Brushing teeth is the primary mode of oral hygiene practice. Toothbrushes may play a significant role in disease transmission and increase the risk of infection since they can serve as a reservoir for micro organisms in healthy, oral diseased and medically ill adults. There is complete lack of awareness among public regarding tooth brush maintenance. So, it is of utmost importance to educate the public about proper storage, replacement and disinfection of tooth brushes. Considering this aspect, the present study was undertaken to evaluate the presence of microorganisms in the tooth brushes and the effect of disinfectants to decontaminate them. The tooth brush samples were randomly collected from apparently healthy individuals and tested for the presence of microbes. The test samples demonstrated the presence of microbes such as E.coli, Pseudomonas aeruginosa and Klebsiella pneumonia. A total of three strains were isolated from the tooth brushes and when the test samples were treated with disinfectants, there was significant reduction in the microbial contamination. Thus it is mandatory for every individual to disinfect the tooth brush at regular intervals thereby maintaining good oral hygiene.