For decades, polyethylene (PE) has been regarded as non-biodegradable. The literature has provided only a limited amount of valid evidence regarding the biodegradation of PE by bacterial cultures, despite the occasional description of this procedure. Two bacterial Bacilli spp. isolates were recovered from the intestine of earthworms and were found to be capable of degrading PE. In addition to microscopic examination, the bacterial isolates were characterized using selective media and a few biochemical assays. The maximal growth rates for the two isolates B1 and B2 were estimated to be 1.8 and 1.3, respectively, three weeks after the experiment commenced, using a Spectrophotometer at a wavelength of 600 nm. During the twenty-one-day incubation period of the two isolates on PE films, viable biofilms were formed, and the hydrophobicity of the PE films decreased. Subsequently, atomic force microscopy (AFM) was employed to detect evident damage, such as fissures and cavities (18.5–21.1 nm in depth), on the surfaces of the PE films for B1 and B2. Fourier transform infrared (FTIR) was employed to confirm the formation of carbonyl groups. The findings indicate that PE-degrading bacteria are present in the guts of earthworms, and they offer optimistic evidence for the biodegradation of PE in the environment.