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) C(10)-C(40)). 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. Microbial diversity in slurries was monitored using DGGE with dominant bands excised and sequenced for identification. Applying the different bioremediation strategies resulted in the formation of four distinct community clusters associated with the activated sludge (inoculum), bioaugmentation strategy at day 0, bioaugmentation strategy at weeks 2-6 and slurries with autoclaved sludge and nutrient additions (bioaugmentation negative control). While hydrocarbon-degrading bacteria genera (e.g. Aquabacterium and Haliscomenobacter) were associated with the hydrocarbon-contaminated soil, bioaugmentation of soil slurries with activated sludge resulted in the introduction of bacteria associated with hydrocarbon degradation (Burkholderiales order and Klebsiella genera) which presumably contributed to the enhanced efficacy for this slurry strategy.

Re-use of remediated soils for the bioremediation of waste oil sludge.

We investigated the possibility of re-using remediated soils for new bioremediation projects by spiking these soils with waste oil sludge in laboratory based microcosms. The level of Total Petroleum Hydrocarbon (TPH) reduction was high (>80%) in naturally attenuated microcosms and was not significantly improved by biostimulation, bioaugmentation and the combined treatment of bioaugmentation and biostimulation by week 12. This indicated that the observed TPH reduction might have been related to the soil's inherent hydrocarbon-degrading potential. Microbial community analysis (16S rDNA and ITS-based Denaturing Gradient Gel Electrophoresis fingerprints) confirmed the dominance of hydrocarbon degrading genera such as Alcanivorax and Scedosporium. Cluster and Shannon diversity analysis revealed similar but stable bacterial and fungal communities in naturally attenuated and amended microcosms indicating that rapid reduction in TPH may not always be accompanied by changes in soil microbial communities. This study has therefore shown that soils previously used for bioremediation can have an improved hydrocarbon degrading potential which was successfully re-harnessed for new projects. This ability to re-harness this potential is attractive because it substantially reduces operational costs as no additional bioremediation treatments are needed. It can also extend a landfill's lifespan as soils can be re-used again before landfill disposal.

The application of a carrier-based bioremediation strategy for marine oil spills.

The application of recycled marine materials to develop sustainable remediation technologies in marine environment was assessed. The remediation strategy consisted of a shell carrier mounted bacterial consortium composed of hydrocarbonoclastic strains enriched with nutrients (Bioaug SC). Pilot scale studies (5000 l) were used to examine the ability of Bioaug-SC to degrade weathered crude oil (10 g l(-1); initially 315,000±44,000 mg l(-1)) and assess the impacts of the introduction and biodegradation of oil. Total petroleum hydrocarbon mass was effectively reduced by 53.3 (±5.75)% to 147,000 (±21,000) mg l(-1) within 27 weeks. 16S rDNA bacterial community profiling using Denaturant Gradient Gel Electrophoresis revealed that cyanobacteria and Proteobacteria dominated the microbial community. Aquatic toxicity assessment was conducted by ecotoxicity assays using brine shrimp hatchability, Microtox and Phaeodactylum tricornutum. This study revealed the importance of combining ecotoxicity assays with oil chemistry analysis to ensure safe remediation methods are developed.

Carrier mounted bacterial consortium facilitates oil remediation in the marine environment.

Marine oil pollution can result in the persistent presence of weathered oil. Currently, removal of weathered oil is reliant on chemical dispersants and physical removal, causing further disruption. In contrast few studies have examined the potential of an environmentally sustainable method using a hydrocarbon degrading microbial community attached to a carrier. Here, we used a tank mesocosm system (50 l) to follow the degradation of weathered oil (10 g l(-1)) using a bacterial consortium mobilised onto different carrier materials (alginate or shell grit). GCMS analysis demonstrated that the extent of hydrocarbon degradation was dependent upon the carrier material. Augmentation of shell grit with nutrients and exogenous hydrocarbon degraders resulted in 75±14% removal of >C32 hydrocarbons after 12 weeks compared to 20±14% for the alginate carrier. This study demonstrated the effectiveness of a biostimulated and bioaugmented carrier material to degrade marine weathered oil.

The importance of weathered crude oil as a source of hydrocarbonoclastic microorganisms in contaminated seawater.

This study investigated the hydrocarbonoclastic microbial community present on weathered crude oil and their ability to degrade weathered oil in seawater obtained from the Gulf St. Vincent (SA, Australia). Examination of the native seawater communities capable of utilizing hydrocarbon as the sole carbon source identified a maximum recovery of just 6.6 × 10(1) CFU/ml, with these values dramatically increased in the weathered oil, reaching 4.1 × 10(4) CFU/ml. The weathered oil (dominated by >C30 fractions; 750,000 +/- 150,000 mg/l) was subject to an 8 week laboratory-based degradation microcosm study. By day 56, the natural inoculums degraded the soluble hydrocarbons (initial concentrations 3,400 +/- 700 mg/l and 1,700 +/- 340 mg/l for the control and seawater, respectively) to below detectable levels, and biodegradation of the residual oil reached 62% (254,000 +/- 40,000 mg/l) and 66% (285,000 +/- 45,000 mg/l) in the control and seawater sources, respectively. In addition, the residual oil gas chromatogram profiles changed with the presence of short and intermediate hydrocarbon chains. 16S rDNA DGGE sequence analysis revealed species affiliated with the genera Roseobacter, Alteromonas, Yeosuana aromativorans, and Pseudomonas, renowned oil-degrading organisms previously thought to be associated with the environment where the oil contaminated rather than also being present in the contaminating oil. This study highlights the importance of microbiological techniques for isolation and characterisation, coupled with molecular techniques for identification, in understanding the role and function of native oil communities.