Biodegradation Of Hydrocarbons Research Articles

A novel strategy combining hydrogenotrophic methanogens' bioaugmentation and biochar biostimulation for simultaneous polycyclic aromatic hydrocarbon biodegradation and bioenergy recovery.

A novel strategy combining bioaugmentation using methanogenic archaea and biostimulation using biochar was proposed for the first time to obtain simultaneous improvement of mixed PAHs' anaerobic biodegradation and bioenergy production. The results showed that the addition of PHAs immediately resulted in inhibition in methane production and accumulation of VFA, indicating that PHAs are more toxic to methanogens than the acetogenic bacteria. The coupling of biochar with hydrogenotrophic methanogen alleviated the inhibitory effects of PAHs, allowing the anaerobic fermentation system to recover its methane production capability rapidly. Compared to the Fe3+ + bioaugmentation group, the biochar + bioaugmentation group exhibited a 7.5% higher restored cumulative methane production. This coupling strategy ultimately facilitated the degradation of most PAHs, achieving a removal rate of over 90%. Moreover, the coupled biochar and bioaugmentation induced significant changes in the archaeal community structure. Direct interspecies electron guilds (i.e., Streptococcus and Methanosarcina) were enriched in the presence of biochar and bioaugmentation, responsible for prominent PAH removal and methane recovery. This study demonstrated the feasibility of simultaneous PAH biodegradation and bioenergy production using electron acceptor and enriched microorganisms.

Simultaneous biodegradation of polycyclic aromatic hydrocarbons and phthalates by bacterial consortium and its bioremediation for complex polluted soil and sewage sludge

Simultaneous biodegradation of multiple micropollutantslike polycyclic aromatic hydrocarbons (PAHs) and phthalates (PAEs) by microbial consortia remain unclear. Here, four distinct bacterial consortia capable of degrading PAHs and PAEs were domesticated from sludge and its composts. PAH-degrading consortium HS and PAE-degrading consortium EC2 displayed the highest degradation efficiencies for PAHs (37 %–99 %) and PAEs (98 %–99 %), respectively, being significantly higher than those of individual member strains. Consortia HS and EC2 could simultaneously degrade both PAHs and PAEs. Remarkably, a synthetic consortium Syn by co-culturing consortia HS and EC2 demonstrated proficient simultaneous biodegradation for both PAHs (65 %–98 %) and PAEs (91 %–97 %). These consortia changed their community structure with enriching pollutant-degrading genera and extracellular polymeric substance contents to promote simultaneous biodegradation of multiple pollutants. Moreover, consortium Syn significantly enhanced degradation of both PAHs and PAEs in soil and sludge. This study provides strong candidates for simultaneous bioremediation of complex polluted environments by PAHs and PAEs.

Biodegradation of petroleum hydrocarbon and related molecular microbiology analysis of some fungal species

Biodegradation of petroleum hydrocarbon and related molecular microbiology analysis of some fungal species

Synergistic Actions of Biostimulation and Bioaugmentation in Microbial Biodegradation of Total Petroleum Hydrocarbons (TPH) Using Keratinaceous Materials

Synergistic Actions of Biostimulation and Bioaugmentation in Microbial Biodegradation of Total Petroleum Hydrocarbons (TPH) Using Keratinaceous Materials

Effect of Multi-Walled Carbon Nanotubes, Salicylic Acid, and Drought Stress on Mazut Biodegradation Efficiency in a Pb- and Zn-Polluted Soil Under Cultivation of the Pinto Bean Inoculated with Piriformospora indica

Abstract Aims: Today, remediation of soils contaminated with heavy metals and petroleum compounds is one of the major environmental problems. This research was done to investigate the effect of multiwall carbon nanotubes (MWCNs) and foliar application of salicylic acid (SA) on biodegradation of mazut in the Pb- and Zn-polluted soil under cultivation of the pinto bean inoculated with Piriformospora indica (P.indica). Methods: Treatments included applying MWCNs (0 and 2 % (W/W)) in the soil that was naturally polluted with Pb and Zn, foliar application of salicylic acid at the rate of 0 and 1.5 mmol/lit on the plants inoculated with P.indica under drought stress. In addition, the studied soil was polluted with mazut at the rate of 0% and 6% (w/w). After 3 months, plants were harvested and the plant Pb and Zn, the soil Pb and Zn, and the biodegradation of mazut in the soil were also measured. Results: Addition of MWCNS and foliar application of salicylic acid at the rate of 1.5 mmo/lit significantly decreased the soil Pb by 14.3 and 13.3%, respectively. Plant inoculation with P. indica significantly increased the biodegradation of mazut in the Pb- and Zn-polluted soil by 15.9%. However, drought stress had a negative effect on degradation of mazut in the soil. Conclusion: Regardless of plant expose to drought stress, using MWCNs and foliar application of SA has a positive effect on biodegradation of petroleum hydrocarbon in the soil.

Enrichment of Polycyclic Aromatic Hydrocarbon (PAH)-Degrading Strictly Anaerobic Sulfate-Reducing Cultures from Contaminated Soil and Sediment.

Sulfate-reducing bacteria (SRB) are crucial players in global biogeochemical cycling and some have been implicated in the anaerobic biodegradation of organic pollutants, including recalcitrant and hazardous polycyclic aromatic hydrocarbons (PAHs). Obtaining PAH-degrading SRB cultures for laboratories is of paramount importance in the development of the young field of anaerobic biodegradation of PAHs. SRB grow exceptionally slowly on PAH substrates and are highly sensitive to oxygen. Consequently, enrichment and maintenance of PAH-degrading SRB cultures and characterization of the biodegradation process remain a tedious and formidable task, especially for new researchers. To address these technical constraints, we have developed robust and effective protocols for obtaining and characterizing PAH-degrading SRB cultures. In this set of protocols, we describe step-by-step procedures for preparing inocula from contaminated soil or sediment, preparing anoxic medium, establishing enrichment cultures with PAHs as substrates under completely anaerobic sulfate-reducing conditions, successive culture transfers to obtain highly enriched cultures, rapid verification of the viability of SRB in slow-growing cultures, assessment of PAH degradation by extracting residuals using organic solvent and subsequent analysis by gas chromatography-mass spectrometry, and spectrophotometric determination of sulfate and sulfide in miniaturized, medium-throughput format. These protocols are expected to serve as a comprehensive manual for obtaining and characterizing PAH-degrading sulfate-reducing cultures. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Obtaining PAH-degrading strictly anaerobic sulfate-reducing enrichment cultures from contaminated soil and sediment Support Protocol 1: Operation and maintenance of an anaerobic workstation Support Protocol 2: Setup of gas purging systems for preparing anoxic solutions Support Protocol 3: Verification of viability in slow-growing SRB enrichment cultures Support Protocol 4: Extraction of genomic DNA from low-biomass cultures Basic Protocol 2: Extraction of residual PAH from liquid culture and analysis by GC-MS Basic Protocol 3: Spectrophotometric determination of sulfate concentration in SRB cultures Basic Protocol 4: Spectrophotometric determination of sulfide concentrations in SRB cultures by the methylene blue method Alternate Protocol: Spectrophotometric determination of sulfide concentrations in SRB cultures by the colloidal copper sulfide method.

Evaluating Natural Source Zone Depletion and Enhanced Source Zone Depletion in laboratory columns via soil redox continuous sensing and microbiome characterization

To optimally employ Natural Source Zone Depletion (NSZD) and Enhanced Source Zone Depletion (ESZD) at sites impacted by light non-aqueous phase liquids (LNAPL), monitoring strategies are required. Emerging use of subsurface oxidation-reduction potential (ORP) sensors shows promise for tracking redox evolution, which reflects ongoing biogeochemical processes. However, further understanding of how soil redox dynamics relate to subsurface microbial activity and LNAPL degradation pathways is needed. In this work, soil ORP sensors and DNA and RNA sequencing-based microbiome analysis were combined to elucidate NSZD and ESZD (biostimulation via periodic sulfate addition and biosparging) processes in columns containing LNAPL-impacted soils from a former petroleum refinery. Results show expected relationships between continuous soil redox and active microbial communities. Continuous data revealed spatial and temporal detail that informed interpretation of the hydrocarbon biodegradation data. Redox increases were transient for sulfate addition, and sequencing revealed how hydrocarbon concentration and composition impacted microbiome structure and naphthalene degradation. Periodic biosparging did not result in fully aerobic conditions suggesting observed biodegradation improvements could be explained by alternative anaerobic metabolisms (e.g., iron reduction due to air oxidizing reduced iron). Collectively, data suggest combining continuous redox sensing with microbiome analysis provides insights beyond those possible with either monitoring tool alone.

Bioremediation of Heavily Contaminated Sandstone from Bida Basin: A pre-emptive model for hydrocarbon exploration in the basin using Bacillus Sustilis and Cereus spp

Petroleum, often known as crude oil, is a complex organic mixture that naturally exists beneath the surface. There is often extreme pollution and other dangerous conditions associated with its exploration, production, refining, storage, and distribution activities. Thus, it is vital to carefully consider the best options after hydrocarbons are found in the Bida Basin, especially in regards to oil spills. In order to prevent this, this study created a preventive model by contaminating samples of sandstone from the Doko Member of the Bida Basin with spent engine oil, which is known to contain a higher percentage of nitrogen and sulfur compounds, aromatic and aliphatic hydrocarbons, and metals like manganese, calcium, zinc, and lead than fresh oils. Two (2) bacterial isolates with a high potential for engine oil biodegradability—Bacillus Subtilis and Cereus—were utilized to assess the effectiveness of total petroleum hydrocarbon (TPH) removal, which is a crucial aspect of crude oil detection. A weekly monitoring program was used to track various physicochemical characteristics of the wasted engine oil, including pH, bacterial biodegradation rate, total petroleum hydrocarbon, nitrate, phosphate, and total organic carbon content. After fourteen and twenty-eight days of contamination, respectively, the test results showed ~50% to <70% biodegradation rate. So, when hydrocarbon development in the Bida Basin begins in earnest, NNPC will have access to first-hand technology for environmental cleanup at a very cheap cost, demonstrating the effectiveness of Bacillus Cereus and substilis as hydrocarbon biodegradation tools.

Determination of soil phenanthrene degradation through a fungal-bacterial consortium.

Fungal-bacterial consortia enhance organic pollutant removal, but the underlying mechanisms are unclear. We used stable isotope probing (SIP) to explore the mechanism of bioaugmentation involved in polycyclic aromatic hydrocarbon (PAH) biodegradation in petroleum-contaminated soil by introducing the indigenous fungal strain Aspergillus sp. LJD-29 and the bacterial strain Pseudomonas XH-1. While each strain alone increased phenanthrene (PHE) degradation, the simultaneous addition of both strains showed no significant enhancement compared to treatment with XH-1 alone. Nonetheless, the assimilation effect of microorganisms on PHE was significantly enhanced. SIP revealed a role of XH-1 in PHE degradation, while the absence of LJD-29 in 13C-DNA indicated a supporting role. The correlations between fungal abundance, degradation efficiency, and soil extracellular enzyme activity indicated that LJD-29, while not directly involved in PHE assimilation, played a crucial role in the breakdown of PHE through extracellular enzymes, facilitating the assimilation of metabolites by bacteria. This observation was substantiated by the results of metabolite analysis. Furthermore, the combination of fungus and bacterium significantly influenced the diversity of PHE degraders. Taken together, this study highlighted the synergistic effects of fungi and bacteria in PAH degradation, revealed a new fungal-bacterial bioaugmentation mechanism and diversity of PAH-degrading microorganisms, and provided insights for in situ bioremediation of PAH-contaminated soil.IMPORTANCEThis study was performed to explore the mechanism of bioaugmentation by a fungal-bacterial consortium for phenanthrene (PHE) degradation in petroleum-contaminated soil. Using the indigenous fungal strain Aspergillus sp. LJD-29 and bacterial strain Pseudomonas XH-1, we performed stable isotope probing (SIP) to trace active PHE-degrading microorganisms. While inoculation of either organism alone significantly enhanced PHE degradation, the simultaneous addition of both strains revealed complex interactions. The efficiency plateaued, highlighting the nuanced microbial interactions. SIP identified XH-1 as the primary contributor to in situ PHE degradation, in contrast to the limited role of LJD-29. Correlations between fungal abundance, degradation efficiency, and extracellular enzyme activity underscored the pivotal role of LJD-29 in enzymatically facilitating PHE breakdown and enriching bacterial assimilation. Metabolite analysis validated this synergy, unveiling distinct biodegradation mechanisms. Furthermore, this fungal-bacterial alliance significantly impacted PHE-degrading microorganism diversity. These findings advance our understanding of fungal-bacterial bioaugmentation and microorganism diversity in polycyclic aromatic hydrocarbon (PAH) degradation as well as providing insights for theoretical guidance in the in situ bioremediation of PAH-contaminated soil.

Biodegradation of high molecular weight polycyclic aromatic hydrocarbons by Sarocladium terricola strain PYR-233 isolated from petrochemical contaminated sediment

Polycyclic aromatic hydrocarbons (PAHs) were frequently found in sediment and were primarily treated through microbial degradation. Thus, efficient management of PAH pollution requires exploring the molecular degradation mechanisms of PAHs and expanding the pool of available microbial resources. A fungus (identified as Sarocladium terricola strain RCEF778) with the remarkable ability to degrade pyrene was screened from sediment near a petrochemical plant, and its growth and pyrene degradation characteristics were comprehensively investigated. The results showed that the fungus exhibited great effectiveness in pyrene degradation, with a degradation ratio of 88.97% at 21 days at the conditions: 35 °C, pH 7, 10 mg L−1 initially pyrene concentration, 3% supplementary salt, and glucose supplementation. The generation and concentration variation of the intermediate products were identified, and the results revealed that the fungus degraded pyrene through two pathways: by salicylic acid and by phthalic acid. Three sediments (M1, M2, M3), each exhibiting different levels of PAH pollution, were employed to examine the effectiveness of fungal degradation of PAHs in practical sediment samples. These data showed that with the fungus, the degradation ratios ranged from 13.64% to 23.50% for 2–3 rings PAHs, 40.93%–49.41% for 4 rings PAHs, and 39.59%–48.07% for 5–6 rings PAHs, which were significantly higher than those for the sediment without the fungus and confirmed the excellent performance of the fungal. Moreover, the Gompertz model was employed to analyze the degradation kinetics of 4-rings and 5–6 rings PAHs in these sediments, and the results demonstrated that the addition of the fungus could significantly increase the maximum degradation ratio, degradation start-up rate and maximum degradation rate of 4-rings and 5–6 rings PAHs and shorten the time required to reach the maximum degradation rate. This study not only supplied fungal materials but also established crucial theoretical foundations for the development of bioremediation technologies aimed at high molecular weight PAH-contaminated sediments.

PAHs Biodegradation by Locally Isolated Phanerochaete chrysosporium and Penicillium citrinum from Liquid and Spiked Soil

In the present study, biodegradation of polycyclic aromatic hydrocarbons (PAHs) was examined using two fungal strains, namely P. chrysosporium and P. citrinum, isolated from locally contaminated soil. These two fungal strains were compared based on degradation properties under standardized conditions (pH 7.0, temperature 30oC, carbon source yeast extract) using PAH sole and a mixture of five different PAHs. In liquid media, PAH degradation was higher as compared to spiked soil by P. chrysosporium, followed by P. citrinum. In liquid culture, maximum degradation was 96.13% phenanathrene, 86.34% fluoranthene, 72.75% pyrene, 52.25% chrysene, and 40.16% benzo(a)pyrene by P. chrysosporium. PAH degradation in spiked soil was 78.5% phenanthrene, 65.91% fluoranthene, 61.73% pyrene, 48.2% chrysene, and 26.82% benzo(a)pyrene within 28 days by P. chrysosporium. Both local fungal isolates showed potential for degradation of PAHs alone and in PAH mixtures.

REMEDIATION OF OIL-CONTAMINATED SOILS WITH COMPOSITIONS BASED ON HUMIC ACIDS

In addition to conventional methods, nowadays new approaches and technologies based on combined application of sorbents and microorganisms are intensively developed for efficient control of oil contaminations. As a biotechnological solution for remediation of various oil-contaminated soils, biocompositions based on an organic matrix – humic acids of peat, and a biological component – a single strain/or an association of oil degrading microorganisms of the genera Rhodococcus and Pseudomonas, have been studied. The efficiency assessment of oil contamination biodegradation using biocompositions was carried out on different types of model oil-contaminated soils with different levels of contamination: grey forest soil, Black Sea pebbles (fraction 20-50mm), constructional broken stone (fraction 10-20mm). Natural soil contaminated with oil products and samples of railroad broken stone (fractions 5-30mm and 35-50mm) were also used. It has been shown that the biocompositions based on three strains of microorganisms – ‘HA R. X5 R. S67 Ps. NF142' – possessed the highest capability of degrading oil hydrocarbons in all systems under study. Biodegradation of oil hydrocarbons upon addition of this biocompositions varied between 62 and 82% depending on the type of oil-contaminated soil, the level of contamination being very high. Monobacterial biocomposition ‘HA Ps.NF142’ has proven to be promising for remediation of oil-contaminated soil ecosystems with a high and very high level of contamination. The increase of the level of biodegradation of oil contamination of soils appears to be due to the combined action of humic acids comprising both the stimulating effect of humic acids on microorganisms – the components of biocompositions and indigenous microflora of model systems, and the detoxifying effect of humic acids in respect of oil hydrocarbons. For citation: Grechishcheva N.Yu., Dmitrieva E.D., Starodubtseva K.A. Remediation of oil-contaminated soils with compositions based on humic acids. ChemChemTech [Izv. Vyssh. Uchebn. Zaved. Khim. Khim. Tekhnol.]. 2024. V. 67. N 7. P. 136-143. DOI: 10.6060/ivkkt.20246707.6927.

Assessment of Biosurfactant Production and Petroleum Hydrocarbons Biodegradation Capability of Actinomycetes Isolated from Soils

Assessment of Biosurfactant Production and Petroleum Hydrocarbons Biodegradation Capability of Actinomycetes Isolated from Soils

Novel enzymes for biodegradation of polycyclic aromatic hydrocarbons identified by metagenomics and functional analysis in short-term soil microcosm experiments

Polycyclic aromatic hydrocarbons (PAHs) are highly toxic, carcinogenic substances. On soils contaminated with PAHs, crop cultivation, animal husbandry and even the survival of microflora in the soil are greatly perturbed, depending on the degree of contamination. Most microorganisms cannot tolerate PAH-contaminated soils, however, some microbial strains can adapt to these harsh conditions and survive on contaminated soils. Analysis of the metagenomes of contaminated environmental samples may lead to discovery of PAH-degrading enzymes suitable for green biotechnology methodologies ranging from biocatalysis to pollution control. In the present study, our goal was to apply a metagenomic data search to identify efficient novel enzymes in remediation of PAH-contaminated soils. The metagenomic hits were further analyzed using a set of bioinformatics tools to select protein sequences predicted to encode well-folded soluble enzymes. Three novel enzymes (two dioxygenases and one peroxidase) were cloned and used in soil remediation microcosms experiments. The experimental design of the present study aimed at evaluating the effectiveness of the novel enzymes on short-term PAH degradation in the soil microcosmos model. The novel enzymes were found to be efficient for degradation of naphthalene and phenanthrene. Adding the inorganic oxidant CaO2 further increased the degrading potential of the novel enzymes for anthracene and pyrene. We conclude that metagenome mining paired with bioinformatic predictions, structural modelling and functional assays constitutes a powerful approach towards novel enzymes for soil remediation.

Synthesis and characterization of iron oxide nanoparticles from Lawsonia inermis and its effect on the biodegradation of crude oil hydrocarbon.

Crude oil hydrocarbons are considered major environmental pollutants and pose a significant threat to the environment and humans due to having severe carcinogenic and mutagenic effects. Bioremediation is one of the practical and promising technology that can be applied to treat the hydrocarbon-polluted environment. In this present study, rhamnolipid biosurfactant (BS) produced by Pseudomonas aeruginosa PP4 and green synthesized iron nanoparticles (G-FeNPs) from Lawsonia inermis was used to evaluate the biodegradation efficiency (BE) of crude oil. The surface analysis of G-FeNPs was carried out by using FESEM and HRTEM to confirm the size and shape. Further, the average size of the G-FeNPs was observed around 10nm by HRTEM analysis. The XRD and Raman spectra strongly confirm the presence of iron nanoparticles with their respective peaks. The BE (%) of mixed degradation system-V (PP4+BS+G-FeNPs) was obtained about 82%. FTIR spectrum confirms the presence of major functional constituents (C=O, -CH3, C-O, and OH) in the residual oil content. Overall, this study illustrates that integrated nano-based bioremediation could be an efficient approach for hydrocarbon-polluted environments. This study is the first attempt to evaluate the G-FeNPs with rhamnolipid biosurfactant on the biodegradation of crude oil.

A combination of microbial electrolysis cells and bioaugmentation can effectively treat synthetic wastewater containing polycyclic aromatic hydrocarbon.

The anaerobic biodegradation of polycyclic aromatic hydrocarbons (PAHs) is challenging due to its toxic effect on the microbes. Microbial electrolysis cells (MECs), with their excellent characteristics of anodic and cathodic biofilms, can be a viable way to enhance the biodegradation of PAHs. This work assessed different cathode materials (carbon brush and nickel foam) combined with bioaugmentation on typical PAHs-naphthalene biodegradation and analyzed the inhibition amendment mechanism of microbial biofilms in MECs. Compared with the control, the degradation efficiency of naphthalene with the nickel foam cathode supplied with bioaugmentation dosage realized a maximum removal rate of 94.5 ± 3.2%. The highest daily recovered methane yield (227 ± 2 mL/gCOD) was also found in the nickel foam cathode supplied with bioaugmentation. Moreover, the microbial analysis demonstrated the significant switch of predominant PAH-degrading microorganisms from Pseudomonas in control to norank_f_Prolixibacteraceae in MECs. Furthermore, hydrogentrophic methanogenesis prevailed in MEC reactors, which is responsible for methane production. This study proved that MEC combined with bioaugmentation could effectively alleviate the inhibition of PAH, with the nickel foam cathode obtaining the fastest recovery rate in terms of methane yield.

Importance of soil ecoenzyme stoichiometry for efficient polycyclic aromatic hydrocarbon biodegradation

Efficient remediation of soil contaminated by polycyclic aromatic hydrocarbons (PAHs) is challenging. To determine whether soil ecoenzyme stoichiometry influences PAH degradation under biostimulation and bioaugmentation, this study initially characterized soil ecoenzyme stoichiometry via a PAH degradation experiment and subsequently designed a validation experiment to answer this question. The results showed that inoculation of PAH degradation consortia ZY-PHE plus vanillate efficiently degraded phenanthrene with a K value of 0.471 (depending on first-order kinetics), followed by treatment with ZY-PHE and control. Ecoenzyme stoichiometry data revealed that the EEAC:N, vector length and angle increased before day five and decreased during the degradation process. In contrast, EEAN:P decreased and then increased. These results indicated that the rapid PAH degradation period induced more C limitation and organic P mineralization. Correlation analysis indicated that the degradation rate K was negatively correlated with vector length, EEAC:P, and EEAN:P, suggesting that C limitation and relatively less efficient P mineralization could inhibit biodegradation. Therefore, incorporating liable carbon and acid phosphatase or soluble P promoted PAH degradation in soils with ZY-PHE. This study provides novel insights into the relationship between soil ecoenzyme stoichiometry and PAH degradation. It is suggested that soil ecoenzyme stoichiometry be evaluated before designing bioremeiation stragtegies for PAH contanminated soils.

Symbiotic virus-bacteria interactions in biological treatment of coking wastewater manipulating bacterial physiological activities

Biological treatment is commonly used in coking wastewater (CWW) treatment. Prokaryotic microbial communities in CWW treatment have been comprehensively studied. However, viruses, as the critical microorganisms affecting microbial processes and thus engineering parameters, still remain poorly understood in CWW treatment context. Employing viromics sequencing, the composition and function of the viral community in CWW treatment were discovered, revealing novel viral communities and key auxiliary metabolic functions. Caudovirales appeared to be the predominant viral order in the oxic-hydrolytic-oxic (OHO) CWW treatment combination, showing relative abundances of 62.47 %, 56.64 % and 92.20 % in bioreactors O1, H and O2, respectively. At the family level, Myoviridae, Podoviridae and Siphoviridae mainly prevailed in bioreactors O1 and H while Phycodnaviridae dominated in O2. A total of 56.23–92.24% of novel viral contigs defied family-level characterization in this distinct CWW habitat. The virus-host prediction results revealed most viruses infecting the specific functional taxa Pseudomonas, Acidovorax and Thauera in the entire OHO combination, demonstrating the viruses affecting bacterial physiology and pollutants removal from CWW. Viral auxiliary metabolic genes (AMGs) were screened, revealing their involvement in the metabolism of contaminants and toxicity tolerance. In the bioreactor O1, AMGs were enriched in detoxification and phosphorus ingestion, where glutathione S-transferase (GSTs) and beta-ketoadipyl CoA thiolase (fadA) participated in biodegradation of polycyclic aromatic hydrocarbons and phenols, respectively. In the bioreactors H and O2, the AMGs focused on cell division and epicyte formation of the hosts, where GDPmannose 4,6-dehydratase (gmd) related to lipopolysaccharides biosynthesis was considered to play an important role in the growth of nitrifiers. The diversities of viruses and AMGs decreased along the CWW treatment process, pointing to a reinforced virus-host adaptive strategy in stressful operation environments. In this study, the symbiotic virus-bacteria interaction patterns were proposed with a theoretical basis for promoting CWW biological treatment efficiency. The findings filled the gaps in the virus-bacteria interactions at the full-scale CWW treatment and provided great value for understanding the mechanism of biological toxicity and sludge activity in industrial wastewater treatment.

Evaluation of surfactant-aided polycyclic aromatic hydrocarbon biodegradation by molecular docking and molecular dynamic simulation in the marine environment

Marine oil spills directly cause polycyclic aromatic hydrocarbons (PAHs) pollution and affect marine organisms due to their toxic property. Chemical and bio-based dispersants composed of surfactants and solvents are considered effective oil spill-treating agents. Dispersants enhance oil biodegradation in the marine environment by rapidly increasing their solubility in the water column. However, the effect of dispersants, especially surfactants, on PAHs degradation by enzymes produced by microorganisms has not been studied at the molecular level. The role of the cytochrome P450 (CYP) enzyme in converting contaminants into reactive metabolites during the biodegradation process has been evidenced, but the activity in the presence of surfactants is still ambiguous. Thus, this study focused on the evaluation of the impact of chemical and bio-surfactants (i.e., Tween 80 (TWE) and Surfactin (SUC)) on the biodegradation of naphthalene (NAP), chrysene (CHR), and pyrene (PYR), the representative components of PAHs, with CYP enzyme from microalgae Parachlorella kessleri using molecular docking and molecular dynamics (MD) simulation. The molecular docking analysis revealed that PAHs bound to residues at the CYP active site through hydrophobic interactions for biodegradation. The MD simulation showed that the surfactant addition changed the enzyme conformation in the CYP-PAH complexes to provide more interactions between the enzyme and PAHs. This led to an increase in the enzyme's capability to degrade PAHs. Binding free energy (ΔG‌‌Bind) calculations confirmed that surfactant treatment could enhance PAHs degradation by the enzyme. The SUC gave a better result on NAP and PYR biodegradation based on ΔG‌‌Bind, while TWE facilitated the biodegradation of CHR. The research outputs could greatly facilitate evaluating the behaviors of oil spill-treating agents and oil spill response operations in the marine environment.

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.