Degradation Of Petroleum Hydrocarbons Research Articles

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 1689.2mg/L. At a critical micelle concentration of 200mg/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.

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.

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.

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.

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.

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.

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.

Differential degradation of petroleum hydrocarbons by Shewanella putrefaciens under aerobic and anaerobic conditions.

The complexity of crude oil composition, combined with the fluctuating oxygen level in contaminated environments, poses challenges for the bioremediation of oil pollutants, because of compound-specific microbial degradation of petroleum hydrocarbons under certain conditions. As a result, facultative bacteria capable of breaking down petroleum hydrocarbons under both aerobic and anaerobic conditions are presumably effective, however, this hypothesis has not been directly tested. In the current investigation, Shewanella putrefaciens CN32, a facultative anaerobic bacterium, was used to degrade petroleum hydrocarbons aerobically (using O2 as an electron acceptor) and anaerobically (using Fe(III) as an electron acceptor). Under aerobic conditions, CN32 degraded more saturates (65.65 ± 0.01%) than aromatics (43.86 ± 0.03%), with the following order of degradation: dibenzofurans > n-alkanes > biphenyls > fluorenes > naphthalenes > alkylcyclohexanes > dibenzothiophenes > phenanthrenes. In contrast, under anaerobic conditions, CN32 exhibited a higher degradation of aromatics (53.94 ± 0.02%) than saturates (23.36 ± 0.01%), with the following order of degradation: dibenzofurans > fluorenes > biphenyls > naphthalenes > dibenzothiophenes > phenanthrenes > n-alkanes > alkylcyclohexanes. The upregulation of 4-hydroxy-3-polyprenylbenzoate decarboxylase (ubiD), which plays a crucial role in breaking down resistant aromatic compounds, was correlated with the anaerobic degradation of aromatics. At the molecular level, CN32 exhibited a higher efficiency in degrading n-alkanes with low and high carbon numbers relative to those with medium carbon chain lengths. In addition, the degradation of polycyclic aromatic hydrocarbons (PAHs) under both aerobic and anaerobic conditions became increasingly difficult with increased numbers of benzene rings and methyl groups. This study offers a potential solution for the development of targeted remediation of pollutants under oscillating redox conditions.

Biosurfactant production by a crude oil‐utilizing bacterium towards petroleum hydrocarbon biodegradation applications

AbstractBiosurfactants produced by microorganisms are green amphiphilic biomolecules. They have numerous advantages compared to chemical surfactants including a lower toxicity, better environmental compatibility and effective and stable properties under extreme conditions such as a wide range of temperature, pH and salinity. A highly bio‐surfactant‐producing strain of Bacillus, VTVK15 was selected among the Bacillus isolates from marine environment and oil contaminated sites in Ba Ria‐Vung Tau. Sequence analysis of 16S rDNA showed that the VTVK15 strain was 99% similar with 16s rDNA sequence of Bacillus megaterium. The suitable conditions for the bio‐surfactant production by the strain VTVK15 were 37 °C, pH 7; 2% w/v and 0.4% w/v for temperature, initial solution pH, concentration of carbon substrate (crude oil), and concentration of nitrogen substrate ((NH4)2SO4), respectively. The emulsification index (E24) increased from 51.8% to 67.7% with suitable conditions. The petroleum hydrocarbon degradation (TPH) efficiency by the strain Bacillus megaterium VTVK15 in crude oil estimated using TPH analysis was 72% after 14 days. These results revealed that the strain VTVK15 exhibited a tremendous potential for bioremediation of petroleum hydrocarbon contaminants.

Effects of biochar immobilization of Serratia sp. F4 OR414381 on bioremediation of petroleum contamination and bacterial community composition in loess soil

Petroleum hydrocarbons pose a significant threat to human health and the environment. Biochar has increasingly been utilized for soil remediation. This study investigated the potential of biochar immobilization using Serratia sp. F4 OR414381 for the remediation of petroleum-contaminated soil through a pot experiment conducted over 90 days. The treatments in this study, denoted as IMs (maize straw biochar-immobilized Serratia sp. F4), degraded 82.5% of the total petroleum hydrocarbons (TPH), 59.23% of the aromatic, and 90.1% of the saturated hydrocarbon fractions in the loess soils. During remediation, the soil pH values decreased from 8.76 to 7.33, and the oxidation-reduction potential (ORP) increased from 156 to 229 mV. The treatment-maintained soil nutrients of the IMs were 138.94 mg/kg of NO3- -N and 92.47 mg/kg of available phosphorus (AP), as well as 11.29% of moisture content. The activities of soil dehydrogenase (SDHA) and catalase (CAT) respectively increased by 14% and 15 times compared to the CK treatment. Three key petroleum hydrocarbon degradation genes, including CYP450, AJ025, and xylX were upregulated following IMs treatment. Microbial community analysis revealed that a substantial microbial population of 1.01E+ 09 cells/g soil and oil-degrading bacteria such as Salinimicrobium, Saccharibacteria_genera_incertae_sedis, and Brevundimonas were the dominant genera in IMs treatment. This suggests that the biochar immobilized on Serratia sp. F4 OR414381 improves soil physicochemical properties and enhances interactions among microbial populations, presenting a promising and environmentally friendly approach for the stable and efficient remediation of petroleum-contaminated loess soil.

Activation of persulfate by g-C3N4/nZVI@SBC for degradation of total petroleum hydrocarbon in groundwater

In this study, we synthesized a high removal efficiency catalyst using biochar-supported nanoscale zero-valent iron and g-C3N4, denoted as g-C3N4/nZVI@SBC, to activate persulfate (PS) for the degradation of total petroleum hydrocarbon (TPH) in groundwater. We characterized the morphology and physiochemical properties of g-C3N4/nZVI@SBC with scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform infrared (FTIR), BET surface area analysis, and X-ray photoelectron spectroscopy (XPS). To assess the performance of the g-C3N4/nZVI@SBC catalyst, we investigated various reaction parameters, such as the mass ratio of g-C3N4 to nZVI@SBC, PS concentration, initial pH, initial TPH concentration, and the presence of coexisting ions in the system. The results from batch experiments and repeated use trials indicate that g-C3N4/nZVI@SBC exhibited both excellent catalytic activation capability and impressive durability, making it a promising choice for TPH degradation. Specifically, when the PS concentration reached 1 mM, the catalyst dosage was 0.3 g/L, and the g-C3N4 to nZVI@SBC mass ratio was 2, we achieved a remarkable TPH removal efficiency of 93.8%. Through electron paramagnetic resonance (EPR) testing and quenching experiments, we identified sulfate radicals, hydroxyl radicals, and superoxide radicals as the primary active substance involved in the TPH degradation process. Moreover, the g-C3N4/nZVI@SBC composite proved highly effective for in-situ TPH removal from groundwater and displayed an 86% removal rate, making it a valuable candidate for applications in permeable reactive barriers (PRB) aimed at enhancing environmental remediation. In summary, by skillfully utilizing g-C3N4/nZVI@SBC, this study has made notable advancements in synthesis and characterization, presenting a feasible and innovative approach to addressing TPH pollution in groundwater.

The Effect of Pyrolysis Temperature of calotropis procera Biochar on Dynamics of Petroleum Hydrocarbons Degradation, Carbon Emission, and Ammonia Volatilization in Artificial Petroleum-Contaminated Soil

AbstractPollution by petroleum hydrocarbons (PHCs) is a current environmental challenge all over the world. Therefore, this study was established to evaluate the impact of calotropis procera biochar (CPB) produced at various pyrolysis temperatures on carbon emission, ammonia volatilization, degradation of PHCs, and properties of artificial petroleum-contaminated soil. An incubation experiment was performed with CPB produced at 400 °C (CPB400) and 650 °C (CPB650). The artificial petroleum-contaminated soil was amended with biochar at doses of 1, 2.5, and 5% along with the unamended soil (control) and incubated for 78 days. The results demonstrate that the CO2-C emission rate in petroleum-contaminated soil increases significantly with all biochar applications compared to the control treatment. Adding CPB400 into the soil, at all doses, decreased cumulative NH3 emission significantly, in comparison with control and CPB650 treatments during all incubation periods. Total PHCs concentration in the soil reduced significantly with CPB400 application at 2.5% compared to the control treatment. CPB400 was more effective in declining total PHCs content in the soil than biochar produced at 650 °C. Available phosphorus and potassium in petroleum-contaminated soil increased significantly under biochar applications compared with the control treatment. Using Calotropis procera biochar produced by low-temperature pyrolysis as an amendment can play a vital role in the remediation of PHCs in polluted soils, it is also a promising way to improve the quality indicators of petroleum-contaminated soil.

Biodegradation of petroleum hydrocarbons using a novel bacterial strain isolated from hydrocarbons contaminated soil of Saudi Arabia

Petroleum spills pose a significant environmental threat, underscoring the need for efficient pollution removal methods. Bioremediation, employing bacterial cultures, emerges as a cost-effective and eco-friendly technique. The success of this method hinges on whether petroleum pollution is eliminated or converted into a harmless form. This study seeks to isolate a bacterial strain exhibiting excellent capacity for degrading petroleum hydrocarbons. Biochemical tests and 16S rRNA results identified various Microbacterium spp., namely Microbacterium barkeri (SA9) and Microbacterium deserti (SA2, SA3, SA5, and SA7), isolated from hydrocarbon-contaminated soil. Within five days, these strains demonstrated degradation percentages of 20.91%, 38.51%, 77.68%, 22.51%, and 31.11% for petroleum, respectively. Phylogenetic and sequence analyses targeting the 16S rRNA gene revealed that M. barkeri (accession number ON 421582) is closely related to strains from India and Saudi Arabia, while M. deserti (ON 421525, ON 421524, ON 421523, and ON 421522) shares affinity with strains from Saudi Arabia, China, and India. Furthermore, these isolates produced a stable emulsion with E−24% of 25% for 72 h. Notably, this marks the first study exploring the use of these bacterial strains for petroleum bioremediation in Saudi Arabia.

Screening biosurfactant-producing actinomycetes: Identification of Streptomyces sp. RP1 as a potent species for bioremediation.

This study aimed to isolate biosurfactant-producing and hydrocarbon-degrading actinomycetes from different soils using glycerol-asparagine and starch-casein media with an antifungal agent. The glycerol-asparagine agar exhibited the highest number of actinomycetes, with a white, low-opacity medium supporting pigment production and high growth. Biosurfactant analyses, such as drop collapse, oil displacement, emulsification, tributyrin agar test, and surface tension measurement, were conducted. Out of 25 positive isolates, seven could utilize both olive oil and black oil for biosurfactant production, and only isolate RP1 could produce biosurfactant when grown in constrained conditions with black oil as the sole carbon source and inducer, demonstrating in situ bioremediation potential. Isolate RP1 from oil-spilled garden soil is Gram-staining-positive with a distinct earthy odor, melanin formation, and white filamentous colonies. It has a molecular size of ~621 bp and 100% sequence similarity to many Streptomyces spp. Morphological, biochemical, and 16 S rRNA analysis confirmed it as Streptomyces sp. RP1, showing positive results in all screenings, including high emulsification activity against kerosene (27.2%) and engine oil (95.8%), oil displacement efficiency against crude oil (7.45 cm), and a significant reduction in surface tension (56.7 dynes/cm). Streptomyces sp. RP1 can utilize citrate as a carbon source, tolerate sodium chloride, resist lysozyme, degrade petroleum hydrocarbons, and produce biosurfactant at 37°C in a 15 mL medium culture, indicating great potential for bioremediation and various downstream industrial applications with optimization.

Construction of Shale Gas Oil-Based Drilling Cuttings Degrading Bacterial Consortium and Their Degradation Characteristics.

Oil-based drilling cuttings (OBDCs) contain petroleum hydrocarbons with complex compositions and high concentrations, which have highly carcinogenic, teratogenic, and mutagenic properties. In this study, three highly efficient petroleum hydrocarbon-degrading bacteria were screened from OBDCs of different shale gas wells in Chongqing, China, and identified as Rhodococcus sp. and Dietzia sp. Because of their ability to degrade hydrocarbons of various chain lengths, a new method was proposed for degrading petroleum hydrocarbons in shale gas OBDCs by combining different bacterial species. Results showed that the bacterial consortium, consisting of the three strains, exhibited the highest degradation rate for petroleum hydrocarbons, capable of degrading 74.38% of long-chain alkanes and 93.57% of short-chain alkanes, respectively. Moreover, the petroleum hydrocarbon degradation performance of the bacterial consortium in actual OBDCs could reach 90.60% in the optimal conditions, and the degradation kinetic process followed a first-order kinetic model. This study provides a certain technical reserve for the bioremediation of shale gas OBDCs.

Unravelling Diatoms’ Potential for the Bioremediation of Oil Hydrocarbons in Marine Environments

The search for practical solutions to alleviate the destructive impact of petroleum hydrocarbons in marine environments is contributing to the implementation of prospecting strategies for indigenous microorganisms with biodegradative and bioremediation potential. The levels of petroleum contamination entering the marine environment each year have been estimated at around 1.3 million tonnes, a figure that is expected to increase by 1.9% annually over the next decade. The recent interest in decarbonizing our energy system and accelerating the clean energy transition has created a demand for greener technologies and strategies to find innovative, sustainable, and cost-effective treatments for the marine environment. Diatoms (Bacillariophyta) are one of the most diverse and successful taxa in coastal–marine environments and are a relatively untapped pool of biodiversity for biotechnological applications. Recent reports have revealed the significant presence of diatoms associated with oil spills and petroleum hydrocarbon degradation. Most diatoms can secrete substantial amounts of exopolysaccharides (EPSs) into their environment, which can act as biosurfactants that, in addition to oxygen and other enzymes produced by diatoms, create suitable conditions to enhance hydrocarbon solubility and degradation into less toxic compounds in seawater. Recent reports on the biodegradation of aliphatic and aromatic hydrocarbons by diatoms are indicative of the potential of these taxa to achieve success in the bioremediation of hydrocarbons in marine environments. This review highlights the main attributes and roles that diatoms could play in integrated strategies for biodegradation and bioremediation of petroleum hydrocarbon pollutants and as such represent a green, eco-friendly, and sustainable contribution to mitigate damage to biodiversity and value chains of marine ecosystems.

Bio-augmentation and bio-stimulation with kenaf core enhanced bacterial enzyme activities during bio-degradation of petroleum hydrocarbon in polluted soil

Indigenous micro-organisms often possess the ability to degrade petroleum hydrocarbon (PHC) in polluted soil. However, this process can be improved by supplementing with nutrients or the addition of more potent microbes. In this study, the ability of kenaf-core to stimulate the PHC degradation capability of microbial isolates from PHC polluted soil samples was evaluated. The standard experimental methods used in this study include: the digestion and analysis of the physico-chemical properties of petroleum hydrocarbon contaminated and non-contaminated soil samples; evaluation of petroleum hydrocarbon biodegradation using bio-augmentation and bio-stimulation (with kenaf-core) treatments; and, determination of soil microbial enzyme activities. Results from this study show that K, Na, total nitrogen, organic carbon, exchangeable cations, and heavy metals were found to be significantly (P < 0.05) higher in the polluted soil than in the non-polluted soil. Also, the polluted samples had pH values ranging from 5.5 to 6.0 while the non-polluted samples had a pH of 7.6. The microbial enzyme activities were comparatively lower in the polluted soils as compared to the non-polluted soil. The percentage degradation in the kenaf-core treated samples (AZ1T2—78.38; BN3T2—70.69; OL1T2—71.06; OT1T2—70.10) were significantly (P < 0.05) higher than those of the untreated (AZ1T1—13.50; BN3T1—12.50; OL1T1—10.55; OT1T1—9.50). The degradation of petroleum hydrocarbon in the bio-augmented and bio-stimulated treatments increased with increasing time of incubation, and were higher than that of the untreated sample. Comparatively, the treatment with a combination of kenaf-core and rhamnolipid exhibited a significantly (P < 0.05) higher degradation rate than that of the treatment with only kenaf core or rhamnolipid. While, the bio-stimulated and bio-augmented treatments had appreciable microbial counts that are higher than that of the untreated. In conclusion, the nutrient-supplement with kenaf-core significantly enhanced microbial growth and activities in the soil, thus improving their ability to biodegrade petroleum hydrocarbons in the polluted soils. Thus, supplementing with Kenaf core to encourage microbiological degradation of petroleum hydrocarbon is recommended.

Diesel degradation capability and environmental robustness of strain Pseudomonas aeruginosa WS02

Petroleum hydrocarbon (PHC) degrading bacteria have been frequently discovered. However, in practical application, a single species of PHC degrading bacterium with weak competitiveness may face environmental pressure and competitive exclusion due to the interspecific competition between petroleum-degrading bacteria as well as indigenous microbiota in soil, leading to a reduced efficacy or even malfunction. In this study, the diesel degradation ability and environmental robustness of an endophytic strain Pseudomonas aeruginosa WS02, were investigated. The results show that the cell membrane surface of WS02 was highly hydrophobic, and the strain secreted glycolipid surfactants. Genetic analysis results revealed that WS02 contained multiple metabolic systems and PHC degradation-related genes, indicating that this strain theoretically possesses the capability of oxidizing both alkanes and aromatic hydrocarbons. Gene annotation also showed many targets which coded for heavy metal resistant and metal transporter proteins. The gene annotation-based inference was confirmed by the experimental results: GC-MS analysis revealed that short chain PHCs (C10–C14) were completely degraded, and the degradation of PHCs ranging from C15–C22 were above 90% after 14 d in diesel-exposed culture; Heavy metal (Mn2+, Pb2+ and Zn2+) exposure was found to affect the growth of WS02 to some extent, but not its ability to degrade diesel, and the degradation efficiency was still maintained at 39–59%. WS02 also showed a environmental robustness along with PHC-degradation performance in the co-culture system with other bacterial strains as well as in the co-cultured system with the indigenous microbiota in soil fluid extracted from a PHC-contaminated site. It can be concluded that the broad-spectrum diesel degradation efficacy and great environmental robustness give P. aeruginosa WS02 great potential for application in the remediation of PHC-contaminated soil.

Degrading characteristics of oil‐degrading bacteria and its study of petroleum hydrocarbon metabolites

AbstractExploring the community characteristics of petroleum degrading bacteria and revealing the types of petroleum metabolites produced by degrading bacteria play an important role in solving the problem of oil pollution. In this study, the bacterial suspension was extracted from the sludge of the sewage treatment plant, and the efficient dominant bacteria were enriched and cultured by microbial screening. The optimal environmental conditions and kinetic behavior of microbial degradation of petroleum hydrocarbons in groundwater were discussed, and the metabolites of microbial degradation of petroleum hydrocarbons were analyzed. The results showed that the optimum degradation conditions of the strain were as follows: bacterial suspension inoculum: 2%, pH: 7, temperature: 30°C, initial concentration of contaminated water sample: 500 mg/L. Under these conditions, the microbial petroleum hydrocarbon degradation efficiency was 84.7% and followed the first‐order kinetics; qualitative and quantitative statistical analysis of the metabolites of the microbial degradation of petroleum hydrocarbons by gas chromatography‐time‐of‐flight mass spectrometry was performed. A total of 10 significantly upregulated products and 12 significantly downregulated products were screened, which were significantly different from the metabolites of the control group. Further microbial community analysis showed that Pseudomonas was dominant in the bacterial suspension, more than 99.5%, which was significantly different from the sludge sample, providing data support for the degradation of petroleum hydrocarbons by Pseudomonas. This study has provided a scientific basis for in situ remediation of petroleum pollution in groundwater.

Draft Genome of Nocardia canadensis sp. nov. Isolated from Petroleum-Hydrocarbon-Contaminated Soil.

The bacterial strain WB46 was isolated from the rhizosphere of willow plants (Salix purpurea L.) growing in soil contaminated with petroleum hydrocarbons. The strain was subjected to whole-genome shotgun sequencing using Illumina HiSeq. Its draft genome is 7.15 Mb, with a 69.55% GC content, containing 6387 protein-coding genes and 51 tRNA and 15 rRNA sequences. The quality and reliability of the genome were assessed using CheckM, attaining an estimated genome completeness of 98.75% and an estimated contamination of 1.68%. These results indicate a high-quality genome (>95%) and low contamination (<5%). Many of these genes are responsible for petroleum hydrocarbon degradation, such as alkane 1-monooxygenase (alkB) and naphthalene dioxygenase (ndo). 16S rRNA gene analysis, including in silico DNA-DNA hybridization (DDH) and average nucleotide identity (ANI), showed that strain WB46 belongs to the genus Nocardia, and the most closely related species is Nocardia asteroides. The strain WB46 showed a distance of 63.4% and sequence identity of 88.63%, respectively. These values fall below the threshold levels of 70% and 95%, respectively, suggesting that the strain WB46 is a new species. We propose the name of Nocardia canadensis sp. nov. for this new species. Interestingly, the sequence divergence of the 16S rRNA gene showed that the divergence only occurred in the V2 region. Therefore, the conventional V3-V4, V5-V7, or V8-V9 targeting metabarcoding, among others, would not be able to assess the diversity related to this new species.