Bioaugmentation Treatment of a PAH-Polluted Soil in a Slurry Bioreactor

A bioslurry reactor was designed and used to treat loamy clay soil polluted with polycyclic aromatic hydrocarbons (PAHs). To this end, biostimulation alone, or combined with bioaugmentation with two bacterial strains (Rhodocccus erythropolis and Pseudomonas stuzeri) previously isolated from the polluted site, was applied. The PAH concentrations decreased notably after 15 days in all of the treatments. The concentrations of the two- and three-ring compounds fell by >80%, and, remarkably, the four- to six-ring PAHs also showed a marked decrease (>70%). These results thus indicate the capacity of bioslurry treatments to improve, notably, the degradation yields obtained in a previous real-scale remediation carried out using biopiles. In this sense, the remarkable results for recalcitrant PAHs can be attributed to the increase pollutants’ bioavailability achieves in the slurry bioreactors. Regarding bioaugmentation, although treatment with R. erythropolis led to a somewhat greater reduction of lighter PAHs at 15 days, the most time-effective treatment was achieved using P. stutzeri, which led to an 84% depletion of total PAHs in only three days. The effects of microbial degradation of other organic compounds were also monitored by means of combined qualitative and quantitative gas chromatography mass spectrometry (GC–MS) tools, as was the evolution of microbial populations, which was analyzed by culture and molecular fingerprinting experiments. On the basis of our findings, bioslurry technology emerges as a rapid and operative option for the remediation of polluted sites, especially for fine soil fractions with a high load of recalcitrant pollutants.

A Comparison of the Microbial Community and Functional Genes Present in Free-Living and Soil Particle-Attached Bacteria from an Aerobic Bioslurry Reactor Treating High-Molecular-Weight PAHs

High-molecular-weight (HMW) polycyclic aromatic hydrocarbons (PAHs) contaminate a wide range of ecosystems, including soils, groundwater, rivers and harbor sediments. The effective removal of HMW PAHs is a difficult challenge if a rapid remediation time and low economic cost are required. Bioremediation provides a cheap and eco-friendly cleanup strategy for the removal of HMW PAHs. Previous studies have focused on removal efficiency during PAHs bioremediation. In such studies, only limited research has targeted the bacterial communities and functional genes present in such bioremediation systems, specifically those of free-living (aqueous) bacteria and soil particle-attached bacteria present. In this study, a high-level of HMW PAH (1992 mg/kg pyrene) was bioremediated in an aerobic bioslurry reactor (ABR) for 42 days. The results showed a pseudo first order constant rate for pyrene biodegradation of 0.0696 day−1. The microbial communities forming free-living bacteria and soil-attached bacteria in the ABR were found to be different. An analysis of the aqueous samples identified free-living Mycobacterium spp., Pseudomonas putida, Rhodanobacter spp. and Burkholderia spp.; these organisms would seem to be involved in pyrene biodegradation. Various biointermediates, including phenanthrene, catechol, dibenzothiophene, 4,4′-bipyrimidine and cyclopentaphenanthrene, were identified and measured in the aqueous samples. When a similar approach was taken with the soil particle samples, most of the attached bacterial species did not seem to be involved in pyrene biodegradation. Furthermore, community level physiological profiling resulted in significantly different results for the aqueous and soil particle samples. Nevertheless, these two bacterial populations both showed positive signals for the presence of various dioxygenases, including PAHs-RHDα dioxygenases, riesk iron-sulfur motif dioxygenases and catechol 2,3-dioxygenases. The present findings provide a foundation that should help environmental engineers when designing future HMW PAH bioremediation systems that use the ABR approach.