Evidence suggests that doffing and possibly disposal of used personal protective equipment (PPE) can lead to environmental contamination.To ascertain the potential for site and floor contamination when medical gloves are inappropriately disposed.Fifteen healthcare workers (HCWs) disposed of gloves inoculated with bacteriophage and a chemical dye into a wastebasket, located 1.22 m away. Following each trial, designated sample areas were visually inspected with a blacklight for fluorescent dye stains and swabbed with a 3M Letheen Broth sponge to quantify the bacteriophage.The area closest to the participant (<0.30 m) had the highest bacteriophage concentrations (geomean: 6.9 × 10 pfu/100 cm; range: 8.07 to 3.93 × 10 pfu/100 cm). Bacteriophage concentrations were significantly higher (P < 0.05) in areas ≤0.61 m compared to >0.61 m from the HCWs. Although the farthest distances (1.22-1.52 m) resulted in 14% bacteriophage- and 4% fluorescent dye-positive occurrences, there was no significant difference (P = 0.069) between the tracers. The bacteriophage and chemical dye indicate highest environmental contamination nearest the HCWs and both tracers could be appropriate for PPE disposal training.HCWs use gloves every workday and potentially could contaminate surrounding surfaces and floors, during improper disposal practices. Therefore, proper disposal techniques are required to minimize pathogen transmission by establishing industry-wide policies, adequate training, and education to HCWs.