This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.Objectives: Bacterial colonization on toothbrushes can cause stomatitis and other oral diseases such as dental caries and periodontitis. In this study we tested 18 toothbrushes, of which six were left exposed to the air, six were stored in a plastic bag, and six in a toothbrush case. Methods: ATP (RLU: relatively light unit) values measured by ATP Luminometer were compared with APC (CFU: colony forming unit) by swabbing culture method using 3M TM petrifilm TM aerobic count plates.Results: Swabs from toothbrushes left exposed in the air developed an average of 651.67±261.94 CFU, while the "In plastic bag" and the "In toothbrush case" specimens developed 593.17±354.33 and 120.83±68.87 CFU, respectively. Conclusion: The best option is to rinse the toothbrush well with water and to keep them dry in a wellventilated area in order to reduce the growth of bacteria in a public bath.