The antimicrobial activity of 19 propolis extracts prepared in different solvents (ethanol and propylene glycol) (EEP/PEP), was evaluated against some bacterial and fungal isolates using the agar-well diffusion method. It was verified that all the samples tested showed antimicrobial activity, although results varied considerably between samples. Results revealed that both types of propolis extracts showed highly sensitive antimicrobial action against Gram-positive bacteria and fungi at a concentration of 20% (Staphylococcus aureus, Streptococcus mutans, Candida albicans and Saccharomyces cerevisae) with a minimal inhibitory concentration (MIC) ranging from 0.5 to 1.5 mg/ml, with a moderate effect against Streptococcus pyogenes (MIC from 17 to 26 mg/ml). To our knowledge, this is the first study showing elevated antimicrobial activity against Gram-negative bacteria [Salmonella enterica (MIC from 0.6 to 1.4 mg/ml)] and lesser activity against Helicobacter pylori (MIC from 6 to 14 mg/ml), while Escherichia coli was resistant. This concluded that the Basque propolis had a strong and dose-dependent activity against most of the microbial strains tested, while database comparison revealed that phenolic substances were responsible for this inhibition, regardless of their geographical origin and the solvent employed for extraction. Statistical analysis showed no significant differences (P ≤ 0.05) between EEP and PEP extracts.