CN107828684B - Mixed microbial inoculum suitable for electric-microbial synergistic remediation of petroleum hydrocarbon contaminated soil and preparation and application thereof - Google Patents
Mixed microbial inoculum suitable for electric-microbial synergistic remediation of petroleum hydrocarbon contaminated soil and preparation and application thereof Download PDFInfo
- Publication number
- CN107828684B CN107828684B CN201711116796.7A CN201711116796A CN107828684B CN 107828684 B CN107828684 B CN 107828684B CN 201711116796 A CN201711116796 A CN 201711116796A CN 107828684 B CN107828684 B CN 107828684B
- Authority
- CN
- China
- Prior art keywords
- microbial inoculum
- soil
- mixed
- remediation
- microbial
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
Abstract
The invention belongs to the technical field of electric-microbial cooperative remediation of petroleum hydrocarbon contaminated soil, and particularly relates to a mixed microbial inoculum suitable for electric-microbial cooperative remediation of high-salinity petroleum hydrocarbon contaminated soil, and preparation and application thereof. The microbial inoculum is obtained by respectively fermenting and mixing halomonas xianhensis, bacillus licheniformis and bacillus cereus; the combined microbial inoculum is applied to an electric-microbial synergistic remediation technology, breaks the bottleneck of remediation of a high-salinity state of soil formed in the remediation process, reasonably utilizes functional microbial inoculum resources, effectively realizes the degradation of petroleum hydrocarbon in petroleum soil, and reduces the toxicity of organic pollutants of the petroleum hydrocarbon in the soil.
Description
Technical Field
The invention belongs to the technical field of electric-microbial cooperative remediation of petroleum hydrocarbon contaminated soil, and particularly relates to a mixed microbial inoculum suitable for electric-microbial cooperative remediation of high-salinity petroleum hydrocarbon contaminated soil, and preparation and application thereof.
Background
The continuous development of petroleum economy leads to the increasing demand for petroleum hydrocarbon exploitation year by year, and the quantity of petroleum hydrocarbon substances entering soil due to the ground exploitation, transportation leakage and the like is increased continuously, thus harming the natural environment and ecological safety. In contrast, the work of treating and repairing petroleum-contaminated soil is rapidly carried out and is gaining more and more attention. At present, petroleum hydrocarbon contaminated soil remediation technologies mainly based on biological and chemical methods are gaining favor. However, in view of the strong hydrophobic property of petroleum hydrocarbon pollutants, it is difficult for a single microbial remediation technology to achieve efficient and stable petroleum hydrocarbon degradation effects, and for this reason, a combined remediation technology formed by organic combination of single treatment technologies is a powerful direction for realizing breakthrough of petroleum-contaminated soil remediation technologies.
The electrokinetic remediation technology is a new hope for remediation of petroleum hydrocarbon contaminated soil as an organic contaminated soil remediation technology which is increasingly concerned in recent years. However, electrokinetic repair techniques also have the adverse effect of "tailing" the repair efficiency. In contrast, the electric-microbial cooperative repair technology is adopted, microbial repair is taken as a main body, microbial metabolism is enhanced through the electric repair process, and the electric-microbial cooperative repair technology is an effective means for quickly starting the petroleum hydrocarbon pollutant degradation process and maintaining the quick degradation period. However, at present, the cooperative repair technology of electrokinetic-microbe still has the tendency of polarizing the soil properties, wherein the high salt characteristic caused by the phenomenon of 'ion wall' at the electrode end is one of the remarkable characteristics of the application of the technology to the change of the soil properties. The high salt characteristic produces salt stress, which causes dehydration of many microorganisms to lose reproductive metabolic function, even death in large quantities, and loss of petroleum hydrocarbon metabolic activity. Therefore, the selection of the bacterial strain with high salt tolerance is one of the feasible methods for repairing the petroleum hydrocarbon pollution of the saline soil.
Disclosure of Invention
The invention aims to provide a mixed microbial inoculum suitable for electric-microbial synergistic remediation of petroleum hydrocarbon contaminated soil, and preparation and application thereof.
In order to achieve the purpose, the invention adopts the technical scheme that:
a mixed microbial inoculum suitable for electric-microbial synergistic remediation of high-salinity petroleum hydrocarbon polluted soil is obtained by respectively fermenting and mixing Halomonas xianhensis, Bacillus licheniformis and Bacillus cereus; halomonas xianhensis is preserved in China general microbiological culture collection center with the address of No. 3 Hospital No.1 of Xilu-Chen-Yang district, Beijing, the preservation number of CGMCC NO.1.6848, the preservation date of 2008, 9 months and 15 days, and the taxonomic name of Halomonas xianhensis; the bacillus licheniformis and the bacillus cereus are both preserved in China center for type culture Collection, the addresses are Wuchang Lodojia mountain in Wuhan city, and the preservation registration numbers are CCTCC NO: m2011261 and M2015705, the preservation days are 22 days 7 and 3 days 12 and 2015 2011 respectively, and the taxonomic nomenclature is Bacillus licheniformis and Bacillus cereus respectively.
The three strains have salt tolerance and can grow in a liquid environment with the salinity of 3-5% (w/v), wherein the Halomonas xianhensis, the Bacillus licheniformis and the Bacillus cereus can respectively utilize PAHs (phenanthrene, anthracene or fluoranthene), alkane (C16-C24) and fatty acid (palmitic acid and stearic acid) as unique carbon sources for growth and metabolism.
The microbial inoculum is obtained by respectively fermenting and mixing halomonas xianhensis, bacillus licheniformis and bacillus cereus and then immobilizing the mixture by a carrier.
Mixing the Halomonas xianhensis, the Bacillus licheniformis and the Bacillus cereus suspension according to the proportion of 2-3: 5-6: 1(v: v) and fixing to obtain an initial mixed microbial inoculum;
the Halomonas xianhensis, the Bacillus licheniformis and the Bacillus cereus suspension are mixed according to the proportion of 1: 3-5: 2-3 (v: v) and are immobilized to obtain the reinforced mixed microbial inoculum.
The application of the mixed microbial inoculum in the electric-microbial synergistic remediation of the high-salinity petroleum hydrocarbon contaminated soil is to apply the microbial inoculum to the remediation of the high-salinity petroleum hydrocarbon contaminated soil under the synergistic action of an electric field.
Mixing the Halomonas xianhensis, the Bacillus licheniformis and the Bacillus cereus suspension according to the proportion of 2-3: 5-6: 1(v: v) and fixing to obtain an initial mixed microbial inoculum;
the Halomonas xianhensis, the Bacillus licheniformis and the Bacillus cereus suspension are mixed according to the proportion of 1: 3-5: 2-3 (v: v) and are immobilized to obtain the reinforced mixed microbial inoculum.
The electric field condition is that a direct current electric field of 0.5-2.0 v/cm is applied, an initial mixed microbial inoculum is used at the beginning of electric restoration, and the number of bacterial microorganisms reaches 2-8 multiplied by 108CFU·g-1When the degradation rate of alkane components in the polluted soil reaches 40-50%, a reinforcing mixed microbial inoculum is applied to maintain the total amount of bacteria in the soil to be 90-110% of the initial content, and the whole repairing process is carried out in an electric-microbial synergistic repairing mode.
Mixing Halomonas xianhensis, Bacillus licheniformis and Bacillus cereus according to the proportion of 2-3: 5-6: 1(v: v) and 1: 3-5: 2-3 (v: v), respectively preparing an initial mixed microbial inoculum and a reinforcing mixed microbial inoculum after immobilization, and applying the initial mixed microbial inoculum and the reinforcing mixed microbial inoculum to the remediation process of petroleum-polluted soil by an electric-microbial synergistic remediation technology; the mixed bacterial liquid is immobilized by a carrier prepared from rice hulls with acidification and mineralization degree of 10-20% (w/w), but the mixed bacterial liquid is not limited to the strain immobilization by adopting carbonized rice hulls.
The method for restoring the soil polluted by the high-salinity petroleum hydrocarbon is to restore the soil polluted by the petroleum by the synergistic effect of the microbial inoculum and an electric field.
The initial mixed microbial inoculum is used at the beginning of electrically repairing the polluted soil to be treated by adopting a direct current electric field of 0.5-2.0 v/cm, so that the number of bacterial microorganisms in the soil reaches 2-8 multiplied by 108CFU·g-1When the degradation rate of alkane components in the polluted soil reaches 40-50%, a reinforcing mixed microbial inoculum is applied, the total amount of bacteria in the soil is maintained to be 90-110% of the total content of the bacteria in the initial soil, and the whole repairing process is carried out in an electric-microbial synergistic repairing mode.
The mixed bacterial agent can grow in soil with salt content of 0.5-2% (w/w), and has metabolic degradation activity on petroleum hydrocarbon pollutants.
The invention has the advantages that:
in view of the complex components of petroleum hydrocarbon, the invention organically combines a plurality of different petroleum hydrocarbon component metabolism functional strains with high salt tolerance characteristic to prepare a mixed bacterial liquid and an immobilized bacterial agent by screening different single component functional degradation bacterial strains, can effectively realize functional complementation, degrade the toxicity of pollutants and realize the synergistic co-metabolism among the bacterial strains, prepares a special bacterial agent by a special formula to realize the continuous and efficient process of electric-microorganism synergistic remediation of the petroleum hydrocarbon polluted soil so as to solve the defect of tailing of the application efficiency of the remediation technology, and concretely comprises the following steps:
1) the halomonas xianhensis, the bacillus licheniformis and the bacillus cereus all have high-salt resistance characteristics, belong to moderately halophilic bacteria, can survive in a liquid environment with the salinity of 3-5% (w/v), and the prepared mixed microbial inoculum also has the high-salt resistance characteristic, so that the mixed microbial inoculum has good adaptability to the local ion accumulation effect generated in the electric restoration process, the growth and metabolism activity of the mixed microbial inoculum cannot be influenced, and the mixed microbial inoculum is more suitable for the restoration application of petroleum polluted soil in saline-alkali soil of an oil field area.
2) The halomonas xianhensis, the bacillus licheniformis and the bacillus cereus respectively have the metabolic capacities of PAHs (phenanthrene, anthracene or fluoranthene), alkane (C16-C24) and fatty acid (palmitic acid and stearic acid), and are all screened from petroleum hydrocarbon polluted soil, so that the halomonas xianhensis, the bacillus licheniformis and the bacillus cereus can specifically metabolize and degrade each specific component in the petroleum hydrocarbon while tolerating a high-salinity soil environment, and have the immune characteristic of non-specific degradation substrates, thereby ensuring the sustainable metabolic activity of the halomonas xianhensis, the bacillus licheniformis and the bacillus cereus under the condition of electric field strengthening treatment.
3) The initial mixed microbial inoculum and the reinforcing mixed microbial inoculum are prepared by three different functional metabolic strains according to a proportion, wherein the mixed microbial inoculum which takes metabolic PAHs and alkane as main components is adopted at the beginning of repair, the degradation effect is carried out on initial petroleum components, when the degradation rate of the alkane reaches 40-50%, fatty acid substances are accumulated to a certain degree, the mixed microbial inoculum which increases the content proportion of the fatty acid metabolic strains is supplemented for continuous treatment, the accumulated fatty acid substances can be removed in a targeted manner, the metabolic bottleneck is effectively crossed, the toxic effect of the fatty acid with higher accumulation is improved, and the composition stability and the functional efficacy of the functional degradation microbial inoculum are promoted.
Drawings
Fig. 1 is a schematic diagram of an electric apparatus used in an electric process participation group according to embodiment 2 of the present invention.
FIG. 2 is a graph showing the change in total salt content at each site of soil in the electrokinetic-microbial remediation treatment A (EK-Bio-A) according to test treatment 35d provided in example 2 of the present invention.
FIG. 3 is a graph showing the change in soil pH and bacterial microorganism counts in soil at the anode end of each experimental treatment provided in example 2 of the present invention.
FIG. 4 is a graph of the total petroleum hydrocarbon degradation rate over time for each of the experimental treatments provided in example 2 of the present invention.
Detailed Description
The mixed microbial inoculum of the invention is prepared from Halomonas xianhensis (Halomonas xianhensis) A-1 CGMCC NO.1.6848, Bacillus licheniformis (Bacillus licheniformis) CCTCC NO: m2011261 and Bacillus cereus (Bacillus cereus) CCTCC NO: m2015705 is prepared by mixing in different proportions and immobilizing, the three strains have high salt resistance, and have high-efficiency metabolic capability respectively for PAHs (phenanthrene, anthracene or fluoranthene), alkane (C16-C24) and fatty acid (palmitic acid or stearic acid) in petroleum hydrocarbon components. Mixing Halomonas xianhensis, Bacillus licheniformis and Bacillus cereus according to the proportion of 2-3: 5-6: 1(V: V), immobilizing to prepare an initial mixed microbial inoculum, and applying the initial mixed microbial inoculum to initial electrokinetic-microbial synergistic remediation of petroleum hydrocarbon contaminated soil, wherein the number of bacterial microorganisms reaches 4-8 multiplied by 108CFU·g-1. When the degradation rate of alkane components reaches 40-50%, a reinforcing mixed microbial inoculum prepared by mixing and immobilizing the halomonas xianhensis, the bacillus licheniformis and the bacillus cereus according to the proportion of 1: 3-5: 2-3 (V: V) is supplemented, and the total amount of bacteria in the soil is maintained to 90-110% of the initial content. The reinforcing microbial inoculum is applied and simultaneously supplied with an organic carbon source and inorganic nutrient ions. The combined microbial inoculum is applied to an electric-microbial synergistic remediation technology, breaks the bottleneck of remediation of a high-salinity state of soil formed in the remediation process, reasonably utilizes the resources of the functional microbial inoculum, effectively realizes the degradation of petroleum hydrocarbon in the petroleum soil, and reduces the toxicity of organic pollutants of the petroleum hydrocarbon in the soil.
The present invention will be described in further detail with reference to the accompanying drawings and examples.
Example 1 preparation of Mixed inoculum suitable for electrokinetic-microbial synergistic remediation of Petroleum Hydrocarbon contaminated soil
The bacteria source sample used in the embodiment is collected from high-salt-content soil polluted by petroleum hydrocarbon in an oil field area for a long time, the total salt content in the soil reaches 1.6% (w/w), 5.0g of petroleum hydrocarbon polluted saline soil is suspended by shaking in 30ml of inorganic salt culture medium, shaking culture is carried out at 160rpm/min for 20min, standing is carried out for 3min, 1ml of upper suspension is absorbed to be enriched, domesticated and cultured for 15d in inorganic salt liquid culture medium respectively containing three inorganic salts of phenanthrene 100mg/L, n-hexadecane 300mg/L and palmitic acid 100mg/L, and the saxiella saxifraga, bacillus licheniformis and the bacillus cereus which take phenanthrene in PAHs as metabolizable carbon sources, the bacillus licheniformis and the palmitic acid as metabolizable carbon sources are obtained after streaking, separating, purifying and identifying by using the solid inorganic salt culture medium.
The Halomonas xianhensis is preserved in China general microbiological culture collection center, the address is No. 3 Siro No.1 Bichen Xilu in the area of rising of Beijing, the preservation number is CGMCC NO.1.6848, the preservation date is 2008, 9 months and 15 days, and the Halomonas xianhensis is obtained by taxonomic nomenclature; the bacillus licheniformis and the bacillus cereus are both preserved in China center for type culture Collection, the addresses are Wuchang Lodojia mountain in Wuhan city, and the preservation registration numbers are CCTCC NO: m2011261 and M2015705, the preservation days are 22 days 7 and 3 days 12 and 2015 2011 respectively, and the taxonomic nomenclature is Bacillus licheniformis and Bacillus cereus respectively.
The inorganic salt liquid culture medium comprises the following components: 1.5 g.L-1NaCl，0.2g·L-1MgSO4，1.5g·L- 1NH4NO3，1.0g·L-1K2HPO4，0.5g·L-1KH2PO4，0.02g·L-1CaCl2，0.02g·L-1FeSO41L of distilled water, pH value of 7.0-7.3, sterilizing at 121 deg.C for 20 min.
The inorganic salt solid culture medium comprises the following components: 1.5 g.L-1NaCl，0.2g·L-1MgSO4，1.5g·L- 1NH4NO3，1.0g·L-1K2HPO4，0.5g·L-1KH2PO4，0.02g·L-1CaCl2，0.02g·L-1FeSO4，15g·L-1Agar and 1L of distilled water are added, the pH value is 7.0-7.3, sterilization is carried out for 20min at 121 ℃, 1ml of ethyl ether solution with the concentration of 1mg/ml phenanthrene, 10ul of n-hexadecane and 1ml of n-hexane solution of palmitic acid with the concentration of 1mg/ml are respectively coated on the surface of the solid culture medium, and the solid culture medium is reserved after the organic solvent is volatilized.
Single colony of each of the three preserved strains is picked and inoculated in 5ml beef extract peptone liquid medium containing 3% (w/v) NaCl, and shake culture is carried out for 16h at 37 ℃ to obtain seed liquid. Respectively sucking 300ul strain culture seed liquid, further inoculating into beef extract peptone amplification medium containing 3% (w/v) NaCl, shake culturing at 37 deg.C for 48 hr, 6000rpm min-1The cells were collected by centrifugation for 10 min. The collected thalli are respectively cleaned by adopting an inorganic salt culture medium, and are prepared into thalli suspension by using the inorganic salt culture medium. Mixing Halomonas xianhensis, Bacillus licheniformis and Bacillus cereus suspensions according to the proportion of 2.5:5:1(v: v) and 1:4:3(v: v) respectively to prepare an initial mixed bacterial liquid and a reinforced mixed bacterial liquid.
The inorganic salt liquid culture medium comprises the following components: 3.0g/L beef extract, 10g/L peptone, 5.0g/L NaCl, 1L distilled water, pH 7.0.
And (2) carbonizing the complete rice hulls by using 50% (w/w) sulfuric acid, neutralizing a sulfuric acid system to be neutral by using ammonia water when the mineralization rate of the rice hulls reaches 15%, washing and air-drying by using clear water, adding the mixture into the mixed bacteria liquid according to the proportion of 20% (w/v, g/ml), and carrying out immobilization treatment at the temperature of 4 ℃ to prepare the initial mixed bacteria agent and the reinforcing mixed bacteria agent. The initial mixed microbial inoculum has the degradation characteristic mainly based on the metabolism of alkane and aromatic hydrocarbon in petroleum hydrocarbon components, and the reinforced mixed microbial inoculum has the characteristic of stronger fatty acid degradation.
Example 2 application of mixed microbial inoculum in electrokinetic-microbial synergistic remediation of petroleum hydrocarbon contaminated soil
The contaminated soil used in the test is collected from petroleum hydrocarbon contaminated soil in an oil field area, the content of petroleum hydrocarbon in the soil reaches 4.6% (w/w), the soil to be treated is naturally air-dried after removing visible weed impurities and the like, and the soil is screened by a 2mm sieve for later use. The initial properties of the soil include pH 7.2, total ion content of the soil of 1.2% (w/w), and soil moisture content adjusted to 23% (w/w) for treatment.
As shown in Table 1, six sets of treatments were set, and they were designated as Control (CK), electrokinetic treatment A (EK-A), electrokinetic treatment B (EK-B), microbial treatment (Bio), electrokinetic-microbial synergistic treatment A (EK-Bio-A) and electrokinetic-microbial synergistic treatment B (EK-Bio-B).
Microorganism-containing remediation treatment groups (Bio, EK-Bio-A, EK-Bio-B) treatment starts with the application of the initial mixed inoculum prepared as in example 1, after which the bacterial microorganism count in the soil reaches 5.2X 108CFU·g-1. The electric restoration device for the petroleum hydrocarbon polluted soil is a box body (figure 1) with the length of 28cm, the width of 15cm and the height of 14cm, and 1600g of petroleum hydrocarbon polluted soil is filled in each box body; the electrode used for electric repair is a stainless steel bar electrode, the diameter of the electrode is 1.2cm, the height of the electrode is 18cm, the distance between the electrodes on two sides is 24cm, and 24V direct current voltage is applied.
Table 1 basic conditions and test parameter tables for each test treatment set-up.
Every 7d of the test is a sampling period, the contents of petroleum hydrocarbon and petroleum hydrocarbon components are monitored after the samples are collected, and when the total degradation rate of alkane in the petroleum hydrocarbon components reaches 42 percent, the reinforcing mixed microbial inoculum is applied to ensure that the total amount of bacterial microorganisms in the soil reaches 6.1 multiplied by 108CFU·g-1And continuing to perform restoration treatment, regularly tracking and monitoring the change of the soil pH value parameter in the electric treatment group B (EK-B) and the electric-microorganism treatment groups (EK-Bio-A and EK-Bio-B), and supplying 1mol/L HCl or 1mol/L NaOH to regulate and control the pH value to be within a range of 6-8 when the pH value of the anode end is less than 6 or the pH value of the cathode end is more than 8, wherein the total restoration period reaches 63 d.
Test results show that with the continuous electric treatment, the pH value of the soil is extremely acid-alkalized, and the number of bacterial microorganisms is influenced (figure 3); however, the soil pH is effectively regulated and controlled to be 6-7, but the total salt content on two sides of the electrode in the soil is obviously improved in the later stage of the electric treatment, the soil salinization is obvious (figure 2), and the number of indigenous microorganisms in the electric treatment B (EK-B) with the soil pH effectively regulated is still obviously reduced. After the initial mixed microbial inoculum B is applied, the microbial number is effectively maintained (figure 3), but in the later treatment period, the accumulation of organic acid limits the activity of the microorganisms, and after the reinforcing mixed microbial inoculum is applied in the electric-microbial synergistic treatment B, the microbial number is supplemented, and the metabolic activity is effectively continued. The total degradation rate of the petroleum hydrocarbon is determined, the application of the initial mixed microbial inoculum remarkably accelerates the degradation and removal of the petroleum hydrocarbon at the initial stage, the addition of the reinforcing microbial inoculum further optimizes the efficient continuous degradation process of the petroleum hydrocarbon, and after 63d of treatment, the total degradation rate of the petroleum hydrocarbon reaches 40.1 percent (shown in figure 4), and is improved by 8 percent compared with the degradation rate of the petroleum hydrocarbon in a treatment group without the reinforcing mixed microbial inoculum. Therefore, the mixed microbial inoculum is suitable for application in the treatment of the electric-microbial cooperative repair technology, and the purpose of overcoming the adverse conditions of the ion accumulation at the electrode end is achieved; and aiming at the function complementary action of alkane, PAHs and fatty acid metabolism, the combined application mode of the initial mixed microbial inoculum and the reinforcing mixed microbial inoculum is more favorable for playing the repair function of the functional microbial inoculum.
Claims (8)
1. A mixed microbial inoculum suitable for electric-microbial synergistic remediation of high-salinity petroleum hydrocarbon contaminated soil is characterized by comprising the following components in parts by weight: the microbial inoculum is made of halomonas xianhensis (Halomonasxianhensis) Bacillus licheniformis (B), (B)Bacillus licheniformis) And Bacillus cereus (B.cereus)Bacillus cereus) Respectively fermenting and mixing to obtain; wherein Halomonas xianhehensis is preserved in China general microbiological culture Collection center with the address of No. 3 Xilu-Beijing-oriented region of Beijing, the preservation number of CGMCC NO.1.6848, the preservation date of 2008-9.15, and taxonomic nomenclatureHalomonasxianhensis(ii) a The bacillus licheniformis and the bacillus cereus are both preserved in China center for type culture Collection, the addresses are Wuchang Lodojia mountain in Wuhan city, and the preservation registration numbers are CCTCC NO: m2011261 and M2015705, wherein,the collection days were 22/2011/7 and 3/2015/12, and taxonomical designations were Bacillus licheniformis PS5 and Bacillus cereus W1.
2. The mixed microbial inoculum in the soil suitable for electro-microbial cooperative remediation of high salinity petroleum hydrocarbon contamination according to claim 1, wherein: the microbial inoculum is obtained by respectively fermenting and mixing halomonas xianhensis, bacillus licheniformis and bacillus cereus and then immobilizing the mixture by a carrier.
3. The mixed microbial inoculum suitable for electrokinetic-microbial cooperative remediation of high salinity petroleum hydrocarbon contaminated soil as defined in claim 1 or 2, wherein:
mixing the suspension of the halomonas xianhensis, the bacillus licheniformis and the bacillus cereus according to the volume ratio of 2 ~ 3:5 ~ 6:1 and fixing to obtain an initial mixed microbial inoculum;
the halomonas xianhensis, the bacillus licheniformis and the bacillus cereus suspension are mixed according to the volume ratio of 1:3 ~ 5:2 ~ 3 and are immobilized to obtain the reinforced mixed microbial inoculum.
4. The use of the mixed microbial inoculum suitable for the electro-microbial synergistic remediation of high salinity petroleum hydrocarbon contaminated soil of claim 1, wherein: the microbial inoculum is applied to the remediation of the soil polluted by the high-salinity petroleum hydrocarbon under the synergistic action of the microbial inoculum and an electric field.
5. The use of the mixed bacterial preparation suitable for electro-microbial synergistic remediation of soil contaminated with high salinity petroleum hydrocarbon according to claim 4, wherein the suspensions of Halomonas xianhensis, Bacillus licheniformis and Bacillus cereus are mixed in a volume ratio of 2 ~ 3:5 ~ 6:1 and fixed to obtain an initial mixed bacterial preparation;
the halomonas xianhensis, the bacillus licheniformis and the bacillus cereus suspension are mixed according to the volume ratio of 1:3 ~ 5:2 ~ 3 and are immobilized to obtain the reinforced mixed microbial inoculum.
6. According to claim 4 or 5The application of the mixed microbial inoculum suitable for electric-microbial synergistic remediation of the high-salinity petroleum hydrocarbon contaminated soil is characterized in that the electric field condition is that a direct current electric field of 0.5 ~ 2.0.0 v/cm is applied, the initial mixed microbial inoculum is used at the beginning of electric remediation, the number of bacteria and microbes reaches 2 ~ 8 multiplied by 108CFU·g-1When the degradation rate of alkane components in the polluted soil reaches 40 ~ 50%, a reinforcing mixed microbial inoculum is applied to maintain the total amount of bacteria in the soil to 90 ~ 110% of the initial content, and the whole restoration process is carried out in an electric-microbial synergistic restoration mode.
7. A method for remediating high salinity petroleum hydrocarbon contaminated soil is characterized by comprising the following steps: the microbial inoculum according to any one of claims 1 to 3 is used for repairing petroleum polluted soil under the synergistic action of an electric field.
8. The method for remediating petroleum-contaminated soil as set forth in claim 7, wherein the initial mixed microbial inoculum is used at the beginning of the electric remediation of the contaminated soil to be treated using a DC electric field of 0.5 ~ 2.0.0 v/cm to increase the number of bacterial microorganisms in the soil to 2 ~ 8X 108CFU·g-1When the degradation rate of alkane components in the polluted soil reaches 40 ~ 50%, a reinforcing mixed microbial inoculum is applied to maintain the total amount of bacteria in the soil to 90 ~ 110% of the total amount of bacteria in the initial soil, and the whole restoration process is carried out in an electric-microbial synergistic restoration mode.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711116796.7A CN107828684B (en) | 2017-11-13 | 2017-11-13 | Mixed microbial inoculum suitable for electric-microbial synergistic remediation of petroleum hydrocarbon contaminated soil and preparation and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711116796.7A CN107828684B (en) | 2017-11-13 | 2017-11-13 | Mixed microbial inoculum suitable for electric-microbial synergistic remediation of petroleum hydrocarbon contaminated soil and preparation and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107828684A CN107828684A (en) | 2018-03-23 |
| CN107828684B true CN107828684B (en) | 2020-01-24 |
Family
ID=61654251
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711116796.7A Active CN107828684B (en) | 2017-11-13 | 2017-11-13 | Mixed microbial inoculum suitable for electric-microbial synergistic remediation of petroleum hydrocarbon contaminated soil and preparation and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107828684B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111440741B (en) * | 2020-04-03 | 2022-05-31 | 中国科学院沈阳应用生态研究所 | Functional bacterium suitable for restoring petroleum-polluted soil in direct-current electric field anode region and application |
| CN112692051B (en) * | 2021-01-25 | 2022-09-30 | 明志科技大学 | Method and equipment for removing pollutants by using beneficial microorganism mixed bacterial liquid |
| CN113388542B (en) * | 2021-06-09 | 2022-05-31 | 中国科学院沈阳应用生态研究所 | Polyhalogenated aromatic hydrocarbon degrading bacteria and application thereof |
| CN114231439B (en) * | 2021-11-22 | 2023-11-14 | 中化珠海石化储运有限公司 | Microbial preparation for degrading oil in high-salinity water |
| CN114480189B (en) * | 2022-01-23 | 2023-09-08 | 北方民族大学 | Microbial composite microbial agent and preparation method and application thereof |
| CN116286461B (en) * | 2022-12-08 | 2023-09-08 | 山东碧蓝生物科技有限公司 | Bacillus salicinus with soil improvement effect, screening method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101838616A (en) * | 2009-03-17 | 2010-09-22 | 清华大学 | Halomonas capable of degrading polyaromatic hydrocarbon and application thereof |
| CN103160448A (en) * | 2011-12-13 | 2013-06-19 | 中国科学院沈阳应用生态研究所 | Petroleum hydrocarbon degradation bacteria being suitable for electric field condition and application thereof |
| CN105602869A (en) * | 2016-02-01 | 2016-05-25 | 中国科学院沈阳应用生态研究所 | Intermediate substrate degrading bacterium suitable for electrically repairing long-chain-alkane polluted soil and application thereof |
-
2017
- 2017-11-13 CN CN201711116796.7A patent/CN107828684B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101838616A (en) * | 2009-03-17 | 2010-09-22 | 清华大学 | Halomonas capable of degrading polyaromatic hydrocarbon and application thereof |
| CN103160448A (en) * | 2011-12-13 | 2013-06-19 | 中国科学院沈阳应用生态研究所 | Petroleum hydrocarbon degradation bacteria being suitable for electric field condition and application thereof |
| CN105602869A (en) * | 2016-02-01 | 2016-05-25 | 中国科学院沈阳应用生态研究所 | Intermediate substrate degrading bacterium suitable for electrically repairing long-chain-alkane polluted soil and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| Chunli Wan等.Electrokinetic remediation and microbial community shift of β-cyclodextrin-dissolved petroleum hydrocarbon-contaminated soil.《Appl Microbiol Biotechnol》.2010,第89卷第2019-2025页. * |
| 王菲等.土壤中多环芳烃的微生物降解及土壤细菌种群多样性.《应用生态学报》.2009,第20卷(第12期),第3020-3026页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107828684A (en) | 2018-03-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107828684B (en) | Mixed microbial inoculum suitable for electric-microbial synergistic remediation of petroleum hydrocarbon contaminated soil and preparation and application thereof | |
| CN102994404B (en) | Bacterial strain for degrading polycyclic aromatic hydrocarbon and application of bacterial strain cooperated with surface active agent in soil remediation | |
| CN103160448B (en) | Petroleum hydrocarbon degradation bacteria being suitable for electric field condition and application thereof | |
| CN103045507B (en) | Bacteria separation method for efficiently degrading high molecular weight polycyclic aromatic hydrocarbons and application thereof | |
| CN102864112B (en) | Heavy metal resistant strain for degrading polycyclic aromatic hydrocarbons (PAHs) and application thereof in combined contaminated soil remediation | |
| CN104946620A (en) | Immobilized microbial inoculant for restoring petroleum-hydrocarbon-polluted alkaline-saline soil and preparation method thereof | |
| CN102943052B (en) | Heavy metal-resistant polycyclic aromatic hydrocarbon (PAHs) degrading bacteria and application thereof in remediation of composite contaminated soil | |
| CN111748483B (en) | Bacillus for degrading petroleum hydrocarbon and application thereof | |
| Deng et al. | Removal of phenanthrene in contaminated soil by combination of alfalfa, white-rot fungus, and earthworms | |
| CN106635908A (en) | Advenella kashmirensis, microbial agent and application of advenella kashmirensis and microbial agent | |
| Ambaye et al. | Insights into rhamnolipid amendment towards enhancing microbial electrochemical treatment of petroleum hydrocarbon contaminated soil | |
| CN108300674A (en) | A kind of oil degradation bacteria and its preparation method and the application in degrading crude oil | |
| CN114292764B (en) | Achromobacter strain JD417 and application thereof | |
| CN111440741B (en) | Functional bacterium suitable for restoring petroleum-polluted soil in direct-current electric field anode region and application | |
| CN1803228A (en) | Biological restoration method for p-chloronitrobenzene compound polluted environment | |
| Beolchini et al. | Bioremediation of sediments contaminated with polycyclic aromatic hydrocarbons: the technological innovation patented review | |
| Olszanowski et al. | The Use of an Electric Field to Enhance Bacterial Movement and Hydrocarbon Biodegradation in Soils. | |
| CN105802884B (en) | A kind of DDT degradation bacteria and its utilize DDT in-situ immobilization contaminated soil method | |
| CN102864086B (en) | Strain capable of degrading polycyclic aromatic hydrocarbons (PAHs) and application thereof in soil remediation | |
| CN101343616B (en) | One-strain high-ring polycyclic aromatic hydrocarbon degradation bacterium and uses thereof | |
| CN101654656A (en) | Separation and purification method and application of aerobic degradation pure culture of polycyclic aromatic hydrocarbon | |
| CN100439491C (en) | Globular bacillus and its use | |
| Vineetha et al. | Bioremediation of oil contaminated soil | |
| Guo et al. | Study on remediation technologies of organic and heavy metal contaminated soils | |
| CN105420163A (en) | Dibenzanthracene degrading bacterium and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |