To select strains capable of degrading aflatoxin B1 (AFB1) efficiently, 32 bacteria strains were isolated from different sources which display AFB1 reduction activity, using coumarin as the sole carbon source and biodegradation technology. AFB1 degradation effectiveness was detected by high-performance liquid chromatography (HPLC) after AFB1 standard was added (final concentration of culture supernatant was 2.5 μg/mL). The bacteria strain isolated from chicken manure was named as F6, which could degrade AFB1 by 83.3%. Based on the morphologic, physiological, biochemical characteristics, the isolated strain F6 was preliminary identified as Bacillus. The analysis of 16S rRNA gene sequence homology revealed that, the nucleotide identity between strain F6 and Bacillus oleronius international standard strain ATCC700005 (accession number NR043325.1) was 99.3%. Thus, strain F6 was determined as B. oleronius and named as B. oleronius GX01 (Accession No. KP297896). The components of the fermentation broth of F6 were then isolated and detected. The results showed that the culture supernatant could degrade 83% AFB1, while the degradation rates of bacterial suspension and intracellular were 22.8% and 17.4%, respectively, suggesting that the degradation activity of B. oleronius strain F6 was exoenzyme.