Petroleum-contaminated Soil Research Articles

Oxygen atmosphere enhances ball milling remediation of petroleum-contaminated soil and reuse as adsorptive/catalytic materials for wastewater treatment

Ball milling is an environmentally friendly technology for the remediation of petroleum-contaminated soil (PCS), but the cleanup of organic pollutants requires a long time, and the post-remediation soil needs an economically viable disposal/reuse strategy due to its vast volume. The present paper develops a ball milling process under oxygen atmosphere to enhance PCS remediation and reuse the obtained carbonized soil (BCS-O) as wastewater treatment materials. The total petroleum hydrocarbon removal rates by ball milling under vacuum, air, and oxygen atmospheres are 39.83%, 55.21%, and 93.84%, respectively. The Langmuir and pseudo second-order models satisfactorily describe the adsorption capacity and behavior of BCS-O for transition metals. The Cu2+, Ni2+, and Mn2+ adsorbed onto BCS-O were mainly bound to metal carbonates and metal oxides. Furthermore, BCS-O can effectively activate persulfate (PDS) oxidation to degrade aniline, while BCS-O loaded with transition metal (BCS-O-Me) shows better activation efficiency and reusability. BCS-O and BCS-O-Me activated PDS oxidation systems are dominated by 1O2 oxidation and electron transfer. The main active sites are oxygen-containing functional groups, vacancy defects, and graphitized carbon. The oxygen-containing functional groups and vacancy defects primarily activate PDS to generate 1O2 and attack aniline. Graphitized carbon promotes aniline degradation by accelerating electron transfer. The paper develops an innovative strategy to simultaneously realize efficient remediation of PCS and sequential reuse of the post-remediation soil.

Potential biodegradation of polycyclic aromatic hydrocarbons (PAHs) and petroleum hydrocarbons by indigenous fungi recovered from crude oil-contaminated soil in Iran

A total of 265 fungal individuals were isolated from soils exposed to heavy oil spills in the Yadavaran oil field in Iran to discover indigenous fungal species with a high potential to biodegrade petroleum hydrocarbon pollutants. Morphological and molecular identification of obtained fungal species led to their assignment into 16 genera and 25 species. Alternaria spp. (78%), Fusarium spp. (5%), and Cladosporium spp. (4%) were the most common genera, along with Penicillium spp., Neocamarosporium spp., Epicoccum sp., Kotlabaea sp., Aspergillus sp., Mortierella sp., and Pleurotus sp. A preliminary screening using the DCPIP indicator revealed that approximately 35% of isolates from Alternaria, Epicoccum, Neocamarosporium, Cladosporium, Fusarium, Stachybotrys, Penicillium, and Stemphylium demonstrated promising tolerance to crude oil. The best-performing isolates (12 fungal individuals) were further investigated for their capacity to mineralize a mixture of four polycyclic aromatic hydrocarbons (PAH) for 47 days, quantified by GC–MS. Eventually, two top-performing isolates, namely 5c-12 (Alternaria tenuissima) and 3b-1 (Epicoccum nigrum), were applied to petroleum-contaminated soil. The GC–MS analysis showed that 60 days after inoculation, these isolates successfully degraded more than 70% of the long-chain hydrocarbons in the soil, including C8-C16 n-alkanes, C36 n-alkane, and Pristane. This study introduces two fungal species (5c-12 and 3b-1) with high potential for biodegrading petroleum compounds and PAHs, offering promising prospects for the decontamination of oil-contaminated soil.

Utilizing blast-furnace dust as a novel persulfate catalyst for the efficient removal of petroleum contaminants from soil

Petroleum-contaminated soil (PCS) poses a significant health risk. Catalyst-activated persulfate presents a promising technology for efficient treatment, while magnetic activated carbon catalysts are increasingly recognized for their easy recovery and robust performance in advanced oxidation processes (AOPs). However, the complex and costly preparation process has limited its widespread implementation. In this work, blast-furnace dust (BFD), a solid waste from steel mill, was used as a novel persulfate catalyst for the total petroleum hydrocarbons (TPHs) removal from PCS. With the BFD/SP system (0.1 g BFD and 1 mM PS for per gram soil, 4:1 water-to-soil ratio, and 40 °C), removal rates of 88.10% and 84.39% were achieved for TPHs in 0.7% and 3.0% PCS, respectively, surpassing most reported persulfate AOPs, highlighting its superior efficacy. The main reason was that Fe/C components on BFD surface could cooperate to cyclically produce diverse reactive oxygen species, including SO4•−, OH•, O2•− and 1O2. Furthermore, the amphiphilicity of BFD could promote both pollutants and oxidants to converge to its surface simultaneously, effectively utilizing short-lived ROS to degrade TPHs. Our findings implies that BFD is an excellent SP activator, providing an innovative approach for PCS treatment and promoting sustainable utilization of BFD.

Effect of Rhodococcus bioaugmentation and biostimulation on dibenzothiophene biodegradation and bacterial community interaction in petroleum-contaminated soils

Bioremediation can be effective method for achieving polycyclic aromatic hydrocarbons (PAHs) degradation in soil contaminated with petroleum. The aim of this study was to compare the effect of Rhodococcus bioaugmentation (BIOE) and biostimulation (BIOS) on dibenzothiophene biodegradation and bacterial community interaction in petroleum-contaminated soils. The findings revealed that compared to natural degradation treatment (NAT) and BIOS, BIOE had the highest dibenzothiophene (DBT) and the majority of DBT degradation occurred within the first 30 days. BIOS had a positive impact in the early stage but an opposite effect in the later stages for degrading DBT. Beta diversity analysis revealed significant differences of bacterial composition among NAT, BIOS, and BIOE. Sequencing results indicated that Bacillus and Paenibacillus were dominant genera involved in DBT degradation. Network analysis revealed co-occurrence patterns and connectivity, with BIOE exhibiting higher connectivity and the highest number of links in BIOS. In summary, Rhodococcus bioaugmentation was the simplest and effective method to enhance the clustering degree of bacterial network and DBT degradation in petroleum-contaminated soil, compared to NAT and BIOS.

Water conditions and arbuscular mycorrhizal symbiosis affect the phytoremediation of petroleum-contaminated soil by Phragmites australis

Phytoremediation of petroleum-contaminated soils using the synergistic functions of plants and rhizosphere microorganisms is a promising technology. However, successfully applying this approach presents challenges under certain conditions (submerged environments). This study analyzed the potential role of Phragmites australis in symbiosis with arbuscular mycorrhizal (AM) fungi during petroleum remediation at two water levels. AM inoculation promoted P. australis aboveground growth under non-flooded conditions, whereas flooding significantly increased P. australis biomass. The highest total petroleum hydrocarbon (TPH) degradation efficiency was observed in non-flooded soils, whereas submergence severely inhibited TPHs dissipation. Plants with AM inoculation treatments substantially enhanced the removal of TPHs under flooded conditions. TPH removal was positively correlated with dehydrogenase activity but negatively correlated with easily extracted glomalin-related soil proteins. Moreover, different petroleum-hydrocarbon-decaying candidates contributed to TPH removal in these two cultured soils. These findings provide valuable information for the remediation of future TPH-contaminated soils, especially applied in intermittently submerged environments.

In Situ Discrimination and Cultivation of Active Degraders in Soils by Genome-Directed Cultivation Assisted by SIP-Raman-Activated Cell Sorting.

The identification and in situ cultivation of functional yet uncultivable microorganisms are important to confirm inferences regarding their ecological functions. Here, we developed a new method that couples Raman-activated cell sorting (RACS), stable-isotope probing (SIP), and genome-directed cultivation (GDC)─namely, RACS-SIP-GDC─to identify, sort, and cultivate the active toluene degraders from a complex microbial community in petroleum-contaminated soil. Using SIP, we successfully identified the active toluene degrader Pigmentiphaga, the single cells of which were subsequently sorted and isolated by RACS. We further successfully assembled the genome of Pigmentiphaga based on the metagenomic sequencing of 13C-DNA and genomic sequencing of sorted cells, which was confirmed by gyrB gene comparison and average nucleotide identity determination. Additionally, the genotypes and phenotypes of this degrader were directly linked at the single-cell level, and its complete toluene metabolic pathways in petroleum-contaminated soil were reconstructed. Based on its unique metabolic properties uncovered by genome sequencing, we modified the traditional cultivation medium with antibiotics, amino acids, carbon sources, and growth factors (e.g., vitamins and metals), achieving the successful cultivation of RACS-sorted active degrader Pigmentiphaga sp. Our results implied that RACS-SIP-GDC is a state-of-the-art approach for the precise identification, targeted isolation, and cultivation of functional microbes from complex communities in natural habitats. RACS-SIP-GDC can be used to explore specific and targeted organic-pollution-degrading microorganisms at the single-cell level and provide new insights into their biodegradation mechanisms.

Remediation of hydrocarbons and metals by hydrocarbon utilizing Pseudomonas taiwanensis YSA-17 isolated from soil contaminated with petroleum.

Careless handling of petroleum in petrochemical industries releases toxic hydrocarbons and metals to soil and water. The aim of the present study was to isolate hydrocarbon-utilizing and metal-tolerant bacteria. Hydrocarbon-utilizing bacteria from petroleum-contaminated soils were isolated on the Bushnell Hass medium. Hydrocarbon degradation by Pseudomonas taiwanensis strain YSA-17 was observed by gas chromatography-mass spectrometry. Bioaccumulation of metals by strain YSA-17 was assessed in nutrient broth. Among different strains, YSA-17 showed the highest potential for hydrocarbon utilization. After 20 days of incubation, YSA-17 completely degraded one compound and during its degradation, there was the formation of 13 new compounds which were absent in uninoculated control. Results of scanning electron microscope and Fourier transmission infrared (FTIR) indicated degradation of hydrocarbons. FTIR showed the formation of new functional groups in YSA-17 inoculated medium. Expression of the total quantity of hydrocarbon-degrading gene (AlkB and NehAc) in petroleum-amended nutrient broth inoculated with strain YSA-17 enhanced significantly during 20 days of incubation compared to control. YSA-17 also significantly removed metals. This study concluded that bioinoculant can be utilized for the bioremediation of pollutants cocontaminated with hydrocarbons and metals. Petroleum-contaminated soil will be remediated for pollutants.

Sequential reuse of remediated soil from petroleum-contaminated site to recover transition heavy metals and activate peroxysulphate oxidation

Pyrolysis can rapidly remediate petroleum-contaminated soil (PCS) and recover some liquid oil, but less attention has been paid to the reuse potential and strategy of the remediated carbonized soil (RCS). In this study, RCS was used as adsorbent to recover Mn2+, Ni2+, and Cu2+ from wastewater. The solid residue (RCS-Me) was reused to activate peroxysulphate oxidation. The total petroleum hydrocarbons and dissolved organic matter can be completely removed by pyrolysis at 500 °C for 3 h. The maximum adsorption capacities of RCS for Cu2+, Ni2+, and Mn2+ are 65.96, 50.75, and 57.95 mg/g, respectively. Sequential extraction analysis shows that transition heavy metals are mainly bound to carbonate and Fe/Mn oxides. RCS and RCS-Me can effectively activate peroxysulphate to degrade aniline, and the activation performance and reusability of RCS-Me are remarkably better than RCS. The nonradical path dominates in the oxidation process, and the main species are hole and 1O2. Electron paramagnetic resonance and linear sweep voltammetry analysis show that hole improves the activation efficiency mainly by accelerating direct electron transfer, while the oxygen-containing groups and oxygen vacancies may activate peroxysulphate to generate 1O2.The strategy can simultaneously realize efficient remediation of PCS and sequential resource utilization of the remediated soil.

Soil Washing to Eliminate Polycyclic Aromatic Hydrocarbons from Petroleum Contaminated Soil - Temperature Effect

This study’s goals were to identify the components of polycyclic aromatic hydrocarbons (PAHs) in soil samples polluted with petroleum and to ascertain how temperature affected the removal of PAHs pollutants from petroleum-contaminated soil. The concentration of contaminants in the soil samples decreased from 46.22-12.07 ppm to 77.01-30.06 ppm at a temperature increase of 20–160°C, as shown by the results of this study, which are also supported by the percentage removal efficiencies of 22.22–79.69% and 32.90–73.81% for samples M and S, respectively. It was further discovered that 160℃ was the ideal temperature range for soil washing to eliminate the pollutants. High removal efficiency values demonstrated the efficacy of the soil washing approach in removing pollutants with high PAH concentrations. This information can help in the creation of more successful and affordable soil remediation techniques.

CHARACTERIZATION OF A NEWLY ISOLATED AZO-DYE DEGRADING KLEBSIELLA SP. STRAIN RGUDBI01

Azo-dyes such as crystal violet are commonly used ingredient in all types of dying industries which more often eventually reach water and soil due to washing and disposal practices. They are non-biodegradable inside higher organisms and are less biodegradable due to their xenobiotic character. However, some of the microbes exhibit their extensive potential to degrade such recalcitrants. This study reports the isolation and characterization of a newly isolated Klebsiella sp. strain RGUDBI01 from petroleum-contaminated soil samples. The species-level identification was done by 16S rDNA gene sequencing (ON945611.1). The isolate exhibited 88% azo-dye degradation just after 3 days of incubation under optimized conditions. The treated samples exhibited a significant increase in the growth of Cicer arietinum seeds compared to the control group. This result suggests that the dye-contaminated samples, after treatment, displayed non-cytotoxic behavior. The strain also showed its potential to tolerate and withstand a wide range of pH, salinity, and substrate concentration which supports its futuristic environmental application. In addition to the above, the strain could also produce biosurfactants with ʋC=C, ʋC=C-H, and ʋC-C functional groups which were evaluated based on FTIR spectral analysis.

Exploring the bioremediation capability of petroleum-contaminated soils for enhanced environmental sustainability and minimization of ecotoxicological concerns.

The bioremediation of soils contaminated with petroleum hydrocarbons (PHCs) has emerged as a promising approach, with its effectiveness contingent upon various types of PHCs, i.e., crude oil, diesel, gasoline, and other petroleum products. Strategies like genetically modified microorganisms, nanotechnology, and bioaugmentation hold potential for enhancing remediation of polycyclic aromatic hydrocarbon (PAH) contamination. The effectiveness of bioremediation relies on factors such as metabolite toxicity, microbial competition, and environmental conditions. Aerobic degradation involves enzymatic oxidative reactions, while bacterial anaerobic degradation employs reductive reactions with alternative electron acceptors. Algae employ monooxygenase and dioxygenase enzymes, breaking down PAHs through biodegradation and bioaccumulation, yielding hydroxylated and dihydroxylated intermediates. Fungi contribute via mycoremediation, using co-metabolism and monooxygenase enzymes to produce CO2 and oxidized products. Ligninolytic fungi transform PAHs into water-soluble compounds, while non-ligninolytic fungi oxidize PAHs into arene oxides and phenols. Certain fungi produce biosurfactants enhancing degradation of less soluble, high molecular-weight PAHs. Successful bioremediation offers sustainable solutions to mitigate petroleum spills and environmental impacts. Monitoring and assessing strategy effectiveness are vital for optimizing biodegradation in petroleum-contaminated soils. This review presents insights and challenges in bioremediation, focusing on arable land safety and ecotoxicological concerns.

Unveiling the novel role of ryegrass rhizospheric metabolites in benzo[a]pyrene biodegradation

Rhizoremediation is a promising remediation technology for the removal of soil persistent organic pollutants (POPs), especially benzo[a]pyrene (BaP). However, our understanding of the associations among rhizospheric soil metabolites, functional microorganisms, and POPs degradation in different plant growth stages is limited. We combined stable-isotope probing (SIP), high-throughput sequencing, and metabolomics to analyze changes in rhizospheric soil metabolites, functional microbes, and BaP biodegradation in the early growth stages (tillering, jointing) and later stage (booting) of ryegrass. Microbial community structures differed significantly among growth stages. Metabolisms such as benzenoids and carboxylic acids tended to be enriched in the early growth stage, while lipids and organic heterocyclic compounds dominated in the later stage. From SIP, eight BaP-degrading microbes were identified, and most of which such as Ilumatobacter and Singulisphaera were first linked with BaP biodegradation. Notably, the relationship between the differential metabolites and BaP degradation efficiency further suggested that BaP-degrading microbes might metabolize BaP directly to produce benzenoid metabolites (3-hydroxybenzo[a]pyrene), or utilize benzenoids (phyllodulcin) to stimulate the co-metabolism of BaP in early growth stage; some lipids and organic acids, e.g. 1-aminocyclopropane-1-carboxylic acid, might provide nutrients for the degraders to promote BaP metabolism in later stage. Accordingly, we determined that certain rhizospheric metabolites might regulate the rhizospheric microbial communities at different growth stages, and shift the composition and diversity of BaP-degrading bacteria, thereby enhancing in situ BaP degradation. Our study sheds light on POPs rhizoremediation mechanisms in petroleum-contaminated soils.

Characterization of three rapidly growing novel Mycobacterium species with significant polycyclic aromatic hydrocarbon bioremediation potential.

Mycobacterium species exhibit high bioremediation potential for the degradation of polycyclic aromatic hydrocarbons (PAHs) that are significant environmental pollutants. In this study, three Gram-positive, rapidly growing strains (YC-RL4T, MB418T, and HX176T) were isolated from petroleum-contaminated soils and were classified as Mycobacterium within the family Mycobacteriaceae. Genomic average nucleotide identity (ANI; < 95%) and digital DNA-DNA hybridization (dDDH; < 70%) values relative to other Mycobacterium spp. indicated that the strains represented novel species. The morphological, physiological, and chemotaxonomic characteristics of the isolates also supported their affiliation with Mycobacterium and their delineation as novel species. The strains were identified as Mycobacterium adipatum sp. nov. (type strain YC-RL4T = CPCC 205684T = CGMCC 1.62027T), Mycobacterium deserti sp. nov. (type strain MB418T = CPCC 205710T = KCTC 49782T), and Mycobacterium hippophais sp. nov. (type strain HX176T = CPCC 205372T = KCTC 49413T). Genes encoding enzymes involved in PAH degradation and metal resistance were present in the genomes of all three strains. Specifically, genes encoding alpha subunits of aromatic ring-hydroxylating dioxygenases were encoded by the genomes. The genes were also identified as core genes in a pangenomic analysis of the three strains along with 70 phylogenetically related mycobacterial strains that were previously classified as Mycolicibacterium. Notably, strain YC-RL4T could not only utilize phthalates as their sole carbon source for growth, but also convert di-(2-ethylhexyl) phthalate into phthalic acid. These results indicated that strains YC-RL4T, MB418T, and HX176T were important resources with significant bioremediation potential in soils contaminated by PAHs and heavy metals.

Screening of Aflatoxin Production by Aspergillus flavus Isolates from Petroleum-contaminated Soil

Fungi are eukaryotic, heterogeneous, unicellular to filamentous, spore-bearing, and chemoorganotrophic organisms which lack chlorophyll. This present study was carried out to isolate and identify fungi from petroleum-contaminated soil. Several fungal genera which included Rhizopus spp., Mucor spp., Penicillium spp., Rhizoctonia spp., Aspergillus spp., Alternaria spp., and Cladosporiumspp. were isolated using potatoes dextrose agar, Czapek-Dox Agar, and Aspergillus flavus Differentiation Agar culture media that comparable with co-amoxiclav (1g) and chloramphenicol to prevent the growth of any bacteria. The direct plate and serial dilution agar plate methods were used for the isolation of fungi. Based on results, Aspergillusand Mucor spp. were the most predominant genera and had the highest number of colonies in the soil samples. In this investigation, seven out of 27 soil samples were morphologically (macroscopically and microscopically) identified, such as A. flavus. Aflatoxigenicity of A. flavus was detected using characteristics in Aspergillus differentiation agar and colony fluorescence on exposure to ultraviolet light. Moreover, molecular approaches were used for the detection of aflatoxigenic of the A. flavus isolates. Three structural (aflD, aflO, and aflP) and one regulatory (aflR) gene of the aflatoxin gene cluster of A. flavus were targeted for amplification by the polymerase chain reaction method. The aflatoxigenic of all six A. flavus isolates was detected molecularly which contained two structural (aflD, aflP)genes out of three structural genes, while there was no specific amplification of the aflO gene in the fourth, fifth, and sixth A. flavus which issimilar to the aflR gene in the first and second A. flavus.

Influence of a Hydrocarbon Biodestructor on the Growth and Content of Phytohormones in Secale cereale L. Plants under Petroleum Pollution of the Soil

The phytoremediation of soil contaminated with petroleum depends on the association of plants with rhizosphere bacteria capable of promoting plant growth and destroying petroleum hydrocarbonates. In the present work, we studied the effects of “Lenoil” biological product containing bacteria Pseudomonas turukhanskensis IB 1.1, capable of destroying petroleum hydrocarbons on Secale cereale L plants, which previously proved to be resistant to the weak oil pollution of gray forest soil and to the composition of microorganisms in their rhizosphere. The composition of microorganisms in the rhizosphere of rye roots was studied, morphometric parameters of shoots and roots of rye plants were estimated, and hormone concentration was immunoassayed under conditions of 4% petroleum pollution of the soil. Addition of petroleum to the soil increased the content of oligonitrophilic (by 24%) and hydrocarbon-oxidizing (by 33%) microorganisms; however, the content of cellulolytic (by 12.5 times) microorganisms in the rhizosphere decreased. The use of Lenoil led to a further increase in the number of cellulolytic (by 5.6 times) and hydrocarbon-oxidizing (by 3.8 times) microorganisms and a decrease in the number of oligonitrophilic (by 22.7%) microorganisms in the rhizosphere. Under petroleum pollution, the content of auxins (IAA), zeatin riboside, zeatin nucleotide, and zeatin decreased, while the content of abscisic acid (ABA) increased in the shoots of rye plants. Lenoil treatment led to an eight-fold increase in the IAA content in the roots and a decrease in the ABA content in the aerial part and in the roots. It was shown for the first time that the treatment of petroleum-contaminated soil with “Lenoil” increased root mass due to the development of lateral roots, concurrent with high root IAA content. Petroleum pollution increased the deposition of lignin and suberin in the roots, which strengthened the apoplastic barrier and, thus, reduced the infiltration of toxic components. The deposition of suberin and lignin decreased under ”Lenoil” treatment, indicating a decrease in the concentration of toxic petroleum components in the soil degraded by the bacteria. Thus, the biological preparation reduced the growth-inhibiting effect of petroleum on rye plants by increasing the content of growth-stimulating phytohormones (IAA and cytokinins) and reducing the content of ABA, justifying the importance of further study of relevant hormones for the improvement of phytoremediation.

Genomic and biotechnological potential of a novel oil-degrading strain Enterobacter kobei DH7 isolated from petroleum-contaminated soil

In this study, a novel oil-degrading strain Enterobacter kobei DH7 was isolated from petroleum-contaminated soil samples from the industrial park in Taolin Town, Lianyungang, China. The whole genome of the strain was sequenced and analyzed to reveal its genomic potential. The oil degradation and growth conditions including nitrogen, and phosphorus sources, degradation cycle, biological dosing, pH, and oil concentration were optimized to exploit its commercial application. The genome of the DH7 strain contains 4,705,032 bp with GC content of 54.95% and 4653 genes. The genome analysis revealed that there are several metabolic pathways and enzyme-encoding genes related to oil degradation in the DH7 genome, such as the paa gene cluster which is involved in the phenylacetic acid degradation pathway, and complete degradation pathways for fatty acid and benzoate, genes related to chlorinated alkanes and olefins degradation pathway including adhP, frmA, and adhE, etc. The strain DH7 under the optimized conditions has demonstrated a maximum degradation efficiency of 84.6% after 14 days of treatment using synthetic oil, which comparatively displays a higher oil degradation efficiency than any Enterobacter species known to date. To the best of our knowledge, this study presents the first-ever genomic studies related to the oil degradation potential of any Enterobacter species. As Enterobacter kobei DH7 has demonstrated significant oil degradation potential, it is one of the good candidates for application in the bioremediation of oil-contaminated environments.

Exploring the existing state of petroleum based on the adsorption capacity of carbon tetrachloride in petroleum-contaminated soil

Exploring the existing state of petroleum based on the adsorption capacity of carbon tetrachloride in petroleum-contaminated soil

Remediation of Petroleum-contaminated Soil by Highly Efficient Oil-degrading Bacteria and Analysis of Its Enhancement Mechanism

A 120-day in situ remediation of oil-contaminated soil was carried out by using highly efficient oil-degrading bacteria. The effects of bio-enhanced remediation and changes in soil physicochemical properties and enzyme activities were investigated. Combined with metagenomic sequencing and bioinformatics analysis, the strengthening mechanism was revealed. The results showed that compared with the blank control group (Ctrl), the degradation rate of total petroleum hydrocarbons in the bioremediation group (Exp-BT) was significantly increased, reaching 81.23%. During enhanced bioremediation by highly efficient oil-degrading bacteria, the pH of the soil was stable, the oxidation capacity of the system was improved, and the electrical conductivity was in the range suitable for agricultural activities. Lipase and dehydrogenase maintained high activity during repair. In addition, the analysis of the initial contaminated soil (B0), the highly efficient oil-degrading bacteria obtained from domestication (GZ), and the soil samples after bioremediation (BT) in the obtained samples showed that, at the phylum level, the total proportion of Proteobacteria and Actinobacteria increased by 17.1%. At the genus level, the abundance of Nocardioides, Achromobacter, Gordonia, and Rhodococcus increased significantly. The species and function contribution analysis of COG and KEGG proved that the above bacterial genera had important contributions to the degradation of petroleum hydrocarbons. A high abundance of petroleum hydrocarbon-related metabolic enzymes and five petroleum hydrocarbon-related degradation genes was found in the soil after remediation:alkM, tamA, rubB, ladA, and alkB. The analysis showed that the introduction of the exogenous petroleum hydrocarbon-degrading bacteria group enhanced the metabolic activity of microorganism-related enzymes and the expression of corresponding functional genes.

Isolation and Characterization of the Zearalenone-Degrading Strain, Bacillus spizizenii B73, Inspired by Esterase Activity.

Zearalenone (ZEN) is a widespread mycotoxin found in grain and feed, presenting a serious threat to animal and human health. This study investigated the ability of the novel strain B73, isolated from petroleum-contaminated soil, to detoxify ZEN. B73 was identified as Bacillus spizizenii through physiological and biochemical tests, and further confirmed based on the 16S rRNA gene sequence and the complete genome sequence. B. spizizenii B73 was capable of degrading up to 99.3% of ZEN at a concentration of 10 μg/mL in a minimal medium (pH = 7.0) within 8 h at 37 °C via HPLC-UV. In addition, B. spizizenii B73 was used to treat ZEN-contaminated wheat bran, dried distillers grains (DDGS), and corn meal, whereby the respective degradation rates reached 96.32%, 98.73%, and 80.31% after 36 h of treatment. HPLC-Q-Exactive-MS/MS analysis revealed one of the degradation products to have the formula C17H24O4. B. spizizenii B73 is a novel strain isolated from petroleum-contaminated soil, and the extracellular enzymes secreted by this strain show a remarkable ability to degrade ZEN.

Exploring the bacterial genetic diversity and community structure of crude oil contaminated soils using microbiomics

The impact of environmental pollution in air and water is reflected mainly in the soil ecosystem as it impairs soil functions. Also, since the soil is the habitat for billions of organisms, the biodiversity is in turn altered. Microbes are precise sensors of ecological contamination, and bacteria have a key and important function in terms of bioremediation of the contaminated soil. Hence in the current work, we aimed at assessing the unidentified bacterial population through Illumina MiSeq sequencing technology and their community structural changes in different levels of petroleum-contaminated soil and sludge samples (aged, sludge, and leakage soil) to identify unique bacteria for their potential application in remediation. The studies showed that major bacterial consortiums namely, Proteobacteria (57%), Alphaproteobacteria (31%), and Moraxellaceae (23%) were present in aged soil, whereas Proteobacteria (52%), Alphaproteobacteria (33%), and Rhodobacteraceae (28%) were dominantly found in sludge soil. In leakage soil, Proteobacteria (59%), Alphaproteobacteria (33%), and Rhodobacteraceae (29%) were abundantly present. The Venn diagrams are used to analyze the distribution of abundances in individual operational taxonomic units (OTUs) within three soil samples. After data filtering, they were grouped into OTU clusters and 329 OTUs were identified from the three soil samples. Among the 329, 160 OTUs were common in the three soil samples. The bacterial diversity is estimated using alpha diversity indices and Shanon index and was found to be 4.490, 4.073 and 4.631 in aged soil, sludge soil and leakage soil, respectively and similarly richness was found to be 618, 417 and 418. The heat map was generated by QIIME software and from the top 50 enriched genera few microbes such as Pseudomonas, Bacillus, Mycobacterium, Sphingomonas and Paracoccus, were shown across all the samples. In addition, we also analyzed various physicochemical properties of soil including pH, temperature, salinity, electrical conductivity, alkalinity, total carbon, total organic matter, nitrogen, phosphorus and potassium to calculate the soil quality index (SQI). The SQI of aged, sludge and leakage soil samples were 0.73, 0.64, and 0.89, respectively. These findings show the presence of unexplored bacterial species which could be applied for hydrocarbon remediation and further they can be exploited for the same.