Inappropriate use of antibiotics is a significant public health problem. Nevertheless, only few studies have been conducted in Egypt in this area. The aim of this study was to assess knowledge, believes and practice regarding antibiotic use and misuse among rural mothers in Gharbia governorate. Material and Method: a cross-sectional design was used. The study was conducted at twenty rural health units in Gharbia governorate. An interview questionnaire was held with 1000 convenient subjects exploring their knowledge, believes, and practices regarding antibiotic use and misuse in a twenty weeks-period. Results: It was found that the majority of mothers had poor knowledge and bad believes about antibiotics. Nearly about two thirds of them used antibiotic inappropriately. More than half of the studied mothers believed that antibiotics will relive influenza manifestation and may change the doctor if not prescribe antibiotic for them. There was a significant statistical relationship found between mothers’ level of education and their score of knowledge, believes, and practice. Conclusion and Recommendation: Mothers use and misuse of antibiotics was positively correlated with their knowledge and believes. Therefore, addressing this problem will require a combined approach including improved access, legislation, education, and culturally relevant client provider interactions.