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Mumijo is a traditional medicine used in Central Asia thousands of years. Its beneficial effect is exerted on metabolic processes and on the human immune system. It is also used as an antiseptic agent. The in vitro antibacterial activity of a mumijo preparation was investigated with agar diffusion technique against 25 Gram-positive and 6 Gram-negative bacterial isolates representing 13 different genera. Effect of different media and pH on the antibacterial action was also studied. Different concentrations of mumijo exerted bactericid and/or bacteriostatic effect against 3 Gram-negative (Acidovorax delafieldii, Serratia marcescens, Variovorax paradoxus) and 15 Gram-positive bacterial isolates (Agrococcus jenensis, Arthrobacter globiformis, Bacillus benzoevorans, Bacillus cereus var. mycoides, Bacillus megaterium,Bacillus subtilis, Enterococcus faecalis, Staphylococcus aureus) on the complete media. The maximum inhibitory potential of mumijo was observed at neutral pH (pH=7.4). Antibacterial effect of mumijo was not observed when minimal media were used for cultivation. The experimental data revealed that mumijo has remarkable growth inhibition effect against Gram-positive bacteria on complete medium at neutral pH. Key words: Mumijo, Gram-positive bacteria, Gram-negative bacteria, bacteriostatic effect, bactericid effect.