The purpose of this study was to analyze the environment microbial contamination produced by Er:YAG laser irradiation in infected root canals. A total of 20 human anterior teeth were prepared, sterilized and, then, inoculated with a mixture of the following microorganisms: S. Aureus, E. Faecalis, P. Aeruginosa, B. Subtilis and C. Albicans. After the contamination period (28 days), the teeth were irrigated with sterile distilled water or 1% sodium hypochlorite and, then, irradiated with an Er:YAG laser with two different laser parameters: 52 mJ or 110 mJ output at the fiber tip. Eighteen Petri dishes with 20 ml of BHI Agar were used in the study. For each group, 3 plates with BHIA were used for the analysis of the microbial contamination of the environment during the activation of the laser in infected root canals. The plates were positioned in differing distances away from the irradiated tooth (plate 1 - distance of 15 cm, plate 2 - distance of 50 cm and plate 3 - distance of 3 meters). After the analysis of the results, it was observed that the larger microbial contamination occurred in Group 1 (teeth irrigated with sterile distilled water and irradiated with Er:YAG laser with 52 mJ output at the fiber tip), plate 1 (positioned 15 cm away from the irradiated tooth), with values greater than 30 Colony-Forming Units (CFU).