Background: Natural oil is an aromatic by-product of highly volatile plants. It can be easily extracted from a variety of natural sources using a variety of distillation processes due to its volatility. Oregano oil is a substance derived from the oregano plant, also known as Origanum vulgare. Objectives: The purpose of this study was to determine the physicochemical properties of oregano oil and to relate them to its chemical composition. Moreover, to examine its antibacterial activity against urinary tract pathogens. Materials and Methods: Commercial natural oregano oil (purity ≥98%) was obtained from Bluray food products operating under the brand name Bluray Food Products. The physico-chemical characteristics of oregano oil were determined using different techniques. GC-MS was used to analyze the chemical properties of oregano oil as well as analysis of saponification value, peroxide value, p- Anisidine value, pH value, and organoleptic properties (color, odor, consistency, nature, and solubility) were performed. Results: According to the results, GC-MS identified 66 different chemicals in oregano oil, accounting for 100% of the overall composition. It was found that oregano oil has a saponification value of 187.94 mg KOH/g, a peroxide value of 6.22, and a P-Anisidine value of 3.645. In addition, the oil of oregano was found to have inhibitory activity against all uropathogenic bacteria tested. E. coli has an inhibitory zone of 29 mm, P. aeruginosa 27 mm, K. pneumoniae 20 mm, P. mirabilis 22 mm, E. aerogenes 21 mm, E. faecalis 21-mm, A. baumannii 22-mm, N. gonorrhoeae 24-mm, S. aureus 26 mm, and S. epidermis 20-mm. Conclusion: Based on these results, it can be concluded that oregano oil was found to have optimal antibacterial activities against all tested urinary pathogenic bacteria.