The goal of this study was to isolate PAHs degraders that can utilize PAHs associated with soil particulates and investigate the biodegradation of PAHs on agar plate, in liquid culture and soil. Two Mycobacterium strains (NJS-1 and NJS-P) were isolated from PAHs-contaminated farmland soil using enrichment based on soil slurry. The isolates could degrade five test PAHs including pyrene, phenanthrene, fluoranthene, anthracene and benzo[a]pyrene on plate, but showed different effects in liquid culture, especially for fluoranthene. Isolate NJS-1 was capable of utilizing benzo[a]pyrene as a sole carbon and energy source, and an enhanced degradation was observed when pyrene was supplied as cometabolic substrate. Reintroduction of the isolates into sterile contaminated soil resulted in a significant removal of aged pyrene and fluoranthene (over 40%) in 2-months incubation. In pyrene-spiked soil, the degradation of pyrene and fluoranthene increased to 90% and 50%, respectively. Comparing PAHs degradation on plate, in liquid culture and soil, we can conclude that there was corresponding degradation in different test systems. In addition, the degradation of aged PAHs in soil suggested the potential application of two isolates in further bioremediation. (c) 2010 Elsevier B.V. All rights reserved.