The present study was carried out to shed light upon the some materials that used in increasing antibiotic activity against the isolated bacteria from otitis media patients. A total of (84) samples were collected from different age group male and female patients with otitis media. The patients were either hospitalized at Al-Kadhumia Educational Hospital or attending the Central Public Health Laboratory of Ministry of Health during the period from 5/12/2006 - 28/5/2007. A total 59 samples gave positive results, of which 25 were identified as Pseudomonas areuginosa, 15 Staphylococcus epidermidis, 6 Staphylococcus aureus, 5 Escherichia coli, 3 Proteus maribilis and 5 fungi. Pseudomonas areuginosa was the most common pathogen in the studied samples of otitis media. Patients in the age range between (1-10) years were more infected than other age groups. Male patients were more infected than female patients with otitis media, and right ear was subjected to infection more than left ear. During winter, the frequency of infection was more than other seasons. The obtained results showed that single infection among patients with otitis media was more common mixed infection. When susceptibility of bacterial isolates to 12 antibiotics was tested using disk diffusion assay. The results revealed that Ciprofloxacin was the most effective antibiotic against bacterial isolates followed by amikacin and then by gentamycin. Nine isolates were selected according to their pattern of resistance as those showing muli-drug resistance and tested to specify their minimum inhibitory concentration for (cephalexin, ciprofloxacin, streptomycin, amikacin, cefotaxim, tetracyclin, erythromycin, ampicillin, gentamycin, nalidixic acid). Ciprofloxacin was found to be the most effective one through its lowest MIC as compared to other antibiotics. The effect of Tris-EDTA and ascorbic acid when were combined separately with antibiotics and tested against the nine isolates by using disk diffusion assay. The obtained results showed that there was a synergistic effect between Tris-EDTA or (ascorbic acid) and each of â- lactam, tetracycline, aminoglycosides, SXT, quinolone and erythromycin against Gram negative and positive bacteria. Adversely, there was an antagonistic effect between Tris-EDTA or ascorbic acid and quinolone or aminoglycosides against gram negative bacteria, only.