American Foulbrood (AFB), a severe disease that affects larvae of the honeybees. Due to the serious effects associated with AFB and the problems related to the use of antibiotics, it is necessary to develop alternative strategies for the control of the disease. The aim of this study was to determine, under field conditions, the effectiveness of tylosin and three kind of Propolis Ethanolic Extract (Chinese, Egyptian and old wax comb extract propolis) for controlling AFB in honeybee colonies. Identification of individual phenolic compounds of the PEE (Propolis Ethanolic Extract) was performed on a HPLC (High Performance Liquid Chromatography) instrument and the result obtained that there is different in phenolic compounds content among the three kinds of PEE. Laboratory studies were conducted to determine the LC50 (half lethal concentration) and LT50 (half lethal time) values for acute oral toxicity of 2, 1, 0.5, 0.1, 0.05, 0.025% in three kind of PEE (Chinese, Egyptian and old wax comb extract propolis) on worker honeybees to chose the safely concentration of PEE on honeybees. The result showed that the 0.1, 0.05 and 0.025% were non-toxic on honeybee workers but 2, 1 and 0.5% were toxic. In field trials the colonies were inoculated by AFB for two weeks before initiation of trial and treated with tylosin (as a dust in confectioners sugar), 0.1, 0.05 and 0.025% PEE (as soluble in sugar solution 50%) which extracted from Chinese propolis (C.), Egyptian propolis (E.), old wax comb extract (W.) and fed with sugar syrup only (Control) for three weeks at one week intervals. The result indicated that tylosin, 0.1 and 0.05 % of E.PEE (Egyptian Propolis Ethanolic Extract) had elimination of AFB clinical symptoms at 100% of reduction rate.