Triclosan (TCS), a synthetic and broad-spectrum antimicrobial agent, is frequently detected in various environmental matrices. A novel TCS degrading bacterial strain, Burkholderia sp. L303, was isolated from local activated sludge. The strain could metabolically degrade TCS up to 8 mg/L, and optimal conditions for TCS degradation were at temperature of 35 degrees C, pH 7, and an increased inoculum size. During TCS degradation, several intermediates were identified, with the initial degradation occurring mainly through hydroxylation of aromatic ring, followed by dechlorination. Further intermediates such as 2-chlorohydroquinone, 4-chlorocatechol, and 4-chlorophenol were produced via ether bond fission and C-C bond cleavage, which could be further transformed into unchlorinated compounds, ultimately resulting in the complete stoichiometric free chloride release. Bioaugmentation of strain L303 in non-sterile river water demonstrated better degradation than in sterile water. Further exploration of the microbial communities provided insights into the composition and succession of the microbial communities under the TCS stress as well as during the TCS biodegradation process in real water samples, the key microorganisms involved in TCS biodegradation or showing resistance to the TCS toxicity, and the changes in microbial diversity related to exogenous bioaugmentation, TCS input, and TCS elimination. These findings shed light on the metabolic degradation pathway of TCS and highlight the significance of microbial communities in the bioremediation of TCS-contaminated environments.