A mathematical model for bioremediation of hydrocarbon-contaminated soils

The AQUASIM executable file is available in DataRepositoriUM (https://doi.org/10.34622/datarepositorium/2BUM2J, accessed on 15 September 2022).Bioremediation of hydrocarbons in soil is a highly complex process, involving a multiplicity of physical, chemical and biological phenomena. Therefore, it is extremely difficult to control and boost the bioremediation of these systems after an oil spill. A mathematical model was developed to assist in the prediction and decision-making regarding the in situ bioremediation of hydrocarbon-contaminated soils. The model considered the most relevant processes involved in the mass transfer and biodegradation of alkanes over time and along the depth of a flooded soil column. Aliphatic hydrocarbons were chosen since they are less water soluble than aromatics and account for 50–90% of the hydrocarbon fraction in several petroleum products. The effect of adding oxygen, nitrate, iron (III) or sulfate as electron acceptors was then simulated (bioremediation scenarios). Additionally, and to feed the model, batch assays were performed to obtain experimental data on hydrocarbon adsorption to soil particles (more than 60% of hydrocarbons tends to be adsorbed to soil particles), as well as hydrocarbon biodegradation rates in the presence of nitrate (0.114 d−1) and oxygen (0.587 d−1). The model indicates that saturated hydrocarbon removal occurs mainly with adsorption/desorption and transport processes in the upper layers of soil due to methanogenic biodegradation in deeper layers, since the other microbial processes are soon limited by the lack of electron acceptors. Simulation results show that higher initial electron acceptor concentrations led to higher hydrocarbon removal, confirming that the model is performing in accordance with the expected. Close to the surface (at 0.1 m depth), all scenarios predicted more than 83% hydrocarbon removal after two years of simulation. Soil re-aeration results in faster hydrocarbon removal (more than 20% after one year) and surfactants addition (around 15% after one year) may also accelerate soil bioremediation. With this model, the simultaneous contributions of the various physicochemical and biological processes are integrated, facilitating the simulation and comparison of different bioremediation scenarios. Therefore, it represents a useful support tool for the management of contaminated sites.This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of project MORE (PTDC/AAG-TEC/3500/2014; POCI-01-0145-FEDER-016575), the strategic funding of UIDB/04469/2020 unit and by LABBELS—Associate Laboratory in Biotechnology, Bioengineering and Microelectromechanical Systems, LA/P/0029/2020.info:eu-repo/semantics/publishedVersio

Iron compounds in anaerobic degradation of petroleum hydrocarbons: a review

Waste and wastewater containing hydrocarbons are produced worldwide by various oil-based industries, whose activities also contribute to the occurrence of oil spills throughout the globe, causing severe environmental contamination. Anaerobic microorganisms with the ability to biodegrade petroleum hydrocarbons are important in the treatment of contaminated matrices, both in situ in deep subsurfaces, or ex situ in bioreactors. In the latter, part of the energetic value of these compounds can be recovered in the form of biogas. Anaerobic degradation of petroleum hydrocarbons can be improved by various iron compounds, but different iron species exert distinct effects. For example, Fe(III) can be used as an electron acceptor in microbial hydrocarbon degradation, zero-valent iron can donate electrons for enhanced methanogenesis, and conductive iron oxides may facilitate electron transfers in methanogenic processes. Iron compounds can also act as hydrocarbon adsorbents, or be involved in secondary abiotic reactions, overall promoting hydrocarbon biodegradation. These multiple roles of iron are comprehensively reviewed in this paper and linked to key functional microorganisms involved in these processes, to the underlying mechanisms, and to the main influential factors. Recent research progress, future perspectives, and remaining challenges on the application of iron-assisted anaerobic hydrocarbon degradation are highlighted.This research was funded by the Portuguese Foundation for Science and Technology (FCT) under the scope of project MORE (POCI-01-0145-FEDER-016575) and of the strategic funding of UIDB/04469/2020 unit. It was also funded by LABBELS—Associate Laboratory in Biotechnology, Bioengineering and Microelectromechanical Systems, LA/P/0029/2020, and by the European Regional Development Fund under the scope of Norte2020—Programa Operacional Regional do Norte—BioEcoNorte project (NORTE-01-0145-FEDER-000070).info:eu-repo/semantics/publishedVersio

Long-term acclimation of anaerobic sludges for high-rate methanogenesis from LCFA

Inhibition of methanogens by long chain fatty acids (LCFA) and the low numbers of LCFA-degrading bacteria are limitations to exploit biogas production from fat-rich wastewaters. Generally reactors fail due to excessive LCFA accumulation onto the sludge. Here, long-term acclimation and bioaugmentation with a LCFA-degrading coculture were hypothesized as strategies to enhance methanogenic conversion of these compounds. Anaerobic sludges previously exposed to LCFA for more than 100 days converted a specific biomass-associated substrate of (3.2 ± 0.1) kg·kg−1 with very short lag phases (<1 day), whereas non-acclimated sludges showed lag phases of 11–15 days for metabolizing (1.6–1.8) kg·kg−1. Addition of a coculture of Syntrophomonas zehnderi and Methanobacterium formicicum to sludges previously loaded with LCFA and containing different amounts of biomass-associated substrate (from (0.5–3.2) kg·kg−1) did not improve methane production neither lag phases were shortened, indicating that the endogenous microbiota are not a limiting factor. Clearly, we show that long-term sludge acclimation to LCFA is essential for high rate methanogenesis from LCFA.The authors acknowledge the financial support by the European Regional Development Fund - ERDF, through the Operational Program Thematic Factors of Competitiveness - COMPETE, and by Portuguese funds, through the Portuguese Foundation for Science and Technology (FCT), in the frame of the project FCOMP-01-0124-FEDER-014784. FCT Strategic Project PEst-OE/EQB/LA0023/2013 is also acknowledged. A.J. Cavaleiro thanks FCT for the post-doctoral fellowship ref. SFRH/BPD/75247/2010. A.J.M. Stams has received funding from the European Research Council under the European Union's Seventh Framework Programme (FP/2007-2013)/ERC Grant Agreement n. [323009]

Addition of electron acceptors stimulates methanogenesis from lipids by anaerobic sludge

Incubation of anaerobic sludge with triolein or oleate in the presence of nitrate or sulphate led to an increased methane production, relatively to incubations without inorganic electron acceptor. Faster methane production was obtained in assays amended with nitrate. Methanogenesis occurred after the reduction of alternative electron acceptors

Exploring syntrophic relationships in the anaerobic biodegradation of lipids and long chain fatty acids

ICBM-3 - 3rd International Conference on Biogas Microbiology (Abstract Book)[Excerpt] Practical knowledge on anaerobic digestion of waste lipids has been improving for several decades, but the microbiology of these processes remains partially undisclosed, with non-cultivated taxonomic groups often detected in anaerobic communities degrading lipids. This work studies the diversity and physiology of anaerobic microorganisms involved in the metabolism of lipids and long chain fatty acids. Anaerobic culturing procedures were applied for the development of enrichment cultures, and combined with next generation sequencing techniques. Enriched microbial communities specialized in the degradation of triolein (0.3 mmol·L-1) and oleate (1 mmol·L-1) were obtained under methanogenic conditions. Oleatedegrading cultures were also developed in the presence of the external electron acceptors ferric hydroxide (75 mmol·L-1) or sulfate (15 mmol·L-1). Three mesophilic sludges from different origins were used as inocula. [...]info:eu-repo/semantics/publishedVersio

Anaerobic co-digestion of cork based oil sorbent and cow manure or sludge

Cork, a material with great economic, social and environmental importance in Portugal, is also a good oil sorbent that can be used in the remediation of oil spills. The oil-impregnated cork can be easily removed, but requires further treatment. In the case of vegetable oil spills, anaerobic digestion may be a potential solution. This study aims to evaluate the effect of adding cork contaminated with sunflower oil as co-substrate in anaerobic digestion processes. Biodegradability assays were prepared with cow manure or sludge from a wastewater treatment plant, in the presence of five concentrations of oil-contaminated cork, between 200 and 1000 mg· L-1 as COD. Maximum cumulative methane production increased with the amount of oily cork up to 41 % and 101 % in the assays with manure and sludge, respectively. Sporadic addition of cork contaminated with vegetable oil during anaerobic digestion of manure or sludge increases significantly the methane production of these processes.Programa Operacional Regional do Norte (ON.2 - 0 Novo Norte), QREN, FEDERPortuguese Foundation for Science and Technology (FCT, in the frame of projects FCOMPO 1-0124-FEDER-014784 (FCT: PTDC/EBBEBI/114364/2009

Conversion of Cn-unsaturated into Cn-2-saturated LCFA can occur uncoupled from methanogenesis in anaerobic bioreactors

Fat, oils, and grease present in complex wastewater can be readily converted to methane, but the energy potential of these compounds is not always recyclable, due to incomplete degradation of long chain fatty acids (LCFA) released during lipids hydrolysis. Oleate (C18:1) is generally the dominant LCFA in lipid-containing wastewater, and its conversion in anaerobic bioreactors results in palmitate (C16:0) accumulation. The reason why oleate is continuously converted to palmitate without further degradation via β-oxidation is still unknown. In this work, the influence of methanogenic activity in the initial conversion steps of unsaturated LCFA was studied in 10 bioreactors continuously operated with saturated or unsaturated C16- and C18-LCFA, in the presence or absence of the methanogenic inhibitor bromoethanesulfonate (BrES). Saturated Cn-2-LCFA accumulated both in the presence and absence of BrES during the degradation of unsaturated Cn-LCFA, and represented more than 50\% of total LCFA. In the presence of BrES further conversion of saturated intermediates did not proceed, not even when prolonged batch incubation was applied. As the initial steps of unsaturated LCFA degradation proceed uncoupled from methanogenesis, accumulation of saturated LCFA can be expected. Analysis of the active microbial communities suggests a role for facultative anaerobic bacteria in the initial steps of unsaturated LCFA biodegradation. Understanding this role is now imperative to optimize methane production from LCFA.European Research Council under the European Union’s Seventh Framework Programme (FP/2007-2013)/ERC Grant Agreement No 323009, and the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684), and Project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462). We also thank the Gravitation grant (project 024.002.002) of the Netherlands Ministry of Education, Culture and Science and the Netherlands Science Foundation (NWO

Effect of sulfate and iron (III) on LCFA degradation by a methanogenic community

[Excerpt] Under anaerobic conditions long chain fatty acids (LCFA) can be converted to methane by syntrophic bacteria and methanogenic archaea. LCFA degradation was also reported in the presence of alternative hydrogenotrophic partners, such as sulfate-reducing bacteria (SRB) and iron-reducing bacteria (IRB), which generally show higher affinity for H2 than methanogens and are more resistant to LCFA [1,2,3]. Their presence in a microbial culture degrading LCFA can be advantageous to reduce LCFA toxicity towards methanogens, although high concentrations of external electron acceptor (EEA) can lead to outcompetition of methanogens and cease methane production. In this work, we tested the effect of adding sub-stoichiometric concentrations of sulfate and iron(III) to methanogenic communities degrading LCFA. (...

Corksorb enhances alkane degradation by hydrocarbonoclastic bacteria

Biosorbent materials are effective in the removal of spilled oil from water, but their effect on hydrocarbonoclastic bacteria is not known. Here, we show that corksorb, a cork-based biosorbent, enhances growth and alkane degradation by Rhodococcus opacus B4 (Ro) and Alcanivorax borkumensis SK2 (Ab). Ro and Ab degraded 96 ± 1% and 72 ± 2%, respectively, of a mixture of n-alkanes (2 g L1) in the presence of corksorb. These values represent an increase of 6 and 24%, respectively, relative to the assays without corksorb. The biosorbent also increased the growth of Ab by 51%. However, no significant changes were detected in the expression of genes involved in alkane uptake and degradation in the presence of corksorb relative to the control without the biosorbent. Nevertheless, transcriptomics analysis revealed an increased expression of rRNA and tRNA coding genes, which confirms the higher metabolic activity of Ab in the presence of corksorb. The effect of corksorb is not related to the release of soluble stimulating compounds, but rather to the presence of the biosorbent, which was shown to be essential. Indeed, scanning electron microscopy images and downregulation of pili formation coding genes, which are involved in cell mobility, suggest that cell attachment on corksorb is a determinant for the improved activity. Furthermore, the existence of native alkane-degrading bacteria in corksorb was revealed, which may assist in situ bioremediation. Hence, the use of corksorb in marine oil spills may induce a combined effect of sorption and stimulated biodegradation, with high potential for enhancing in situ bioremediation processes. © Copyright © 2021 Martins, Freitas, Castro, Silva, Gudiña, Sequeira, Salvador, Pereira and Cavaleiro.This study was funded by the Portuguese Foundation for Science and Technology (FCT) under the scope of project MORE (POCI-01-0145-FEDER-016575) and Salt Oil+ (POCI-01-0145-FEDER-030180) and of the strategic funding of UIDB/04469/2020 unit. Research of RS and JS was supported by Ph.D. grants SFRH/BD/116154/2016 and SFRH/BD/147271/2019, respectively, funded by FCT.info:eu-repo/semantics/publishedVersio

Addition of co-substrates stimulates hexadecene conversion to methane by an enriched microbial consortium

ICBM-3 - 3rd International Conference on Biogas MicrobiologyLinear olefins with 16 to 18 carbon atoms are frequently used as hydrophobic groups in oil soluble surfactants and as lubricating fluids. The production of olefins in petrochemical plants generates olefin contaminated wastewater that can be treated anaerobically in methanogenic bioreactors, coupling degradation to energy recovery. However, this conversion is generally slow, due to olefins ́ insolubility in water and poor bioavailability for microorganisms. Addition of an easy degradable carbon source may enhance the growth of hydrocarbon degrading methanogenic communities. In this study, hexadecene degradation by a methanogenic enrichment was stimulated by addition of yeast extract (0.5 g·L-1), lactate (4.5 mmol·L-1) or crotonate (4.5 mmol·L-1) as co-substrates. After stimulation with yeast extract or lactate, the microbial communities were able to convert hexadecene to methane 5 and 2.5 times faster, respectively, than non-stimulated cultures. Hexadecene conversion to methane was not enhanced by crotonate addition. Further incubations with fermented yeast extract did not improve methane production from hexadecene, which suggests that the positive stimulatory effect of yeast extract was due to the extra carbon source and not to the supply of essential co-factors. The microbial community composition of the hexadecene degrading enrichments was studied by 16S rRNA sequencing. Bacteria from the Chloroflexi, Firmicutes, Proteobacteria(Deltaproteobacteria), Spirochaetes, Synergistetes and Thermotogaephyla were identified, with Syntrophobacterales, Spirochaetales and Synergistales as the most abundant orders. Hydrogenotrophic methanogens predominated over acetoclastic methanogens. Currently the isolation and identification of key microbial players involved in hexadecene degradation are ongoing. This study can be useful for improving the treatment of olefin contaminated wastewater using methanogenic conditionsinfo:eu-repo/semantics/publishedVersio