This study demonstrated the biodegradation of two different brands of cigarette butts (CBs), which are primarily composed of cellulose acetate, by four distinct microorganisms. These included the white rot fungus Pleurotus ostreatus, the brown rot fungus Lentinus lepideus, and the bacteria Bacillus cereus and Pseudomonas putida. After 31 days of treatment, weight loss measurements revealed a mass loss of 24-34%, where B. cereus exhibited the greatest efficacy in terms of mass loss for both brands of CBs. Fourier-Transform Infrared Spectroscopy (FTIR), confocal microscopy, and scanning electron microscopy (SEM) confirmed changes in the surface of the CBs, attributable to structural wear and material breakdown, indicating effective biodegradation by the evaluated microorganisms. Furthermore, the analyses confirmed changes in the surface of the CBs, attributable to structural wear and material breakdown, indicating effective biodegradation by the evaluated microorganisms.