Background: Objective of the study was to explore exclusive breastfeeding knowledge and practice of among mothers of children less than 2 years in rural areas in Beni-Suef governorate and factors that determine them.Methods: A community-based cross-sectional study was conducted in rural areas, Beni-Suef governorate over 6 months (February 2019-July 2019) using a designed well-structured questionnaire.Results: The mean breastfeeding knowledge score of enrolled mothers was 7.87±2.2 out of 15. (74.5%) of mothers recruited in this study obtained their knowledge from their mothers, families, and friends. Only 40.4% of the mothers have exclusively breastfed their children. While 57% of participants added food or fluids before 6 months and 37.6% added additional artificial feeding. Mothers’ age, education, occupation, marital status, the order of last-child, mode of delivery, place of delivery and monthly family income were independently associated with exclusive breastfeeding in univariate analysis. While in multi-variate analysis, only mothers’ age, education, occupation, marital status and breastfeeding knowledge score was associated.Conclusions: Rural women had suboptimal knowledge about exclusive breastfeeding, justifying the suboptimal practices of the mothers. Hence, it is recommended to develop interventions emphasizing practical education targeted at addressing factors that influence exclusive breastfeeding. And successful infant-feeding interventions aimed at promoting overall infant health.