Canine oocyte nuclear maturation with Nano-ozone (NZS) supplementation: The alterations of antioxidant, and oxidant status and CDK1, cyclin B1 expressions.

This study aims to evaluate the effects of nano-ozone solution (NZS) on canine oocyte nuclear maturation, associated with the alterations of antioxidant and oxidant status and cyclin-dependent kinase 1 (CDK1), cyclin B1 gene expressions. Oocytes were cultured in four distinct concentrations of NZS (0.5, 1, 2, and 5 µg/mL) and parthenogenetically activated. The rates of oocytes arrested at the Germinal Vesicle (GV), Germinal Vesicle Breakdown (GVBD), Metaphase I (MI), and Metaphase II (MII) stages were statistically different among groups (P < 0.05). The oocytes cultured in 1 µg/mL NZS yielded the best oocyte maturation rate at the MI and MII stages; however, the lowest maturation and high degeneration rates were observed in Group E. The measurements of Malondialdehyde (MDA), reduced Glutathione (GSH), Superoxide Dismutase (SOD), and Ferric Reducing/Antioxidant Power assay (FRAP) were performed from IVM culture media. No statistical difference was observed in SOD and MDA results (P > 0.05). GSH levels were statistically significant between Group A-Group E (p = 0.003), Group B-Group E (p = 0.045), and Group E-Group D (p = 0.021). The culture media in Group D and Group E had high FRAP concentrations and significantly differed between groups (P < 0.05). CDK1, and cyclin B1 genes, which are subunits of maturation-promoting factor (MPF), are upregulated in Group B and Group C, while are downregulated in oocytes of Group E. This study showed that low, controlled doses of NZS (1 µg/mL) supplementation could improve the meiotic competence of canine oocytes and lead to positive response in expressions of CDK1 and cyclin B1 on the gene level.

Can Nanobubble Ozone Liposomes Be A New Agent In The Fight Against Foodborne Infections?

Backrounds: In our study, a nanoparticle liposome molecule with patent application number TR2021004032 was used, and the Minimum Inhibitor Concentration (MIC) was found to be 1562 ppm. According to the ASTM F 1980 standard, it has been determined that the nanoparticle liposome solution kept at 37 days and 55 oC in return for one-year stability preserves its effectiveness. Our study aimed to show that the newly developed solution maintains its effectiveness for a long time. METHODS: In this study, a nanobubble ozone liposome solution containing 2% ZnCl2 was used. The aging tests were conducted according to the ASTM F 1980 [1] standards. The minimum inhibitory concentration (MIC) level of the nanobubble ozone liposome solution with 2% ZnCl2 was determined as 1.562 ppm for strains of Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) by the CLSI M07 A9 [2] standard test method. To assess the time-dependent antibacterial effect of the nanobubble liposome solution with 2% ZnCl2, the solution's efficiency at a concentration of 2000 ppm and for different time intervals was tested on strains of Salmonella enterica subsp. enterica (ATCC® 14028™) and Listeria monocytogenes (ATCC® 7644™). RESULTS: The results showed that the antibacterial activity of the strains of S. enterica subsp. enterica started at the end of the 10th minute and the solution was effective after 30 minutes. For strains of L. monocytogenes, it was observed that the activity started at the end of the 2nd minute and the product was effective after the 10th minute. According to the ASTM F 1980 standards, it was found that the nanobubble ozone liposome solution retained its effectiveness in one-year stability tests. CONCLUSION: As a result, the nanoparticle liposome solution, a new product, does not lose its stability and effectiveness for a long time, contrary to what is known. Although the half-life of gaseous ozone is as short as 20 minutes, the stability in the nanoparticle liposome solution has been determined as at least one year. Since nanoparticle liposome solution is a natural and slow-release product, nanobubble ozone liposome solution with 2% ZnCl2 may be used as a newly developed agent against contaminations in food processing facilities caused by biofilm-forming microorganisms through the use in disinfections of surfaces that are in direct contact with food products.