Harnessing the hydrocarbon degrading potential of contaminated soils for the bioremediation of waste engine oil

Waste engine oil pollution is an endemic problem in African countries as waste oil is often discharged into the environment without adequate treatment because waste oil recycling facilities are not readily available. In this study, laboratory-based microcosms (natural attenuation, biostimulation, bioaugmentation and combined treatment of biostimulation-bioaugmentation) were set up with soils (from old hydrocarbon biopiles) spiked with waste engine oil and monitored for 3 months. Total petroleum hydrocarbon analysis showed that biostimulation and biostimulation-bioaugmentation accelerated hydrocarbon degradation with over 84% reduction (< 10,000 mg kg(-1)) by week 8. It took another 2 weeks for other microcosms to get below this classification of low-level contaminated waste and landfill disposal level. The highest degradation rate of 92% was obtained in biostimulated-bioaugmented microcosms (week 10). However, by week 12, there were no significant differences in hydrocarbon levels in naturally attenuated and treated microcosms. 16S rRNA and ITS-based denaturing gradient gel electrophoresis profiling showed diverse bacterial and fungal communities with some dominant members belonging to hydrocarbon-degrading Proteobacteria, Ascomycetes and Basidiomycete

Potential impact of soil microbial heterogeneity on the persistence of hydrocarbons in contaminated subsurface soils

In situ bioremediation is potentially a cost effective treatment strategy for subsurface soils contaminated with petroleum hydrocarbons, however, limited information is available regarding the impact of soil spatial heterogeneity on bioremediation efficacy. In this study, we assessed issues associated with hydrocarbon biodegradation and soil spatial heterogeneity (samples designated as FTF 1, 5 and 8) from a site in which in situ bioremediation was proposed for hydrocarbon removal. Test pit activities showed similarities in FTF soil profiles with elevated hydrocarbon concentrations detected in all soils at 2 m below ground surface. However, PCR-DGGE-based cluster analysis showed that the bacterial community in FTF 5 (at 2 m) was substantially different (53% dissimilar) and 2e3 fold more diverse than communities in FTF 1 and 8 (with 80% similarity). When hydrocarbon degrading potential was assessed, differences were observed in the extent of 14C-benzene mineralisation under aerobic conditions with FTF 5 exhibiting the highest hydrocarbon removal potential compared to FTF 1 and 8. Further analysis indicated that the FTF 5 microbial community was substantially different from other FTF samples and dominated by putative hydrocarbon degraders belonging to Pseudomonads, Xanthomonads and Enterobacteria. However, hydrocarbon removal in FTF 5 under anaerobic conditions with nitrate and sulphate electron acceptors was limited suggesting that aerobic conditions were crucial for hydrocarbon removal. This study highlights the importance of assessing available microbial capacity prior to bioremediation and shows that the site's spatial heterogeneity can adversely affect the success of in situ bioremediation unless area-specific optimizations are performed

Impact of bacterial and fungal processes on 14C-hexadecane mineralisation in weathered hydrocarbon contaminated soil

In this study, the impact of bacterial and fungal processes on 14 C-hexadecane mineralisation was investigated in weathered hydrocarbon contaminated soil. The extent of 14 C-hexadecane mineralisation varied depending on the bioremediation strategy employed. Under enhanced natural attenuation conditions, 14 C-hexadecane mineralisation after 98 days was 8.5± 3.7% compared to b1.2% without nitrogen and phosphorus additions

Assessing impediments to hydrocarbon biodegradation in weathered contaminated soils

In this study, impediments to hydrocarbon biodegradation in contaminated soils were assessed using chemical and molecular methodologies. Two long-term hydrocarbon contaminated soils were utilised which were similar in physico-chemical properties but differed in the extent of hydrocarbon (C10-C40) contamination (S1: 16.5 g kg-1; S2: 68.9 g kg-1). Under enhanced natural attenuation (ENA) conditions, hydrocarbon biodegradation was observed in S1 microcosms (26.4% reduction in C10-C40 hydrocarbons), however, ENA was unable to stimulate degradation in S2. Although eubacterial communities (PCR-DGGE analysis) were similar for both soils, the alkB bacterial community was less diverse in S2 presumably due to impacts associated with elevated hydrocarbons. When hydrocarbon bioaccessibility was assessed using HP-ß-CD extraction, large residual concentrations remained in the soil following the extraction procedure. However, when linear regression models were used to predict the endpoints of hydrocarbon degradation, there was no significant difference (P > 0.05) between HP-ß-CD predicted and microcosm measured biodegradation endpoints. This data suggested that the lack of hydrocarbon degradation in S2 resulted primarily from limited hydrocarbon bioavailability

Assessment of five bioaccessibility assays for predicting the efficacy of petroleum hydrocarbon biodegradation in aged contaminated soils

In this study, the bioaccessibility of petroleum hydrocarbons in aged contaminated soils (1.6-67gkg -1) was assessed using four non-exhaustive extraction techniques (100% 1-butanol, 100% 1-propanol, 50% 1-propanol in water and hydroxypropyl-?-cyclodextrin) and the persulfate oxidation method. Using linear regression analysis, residual hydrocarbon concentrations following bioaccessibility assessment were compared to residual hydrocarbon concentrations following biodegradation in laboratory-scale microcosms in order to determine whether bioaccessibility assays can predict the endpoint of hydrocarbon biodegradation. The relationship between residual hydrocarbon concentrations following microcosm biodegradation and bioaccessibility assessment was linear (r 2=0.71-0.97) indicating that bioaccessibility assays have the potential to predict the extent of hydrocarbon biodegradation. However, the slope of best fit varied depending on the hydrocarbon fractional range assessed. For the C 10-C 14 hydrocarbon fraction, the slope of best fit ranged from 0.12 to 0.27 indicating that the non-exhaustive or persulfate oxidation methods removed 3.5-8 times more hydrocarbons than biodegradation. Conversely, for the higher molecular weight hydrocarbon fractions (C 29-C 36 and C 37-C 40), biodegradation removed up to 3.3 times more hydrocarbons compared to bioaccessibility assays with the resulting slope of best fit ranging from 1.0-1.9 to 2.0-3.3 respectivel

Impact of bacterial and fungal processes on C-14-hexadecane mineralisation in weathered hydrocarbon contaminated soil

In this study, the impact of bacterial and fungal processes on C-14-hexadecane mineralisation was investigated in weathered hydrocarbon contaminated soil. The extent of C-14-hexadecane mineralisation varied depending on the bioremediation strategy employed. Under enhanced natural attenuation conditions, C-14-hexadecane mineralisation after 98 days was 8.5 +/- 3.7% compared to <1.2% without nitrogen and phosphorus additions

Comparison of indigenous and exogenous microbial populations during slurry phase biodegradation of long-term hydrocarbon-contaminated soil

In this study, a number of slurry-phase strategies were trialled over a 42 day period in order to determine the efficacy of bioremediation for long-term hydrocarbon-contaminated soil (145 g kg-1 C10-C40). The addition of activated sludge and nutrients to slurries (bioaugmentation) resulted in enhanced hydrocarbon removal (51.6 ± 8.5 %) compared to treatments receiving only nutrients (enhanced natural attenuation [ENA]; 41.3 ± 6.4 %) or no amendments (natural attenuation; no significant hydrocarbon removal, P < 0.01). This data suggests that the microbial community in the activated sludge inoculum contributed to the enhanced removal of hydrocarbons in ENA slurries

A polyphasic approach for assessing the suitability of bioremediation for the treatment of hydrocarbon-impacted soil

Bioremediation strategies, though widely used for treating hydrocarbon-contaminated soil, suffer from lack of biodegradation endpoint accountability. To address this limitation, molecular approaches of alkB gene analysis and pyrosequencing were combined with chemical approaches of bioaccessibility and nutrient assays to assess contaminant degrading capacity and develop a strategy for endpoint biodegradation predictions. In long-term hydrocarbon-contaminated soil containing 10.3 g C10-C36 hydrocarbons kg− 1, 454 pyrosequencing detected the overrepresentation of potential hydrocarbon degrading genera such as Pseudomonas, Burkholderia, Mycobacterium and Gordonia whilst amplicons for PCR-DGGE were detected only with alkB primers targeting Pseudomonas. This indicated the presence of potential microbial hydrocarbon degradation capacity in the soil. Using non-exhaustive extraction methods of 1-propanol and HP-β-CD for hydrocarbon bioaccessibility assessment combined with biodegradation endpoint predictions with linear regression models, we estimated 33.7% and 46.7% hydrocarbon removal respectively. These predictions were validated in pilot scale studies using an enhanced natural attenuation strategy which resulted in a 46.4% reduction in soil hydrocarbon content after 320 days. When predicted biodegradation endpoints were compared to measured values, there was no significant difference (P = 0.80) when hydrocarbon bioaccessibility was assessed with HP-β-CD. These results indicate that a combination of molecular and chemical techniques that inform microbial diversity, functionality and chemical bioaccessibility can be valuable tools for assessing the suitability of bioremediation strategies for hydrocarbon-contaminated soi