The emergence and spread of bacterial resistance to antibiotics is a growing problem worldwide, which presents a significant threat to public health globally. The present study was designed to determine knowledge, attitude and practice of antibiotic use. A cross-sectional survey was performed using a questionnaire on a sample randomly selected from east Libya for pediatric doctors, pharmacies and parents. A total of 400 participating subjects were approached to be included in the study, 234 agreed to participate giving a response rate of 58.5%. The awareness of antimicrobial resistant was very high among the pharmacist and doctors responder (96.43%) (93.91%) respectively, in contrast participants parents (41.76%). More than one-third (37.36%) were self-medicated with antibiotics to treat mainly common cold and cough. The reasons of antibiotic misuse in the present study according to the opinion of participant doctors were an empirical use, incorrect dose and low level of education with 68.70%, 18. 26%, 13.04% respectively. Pharmacies are the major source of antibiotics reporting more than –quarter 28.57% were sold antibiotic without prescription. These findings presented aid in the assessment of the adequacy of present public educational campaigns to promote specific messages to rationalize antibiotic use, and compensate for knowledge and attitude gaps as an effort towards preventing development of antibiotic resistance.