The aim of this study was to determine the peroxide values, acid numbers, oxidative stability (Rancimat method), antioxidant activity (DPPH method), antimicrobial activity (disc diffusion method), sensory properties (9-point hedonic scale), and fatty acid profiles (FAME) of five edible oils purchased from local Slovakian producers—grape seed oil, flax seed oil, walnut oil, poppy seed oil, and milk thistle seed oil. The peroxide value ranged from 2.27 (milk thistle oil) to 8.51 (flax seed oil) mmol O2/kg. All these values were in accordance with regulations (upper limit of 20 mmol O2/kg). The values of the acid number ranged from 0.11 (walnut oil) to 2.49 (milk thistle oil) mg KOH/g, and were in accordance with regulations as they did not exceed the value of 4 mg KOH/g. The oxidation stability was the lowest in flax seed oil (0.18 h) and the highest in grape seed oil (2.05 h). In milk thistle oil, the highest amounts of oleic and behenic acids, in flax seed oil, the highest amount of α-linolenic acid, and in grape seed oil, the highest amount of linolic acid were determined. Antioxidant activity was the strongest in the sample of grape seed oil—65.53 mg TEAC/L (Trolox equivalent antioxidant capacity). Samples of flax seed oil showed the strongest inhibition of Candida albicans CCM 8186 (4.58 mm) and Bacillus subtilis CCM 2010 (0.31 mm). Poppy seed oil was determined to be the most inhibiting towards Klebsiella pneumoniae CCM 2318 (3.68 mm). Milk thistle oil showed the strongest inhibition of Clostridium perfringens CCM 4435 (6.31 mm). Grape seed oil was the most inhibitory towards Staphylococcus aureus subs. aureus CCM 2461 (5.32 mm). Walnut oil showed the strongest activity towards Yersinia enterocolitica CCM 5671 (6.33 mm). The sensory analysis resulted in the samples of walnut and grape seed oil being awarded the highest scores for smell, taste, and overall acceptability. The tested edible oils are rich in biologically active compounds with antioxidant and antimicrobial activities. Their consumption can have a positive effect on the functioning of the human body and its health. Proper storage conditions are, however, necessary because of the susceptibility of these oils to oxidation.