WO2018164648A1 - Method for bioremediation of petroleum-contaminated soils - Google Patents
Method for bioremediation of petroleum-contaminated soils Download PDFInfo
- Publication number
- WO2018164648A1 WO2018164648A1 PCT/TR2017/050087 TR2017050087W WO2018164648A1 WO 2018164648 A1 WO2018164648 A1 WO 2018164648A1 TR 2017050087 W TR2017050087 W TR 2017050087W WO 2018164648 A1 WO2018164648 A1 WO 2018164648A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- petroleum
- bioremediation
- soil
- contaminated
- contaminated soils
- Prior art date
Links
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
- B09C1/105—Reclamation of contaminated soil microbiologically, biologically or by using enzymes using fungi or plants
Definitions
- the present invention relates to a method which allows performing bioremediation of petroleum-contaminated soils by incorporating bio stimulation and bioaugmentation methods.
- the invention particularly relates to a method which allows subjecting petroleum- contaminated soils to bioremediation using a composition in which Bjerkandera adusta and Trametes versicolor fungi are used together for bioremediation.
- Petroleum wastes cause health issues in human life and ecosystem degradation as contaminants comprising toxic, mutagenic, and carcinogenic compounds.
- Petroleum sludge comprises water, oil emulsion, metal, tar, and many petroleum hydrocarbon compounds and has a very complex structure. A wrong petroleum sludge disposal results in serious soil, water, and groundwater pollution.
- Bioremediation overcomes these problems as an economic and environmentally friendly treatment method. During this process, contaminant compounds are converted into either non-toxic materials or C0 2 and H 2 0 as a whole. At sites where bioremediation is applied, local microorganisms or nutrients used as inoculum may be stimulated for digesting organic contaminants by using oxygen or another additive. Bioaugmentation, as an improved bioremediation process, is to improve the degradation potential of the site at which bioremediation technique is used by adding microbial water. This process has an impact on the degradation of various contaminants.
- biostimulation is a more effective method of soil bioremediation than bioaugmentation.
- One of the important steps while applying biostimulation is to monitor the efficiency of the process and evaluate the degradation level in a contaminated environment. Effective monitoring may reduce bioremediation costs as well as the remediation time.
- NZ519022A filed on 17.05.2002, discloses a composition for accelerating bioremediation process, comprising particularly Trametes versicolor and other Trametes species.
- the composition used in this patent contains cultures A, B and C, said cultures being disclosed in three different patent documents. Nevertheless, the applications described in these documents fail to satisfy bioremediation with the desired efficiency. Therefore, developing a method for subjecting petroleum-contaminated soils to bioremediation according to the invention has been deemed necessary.
- An object of the present invention is to provide an economic method for bioremediation of petroleum-contaminated soils.
- Another object of the present invention is to provide a method for subjecting petroleum-contaminated soils to bioremediation, wherein biostimulation and bioaugmentation techniques are used together.
- Another object of the present invention is to determine the optimum amount of biostimulation, and then to perform a method of successful biostimulation using two different types of fungi specific for biostimulation.
- Yet another object of the present invention is to provide a method by which petroleum wastes are disposed with maximum efficiency.
- Petroleum-derived wastes consist of organic molecules groups differing in terms of molecular weight, structural configuration, solubility, number of aromatic chains, volatility, adsorption coefficient, etc. These components, due to the inhibitory properties and low viscosity solubility thereof, are highly resistant against biodegradation and remain in nature without degradation.
- biostimulation and bioaugmentation methods are incorporated.
- a successful biostimulation has been achieved by first determining the optimum amount of biostimulation (C:N:P ratio, 100: 15: 1), and then using two different types of fungi specific for the determined biostimulation.
- the subject matter of the invention is a method which allows performing bioremediation of petroleum-contaminated soils, wherein bio stimulation and bioaugmentation steps are comprised together.
- the invention comprises the following specific steps: collecting petroleum-contaminated soil samples from petroleum waste lagoons and characterizing the same, determining dominant bacterial species in the collected samples by Illumina sequencing, one of the next-generation sequencing analysis methods, subjecting soil mixtures to a bio stimulation process at a C:N:P ratio of 100: 15: 1, isolating genomic DNA, total RNA, and plasmids from soil samples, measuring the degrading bacteria, functional genes alkB, phnAc and nah using alkane and polyaromatic hydrocarbons (PAH) by quantitative real time PCR method, applying bioaugmentation to the petroleum-contaminated soil, having undergone biostimulation at a C:N:P ratio of 100: 15: 1, using Trametes versicolor and Bjerkandera adusta fungi species, and
- Petroleum-contaminated soil samples were collected from petroleum waste lagoon and they were characterized, and then dominant bacterial species were determined in the collected samples by means of Illumina sequencing method. Petroleum- contaminated samples were mixed with clean forest soil samples at different ratios and placed in glass containers, and then N:P at different ratios was added to the resulting soil mixtures and incubated at 24°C for 90 days. Genomic DNA, total RNA, and plasmids were isolated from the incubated samples. Alkane and polyaromatic hydrocarbons (PAH) were used for measuring the degrading bacteria and functional genes alkB, phnAc and nah by means of quantitative real time PCR performed with TaqMan probe. As a final treatment method, the treated contaminated soil was subject to biostimulation, followed by bioaugmentation method, using Trametes versicolor and Bjerkandera adusta fungi species.
- PHA polyaromatic hydrocarbons
Abstract
The invention relates to a method which allows subjecting petroleum- contaminated soils to bioremediation using a composition in which Bjerkandera adusta and Trametes versicolor fungi are used together for bioremediation. The invention is a method for bioremediation of petroleum-contaminated soils, said method comprising the steps of collecting petroleum-contaminated soil samples from petroleum waste lagoons and characterizing the same; determining dominant bacterial species in the collected samples; adding N:P at different ratios to the resulting soil mixtures and incubating the same; isolating genomic DNA, total RNA, and plasmids; measuring the degrading bacteria, functional genes alkB, phnAc and nah using alkane and polyaromatic hydrocarbons (PAH) by quantitative real time PCR; applying biostimulation to the contaminated soil having been treated using Trametes versicolor and Bjerkandera adusta fungi species; and evaluating the effect of the fungi on local microbial community, functional genes alkB, phnAc and nah by applying bioaugmentation. In the present method, the optimal biostimulation ratio (C:N:P ratio, 100:15:1) was determined and biological cleaning agents suitable for bioaugmentation (Bjerkandera adusta and Trametes versicolor) were detected.
Description
METHOD FOR BIOREMEDIATION OF
PETROLEUM-CONTAMINATED SOILS
Technical Field of the Invention The present invention relates to a method which allows performing bioremediation of petroleum-contaminated soils by incorporating bio stimulation and bioaugmentation methods.
The invention particularly relates to a method which allows subjecting petroleum- contaminated soils to bioremediation using a composition in which Bjerkandera adusta and Trametes versicolor fungi are used together for bioremediation.
Background of the Invention
Although the technological background of the renewable energy generation is highly advanced, petroleum products still remain to be one of the major sources of the energy used for industrial and domestic consumption. Depending on the use of this source, more than three billion tons of petroleum is produced in the world annually.
Petroleum wastes cause health issues in human life and ecosystem degradation as contaminants comprising toxic, mutagenic, and carcinogenic compounds.
Petroleum sludge comprises water, oil emulsion, metal, tar, and many petroleum hydrocarbon compounds and has a very complex structure. A wrong petroleum sludge disposal results in serious soil, water, and groundwater pollution.
Physicochemical methods such as soil vapor extraction, soil washing and dispersion are commonly used for remediation of petroleum-contaminated sites; however, these methods fail to completely eliminate petroleum contamination and also they increase the toxicity of the contaminated sites. Bioremediation overcomes these problems as an economic and environmentally friendly treatment
method. During this process, contaminant compounds are converted into either non-toxic materials or C02 and H20 as a whole. At sites where bioremediation is applied, local microorganisms or nutrients used as inoculum may be stimulated for digesting organic contaminants by using oxygen or another additive. Bioaugmentation, as an improved bioremediation process, is to improve the degradation potential of the site at which bioremediation technique is used by adding microbial water. This process has an impact on the degradation of various contaminants. However, the concerns of genetic engineering about the use of microorganisms and the possible effects of these microorganisms on the environment are common. As an alternative to bioaugmentation and as a digestible bioremediation method, biostimulation is a more effective method of soil bioremediation than bioaugmentation. One of the important steps while applying biostimulation is to monitor the efficiency of the process and evaluate the degradation level in a contaminated environment. Effective monitoring may reduce bioremediation costs as well as the remediation time.
The prior art search revealed the International Patent Application No. WO2014031016A1, filed on 16.10.2012, wherein a method of bioremediation of soil and/or soil-based masses polluted with petroleum-derived compounds under biodegradation by subjecting the same to various processes is disclosed. Another prior art document is the U.S. Patent Application No. US5725885A, filed on 06.06.1995, wherein a composition, as well as a method of manufacturing said composition, for promoting a faster degradation of the bacteria in petroleum and petroleum-derived materials to aid in bioremediation of oil spills on water is disclosed. On the other hand, New Zealand Patent Application No. NZ519022A, filed on 17.05.2002, discloses a composition for accelerating bioremediation process, comprising particularly Trametes versicolor and other Trametes species. The composition used in this patent contains cultures A, B and C, said cultures being disclosed in three different patent documents.
Nevertheless, the applications described in these documents fail to satisfy bioremediation with the desired efficiency. Therefore, developing a method for subjecting petroleum-contaminated soils to bioremediation according to the invention has been deemed necessary. Objects and Summary of the Invention
An object of the present invention is to provide an economic method for bioremediation of petroleum-contaminated soils.
Another object of the present invention is to provide a method for subjecting petroleum-contaminated soils to bioremediation, wherein biostimulation and bioaugmentation techniques are used together.
Another object of the present invention is to determine the optimum amount of biostimulation, and then to perform a method of successful biostimulation using two different types of fungi specific for biostimulation.
And another object of the present invention is to provide a method by which petroleum wastes are disposed with maximum efficiency.
Petroleum-derived wastes consist of organic molecules groups differing in terms of molecular weight, structural configuration, solubility, number of aromatic chains, volatility, adsorption coefficient, etc. These components, due to the inhibitory properties and low viscosity solubility thereof, are highly resistant against biodegradation and remain in nature without degradation. With the method according to the invention, an economic method for bioremediation of petroleum-contaminated soils has been developed. In this method, biostimulation and bioaugmentation methods are incorporated. A successful biostimulation has been achieved by first determining the optimum amount of biostimulation (C:N:P ratio, 100: 15: 1), and then using two different types of fungi specific for the determined biostimulation.
Detailed Description of the Invention
The subject matter of the invention is a method which allows performing bioremediation of petroleum-contaminated soils, wherein bio stimulation and bioaugmentation steps are comprised together. The invention comprises the following specific steps: collecting petroleum-contaminated soil samples from petroleum waste lagoons and characterizing the same, determining dominant bacterial species in the collected samples by Illumina sequencing, one of the next-generation sequencing analysis methods, subjecting soil mixtures to a bio stimulation process at a C:N:P ratio of 100: 15: 1, isolating genomic DNA, total RNA, and plasmids from soil samples, measuring the degrading bacteria, functional genes alkB, phnAc and nah using alkane and polyaromatic hydrocarbons (PAH) by quantitative real time PCR method, applying bioaugmentation to the petroleum-contaminated soil, having undergone biostimulation at a C:N:P ratio of 100: 15: 1, using Trametes versicolor and Bjerkandera adusta fungi species, and
evaluating the effect of the fungi used in bioaugmentation method on local microbial community, functional genes alkB, phnAc and nah.
Petroleum-contaminated soil samples were collected from petroleum waste lagoon and they were characterized, and then dominant bacterial species were determined in the collected samples by means of Illumina sequencing method. Petroleum- contaminated samples were mixed with clean forest soil samples at different ratios and placed in glass containers, and then N:P at different ratios was added to the
resulting soil mixtures and incubated at 24°C for 90 days. Genomic DNA, total RNA, and plasmids were isolated from the incubated samples. Alkane and polyaromatic hydrocarbons (PAH) were used for measuring the degrading bacteria and functional genes alkB, phnAc and nah by means of quantitative real time PCR performed with TaqMan probe. As a final treatment method, the treated contaminated soil was subject to biostimulation, followed by bioaugmentation method, using Trametes versicolor and Bjerkandera adusta fungi species.
The effect of said fungi on local microbial community and functional genes alkB, phnAc and nah was evaluated by performing bioaugmentation at 30°C for 60 days.
Throughout the degradation test, maximum amount of degradation was obtained from the microcosmoses whose C:N:P ratio was adjusted to 100: 15: 1. The amount of total organic carbon (TOC) elimination was found to increase 25% by biostimulation. The results of final treatment showed that Trametes versicolor and Bjerkandera adusta fungi were able to degrade petroleum residues remaining after biostimulation phase.
These two types of fungi were compared and it was shown that Trametes versicolor was much more effective than Bjerkandera adusta during degradation process of petroleum residues and that it eliminated petroleum residues by 36% at 30°C for 60 days.
Claims
A method for bioremediation of petroleum-contaminated soils, characterized by comprising the steps of: collecting petroleum-contaminated soil samples and determining dominant bacterial species in these samples, subjecting soil mixtures to a bio stimulation process at a C:N:P ratio of 100: 15: 1, isolating genomic DNA, total RNA, and plasmids from soil mixture samples having undergone biostimulation, measuring the degrading bacteria, functional genes alkB, phnAc and nah using alkane and polyaromatic hydrocarbons (PAH) by quantitative real time PCR, applying bioaugmentation to the contaminated soil having been treated using Trametes versicolor and Bjerkandera adusta fungi species, and evaluating the effect of the fungi used in bioaugmentation method on local microbial community, functional genes alkB, phnAc and nah present in the soil.
The method for bioremediation of petroleum-contaminated soils according to Claim 1, characterized in that N:P is added to the resulting soil mixtures and incubated at 24°C for 90 days.
The method for bioremediation of petroleum-contaminated soils according to Claim 2, characterized in that the degrading bacteria and functional genes alkB, phnAc and nah were measured using alkane and polyaromatic hydrocarbons (PAH) by quantitative real time PCR performed with TaqMan probe.
The method for bioremediation of petroleum-contaminated soils according to Claim 3, characterized in that the optimum fungus concentration was determined to be 18.7% and 12% for Trametes versicolor and Bjerkandera adusta, respectively.
The method for bioremediation of petroleum-contaminated soils according to Claim 4, characterized in that the maximum amount of degradation was obtained from the microcosmoses whose C:N:P ratio was adjusted to 100: 15: 1 throughout the biostimulation process.
The method for bioremediation of petroleum-contaminated soils according to Claim 5, characterized in that the effect of the fungi on local microbial community, functional genes alkB, phnAc and nah was evaluated by performing bioaugmentation at 30°C for 60 days.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/TR2017/050087 WO2018164648A1 (en) | 2017-03-06 | 2017-03-06 | Method for bioremediation of petroleum-contaminated soils |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/TR2017/050087 WO2018164648A1 (en) | 2017-03-06 | 2017-03-06 | Method for bioremediation of petroleum-contaminated soils |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2018164648A1 true WO2018164648A1 (en) | 2018-09-13 |
Family
ID=58455634
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/TR2017/050087 WO2018164648A1 (en) | 2017-03-06 | 2017-03-06 | Method for bioremediation of petroleum-contaminated soils |
Country Status (1)
| Country | Link |
|---|---|
| WO (1) | WO2018164648A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111922057A (en) * | 2020-07-30 | 2020-11-13 | 北京丰泽绿源环境技术有限公司 | In-situ remediation method for petroleum-polluted soil |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5725885A (en) | 1993-04-22 | 1998-03-10 | Bionutratech | Composition for enhanced bioremediation of oil |
| NZ519022A (en) | 2002-05-17 | 2004-08-27 | Horticulture & Food Res Inst | Fungal bioremediation |
| WO2014031016A1 (en) | 2012-08-21 | 2014-02-27 | Fundacja Uniwesytetu | Method of bioremediation of soil and/or soil-based masses polluted with petroleum-derived compounds |
-
2017
- 2017-03-06 WO PCT/TR2017/050087 patent/WO2018164648A1/en active Application Filing
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5725885A (en) | 1993-04-22 | 1998-03-10 | Bionutratech | Composition for enhanced bioremediation of oil |
| NZ519022A (en) | 2002-05-17 | 2004-08-27 | Horticulture & Food Res Inst | Fungal bioremediation |
| WO2014031016A1 (en) | 2012-08-21 | 2014-02-27 | Fundacja Uniwesytetu | Method of bioremediation of soil and/or soil-based masses polluted with petroleum-derived compounds |
Non-Patent Citations (2)
| Title |
|---|
| AIYOUB SHAHI ET AL: "Reconstruction of bacterial community structure and variation for enhanced petroleum hydrocarbons degradation through biostimulation of oil contaminated soil", CHEMICAL ENGINEERING JOURNAL, vol. 306, 14 July 2016 (2016-07-14), CH, pages 60 - 66, XP055418528, ISSN: 1385-8947, DOI: 10.1016/j.cej.2016.07.016 * |
| AIYOUB SHAHI ET AL: "The effects of white-rot fungi Trametes versicolor and Bjerkandera adusta on microbial community structure and functional genes during the bioaugmentation process following biostimulation practice of petroleum contaminated soil", INTERNATIONAL BIODETERIORATION & BIODEGRADATION, vol. 114, 1 October 2016 (2016-10-01), AMSTERDAM, NL, pages 67 - 74, XP055418356, ISSN: 0964-8305, DOI: 10.1016/j.ibiod.2016.05.021 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111922057A (en) * | 2020-07-30 | 2020-11-13 | 北京丰泽绿源环境技术有限公司 | In-situ remediation method for petroleum-polluted soil |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Haleyur et al. | Influence of bioaugmentation and biostimulation on PAH degradation in aged contaminated soils: response and dynamics of the bacterial community | |
| Dos Santos et al. | Rhizospheric microorganisms as a solution for the recovery of soils contaminated by petroleum: A review | |
| Wu et al. | Bioaugmentation and biostimulation of hydrocarbon degradation and the microbial community in a petroleum-contaminated soil | |
| Menendez-Vega et al. | Engineered in situ bioremediation of soil and groundwater polluted with weathered hydrocarbons | |
| Han et al. | Effects of different agricultural wastes on the dissipation of PAHs and the PAH-degrading genes in a PAH-contaminated soil | |
| Ahmad et al. | Remediation methods of crude oil contaminated soil | |
| Sheppard et al. | Microbial community and ecotoxicity analysis of bioremediated, weathered hydrocarbon-contaminated soil | |
| Pizarro‐Tobías et al. | Field trial on removal of petroleum‐hydrocarbon pollutants using a microbial consortium for bioremediation and rhizoremediation | |
| Xu et al. | Acceleration of polychlorinated biphenyls remediation in soil via sewage sludge amendment | |
| Alvarez et al. | Comparative bioremediation of crude oil-amended tropical soil microcosms by natural attenuation, bioaugmentation, or bioenrichment | |
| Heydarnezhad et al. | Optimizing toluene degradation by bacterial strain isolated from oil-polluted soils. | |
| Baig et al. | Assessment of degradation potential of Pseudomonas species in bioremediating soils contaminated with petroleum hydrocarbons | |
| Dewi et al. | Bioremediation of Indigosol Blue 04B Batik Effluent by Indigenous Fungal Isolates, Aspergillus spp. | |
| Mansur et al. | Assessment of the hydrocarbon degrading abilities of three bioaugmentation agents for the bioremediation of crude oil tank bottom sludge contaminated Libyan soil | |
| Medina et al. | Exploring the effect of composting technologies on the recovery of hydrocarbon contaminated soil post chemical oxidative treatment | |
| Anani et al. | Application of biosurfactant as a noninvasive stimulant to enhance the degradation activities of indigenous hydrocarbon degraders in the soil | |
| Agamuthu et al. | Biodegradation of used lubricating oil by microbes isolated from pristine soil environment | |
| Nozari et al. | Investigation of the effect of co-metabolism on removal of dodecane by microbial consortium from soil in a slurry sequencing bioreactor | |
| Thangarajan et al. | Comparison between different bio-treatments of a hydrocarbon contaminated soil from a landfill site | |
| WO2018164648A1 (en) | Method for bioremediation of petroleum-contaminated soils | |
| Bada et al. | Degradation of total petroleum hydrocarbon in petroleum products-contaminated soil using pig dung | |
| Czarny et al. | Metagenomic analysis of soil bacterial community and level of genes responsible for biodegradation of aromatic hydrocarbons | |
| Riccardi et al. | Characterization of bacterial population coming from a soil contaminated by polycyclic aromatic hydrocarbons (PAHs) able to degrade pyrene in slurry phase | |
| Berde et al. | Bioremediation of Petroleum Contaminated Soils | |
| Khalid et al. | Analysis of diesel hydrocarbon decomposition using efficient indigenous bacterial isolate: Bacterial growth and biodegradation kinetics |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 17714576 Country of ref document: EP Kind code of ref document: A1 |
|
| NENP | Non-entry into the national phase |
Ref country code: DE |
|
| 122 | Ep: pct application non-entry in european phase |
Ref document number: 17714576 Country of ref document: EP Kind code of ref document: A1 |