Dataset Information

Bioremediation of Petroleum-Contaminated Soils with Biosurfactant-Producing Degraders Isolated from the Native Desert Soils.

ABSTRACT: A crude oil spill in 2014 resulted in extensive soil contamination of the hyper arid Evrona Nature Reserve in Israel's Negev Desert. The contaminated soils became highly hydrophobic, threatening the existence of plants in the habitat. We hypothesized that bioaugmenting the soil with indigenous biosurfactant-producing, hydrocarbon-degrading bacteria (HDB) would accelerate the reduction in the soil's hydrophobicity. We aimed to isolate and characterize biosurfactant-producing HDBs from the desert-contaminated soil and test if they can be used for augmenting the soil. Twelve hydrocarbon-degrading strains were isolated, identified as Pseudomonas, and classified as biosurfactants "producing" and "nonproducing". Inoculating 10⁹ CFU/g of "producing" strains into the polluted soil resulted in a 99.2% reduction in soil hydrophobicity within seven days. At the same time, nonproducing strains reduced hydrophobicity by only 17%, while no change was observed in the untreated control. The microbial community in the inoculated soil was dominated by the introduced strains over 28 days, pointing to their persistence. Rhamnolipid biosynthesis gene rhlAB remained persistent in soil inoculated with biosurfactants, indicating in situ production. We propose that the success of the treatment is due to the use of inoculum enriched from the polluted soil.

SUBMITTER: Li Z

PROVIDER: S-EPMC9694877 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature

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Bioremediation of Petroleum-Contaminated Soils with Biosurfactant-Producing Degraders Isolated from the Native Desert Soils.

Li Zheng Z Rosenzweig Ravid R Chen Fengxian F Qin Ji J Li Tianyi T Han Jincheng J Istvan Paula P Diaz-Reck Damiana D Gelman Faina F Arye Gilboa G Ronen Zeev Z

Microorganisms 20221115 11

A crude oil spill in 2014 resulted in extensive soil contamination of the hyper arid Evrona Nature Reserve in Israel's Negev Desert. The contaminated soils became highly hydrophobic, threatening the existence of plants in the habitat. We hypothesized that bioaugmenting the soil with indigenous biosurfactant-producing, hydrocarbon-degrading bacteria (HDB) would accelerate the reduction in the soil's hydrophobicity. We aimed to isolate and characterize biosurfactant-producing HDBs from the desert- ...[more]

PMID: 36422337

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Project description:Petroleum is, up to this date, an inimitable nonrenewable energy resource. Petroleum leakage, which arises during transport, storage, and refining, is the most important contaminant in the environment, as it produces harm to the surrounding ecosystem. Bioremediation is an efficient method used to treat petroleum hydrocarbon-contaminated soil using indigenous microorganisms. The degradation characteristics for a variety of hydrocarbons (hexane, benzene, gasoline, and diesel) were qualitatively and quantitatively investigated using Bacillus isolates. Microbiological and biochemical methods have been used including isolation of oil-degrading bacteria, enzymatic activities, the determination of physicochemical parameters, biosurfactant production and extraction assay, oil displacement assay, antimicrobial assay of the biosurfactants, and bioremediation kinetics. Consequently, of the 60 isolates capable of degrading different hydrocarbons at fast rates, 34 were suspected to be Bacillus isolates capable of growing in 24 h or 48 h on BH medium supplemented with 2% of hexane, benzene, gasoline, diesel, and olive oil, respectively. Among the 34 isolates, 61% (21/34) are capable of producing biosurfactant-like molecules by using gasoline, 70% (24/34) with diesel oil, 85% (29/34) with hexane, and 82% (28/34) with benzene. It was found that biosurfactant-producing isolates are extractable with HCl (100%), ammonium sulphate (95%), chloroform (95%), and ethanol (100%). Biosurfactants showed stability at 20°C, 37°C, 40°C, and 60°C. Biosurfactant secreted by Bacillus strains has shown an antagonistic effect in Escherichia coli, Shigella flexneri 5a M90T, and Bacillus cereus. The selected isolates could therefore be safely used for biodegradation. Substrate biodegradation patterns by individual isolates were found to significantly differ. The study shows that benzene was degraded faster, followed by hexane, gasoline, and finally diesel. The Bacillus consortium used can decrease hydrocarbon content from 195 to 112 (g/kg) in 15 days.

Project description:Biosurfactants are a unique class of compounds that have been shown to have a variety of potential applications in the remediation of organic- and metal-contaminated sites, in the enhanced transport of bacteria, in enhanced oil recovery, as cosmetic additives, and in biological control. However, little is known about the distribution of biosurfactant-producing bacteria in the environment. The goal of this study was to determine how common culturable surfactant-producing bacteria are in undisturbed and contaminated sites. A series of 20 contaminated (i.e., with metals and/or hydrocarbons) and undisturbed soils were collected and plated on R(2)A agar. The 1,305 colonies obtained were screened for biosurfactant production in mineral salts medium containing 2% glucose. Forty-five of the isolates were positive for biosurfactant production, representing most of the soils tested. The 45 isolates were grouped by using repetitive extragenic palindromic (REP)-PCR analysis, which yielded 16 unique isolates. Phylogenetic relationships were determined by comparing the 16S rRNA gene sequence of each unique isolate with known sequences, revealing one new biosurfactant-producing microbe, a Flavobacterium sp. Sequencing results indicated only 10 unique isolates (in comparison to the REP analysis, which indicated 16 unique isolates). Surface tension results demonstrated that isolates that were similar according to sequence analysis but unique according to REP analysis in fact produced different surfactant mixtures under identical growth conditions. These results suggest that the 16S rRNA gene database commonly used for determining phylogenetic relationships may miss diversity in microbial products (e.g., biosurfactants and antibiotics) that are made by closely related isolates. In summary, biosurfactant-producing microorganisms were found in most soils even by using a relatively limited screening assay. Distribution was dependent on soil conditions, with gram-positive biosurfactant-producing isolates tending to be from heavy metal-contaminated or uncontaminated soils and gram-negative isolates tending to be from hydrocarbon-contaminated or cocontaminated soils.

Project description:In this study, we compared the culturability of aerobic bacteria isolated from long-term oil-contaminated soils via enrichment and direct-plating methods; bacteria were cultured at 30°C or ambient temperatures. Two soil samples were collected from two sites in the Shengli oilfield located in Dongying, China. One sample (S0) was close to the outlet of an oil-production water treatment plant, and the other sample (S1) was located 500 m downstream of the outlet. In total, 595 bacterial isolates belonging to 56 genera were isolated, distributed in Actinobacteria, Firmicutes, Bacterioidetes, and Proteobacteria. It was interesting that Actinobacteria and Firmicutes were not detected from the 16S rRNA gene clone library. The results suggested the activation of rare species during culture. Using the enrichment method, 239 isolates (31 genera) and 96 (22 genera) isolates were obtained at ambient temperatures and 30°C, respectively, from S0 soil. Using the direct-plating method, 97 isolates (15 genera) and 163 isolates (20 genera) were obtained at ambient temperatures and 30°C, respectively, from two soils. Of the 595 isolates, 244 isolates (41.7% of total isolates) could degrade n-hexadecane. A greater number of alkane-degraders was isolated at ambient temperatures using the enrichment method, suggesting that this method could significantly improve bacterial culturability. Interestingly, the proportion of alkane degrading isolates was lower in the isolates obtained using enrichment method than that obtained using direct-plating methods. Considering the greater species diversity of isolates obtained via the enrichment method, this technique could be used to increase the diversity of the microbial consortia. Furthermore, phenol hydroxylase genes (pheN), medium-chain alkane monooxygenases genes (alkB and CYP153A), and long-chain alkane monooxygenase gene (almA) were detected in 60 isolates (11 genotypes), 91 isolates (27 genotypes) and 93 isolates (24 genotypes), and 34 isolates (14 genotypes), respectively. This study could provide new insights into microbial resources from oil fields or other environments, and this information will be beneficial for bioremediation of petroleum contamination and for other industrial applications.

Dataset Information

Bioremediation of Petroleum-Contaminated Soils with Biosurfactant-Producing Degraders Isolated from the Native Desert Soils.

Publications

Bioremediation of Petroleum-Contaminated Soils with Biosurfactant-Producing Degraders Isolated from the Native Desert Soils.

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