medium, in vitro culture of embryos were performed. The cleavage and morula rates were assessed on days 2 and 5, respectively. Blastocyst formation and quality were assessed on days 7-8. Results: Statistical analysis showed cleavage and morula rates were significantly higher in the ergothioneine group compared to the quercetin and control groups (P<0.05). While no blastocysts formed in the quercetin group, the blastocyst rate reached to 17.96% with ergothioneine supplementation on day 8. Conclusion: In conclusion, supplementation with 10 ?M ergothioneine enhanced the in vitro development of bovine oocytes. However, 10 ?M quercetin supplementation impaired development, and no blastocyst formation observed. Further studies utilizing different concentrations are warranted to better understand the effects. This study provides insights into modulating oxidative stress during in vitro embryonic production.
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    Effects of Heat-Stress on Oocyte Number and Quality and In Vitro Embryo Production in Holstein Heifers
    (Univ Fed Rio Grande Do Sul, 2022) Cizmeci, Sakine Ulkum; Dinc, Dursun Ali; Yesilkaya, Omer Faruk; Ciftci, Muhammed Furkan; Takci, Abdurrahman; Bucak, Mustafa Numan
    Background: This study aimed to determine the effects of environmental temperature on the number and quality of oocytes and embryo production rates obtained by performing ovum pick up (OPU). Heat stress leads to long-term, short-term, visible, and invisible effects in dairy cows. Its effects on reproduction are evident in all stages, from oocyte development to birth. Disturbance in ovarian follicle development, follicular dominance deficiency, anoestrus, polyspermia, embryonic losses, decreased fetal growth, and abortion are some examples of responses to these effects. The aim of the present study was aimed to determine the effects of ambient temperature on oocyte quality and number and embryo production rates. Materials, Methods & Results: The animal material used in this study comprised 10 Holstein heifers. At the beginning of the study, the heifers were 13-15 months old. OPU was performed at different times of the year, and weather conditions were recorded. Grouping according to ambient temperature was done as < 10 degrees C (group 1), 10-25 degrees C (group 2), and > 25 degrees C (group 3). The veterinary ultrasonography device and a set of compatible intravaginal OPU probe, catheter, and aspiration device were used for OPU application. All antral follicles with diameters of 2-8 mm in the ovaries were aspirated. The aspirated follicle fluids were examined under a stereo microscope, and the cumulus-oocyte complexes (COC) were collected and classified according to their structure. A, B, and C-quality oocytes were included in the in vitro embryo production process. After performing 69 OPUs on random days of the cycle, the number of oocytes per OPU was found to be 8.72, 6.32, and 6.85 in groups 1, 2, and 3, respectively (P < 0.05). The number of viable oocytes per OPU was 6.83, 4.64, and 4.65 in groups 1, 2, and 3, respectively (P < 0.05). The statistical difference between the first group and the other groups was significant for cleavage and blastocyst counts (P < 0.05). Discussion: All the negative effects of heat stress on animals resulted from the increased body temperature. Reproductive performance is adversely affected by high temperatures and humidity during periods of high ambient temperatures. Metabolic heat is released, and the heat load increases due to the metabolism of nutrients in cattle. Internal body temperature is regulated via the dissipation of metabolic heat to the environment. The amount of heat dissipated via conduction and convection depends on the unit body weight, surface area, skin and coat color, difference in temperature gradient of the animal and ambient temperature, and humidity. In the present study, it was determined that the blastocyst development rates of the oocytes obtained in the warm season (>25 degrees C [group 3]) were lower than those of the other groups. It was concluded that this may be because the oocytes developed under chronic heat stress in the animals, and several cycles were required to enhance oocyte quality and developmental potential. Additional studies are needed to investigate the response of oocytes obtained with OPU to heat stress during embryonic developmental stages and to determine the sensitivity and effects of embryonic tissue damage according to developmental stages. Based on the results of the present study, it was concluded that performing OPU and in vitro embryo production (IVEP) when the ambient temperature is close to the thermoneutral limits may increase the blastocyst development rates.
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    Interaction of cumulus-oocyte complex (coc) number, oocyte quality, and blastocyst numbers by repeated ovum pick-up (opu) in in vitro embryo production
    (Chulalongkorn University Printing House, 2024) Cizmeci, Sakine Ulkum; Dinc, Dursun Ali; Bucak, Mustafa Numan; Yesi̇Lkaya, Omer Faruk; Ciftci, Muhammed Furkan; Kivrak, Mehmet Bugra; Sari, Ayse
    Ovum pick-up (OPU) is a repeatable technique that is used for the retrieval of large numbers of immature oocytes from the antral follicles of live animals. The aim of the present study was to determine the effect of repeated OPU application on cumulus-oocyte complex (COC) number, oocyte quality, and blastocyst rates. The animal material for the study consisted of 10 Holstein heifers. Heifers selected as donors were administered OPU a total of 9 times on random days of the cycle at 1-week intervals. All antral follicles with a diameter of 2-8 mm were aspirated during the OPU procedure. In vitro embryo production (IVEP) was performed on the oocytes obtained after quality assessment. Various degrees of adhesion and connective tissue thickening were found in the ovaries of the animals after 9 repeated applications of OPU. It was found that the average number of A-quality oocytes was higher than the number of B, C, and D-quality oocytes. The number of cleavages per OPU was 2.69 (cleaved oocytes/opu session), and the number of blastocysts was 0.72 (number of blastocysts/opu session). It was determined that the general average of cleavage ratios was 73.62%, and blastocyst rates were 18.97%. It was observed that the number of oocytes, cleavage, and blastocysts obtained as a result of repeated OPU applications was not affected by repeated OPU. It was concluded that in order to achieve the target number of oocytes and blastocysts with recurrent OPU, experiments should be carried out with varying frequencies of OPU, and prophylactic measures should be taken to prevent adhesions. © 2024 Chulalongkorn University Printing House. All rights reserved.