An isolate of unkown bacterium was isolated from honeybee larvae infected with European foulbrood. Based on its characteristics of isolation and culturing, colony morphology, physiological-biochemical characteristics and Molecular Biology, this isolate was identified the secondary invader of European foulbrood, Enterococcus faecalis, which was belong to the genus of Enterococcus, and was named Enterococcus faecalis FB102 tentatively. Meanwhile it showed that people could different the bacterium from the pathogen of European foulbrood by the biological characters of isolates and cultivation. The bacterium could not infect honeybee larvae alone, and it could simplify the diagnosis process of European foulbrood by the isolation and culturing of Enterococcus faecalis, furthermore the bacterium from honeybee larvae might threaten human health.