Paenibacillus larvae is the causative agent of American Foulbrood (AFB ), a severe disease that affects the larvae of the honeybees. The use of plant extracts are considered to be an alternative way of controlling the disease. In this study, the in vitro antimicrobial activity of Buddleja thyrsoides Lam. against the Paenibacillus species, including P. larvae, was evaluated. In Mueller-Hinton broth, the minimal inhibitory concentration (MI C) was assessed using the microdilution method. All Paenibacillus species were sensitive to the crude extract and the fractions of B. thyrsoides. The ethyl acetate (EA ) fraction showed a better result with MI C values of 1.68 - 3.36 mg/mL, followed by butanolic (BU) (MI C = 2.18 - 6.54 mg/mL), dichloromethane (DCM) (7.40 - 14.80 mg/mL), and crude extract (CE) (7.51 - 16.90 mg/mL). The toxic effect of the CE and fractions of B. thyrsoides against bees were also evaluated using the spraying application method with the same concentrations of MI Cs. Bee mortality was evident in treatment with DCM fractions only, while CE, EA , and BU extracts showed no toxic effects after 15 days of observation. Furthermore, phenolic acids, tannins, and flavonoids were identified and quantified by highperformance liquid chromatography (HPLC), and may be partially responsible for the antimicrobial properties observed. These results show, for the first time, that B. thyrsoides might be a natural alternative for the prevention/control of AFB .