the aim of this study was to determine the phenolic compounds and antibacterial activity components of pollen grains, obtained from different concentrations extracts of ethanol and water for controlling American foulbrood (AFB) disease. In this study four Egyptian bee pollen grains types were collected from pollen basket, namely Broad bean (Vicia faba), date palm (Phoenix dactylifera), clover (Trifolium alexandrinum) and maize (Zea mays), during spring. Identification of Phenolic composition in pollen grains samples were investigated by high performance liquid chromatography (HPLC) instrument. Two pollen extracts, using ethanol and water by three concentrations 5, 10 and 20% agent's Paenibacillus larvae subspecies larvae, formerly Bacillus larvae and it's potential for the control of American foulbrood disease (AFB) in vitro. Generally the concentration of 5% gave no effect neither pollen ethanol or aqueous extracts. But pollen grain water extracts gave inhibition zone diameter with different pollen grain of types (10%) and (20%) concentrations. The water extract of date palm pollen grains recorded a highly effect of inhibition zone which were 21.30& 28.65mm with 10&20% concentrations respectively. This study considers a primary step for the pollen grains on P. larvae spores in vitro. These products showed efforts no toxicity effects on different stage of honeybee. Suggesting them as perspective, low cost and consumer-acceptable agent for control of AFB disease in honeybee colonies.