Simulation of a surface spill of different diesel/biodiesel mixtures in an ultisol, using natural attenuation and bioaugmentation/biostimulation.

Accidents caused by leaks and/or spills on soils need to be addressed. Natural attenuation, biostimulation and bioaugmentation can be useful bioremediation strategies for decontamination processes in soils of diesel/biodiesel mixtures. The purpose of this study was to evaluate the degradation rate of the different fuels (B0, B20 and B100) in an ultisol under natural attenuation and biostimulation/bioaugmentation during 60 days of incubation in a controlled microcosm simulating a surface spill over soil. The degradation of different diesel/biodiesel mixtures was monitored for up to 60 days by dehydrogenase activity, respirometry by CO2 release, the most probable number of heterotrophic and degrading microorganism and gas chromatography. The bacterial inoculum employed for biostimulation/bioaugmentation strategy consisted of Bacillus megaterium, Bacillus pumilus, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. The two bioremediation strategies have showed great degradation rates. The natural attenuation was effective for B0 and B20 treatments. The addition of the bacterial consortium and macronutrients contributed to the increased degradation of pure biodiesel in relation to natural attenuation, with higher rates for CO2 release, enzymatic and degrading activity. It is suggested that the bacterial consortium has proven effective for presenting significant values for such parameters until the end of the 60-day incubation period.

Comparative metagenomics reveals different hydrocarbon degradative abilities from enriched oil-drilling waste.

The oil drilling process generates large volumes of waste with inadequate treatments. Here, oil drilling waste (ODW) microbial communities demonstrate different hydrocarbon degradative abilities when exposed to distinct nutrient enrichments as revealed by comparative metagenomics. The ODW was enriched in Luria Broth (LBE) and Potato Dextrose (PDE) media to examine the structure and functional variations of microbial consortia. Two metagenomes were sequenced on Ion Torrent platform and analyzed using MG-RAST. The STAMP software was used to analyze statistically significant differences amongst different attributes of metagenomes. The microbial diversity presented in the different enrichments was distinct and heterogeneous. The metabolic pathways and enzymes were mainly related to the aerobic hydrocarbons degradation. Moreover, our results showed efficient biodegradation after 15 days of treatment for aliphatic hydrocarbons (C8-C33) and polycyclic aromatic hydrocarbons (PAHs), with a total of about 50.5% and 46.4% for LBE and 44.6% and 37.9% for PDE, respectively. The results obtained suggest the idea that the enzymatic apparatus have the potential to degrade petroleum compounds.

Biosorption and bioreduction of copper from different copper compounds in aqueous solution.

High copper concentration is toxic for living organisms including humans. Biosorption is a bioremediation technique that can remove copper and other pollutants from aqueous medium and soils, consequently cleaning the environment. The aim of this study was, therefore, to investigate the influence of different copper compounds (Cu(II) as CuCl2; Cu(II) as CuSO4; and Cu(I) as CuCl) on copper bioreduction and biosorption using four copper-resistant bacteria isolated from the rhizosphere of two plants (Avena sativa and Plantago lanceolata) in aqueous matrix. Copper resistance profile, bioreduction, and biosorption after 48 h of incubation were evaluated. The isolates displayed high copper resistance. However, isolate A1 did not grow very well in the CuCl2 and isolate T5 was less resistant to copper in aqueous solutions amended with CuCl (Cu(I)). The best copper source for copper bioreduction and biosorption was CuSO4 and the isolates removed as much as ten times more copper than in aqueous solutions amended with the other copper compounds. Moreover, Cu(I) did not succumb to biosorption, although the microbes were resistant to aqueous solutions of CuCl. In summary, Cu(II) from CuSO4 was furthermost susceptible to bioreduction and biosorption for all isolates. This is an indication that copper contamination of the environment from the use of CuSO4 as an agrochemical is amenable to bioremediation.

Bioprospection and selection of bacteria isolated from environments contaminated with petrochemical residues for application in bioremediation.

The use of microorganisms with hydrocarbon degrading capability and biosurfactant producers have emerged as an alternative for sustainable treatment of environmental passives. In this study 45 bacteria were isolated from samples contaminated with petrochemical residues, from which 21 were obtained from Landfarming soil contaminated with oily sludge, 11 were obtained from petrochemical industry effluents and 13 were originated directly from oily sludge. The metabolization capability of different carbon sources, growth capacity and tolerance, biosurfactant production and enzymes detection were determined. A preliminary selection carried out through the analysis of capability for degrading hydrocarbons showed that 22% of the isolates were able to degrade all carbon sources employed. On the other hand, in 36% of the isolates, the degradation of the oily sludge started within 18-48 h. Those isolates were considered as the most efficient ones. Twenty isolates, identified based on partial sequencing of the 16S rRNA gene, were pre-selected. These isolates showed ability for growing in a medium containing 1% of oily sludge as the sole carbon source, tolerance in a medium containing up to 30% of oily sludge, ability for biosurfactant production, and expression of enzymes involved in degradation of aliphatic and aromatic compounds. Five bacteria, identified as Stenotrophomonas acidaminiphila BB5, Bacillus megaterium BB6, Bacillus cibi, Pseudomonas aeruginosa, and Bacillus cereus BS20 were shown to be promising for use as inoculum in bioremediation processes (bioaugmentation) of areas contaminated with petrochemical residues since they can use oily sludge as the sole carbon source and produce biosurfactants.

Biodegradation potential of oily sludge by pure and mixed bacterial cultures.

The biodegradation capacity of aliphatic and aromatic hydrocarbons of petrochemical oily sludge in liquid medium by a bacterial consortium and five pure bacterial cultures was analyzed. Three bacteria isolated from petrochemical oily sludge, identified as Stenotrophomonas acidaminiphila, Bacillus megaterium and Bacillus cibi, and two bacteria isolated from a soil contaminated by petrochemical waste, identified as Pseudomonas aeruginosa and Bacillus cereus demonstrated efficiency in oily sludge degradation when cultivated during 40 days. The bacterial consortium demonstrated an excellent oily sludge degradation capacity, reducing 90.7% of the aliphatic fraction and 51.8% of the aromatic fraction, as well as biosurfactant production capacity, achieving 39.4% reduction of surface tension of the culture medium and an emulsifying activity of 55.1%. The results indicated that the bacterial consortium has potential to be applied in bioremediation of petrochemical oily sludge contaminated environments, favoring the reduction of environmental passives and increasing industrial productivity.

Biodegradation of commercial gasoline (24% ethanol added) in liquid medium by microorganisms isolated from a landfarming site.

Isolation of soil microorganisms from a landfarming site with a 19-year history of petrochemical residues disposal was carried out. After isolation, the bacteria behavior in mineral medium with 1% commercial gasoline (24% ethanol) was evaluated. Parameters employed for microorganism evaluation and selection of those with the greatest degradation potential were: microbial growth; biosurfactant generation and compound reduction in commercial gasoline. Starting from bacteria that presented the best degradation results, consortiums formed by 4 distinct microorganisms were formed. A microbial growth in the presence of commercial gasoline was observed and, for most of the bacteria, degradations of compounds such as benzene, toluene and xylenes (BTX) as well as biosurfactant production was observed. Ethanol was partially degraded by the bacterial isolates although the data does not allow affirming that it was degraded preferentially to the aromatic hydrocarbons investigated. The analyzed consortiums present an efficiency of 95% to 98% for most of the commercial gasoline compounds and a preferential attack to ethanol under the essay condition was not observed within 72 h.

Improved enrichment and isolation of polycyclic aromatic hydrocarbons (PAH)-degrading microorganisms in soil using anthracene as a model PAH.

Lack of attention to soil and microbial characteristics that influence PAHs degradation has been a leading cause of failures in isolation of efficient PAH degraders and bioaugumentation processes with microbial consortia. This study compared the classic method of isolation of PAHs-degraders with a modified method employing a pre-enrichment respirometric analysis. The modified enrichment of PAH degrading microorganisms using in vitro microcosm resulted to reduced enrichment period and more efficient PAH-degrading microbial consortia. Results indicate that natural soils with strong heterotrophic microbial activity determined through pre-enrichment analysis, are better suited for the isolation of efficient PAH degrading microorganisms with significant reduction of the enrichment period.

Mass spectrometry analysis of surface tension reducing substances produced by a pah-degrading Pseudomonas citronellolis strain / Análise por espectrometria de massa de substâncias redutoras da tensão superficial produzidas por uma cepa de Pseudomonas citronellolis degradadora de hidrocarbonetos aromáticos policíclicos

In this work we investigated the structure of the iron-stimulated surface tension reducing substances produced by P. citronellolis 222A isolated from a 17-years old landfarming used for sludge treatment in petrochemical industries and oil refinery. Its mass spectrum differs from P. aeruginosa spectrum, indicating that the surface tension reducing substances produced by P. citronellolis can be a new kind of biosurfactant.

Neste trabalho é apresentado um estudo a respeito da análise da estrutura de substâncias redutoras de tensão superficial produzidas por Pseudomonas citronellolis 222A estimulado pela presença de ferro. Esta bactéria foi isolada de um solo que há 17 anos vem sendo utilizado para o tratamento de borra oleosa proveniente da indústria petroquímica e de refinaria de petróleo. O espectro de massa difere do espectro de P. aeruginosa, indicando que as substâncias redutoras de tensão superficial produzidas por P. citronellolis podem ser um novo tipo de biosurfactante.

Mass spectrometry analysis of surface tension reducing substances produced by a pah-degrading Pseudomonas citronellolis strain.

In this work we investigated the structure of the iron-stimulated surface tension reducing substances produced by P. citronellolis 222A isolated from a 17-years old landfarming used for sludge treatment in petrochemical industries and oil refinery. Its mass spectrum differs from P. aeruginosa spectrum, indicating that the surface tension reducing substances produced by P. citronellolis can be a new kind of biosurfactant.

Microbial consortium bioaugmentation of a polycyclic aromatic hydrocarbons contaminated soil.

In this study we evaluated the capacity of a defined microbial consortium (five bacteria: Mycobacterium fortuitum, Bacillus cereus, Microbacterium sp., Gordonia polyisoprenivorans, Microbacteriaceae bacterium, Naphthalene-utilizing bacterium; and a fungus identified as Fusarium oxysporum) isolated from a PAHs contaminated landfarm site to degrade and mineralize different concentrations (0, 250, 500 and 1000 mg kg(-1)) of anthracene, phenanthrene and pyrene in soil. PAHs degradation and mineralization was evaluated by gas chromatography and respirometry, respectively. The microbial consortium degraded on average, 99%, 99% and 96% of the different concentrations of anthracene, phenanthrene and pyrene in the soil, in 70 days, respectively. This consortium mineralized 78%, on average, of the different concentrations of the 3 PAHs in soil after 70 days. Contrarily, the autochthonous soil microbial population showed no substantial mineralization of the PAHs. Bacterial and fungal isolates from the consortium, when inoculated separately to the soil, were less effective in anthracene mineralization compared to the consortium. This signifies synergistic promotion of PAHs mineralization by mixtures of the monoculture isolates (the microbial consortium).

Anthracene biodegradation and surface activity by an iron-stimulated Pseudomonas sp.

Iron may enhance polycyclic aromatic hydrocarbons (PAHs) degradation directly by increasing the activity of the enzymes involved in the aerobic biodegradation pathways for hydrocarbons, and indirectly by increasing the PAHs bioavailability due to the stimulation of biosurfactant production. In the present work, the PAH anthracene was used in order to study the effect of different forms and concentrations of iron on its biodegradation and surfactant production by Pseudomonas spp. isolates from a 14-years old petrochemical sludge landfarm site. Among the iron forms, iron nitrate was chosen based on its high solubility and effect on the increase in the growth of the isolate. Iron concentration of 0.1mM was selected as the limit between deficiency and toxicity for isolates growth and anthracene degradation. After 48 days Pseudomonas citronellolis isolate 222A degraded 72% of anthracene related to iron stimulation and surface tension decrease, indicating surfactant production. Pseudomonas aeruginosa isolate 332C was iron-stimulated but did not reduce surface tension while P. aeruginosa isolate 312A exhibited a noniron and surfactant dependence to degrade 72% of anthracene. Isolate 222A showed a direct dependence on iron to stimulate surfactant activity, which probably increased anthracene bioavailability. To our knowledge, this is the first report about the iron effect on anthracene degradation and surfactant production by a Pseudomonas sp. Based on the iron requirement and surfactant activity, the Pseudomonas isolates may be useful for bioremediation of PAHs.

Enhancing bioremediation of diesel oil and gasoline in soil amended with an agroindustry sludge.

This paper presents a study of the bioremediation of diesel oil and gasoline by a series of controlled laboratory tests. Sludge from an agroindustry was used to enhance bioremediation of both gasoline and diesel oil mixed with a soil mass to compare its efficiency with that of a mineral fertilizer. Effects of soil microbiology and soil mixtures were investigated by means of evolution of CO2, microorganism populations at 90 days, pH at 65 and 95 days, mineral nitrogen, and gas chromatographic analysis of the benzene, toluene, methyl tertiary butyl ether, C8, and C9+total aromatics at the end of the experiments. Treatments containing sludge showed better soil conditions after 170 days of treatment (inorganic nitrogen and microbiota activity) compared with gasoline and diesel oil without amendments. Samples had no detectable traces of the measured hydrocarbons at 170 days of treatment.

Comparative bioremediation of soils contaminated with diesel oil by natural attenuation, biostimulation and bioaugmentation.

Bioremediation of diesel oil in soil can occur by natural attenuation, or treated by biostimulation or bioaugmentation. In this study we evaluated all three technologies on the degradation of total petroleum hydrocarbons (TPH) in soil. In addition, the number of diesel-degrading microorganisms present and microbial activity as indexed by the dehydrogenase assay were monitored. Soils contaminated with diesel oil in the field were collected from Long Beach, California, USA and Hong Kong, China. After 12 weeks of incubation, all three treatments showed differing effects on the degradation of light (C12-C23) and heavy (C23-C40) fractions of TPH in the soil samples. Bioaugmentation of the Long Beach soil showed the greatest degradation in the light (72.7%) and heavy (75.2%) fractions of TPH. Natural attenuation was more effective than biostimulation (addition of nutrients), most notably in the Hong Kong soil. The greatest microbial activity (dehydrogenase activity) was observed with bioaugmentation of the Long Beach soil (3.3-fold) and upon natural attenuation of the Hong Kong sample (4.0-fold). The number of diesel-degrading microorganisms and heterotrophic population was not influenced by the bioremediation treatments. Soil properties and the indigenous soil microbial population affect the degree of biodegradation; hence detailed site specific characterization studies are needed prior to deciding on the proper bioremediation method.

Microbial contamination of stored diesel oil in Brasil

As refinarias e sistemas de distribuiçäo tem enfrentado problemas com a contaminaçäo microbiana em óleo diesel armazenado. A atividade microbiana nesses sistemas leva à produçäo de uma biomassa na interface óleo/água, provocando problemas operacionais, como entupimento de filtros, tubulaçöes, alteraçäo na qualidade do combustível. O controle dessa contaminaçäo pode envolver o uso de agentes químicos, como os biocidas. O objetivo desse trabalho foi selecionar biocidas eficientes para o controle de fungos, bactérias aeróbicas e bactérias redutoras de sulfato (BSR), que foram isoladas de óleo diesel comum, urbano e naval, armazenados em tanques de refinaria no Rio Grande do Sul. Dois biocidas (um composto quaternário de amônio e uma mistura de isothiazolona) foram selecionados por apresentarem alta eficácia, em baixas concentraçöes para os principais contaminantes, como fungos do gênero Aspergillus e Hormoconis, BSR e bactérias aeróbias do gênero Pseudomonas e Bacillus