Chronic suppurative otitis media is still a significant health problem. Several topical agents such as antibacterials, steroids and acid media eardrops are widely used alone or in combination to control active otorrhea and obtain a dry ear. Patients frequently ask if there is any benefit or loss in the effect of these due to their use in a successive or staggered manner. This question led us to design this in vitro study to investigate the effect of the acetic acid solution and dexamethasone sodium phosphate on the antimicrobial effects of some ototopical antibiotics during their successive or staggered usage. Staphylococcus aureus, Pseudomonas aeruginosa, Proteus mirabilis and Escherichia coli were used for testing the antimicrobial activities of the selected antibiotic drops. Six microliters of acetic acid solution and dexamethasone sodium phosphate were used alone or together with 6 mu l each of ciprofloxacin HCl, tobramycin and ofloxacin solution. Drops were placed on Whatman no. 1 paper disks with a diameter of 6 mm, and the disks were placed onto Mueller-Hinton agar plates on which bacteria had been swabbed over the entire surface. After overnight incubation of the plates at 35 degrees C, the diameters of the inhibition zones were recorded. There was no antibacterial effect in response to either acid solution or dexamethasone used alone. The inhibition zones of the antibiotics did not change with either steroid or acid media solution added onto the antibiotic disks alone or together. Acid solution or steroids should be preferred to use in combination with antibacterial otic drugs in in-vivo settings; no negative or positive effects were seen in in-vitro conditions. Therefore, prescribed eardrops can be used in a successive or staggered manner without any influence on the treatment results.