Propolis (bee glue), a valuable biological mixture produced by European honey bees (Apis mellifera L.), is widely used as a remedy in traditional and alternative medicine, as an effective food biopreservative and as a nutritional value enhancer. The present study aimed to investigate eighty propolis samples collected from all twenty-eight districts of Bulgaria in the period of 2020–2022 in order to determine their physicochemical properties as well as their antioxidant and antimicrobial potential. The raw propolis samples exhibited brown or green colour depending on the climatic zone from which they are collected and pH values from 4.82 to 5.87, while the moisture content ranged between 0.98% and 2.97%. The total phenolics content (TPC) and total flavonoids content (TFC) varied from 63.14 mg GAE/g to 737.27 mg GAE/g, and from 29.22 mg QE/g to 234.17 mg QE/g, respectively. The propolis samples demonstrated significant antioxidant potential determined by DPPH and FRAP methods, from 18.56 mM TE/g to 1598.66 mM TE/g and from 82.28 mM TE/g to 1208.81 mM TE/g, respectively, whose values showed a positive correlation (r2) with the TPC and TFC. The results from the antimicrobial screening revealed that the methanolic propolis extracts in concentration of 20 mg/ml exhibited the highest inhibitory effect on the Gram-positive bacteria such as Micrococcus luteus 2YC-YT, Bacillus subtilis ATCC 6633, Staphylococcus aureus ATCC 25923, Listeria monocytogenes NBIMCC 8632 and Listeria innocua ATCC 33090.
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