Toothbrush oral cleaning tool is designed to be the most reasonable and easy to use for removing plaque attached to teeth surface physically. Bacterias on toothbrush causes stomatitis oral diseases such as dental caries, periodontitis. To prevent diseases, there are some ways to keep toothbrush to clean. Bacterias present in toothbrush after brushing so drying out or using a toothbrush sterilizer should prevent bacterial growth. In this study we used a total of thirty toothbrushes, ten were stored in a locker, ten were stored in a locker and cover with a toothbrush cap, and ten were kept in a toothbrush sterilizer. Of the toothbrushes stored in the locker two samples had bacteria levels of more than 20,000 CFU/mL, two samples had levels between 10,000-20,000 CFU/mL, and six samples were found to have levels less than 10,000 CFU/mL. Regarding the toothbrushes stored in the locker and covered with a toothbrush cap, three samples had bacteria levels more than 20,000 CFU/mL, two samples had levels of 10,000-20,000 CFU/mL, four samples had levels less than 10,000 CFU/mL, and one sample did not show significant bacterial levels. As for the toothbrushes kept in the toothbrush sterilizer, three samples showed bacteria levels less than 10,000 CFU/mL, while the remaining seven samples were not found to contain significant levels of bacteria. The Gram-positive spherical-shaped bacteria isolated from the toothbrushes included Micrococcus spp. (two different strains), Staphylococcus capitis (two different strains), Staphylococcus aureus (three different strains), Staphylococcus epidermidis (one different strain), Staphylococcus lugdunenesis (one different strain). While the Gram-negative rod-shaped bacteria isolated from the toothbrushes included Enterobacter cloacae (three different strains), Burkholderia cepacia (one different strain), Klebsiella pneumoniae (one different strain), Pantoea spp. (one different strain), Enterobacter amnigenus (one different strain). Due to these findings it is recommended that toothbrushes be kept in a toothbrush sterilizer rather than leaving them unattended, regardless of where, in storage caps. However, due to problems of cost, most people do not have access to a sanitizer. The next best option is to wash the toothbrush well with water and to keep it dry in a well-ventilated area so as to reduce the growth of bacteria.