Hydrocarbon Degradation Research Articles

Bioremediation of petroleum hydrocarbons polluted soil by spent mushroom substrates: Microbiological structure and functionality

Spent mushroom substrate (SMS) holds valuable microbiota that can be useful in remediating polluted soils with hydrocarbons. However, the microorganisms behind the bioremediation process remain uncertain. In this work, a bioremediation assay of total petroleum hydrocarbons (TPHs) polluted soil by SMS application was performed to elucidate the microorganisms and consortia involved in biodegradation by a metabarcoding analysis. Untreated polluted soil was compared to seven bioremediation treatments by adding SMS of Agaricus bisporus, Pleurotus eryngii, Pleurotus ostreatus, and combinations. Soil microbial activity, TPH biodegradation, taxonomic classification, and predictive functional analysis were evaluated in the microbiopiles at 60 days. Different metagenomics approaches were performed to understand the impact of each SMS on native soil microbiota and TPHs biodegradation. All SMSs enhanced the degradation of aliphatic and aromatic hydrocarbons, being A. bisporus the most effective, promoting an efficient consortium constituted by the bacterial families Alcanivoraceae, Alcaligenaceae, and Dietziaceae along with the fungal genera Scedosporium and Aspergillus. The predictive 16 S rRNA gene study partially explained the decontamination efficacy by observing changes in the taxonomic structure of bacteria and fungi, and changes in the potential profiles of estimated degradative genes across the different treatments. This work provides new insights into TPHs bioremediation.

Study of Bacterization of Medicago sativa L. and Cynodon dactylon (L.) Pers. During the Process of Phytoremediation of Oil-contaminated Soils

Recultivation of oil-contaminated soils on the Absheron Peninsula is one of the most important environmental and social problems for this region. One of the promising ways to solve the problem of soil pollution with petroleum hydrocarbons is the development of methods and approaches for their purification and detoxification in situ, and above all, biodetoxification and bioremediation. In a model vegetation experiment using complex systems of a mixture of plants and microorganisms, phytoremediation of soil contaminated with crude oil at a concentration of 15 g/kg was investigated. It has been established that the inoculation of plants with the modified Fermi-Start biological product affects the number of rhizosphere microorganisms that stimulate plant growth, as well as the degree of purification of gray-brown soil from oil. Cynodon dactylon is an effective biosystem for the remediation of oil-contaminated gray-brown soils as it is modified by a culture of an oil-oxidizing microorganism of the Pseudomonas aeruginosa, isolated from oil-contaminated gray-brown soil, the Fermi-Start biological product and belongs to the group of “effective microorganisms”, associated with plants — Medicago sativa. The joint introduction of the Fermi-Start biological product and the culture of P. aeruginosa had the greatest protective effect on seedlings from exposure to crude oil. The height of the shoots increased by 71% compared with the negative control, in parallel, the degree of oil degradation increased to 24%. A significant role of alfalfa in stimulating the number of rhizosphere microorganisms capable of hydrocarbon degradation has been revealed. This is confirmed by data showing that the number of microorganisms in the plant rhizoplane was 1–2 orders of magnitude higher (about 1.1×107 on average) than in the rhizosphere (about 1.4×105 on average), which may be due to secretion of plant root exudates. The results of the research allow us to recommend the use of a plant-microbial biosystem consisting of Medicago sativa and Cynodon dactylon, together with a Fermi-Start modified biological product for phytoremediation of gray-brown soil contaminated with crude oil.

Microbial community response to hydrocarbon exposure in iron oxide mats: an environmental study.

Hydrocarbon pollution is a widespread issue in both groundwater and surface-water systems; however, research on remediation at the interface of these two systems is limited. This interface is the oxic-anoxic boundary, where hydrocarbon pollutant from contaminated groundwaters flows into surface waters and iron mats are formed by microaerophilic iron-oxidizing bacteria. Iron mats are highly chemically adsorptive and host a diverse community of microbes. To elucidate the effect of hydrocarbon exposure on iron mat geochemistry and microbial community structure and function, we sampled iron mats both upstream and downstream from a leaking underground storage tank. Hydrocarbon-exposed iron mats had significantly higher concentrations of oxidized iron and significantly lower dissolved organic carbon and total dissolved phosphate than unexposed iron mats. A strong negative correlation between dissolved phosphate and benzene was observed in the hydrocarbon-exposed iron mats and water samples. There were positive correlations between iron and other hydrocarbons with benzene in the hydrocarbon-exposed iron mats, which was unique from water samples. The hydrocarbon-exposed iron mats represented two types, flocculent and seep, which had significantly different concentrations of iron, hydrocarbons, and phosphate, indicating that iron mat is also an important context in studies of freshwater mats. Using constrained ordination, we found the best predictors for community structure to be dissolved oxygen, pH, and benzene. Alpha diversity and evenness were significantly lower in hydrocarbon-exposed iron mats than unexposed mats. Using 16S rDNA amplicon sequences, we found evidence of three putative nitrate-reducing iron-oxidizing taxa in microaerophile-dominated iron mats (Azospira, Paracoccus, and Thermomonas). 16S rDNA amplicons also indicated the presence of taxa that are associated with hydrocarbon degradation. Benzene remediation-associated genes were found using metagenomic analysis both in exposed and unexposed iron mats. Furthermore, the results indicated that season (summer vs. spring) exacerbates the negative effect of hydrocarbon exposure on community diversity and evenness and led to the increased abundance of numerous OTUs. This study represents the first of its kind to attempt to understand how contaminant exposure, specifically hydrocarbons, influences the geochemistry and microbial community of freshwater iron mats and further develops our understanding of hydrocarbon remediation at the land-water interface.

Biomolecular condensates of Chlorocatechol 1,2-Dioxygenase as prototypes of enzymatic microreactors for the degradation of polycyclic aromatic hydrocarbons

Polycyclic aromatic hydrocarbons (PAHs) are molecules with two or more fused aromatic rings that occur naturally in the environment due to incomplete combustion of organic substances. However, the increased demand for fossil fuels in recent years has increased anthropogenic activity, contributing to the environmental concentration of PAHs. The enzyme chlorocatechol 1,2-dioxygenase from Pseudomonas putida (Pp 1,2-CCD) is responsible for the breakdown of the aromatic ring of catechol, making it a potential player in bioremediation strategies. Pp 1,2-CCD can tolerate a broader range of substrates, including halogenated compounds, than other dioxygenases. Here, we report the construction of a chimera protein able to form biomolecular condensates with potential application in bioremediation. The chimera protein was built by conjugating Pp 1,2-CCD to low complex domains (LCDs) derived from the DEAD-box protein Dhh1. We showed that the chimera could undergo liquid-liquid phase separation (LLPS), forming a protein-rich liquid droplet under different conditions (variable protein and PEG8000 concentrations and pH values), in which the protein maintained its structure and main biophysical properties. The condensates were active against 4-chlorocatechol, showing that the chimera droplets preserved the enzymatic activity of the native protein. Therefore, it constitutes a prototype of a microreactor with potential use in bioremediation.

Degradation of petroleum hydrocarbon contaminants by Rhodococcus erythropolis KB1 synergistic with alfalfa (Medicago sativa L.).

Petroleum hydrocarbons are a stubborn pollutant that is difficult to degrade globally, and plant-microbial degradation is the main way to solve this type of pollutant. In this study, the physiological and ecological responses of alfalfa to petroleum hydrocarbons in different concentrations of petroleum hydrocarbon-contaminated soil with KB1 (Rhodococcus erythropolis) were analyzed and determined by laboratory potting techniques. The growth of alfalfa (CK) and alfalfa with KB1 (JZ) in different concentrations of petroleum hydrocarbons contaminated soil was compared and analyzed. The results of the CK group showed that petroleum hydrocarbons could significantly affect the activity of alfalfa antioxidant enzyme system, inhibit the development of alfalfa roots and the normal growth of plants, especially in the high-concentration group. KB1 strain had the ability to produce IAA, form biofilm, fix nitrogen, produce betaine and ACC deaminase, and the addition of KB1 could improve the growth traits of alfalfa in the soil contaminated with different concentrations of petroleum hydrocarbons, the content of soluble sugars in roots, and the stress resistance and antioxidant enzyme activities of alfalfa. In addition, the degradation kinetics of the strain showed that the degradation rate of petroleum could reach 75.2% after soaking with KB1. Furthermore, KB1 can efficiently degrade petroleum hydrocarbons in advance and significantly alleviate the damage of high concentration of petroleum hydrocarbons to plant roots. The results showed that KB1 strains and alfalfa plants could effectively enhance the degradation of petroleum hydrocarbons, which provided new ideas for improving bioremediation strategies.

Construction of yeast microbial consortia for petroleum hydrocarbons degradation.

Microbial degradation of petroleum hydrocarbons plays a vital role in mitigating petroleum contamination and heavy oil extraction. In this study, a Saccharomyces cerevisiae capable of degrading hexadecane has been successfully engineered, achieving a maximum degradation rate of up to 20.42%. However, the degradation ability of this strain decreased under various pressure conditions such as high temperature, high osmotic pressure, and acidity conditions. Therefore, a S. cerevisiae with high tolerance to these conditions has been constructed. And then, we constructed an "anti-stress hydrocarbon-degrading" consortium comprising engineered yeast strain SAH03, which degrades hexadecane, and glutathione synthetic yeast YGSH10, which provides stress resistance. This consortium was able to restore the degradation ability of SAH03 under various pressure conditions, particularly exhibiting a significant increase in degradation rate from 5.04% to 17.04% under high osmotic pressure. This study offers a novel approach for improving microbial degradation of petroleum hydrocarbons.

Remediation of petroleum hydrocarbon contaminated soils by nZVI coupled with electrokinetic activation of persulfate

This paper explores the use of nanoscale zero-valent iron (nZVI) coupled with electrically activated persulfate (nZVI-EK/PS) for remediating petroleum hydrocarbon-contaminated soil. The effects of persulfate (PS) dosage, nZVI dosage, and soil organic matter content on the remediation of petroleum hydrocarbons were investigated. The study concludes that persulfate (PS) and nanoscale zero-valent iron (nZVI) are the primary factors influencing the remediation of petroleum hydrocarbons in the nZVI-EK/PS system. The optimal reaction conditions of PS = 170 mmol and nZVI = 17 mmol increased the efficiency of petroleum hydrocarbon treatment to 40.45%, with the highest removal efficiency of petroleum hydrocarbons observed at position S2, reaching 48.5%. With the addition of organic matter at 5%, the efficiency of petroleum hydrocarbon treatment reached 50.08%, representing a 9.68% enhancement in the nZVI-EK/PS system. Furthermore, potential mechanisms for the degradation of petroleum hydrocarbons by the nZVI-EK/PS system were identified using gas chromatography-mass spectrometry (GC-MS). The degradation order of different components was found to be esters < alkenes < alkanes < phenols.

Microbial response to a port fuel spill: Community dynamics and potential for bioremediation

Following a fuel leakage inside a Portuguese maritime port, we conducted parallel 30-day experiments using contaminated seawater and fuel, sampled five days after the incident. This study aimed to (i)survey the native microbial community response to the spilled fuel and (ii)evaluate the efficacy of bioremediation, both biostimulation and bioaugmentation with a lyophilized bacterial consortium (Rhodococcus erythropolis, Pseudomonas sp.), in accelerating hydrocarbon degradation. Metabarcoding analysis revealed a shift in microbial communities, with increased abundance of hydrocarbon-degraders (e.g. Alcanivorax, Thalassospira). Ninety-five hydrocarbonoclastic bacteria were isolated, including key groups from the enriched communities. The lyophilized bacteria added in bioaugmentation, enhanced the abundance of hydrocarbon-degraders over time and were recovered throughout time. Bioremediation treatments favoured biodegradation, achieving over 60 % removal of total petroleum hydrocarbons after 15 days, contrasting with natural attenuation where almost no TPH was removed. This work highlights the potential of bioremediation technologies to accelerate hydrocarbon-degrading activity, for oil spills inside ports.

Microbial remediation of petroleum-contaminated soil focused on the mechanism and microbial response: a review.

The environmental pollution caused by petroleum hydrocarbons has received considerable attention in recent years. Microbial remediation has emerged as the preferred method for the degradation of petroleum hydrocarbons, which is experiencing rapid development driven by advancements in molecular biology. Herein, the capacity of different microorganisms used for crude oil bioremediation was reviewed. Moreover, factors influencing the effectiveness of microbial remediation were discussed. Microbial remediation methods, such as bioaugmentation, biostimulation, and bioventilation, are summarized in this review. Aerobic and anaerobic degradation mechanisms were reviewed to elucidate the metabolic pathways involved. The impacts of petroleum hydrocarbons on microorganisms and the environment were also revealed. A brief overview of synthetic biology and a unique perspective of technique combinations were presented to provide insight into research trends. The challenges and future outlook were also presented to stimulate contemplation of the mechanisms involved and the development of innovative techniques.

Fine-tuning an aromatic ring-hydroxylating oxygenase to degrade high molecular weight polycyclic aromatic hydrocarbon

Rieske nonheme iron aromatic ring-hydroxylating oxygenases (RHOs) play pivotal roles in determining the substrate preferences of polycyclic aromatic hydrocarbon (PAH) degraders. However, their potential to degrade high molecular weight PAHs (HMW-PAHs) has been relatively unexplored. NarA2B2 is an RHO derived from a thermophilic Hydrogenibacillus sp. strain N12. In this study, we have identified four “hotspot” residues (V236, Y300, W316, and L375) that may hinder the catalytic capacity of NarA2B2 when it comes to HMW-PAHs. By employing structure-guided rational enzyme engineering, we successfully modified NarA2B2, resulting in NarA2B2 variants capable of catalyzing the degradation of six different types of HMW-PAHs, including pyrene, fluoranthene, chrysene, benzo[a]anthracene, benzo[b]fluoranthene, and benzo[a]pyrene. Three representative variants, NarA2B2W316I, NarA2B2Y300F-W316I, and NarA2B2V236A-W316I-L375F, not only maintain their abilities to degrade low molecular weight PAHs (LMW-PAHs), but also exhibited 2-4 times higher degradation efficiency for HMW-PAHs in comparison to another isozyme, NarAaAb. Computational analysis of the NarA2B2 variants predict that these modifications altering the size and hydrophobicity of the active site pocket making it more suitable for HMW-PAHs. These findings provide a comprehensive understanding of the relationship between three-dimensional structure and functionality, thereby opening up possibilities for designing improved RHOs that can be more effectively used in the bioremediation of PAHs.

Biodegradation of bonny light crude oil by plasmid and non-plasmid borne soil bacterial strains using biostimulation and bioaugmentation techniques

A laboratory scale study was designed and conducted to assess the biodegradation of Bonny light crude oil by plasmid and non-plasmid borne soil bacterial strains using biostimulation and bioaugmentation techniques. The enrichment technique, turbidometric test, plasmid curing test, molecular identification method, biostimulation test, bioaugmentation test and gas chromatographic technique were carried out using standard analytical techniques. The physicochemical analysis result showed that the pH was slightly neutral, the organic carbon content was higher (2.32 to 4.34%), the conductivity was higher (0.41 to 0.44 μS/cm), and the water holding capacity was lower (0.27 percent and 10.11 kg, respectively). Based on their capacity to use crude oil, the results showed that 22 of the 60 isolated bacterial strains had higher pollutant degrading potentials (A600nm &gt; 0.3).The identified potent hydrocarbon degraders includes: Bacillus spp., Pseudomonas aeruginosa KAVK01 and Ochrobacterium E85b strains. The highest degradation efficiency of 91% was found in soil that had been contaminated with 3 % (v/w) crude oil, amended with inorganic salts, and inoculated with plasmid-borne mixed cultures. The result further indicated that the consortium of plasmid borne isolates enhanced the reduction of the crude oil from the initial concentration of 10,318 ppm to 501 ppm (95 %) whereas 64 % decontamination was facilitated by the consortium of plasmid cured isolates. The information gathered from this investigation may be useful in choosing bacterial species, particularly plasmid-borne ones that can be employed to biodegrade soil contaminated by crude oil in Nigeria's Niger Delta region as well as the sample collection locations.

Isolation and identification of hydrocarbon degrading bacteria and fungi from waste engine oil impacted soil, their distribution frequency and hydrocarbon degradation capacity

The isolation and identification of hydrocarbon degrading bacteria and fungi from waste engine oil contaminated soil obtained from Mechanical Village, Uyo, Akwa Ibom State, their distribution frequency and hydrocarbon degradation capacity were investigated using standard method. The results revealed that Bacillus myocoides, Staphylococcus aureus, Listeria murrayi, Actinomycete viscosus, Clostridium sporogenes, Bacillus licheniformis, Corynebacterium ulcerans and Clostridium histolyticum were bacteria identified. Fungi identified were Fusarium oxysporum, Cryptococcus terreus, Penicillium notatum, Aspergillus fumigatus, Rhizopus stolonifer, Rhizopus oryzae and Rhodotorula mulcilaginosa. Their distribution frequency were B. myocoides (27.9 %), S. aureus (8.1 %), L. murrayi (9.9 %), A. viscosus (11.7 %), C. sporogenes (16.2 %), B. licheniformis (7.2%), C. ulcerans (12.6 %) and C. histolyticum (6.3 %) for bacteria. For the fungi, F. oxysporum (15.3 %), C. terreus (4.1%), P. notatum (20.4 %), A. fumigatus (11.2 %), R. stolonifer (10.2 %), R. oryzae (24.5%) and R. mulcilaginosa (14.3%). The efficiency of the microbial isolates to degrade waste engine oil from petrol and diesel driven vehicles as their major source of energy were found to vary among the microorganisms. C. sporogenes degraded waste engine oil from petrol driven vehicle with the highest efficiency (38.09%), whereas S. aureus demonstrated the least efficiency (19.28 %). C. histolyticum utilized waste oil from diesel vehicle with the highest efficiency (35.00%), while C. ulcerans was the least (17.31%). Among the fungi isolates, R. mulcilaginosa showed most degradation potential for spent oil from petrol driven vehicle (34.85 %) whereas C. terreus was the least (20.00 %). A. fumigatus exhibited highest degradation capacity for spent oil from diesel driven vehicle (29.73 %) while R. oryzae demonstrated the least potential (20.76 %). These results implied that microbial consortium could best be used for remediation of hydrocarbon contaminated soil.

Shifts in structure and dynamics of the soil microbiome in biofuel/fuel blend-affected areas triggered by different bioremediation treatments.

The use of biofuels has grown in the last decades as a consequence of the direct environmental impacts of fossil fuel use. Elucidating structure, diversity, species interactions, and assembly mechanisms of microbiomes is crucial for understanding the influence of environmental disturbances. However, little is known about how contamination with biofuel/petrofuel blends alters the soil microbiome. Here, we studied the dynamics in the soil microbiome structure and composition of four field areas under long-term contamination with biofuel/fossil fuel blends (ethanol 10% and gasoline 90%-E10; ethanol 25% and gasoline 75%-E25; soybean biodiesel 20% and diesel 80%-B20) submitted to different bioremediation treatments along a temporal gradient. Soil microbiomes from biodiesel-polluted areas exhibited higher richness and diversity index values and more complex microbial communities than ethanol-polluted areas. Additionally, monitored natural attenuation B20-polluted areas were less affected by perturbations caused by bioremediation treatments. As a consequence, once biostimulation was applied, the degradation was slower compared with areas previously actively treated. In soils with low diversity and richness, the impact of bioremediation treatments on the microbiomes was greater, and as a result, the hydrocarbon degradation extent was higher. The network analysis showed that all abundant keystone taxa corresponded to well-known degraders, suggesting that the abundant species are core targets for biostimulation in soil remediation processes. Altogether, these findings showed that the knowledge gained through the study of microbiomes in contaminated areas may help design and conduct optimized bioremediation approaches, paving the way for future rationalized and efficient pollutant mitigation strategies.

Remediation of diesel contaminated soil by using activated persulfate with Fe3O4 magnetic nanoparticles: effect and mechanisms.

In this study, Fe3O4 magnetic nanoparticles (Fe3O4 MNPs) were assessed for their ability to enhance the activity of persulfate (PS). Various controlling factors including PS dosages, initial pH, water-soil ratio, ratio of Fe2+, and Fe3O4 MNPs to PS were considered in both the Fe2+/PS system and the Fe3O4 MNPs/PS system. Results showed that the Fe3O4 MNP-activated PS system exhibited high processing efficiency owing to the gradual release of Fe2+. This process occurred in a wide pH range (5-11), attributed to the synergistic action of sulfate radicals (SO4-·) and hydroxyl radicals (OH·) under alkaline conditions, effectively mitigating soil acidification. The ratio of Fe3O4 MNPs to PS and water-soil ratio significantly influenced the degradation rate with the highest petroleum hydrocarbon degradation rate exceeding 80% (82.31%). This rate was 3.1% higher than that achieved by the Fe2+/PS system under specific conditions: PS dosage of 0.05 mol/L, Fe3O4 MNPs to PS ratio of 1:10, water-soil ratio of 2:1, and initial pH of 11. Meanwhile, oxidant consumption in the Fe3O4 MNPs/PS system was halved compared to the Fe2+/PS system due to the slow release of Fe2+ and less ineffective consumption of SO4-·. Mechanistically, the possible degradation process was divided into three parts: the initial chain reaction, the proliferating chain reaction, and the terminating chain reaction. The introduction of Fe3O4 MNPs accelerated the degradation rate of pentadecane, heneicosane, eicosane, tritetracontane, and 9-methylnonadecane.

Field-aged rice hull biochar stimulated the methylation of mercury and altered the microbial community in a paddy soil under controlled redox condition changes

Mercury (Hg) contaminated paddy soils are hot spots for methylmercury (MeHg) which can enter the food chain via rice plants causing high risks for human health. Biochar can immobilize Hg and reduce plant uptake of MeHg. However, the effects of biochar on the microbial community and Hg (de)methylation under dynamic redox conditions in paddy soils are unclear. Therefore, we determined the microbial community in an Hg contaminated paddy soil non-treated and treated with rice hull biochar under controlled redox conditions (< 0 mV to 600 mV) using a biogeochemical microcosm system. Hg methylation exceeded demethylation in the biochar-treated soil. The aromatic hydrocarbon degraders Phenylobacterium and Novosphingobium provided electron donors stimulating Hg methylation. MeHg demethylation exceeded methylation in the non-treated soil and was associated with lower available organic matter. Actinobacteria were involved in MeHg demethylation and interlinked with nitrifying bacteria and nitrogen-fixing genus Hyphomicrobium. Microbial assemblages seem more important than single species in Hg transformation. For future directions, the demethylation potential of Hyphomicrobium assemblages and other nitrogen-fixing bacteria should be elucidated. Additionally, different organic matter inputs on paddy soils under constant and dynamic redox conditions could unravel the relationship between Hg (de)methylation, microbial carbon utilization and nitrogen cycling.

Degradation and mechanism of PAHs by Fe-based activated persulfate: Effect of temperature and noble metal

The accumulation of contaminants like PAHs in soil due to industrialization, urbanization, and intensified agriculture poses environmental challenges, owing to their persistence, hydrophobic nature, and toxicity. Thus, the degradation of PAHs has attracted worldwide attention in soil remediation. This study explored the effect of noble metal and temperature on the degradation of various polycyclic aromatic hydrocarbons (PAHs) in soil, as well as the types of reactive radicals generated and mechanism. The Fe-Pd/AC and Fe-Pt/AC activated persulfate exhibited high removal efficiency of 19 kinds of PAHs, about 79.95 % and 83.36 %, respectively. Fe-Pt/AC-activated persulfate exhibits superior degradation efficiency than that on Fe-Pd/AC-activated persulfate, due to the higher specific surface area and dispersity of Pt particles, thereby resulting in increased reactive radicals (·OH, SO4−· and ·OOH). Additionally, thermal activation enhances the degradation of PAHs, with initial efficiencies of 64.20 % and 55.49 % on Fe-Pd/AC- and Fe-Pt/AC-activated persulfate systems respectively, increasing to 76.05 % and 73.14 % with elevated temperatures from 21.5 to 50 °C. Metal and thermal activation facilitate S2O82− activation, generating reactive radicals, crucial for the degradation of PAHs via ring opening and oxygen hydrogenation reactions, yielding low-ring oxygen-containing derivatives such as organic acids, keto compounds, ethers, and esters. Furthermore, understanding the impact of parameters such as activation temperature and the types of noble metals on the degradation of PAHs within the activated persulfate system provides a theoretical foundation for the remediation of PAH-contaminated soil.

Insights into the response of electroactive biofilm with petroleum hydrocarbons degradation ability to quorum sensing signals

Bioelectrochemical technologies based on electroactive biofilms (EAB) are promising for petroleum hydrocarbons (PHs) remediation as anode can serve as inexhaustible electron acceptor. However, the toxicity of PHs might inhibit the formation and function of EABs. Quorum sensing (QS) is ideal for boosting the performance of EABs, but its potential effects on reshaping microbial composition of EABs in treating PHs are poorly understood. Herein, two AHL signals, C4-HSL and C12-HSL, were employed to promote EABs for PHs degradation. The start-times of AHL-mediated EABs decreased by 18–26%, and maximum current densities increased by 28–63%. Meanwhile, the removal of total PHs increased to over 90%. AHLs facilitate thicker and more compact biofilm as well as higher viability. AHLs enhanced the electroactivity and direct electron transfer capability. The total abundance of PH-degrading bacteria increased from 52.05% to 75.33% and 72.02%, and the proportion of electroactive bacteria increased from 26.14% to 62.72% and 63.30% for MFC-C4 and MFC-C12. Microbial networks became more complex, aggregated, and stable with addition of AHLs. Furthermore, AHL-stimulated EABs showed higher abundance of genes related to PHs degradation. This work advanced our understanding of AHL-mediated QS in maintaining the stable function of microbial communities in the biodegradation process of petroleum hydrocarbons.

Origin and microbial degradation of thermogenic hydrocarbons within the sandy gas hydrate reservoirs in the Qiongdongnan Basin, northern South China Sea

Large-scale natural gas hydrate accumulations (generally over one billion cubic meters of natural gas) in coarse-grained marine sediments are the most valuable types for commercial exploration and exploitation. The formation of such a gas hydrate deposit generally requires a sufficient natural gas supply such as deeply buried thermogenic gas source. However, the worldwide discovered thermogenic gas hydrate deposits are rare, leaving relatively a poor understanding of their formation and evolution. Based on the geochemical and microbial analyses of eleven hydrate-related gas and five sediment samples from the thermogenic gas hydrate deposits in the Qiongdongnan Basin in northern South China Sea, we systematically investigated the origin and evolution of thermogenic hydrocarbons within sandy gas hydrate reservoirs. Results show that natural gas within high saturated gas hydrate-filled sandy sediments is predominately of thermogenic (the proportion is 57.1–70.5%), characterized by “dry gas” (99.38–99.7% C1, 0.26–0.47% C2 and <0.5% C3+), relatively lower C1/(C2+C3) ratio (187–376), and heavier δ13C–C1 values (−55.0 to −31.3‰). In addition, typical biomarkers (including the alkanes, tricyclic terpanes, hopanes and steranes) generated from mature source rocks were detected in the hydrate-filled fine sand reservoir, as well as the biomarker indicators (including the unresolved complex mixtures (UCMs) hump and the 25-norhopane homologous series) for severe hydrocarbon biodegradation. Furthermore, large amounts of hydrocarbon degrading bacteria (50.5% of bacteria) and methanogenic archaea (34.3% of archaea) were detected within hydrate-filled sandy sediment. Our results indicated that the deeply buried thermogenic source can indeed provide sufficient gas supply for the formation of a large-scale gas hydrate deposit, but thermogenic liquid hydrocarbons within gas hydrate reservoirs are very susceptible to be degraded and consumed by hydrocarbon degrading bacteria. In addition, we also found that the clay components of argillaceous gas hydrate reservoirs can adsorb and store large amounts of in situ immature organic matter, which can significantly obscure or cover up the original geochemical signatures of thermogenic hydrocarbons therein. These findings suggest that the relative contribution of deeply buried thermogenic source for gas hydrate accumulation worldwide may be underestimated, because we have previously ignored the fact that thermogenic hydrocarbons within gas hydrate reservoirs would be significantly consumed by hydrocarbon degrading microbes or diluted by in situ immature sedimentary organic matter. Our findings are highly significant for the evaluation and exploration of thermogenic gas hydrate accumulations worldwide.

Costus speciosus (Koen ex. Retz.) Sm.: a suitable plant species for remediation of crude oil and mercury-contaminated soil.

The aim of this study was to evaluate the efficiency of Costus speciosus (Koen ex. Retz.) Sm. in the degradation of crude oil and reduction of mercury (Hg) from the contaminated soil in pot experiments in the net house for 180days. C. speciosus was transplanted in soil containing 19150mgkg-1 crude oil and 3.2mgkg-1 Hg. The study includes the evaluation of plant biomass, height, root length, total petroleum hydrocarbon (TPH) degradation, and Hg reduction in soil, TPH, and Hg accumulation in plants grown in fertilized and unfertilized pots, chlorophyll production, and rhizospheric most probable number (MPN) at 60-day interval. The average biomass production and heights of C. speciosus in contaminated treatments were significantly (p < 0.05) lower compared to the unvegetated control. Plants grown in contaminated soil showed relatively reduced root surface area compared to the uncontaminated treatments. TPH degradation in planted fertilized, unplanted, and planted unfertilized pot was 63%, 0.8%, and 38%, respectively. However, compared to unvegetated treatments, TPH degradation was significantly higher (p < 0.05) in vegetated treatments. A comparison of fertilized and unfertilized soils showed that TPH accumulation in plant roots and shoots was relatively higher in fertilized soils. Hg degradation in soil was significantly (p < 0.05) more in planted treatment compared to unplanted treatments. The fertilized soil showed relatively more Hg degradation in soil and its accumulation in roots and shoots of plants in comparison to unfertilized soil. MPN in treatments with plants was significantly greater (p < 0.05) than without plants. The plant's ability to produce biomass, chlorophyll, break down crude oil, reduce Hg levels in soil, and accumulate TPH and Hg in roots and shoots of the plant all point to the possibility of using this plant to remove TPH and Hg from soil.

Autochthonous bioaugmentation strategies for the successful bioremediation of high-salt petrochemical wastewater using a biosurfactant-producing Enterobacter cloacae Z11

Petrochemical wastewater contains a high concentration of refractory aliphatic and aromatic hydrocarbons. Bioremediation of these hazardous materials using autochthonous bioaugmentation (ABA) strategy is attracting increasing interest. Herein, an indigenous biosurfactant-producing bacteria Z11 was isolated and identified as Enterobacter sp. The reintroduction of Z11 into petrochemical wastewater led to a substantial reduction (P < 0.01) in TPHs, decreasing from 8099 to 1723 mg/L, as well as a significant decrease in COD, decreasing from 8064.2 to 1689.2 mg/L. At a critical micelle concentration of 200 mg/L, the biosurfactant synthesized by Z11 exhibited an extraordinary decrease in surface tension from 67.5 to 29 mN/m. The biosurfactant demonstrated favorable surface activity over a wide pH, temperature, and salinity range. Based on a series of chemical analyses, the biosurfactant was classified as phospholipid. The autochthonous bioaugmentation technique developed by Z11 indicated a significant increase (P < 0.05) in the degradation of n-alkanes of varying chain lengths and polycyclic aromatic hydrocarbons. The analysis of the microbial composition demonstrated that the addition of Z11 caused a rise in bacterial populations capable of hydrocarbon decomposition, such as Pseudomonas sp. and Stenotrophomonas sp. The activities of two enzymes involved in degradation, alkane hydroxylase and alcohol dehydrogenase, increased dramatically and showed a significant (P < 0.05) correlation with Enterobacter sp. Z11. The proposed mechanism could be elucidated that functional genes regulate the secretion of biodegradation enzymes and biosurfactants to promote the deep degradation of petroleum hydrocarbons. This study provides technical support for the reintroduction of autochthonous microorganisms capable of producing biosurfactants in implementing in situ ABA strategies.