Succession of bacterial community along with the removal of heavy crude oil pollutants by multiple biostimulation treatments in the Yellow River Delta, China.

Multiple biostimulation treatments were applied to enhance the removal of heavy crude oil pollutants in the saline soil of Yellow River Delta. Changes of the soil bacterial community were monitored using the terminal restriction fragment length polymorphism (T-RFLP) and clone library analyses. The 140-day microcosm experiments showed that low C:N:P ratio, high availability of surfactant and addition of bulking agent significantly enhanced the performance, leading to the highest total petroleum hydrocarbon removal. Meanwhile, the bacterial community was remarkably changed by the multiple biostimulation treatments, with the Deltaproteobacteria, Firmicutes, Actinobacteria, Acidobacteria and Planctomycetes being inhibited and the Alpha- and Beta-proteobacteria and some unknown Gammaproteobacteria bacteria being enriched. In addition, different hydrocarbon-degraders came to power in the following turn. At the first stage, the Alcanivorax-related Gammaproteobacteria bacteria dominated in the biostimulated soil and contributed mainly to the biodegradation of easily degradable portion of the heavy crude oil. Then the bacteria belonging to Alphaproteobacteria, followed by bacteria belonging to Candidate division OD1, became the dominant oil-degraders to degrade the remaining recalcitrant constituents of the heavy crude oil.