Chronic pollution by used lubricant oils (ULOs) poses a serious challenge to the environment. Under stress conditions, microorganisms, including potential degraders, can enter a viable but non-culturable (VBNC) state, complicating the bioremediation of ULO-polluted areas. Resuscitation-promoting factors (Rpfs) can reverse this transition and/or enhance the biodegradation performance of both native and augmented strains. Here, Rpf-containing extracellular organic matter (EOM) from Micrococcus luteus was used to enhance the ex situ ULO removal in biostimulated and bioaugmented (with Rhodococcus qingshengii KAG C, R. erythropolis PR4) soils. ULO bioconversion, microbial activity, and CFUs were significantly higher in EOM-treated soils compared to corresponding control soils. After 60 days, the initial ULO concentration (52,500 mg kg^-1) was reduced by 37% and 45% with EOM-supplemented biostimulation and bioaugmentation, respectively. Based on high-throughput 16S rRNA analysis, the enhancement was attributable both to the reactivation of EOM-responsive hydrocarbonoclastic bacterial genera (e.g., Pseudomonas, Comamonas, Stenotrophomonas, Gordonia) and to the long-term positive effect of EOM on the degradative efficacy of the introduced rhodococci. Ecotoxicological responses revealed that reduced ULO concentration did not correlate with decreased soil toxicity. Our findings provide an insight into the applicability of EOM in bioremediation and its effects on the soil microbial activity and community composition.

Used lubricant oils (ULOs) strongly bind to soil particles and cause persistent pollution. In this study, soil microcosm experiments were conducted to model the ex situ bioremediation of a long term ULO-polluted area. Biostimulation and various inoculation levels of bioaugmentation were applied to determine the efficacy of total petrol hydrocarbon (TPH) removal. ULO-contaminated soil microcosms were monitored for microbial respiration, colony-forming units (CFUs) and TPH bioconversion. Biostimulation with inorganic nutrients was responsible for 22% of ULO removal after 40 days. Bioaugmentation using two hydrocarbon-degrader strains: Rhodococcus quingshengii KAG C and Rhodococcus erythropolis PR4 at a small inoculum size (10⁷ CFUs g⁻¹ soil), reduced initial TPH concentration by 24% and 29%, respectively; the application of a higher inoculum size (10⁹ CFUs g⁻¹ soil) led to 41% and 32% bioconversion, respectively. After 20 days, all augmented CFUs decreased to the same level as measured in the biostimulated cases, substantiating the challenge for the newly introduced hydrocarbon-degrading strains to cope with environmental stressors. Our results not only highlight that an increased number of degrader cells does not always correlate with enhanced TPH bioconversion, but they also indicate that biostimulation might be an economical solution to promote ULO biodegradation in long term contaminated soils.

Petroleum hydrocarbons and derivatives are widespread contaminants in both aquifers and soil, their elimination is in the primary focus of environmental studies. Microorganisms are key components in biological removal of pollutants. Strains capable to utilize hydrocarbons usually appear at the contaminated sites, but their metabolic activities are often restricted by the lack of nutrients and/or they can only utilize one or two components of a mixture. We isolated a novel Rhodococcus sp. MK1 strain capable to degrade the components of diesel oil simultaneously. The draft genome of the strain was determined and besides the chromosome, the presence of one plasmid could be revealed. Numerous routes for oxidation of aliphatic and aromatic compounds were identified. The strain was tested in ex situ applications aiming to compare alternative solutions for microbial degradation of hydrocarbons. The results of bioaugmentation and biostimulation experiments clearly demonstrated that – in certain cases – the indigenous microbial community could be exploited for bioremediation of oil-contaminated soils. Biostimulation seems to be efficient for removal of aged contaminations at lower concentration range, whereas bioaugmentation is necessary for the treatment of freshly and highly polluted sites.