Many biodegradation studies have focused on survival of isolated bacteria to increase the bacteria population and subsequently enhance the efficiency of polycyclic aromatic hydrocarbon (PAH) biodegradation. However, there is limited research on enhancing the performance of isolated bacteria through reinoculation. Thus, this study was designed to evaluate the effects of reinoculation on the performance of Sphingobacterium spiritivorum in degradation of phenanthrene contaminated sand. Experiments were performed in three different reactors. Inoculation was performed once (day 0) in reactor 1. In reactor 2, inoculation was performed twice (day 0 and day 5). The bacteria was isolated from reactor 2 and inoculated into reactor 3. The study results show reactor 3 having the highest degradation rate (13.61 mg/kg/day) and percentage removal (95.36 percent). In contrast, without reinoculation in reactor 1, 68.93 percent of phenanthrene was removed. Thus, the performance of S. spiritivorum in phenanthrene degradation can be enhanced through reinoculation. © 2014 Wiley Periodicals, Inc.