CN108823195A - A kind of preparation method and application method of mushroom bran immobilized microbial inoculum - Google Patents
A kind of preparation method and application method of mushroom bran immobilized microbial inoculum Download PDFInfo
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- CN108823195A CN108823195A CN201810621166.3A CN201810621166A CN108823195A CN 108823195 A CN108823195 A CN 108823195A CN 201810621166 A CN201810621166 A CN 201810621166A CN 108823195 A CN108823195 A CN 108823195A
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- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
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Abstract
The invention discloses a kind of preparation methods of mushroom bran immobilized microbial inoculum, include the following steps：（1）The preparation of bacteria suspension：Petroleum hydrocarbon degradation bacterium addition physiological saline is prepared into bacteria suspension；（2）The preparation of carrier：It is air-dried after mushroom bran is cleaned up, it is spare to cross 20-80 mesh；（3）Absorption：By the step（2）In mushroom bran and the step（1）In bacteria suspension with 1:15-25（v/v）Ratio mixing is subsequently placed in 30-37 DEG C, adsorbs 16-24h in 120-160rpm shaking table, discards upper solution；（4）Centrifugation：The mushroom bran after absorption is transferred in centrifuge tube with physiological saline, is then centrifuged for discarding supernatant liquid；After the above-mentioned centrifugally operated of repetition is multiple, centrifugation gained precipitating is immobilized microbial inoculum.Mushroom bran immobilized microbial inoculum produced by the present invention can be applied to remedying oil-polluted soils, and high to petroleum hydrocarbon removal efficiency, without secondary pollution, more independent mushroom bran, microorganism remediation degradation rate are significantly promoted.
Description
Technical field
The present invention relates to environment remediation technical fields, and in particular to it is a kind of can remedying oil-polluted soils mushroom bran immobilization
The preparation and application of microbial inoculum.
Background technique
With the development of petroleum industry, oil pollution gradually aggravates, and the residence time is long in the environment for organic pollution materials, knot
Structure is not stable, degradable, causes to seriously threaten to environment and human body.Due to microorganism remediation technology have it is at low cost, without secondary
The advantages that pollution and high treating effect, in recent years using than wide in organic contamination reparation.Immobilized microorganism technique because
It is conducive to shield adverse circumstances, improves the advantages such as degradation efficiency, receive more and more attention and answer with strain density height
With.
Mushroom bran is the residual waste of planting edible mushroom industry, and every production 1kg edible mushroom about generates 5kg mushroom bran, every year about
More than 8000 ten thousand tons of waste mushroom leftovers are not only wasted land resource, can also in combustion process by the traditional approach processing such as burning, landfill
Greenhouse gases such as CH4 is generated He the poisonous and harmful substances such as bioxin, will cause different degrees of environmental pollution.Therefore mushroom bran is probed into
Resource utilization mode becomes particularly important.Research find in mushroom bran rich in organic matter, potassium, calcium, magnesium, nitrogen, phosphorus and copper, zinc,
The various nutrient elements such as iron can be used for crop fertilizer and soil conditioner after working process.It is loose porous in its structure, surface compared with
It is coarse, it can be used for adsorbing heavy metal and organic pollutant.Secondly functional group such as hydroxyl, carboxyl, acyl rich in mushroom bran
Amido etc. can with heavy metal, organic pollutant by way of absorption, complexing, ion exchange stable bond, reduce ambient wind
Danger.It is living containing a large amount of fungal myceliums, enzyme material and the organic acid generated through mycelium conversion and biology in mushroom bran simultaneously
Property substance, being capable of enhancing degradation organic pollutant.
Fixed microorganism carrier material most of selects as active carbon, haydite and zeolite etc., these carrier materials at present
Material will receive non-natural existing region and the conditions such as non-renewable and lack the limitation of availability.Therefore select one kind that can hold
Continuous and widely applied carrier material seems especially urgent.For using agricultural wastes as carrier material immobilized microorganism
Renovation of organic pollution object achieves certain effect, but since it only has single absorption property, degradation of organic substances ability phase
To weaker, it is subject to certain restrictions.And mushroom bran is as one of abandoned biomass resource, from a wealth of sources, nutriment is abundant, makes it
Existence place can be not only provided for microorganism, guarantee its sufficient nutrient, while fungal mycelium being capable of secretion laccase, polyphenol
Oxidizing ferment, manganese peroxidase etc. constitute fungi-enzyme system and strengthen bacterial degradation organic pollutant, improve removal efficiency.At present
The research for being used to repair the organic pollutants such as petroleum hydrocarbon about mushroom bran immobilized microorganism is less, therefore probes into mushroom bran immobilized bacterium
The preparation and application of agent become particularly important.
Summary of the invention
In view of this, promoting oil pollution organic matter using fungi, enzyme and bacterium synergistic effect the present invention provides a kind of
The technical solution of degradation, the carrier for using the mushroom bran with absorption and enzyme degradation to grow for bacterium living beings, has greatly promoted
The degradation of machine pollutant improves pollutant remediation efficiency.
To achieve the goals above, the present invention adopts the following technical scheme that：
One of present invention is to provide a kind of preparation method of mushroom bran immobilized microbial inoculum, includes the following steps：
（1）The preparation of bacteria suspension：Petroleum hydrocarbon degradation bacterium addition physiological saline is prepared into bacteria suspension；
（2）The preparation of carrier：It is air-dried after mushroom bran is cleaned up, it is spare to cross 20-80 mesh；
（3）Absorption：By the step（2）In mushroom bran and the step（1）In bacteria suspension with 1:15-25（v/v）Ratio is mixed
It closes, is subsequently placed in 30-37 DEG C, adsorbs 16-24h in 120-160rpm shaking table, discard upper solution；
（4）Centrifugation：The mushroom bran after absorption is transferred in centrifuge tube with physiological saline, is then centrifuged for discarding supernatant liquid；In repetition
State centrifugally operated it is multiple after, centrifugation gained precipitating be immobilized microbial inoculum.
Further, the step（1）In petroleum hydrocarbon degradation bacterium be two plants, respectively anthropi
（Ochrobactrum sp.）And microbacterium（Microbacterium sp.）, specific screening means are with aged petroleum Polluted Soil
Earth is bacterium source, and petroleum hydrocarbon is sole carbon source, filters out two plant heights effect petroleum hydrocarbon degradation bacterium, identifies through 16SrRNA, respectively pale
Bacillus（Ochrobactrum sp.）And microbacterium（Microbacterium sp.）, later by the two bacterium solution with 2:3 volume ratios
Mixing.Bacterium solution absorption values in the case where ultraviolet specrophotometer measures its 600nm wavelength are denoted as OD600, are measured as 0.4-
0.8, i.e., each bacterium solution OD600 adjusted before bacterium solution mixing is between 0.4-0.8.
Further, the step（1）Middle bacteria suspension extinction degree in the case where ultraviolet specrophotometer measures its 600nm wavelength
Value is denoted as OD600, is measured as 0.4-0.8.That is control bacteria suspension strain density OD600 is in 0.4-0.8.
Further, the step（3）Middle mushroom bran and bacteria suspension mixed proportion are 1:20（v/v）.
Further, the step（4）Middle centrifugation is to be centrifuged 10min at 1000rpm；The centrifugally operated number is 3-5 times.
Further, the step（1）In petroleum hydrocarbon degradation bacterium acquisition modes be：By what is cultivated in liquid medium
Supernatant is abandoned after the centrifugation of petroleum hydrocarbon degradation bacterium bacterium solution, then rinses to obtain with sterile saline；The petroleum hydrocarbon degradation bacterium is
It cultivates to the bacterium of late log phase；Further, the fluid nutrient medium is beef extract-peptone fluid nutrient medium, the beef
Cream peptone Liquid Culture based component is：Beef extract 5g, peptone 10g, NaCl 5g, 1000 mL of distilled water；The liquid training
The preparation for supporting base is adjustment pH 7. 0~7.5 after mixing beef extract, peptone, NaCl, distilled water in proportion, is then existed
121 DEG C of 20 min of sterilizing to obtain the final product.
Further, the step（2）Middle mushroom bran is residual waste after planting edible mushroom harvest；The edible bacterium includes
Pleurotus eryngii, oyster mushroom and needle mushroom.
The two of the present invention are to provide a kind of application method of mushroom bran immobilized microbial inoculum, the life applied to petroleum hydrocarbon contaminated soil
Object reparation.
Further, the mushroom bran immobilized microbial inoculum is added with 10% mass ratio and repairs 35- in oil-polluted soils
60 days
Compared with prior art, present invention tool has the advantage that：
1. the present invention is using abandoned biomass resource mushroom bran as carrier material, preparation cost is low, raw material is extensive, full of nutrition, real
Existing waste resource comprehensive utilization；
2. the present invention, using mushroom bran as carrier material, functional group content is high, certain skeleton structure is presented, life can be provided for degradation bacteria
Long breeding place, while fungal mycelium can secrete secretion organic acid, enzyme material in mushroom bran, promote petroleum hydrocarbon class pollutant
Degradation, to reach " treatment of wastes with processes of wastes against one another " double effects；
The length 3. mushroom bran immobilized microbial inoculum activity prepared by the present invention is held time, viable count is more, is readily transported preservation；
4. mushroom bran immobilized microbial inoculum produced by the present invention can be applied to remedying oil-polluted soils, high to petroleum hydrocarbon removal efficiency,
Without secondary pollution, more independent mushroom bran, microorganism remediation degradation rate are significantly promoted.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
The embodiment of invention for those of ordinary skill in the art without creative efforts, can also basis
The attached drawing of offer obtains other attached drawings.
Fig. 1 is the mushroom bran scanning electron microscope map that the present invention uses；
Fig. 2 is mushroom bran immobilization degradation bacteria scanning electron microscope (SEM) photograph produced by the present invention；
Fig. 3 be anthropi and microbacterium individually degrade, mixed degradation and mushroom bran immobilization produced by the present invention in varing proportions
Degradation bacteria is to petroleum hydrocarbon degradation result figure；
Fig. 4 be using mushroom bran immobilization degradation bacteria produced by the present invention in petroleum hydrocarbon contaminated soil repair process experimental group and right
According to group to the degradation results figure of four component of petroleum hydrocarbon；
Fig. 5 be using mushroom bran immobilization degradation bacteria produced by the present invention in polycyclic aromatic hydrocarbon pollution repair process experimental group and
Control group to phenanthrene, pyrene, benzo [ɑ] pyrene degradation results figure.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
Embodiment 1
Using oil from Shengli oil field aging contaminated soil as bacterium source, soil sampling 5g is added sterile water 100ml, is placed in shaking table and vibrates
Soil bacteria suspension is made in 30min, using method of dilution butteron on plate, obtains the bacterial strain of different shape, picking single bacterium is in beef extract-peptone
It is enriched with 18h or so in fluid nutrient medium, then draws 2ml and is placed in crude oil culture medium, method is increased using oil concentration gradient
（5000mg/L,10000mg/L,15000mg/L）Each domestication culture 7d；Take bacterium solution 1ml progress in 15000 mg/L culture mediums dilute
Coating is released, bacterium is chosen, is enriched with, is coated with again, until forming single bacterium colony, the single bacterium colony that will be singled out is put in crude oil culture medium
It cultivates, residual petroleum hydrocarbon content is measured under 225nm wavelength using ultrasonic liquid separation extraction-ultraviolet spectrophotometry after 7d, thus
Petroleum hydrocarbon degradation rate is calculated using following formula, in formula：For petroleum hydrocarbon degradation rate, %；m0It is first
Beginning petroleum hydrocarbon quality, g；m1Residual petroleum hydrocarbonaceous amount, g.
Wherein, crude oil medium component is as follows：
K2HPO4·3H2O 2.266g, KH2PO40.68g, MgSO4·7H2O 0.1g, FeSO4·7H2O 0.03g, MnSO4·
H2O 0.03g, CaCl20.0151g, NH4Cl 1. 325g, NaCl 10g, distilled water 1000mL, pH 7.0~7.5.It will be above-mentioned
Solution 100ml is sub-packed in 250ml conical flask, 121 DEG C of 20 min of sterilizing.Crude oil 0.5g, 1.0g, 1.5g are added separately to cultivate
In base, make its final oil concentration gradient 5000mg/L, 1000mg/L, 15000mg/L.
8 plants of degradation bacterias are filtered out altogether, and wherein Q1, Q5 are preferable to oil degradation efficiency, identify that it is respectively through 16SrRNA
Ochrobactrum and Microbacterium adjust each bacterium solution OD600 between 0.4-0.8, then in varing proportions mix the two,
It was found that 2:3（v/v）Under ratio, decomposing petroleum hydrocarbon effect is best, and 7d degradation rate is 43.78%.
Embodiment 2
A kind of preparation of mushroom bran immobilized microbial inoculum, includes the following steps：
1）The preparation of bacteria suspension：The two plants of equal anthropis filtered out using embodiment 1（Ochrobactrumsp.）Q1 and micro-
Bacillus（Microbacteriumsp.）Q5 is accessed in beef extract-peptone fluid nutrient medium and is cultivated respectively to logarithmic phase latter stage
Afterwards, each bacterium solution OD600 is adjusted between 0.4-0.8, later by the two with 2:3 volume ratios are uniformly mixed, then by efficient stone
Supernatant is abandoned after the centrifugation of petroleum hydrocarbon degradation germ liquid, is rinsed with sterile saline, is added physiological saline and be prepared into highly effective petroleum
Surfactant hydrocarbon degradation bacteria suspension, bacteria suspension control：In the case where ultraviolet specrophotometer measures its 600nm wavelength, absorption values are denoted as OD600,
It is measured as 0.4-0.8.
2）The preparation of carrier：It is air-dried after mushroom bran is cleaned up, it is spare to cross 20-80 mesh.
3）By step 2）In mushroom bran and step 1）In bacteria suspension with 1:20（v/v）Ratio mixing, is placed in 30-37 DEG C,
16-24h is adsorbed in 120-160rpm shaking table, discards upper solution, is transferred in centrifuge tube with physiological saline, in 1000-
It is centrifuged 10-15min under 2000rpm, discards supernatant liquid again, after repeating 3-5 times, centrifugation gained precipitating is immobilized microbial inoculum.
4）Preparation gained immobilized microbial inoculum suitably dry be placed in 4 DEG C of refrigerators is saved.
Wherein, step 1）Middle fluid nutrient medium is beef extract-peptone fluid nutrient medium, and 1L beef extract-peptone liquid
Medium component is：Beef extract 5g, peptone 10g, NaCl 5g, 1000 mL of distilled water, are adjusted after being mixed using aforementioned proportion
PH 7. 0~7. 5,121 DEG C of 20 min of sterilizing can be prepared by.
Step 2）Middle mushroom bran refers to residual waste after the planting edible mushrooms such as Pleurotus eryngii, oyster mushroom, needle mushroom harvest.
Embodiment 3
Mushroom bran immobilized microbial inoculum prepared by embodiment 2 is applied to the reparation of petroleum hydrocarbon contaminated soil.
It is experiment soil with Shengli Oil Field oil-polluted soils, soil sample crushing is stirred to loose, is sieved after air-drying, takes 80
Mesh removes to the soil between 40 mesh and is used for flowerpot reparative experiment after larger impurity and the too small fine earth of diameter, and adjust
Soil C, N, P ratio is 100:10:1, the moisture content in each flowerpot is adjusted using weight reduction in repair process, is allowed to be maintained at
15-30%。
With natural reparation（CK）For blank control group, it is divided into target contaminant with four groups in petroleum hydrocarbon, is separately added into free
Bacterium group（JK1,10% free bacterium）, mushroom bran group（JK2,5% mushroom bran）, mushroom bran and free bacterium group（JK3 ,+10% free bacterium of 5% mushroom bran）,
Mushroom bran immobilized microbial inoculum group（JK4,10% immobilized microbial inoculum）.Every 2-7d is stirred once in repair process, is placed in 20-30 DEG C, humidity
For 50d in the constant temperature and humidity incubator of 50-70%.
Using NB/SH/T0609-2010 measurement four constituent content of petroleum hydrocarbon variation.
After measured, after 50d is repaired, four constituent content removal rate of CK group petroleum hydrocarbon is respectively 9.01%, 5.93%,
3.44%, 2.19%, JK1 group removal rate are 26.27%, 13.11%, 5.41%, and 2.61%, JK2 group removal rate is 20.67%,
12.29%, 8.93%, 4.64%, JK3 group removal rate are 39.89%, 20.77%, 15.56%, and 13.99%, JK4 group removal rate is
62.99%, 40.68%, 30.57%, 24.88%, as a result illustrate that mushroom bran immobilized microbial inoculum repairing effect is best.
And in conjunction with attached drawing 4 as can be seen that compared to reparation naturally, free bacterium is individually repaired, mushroom bran is individually repaired, bacterium
Chaff+free bacterium reparation, the degradation effect of immobilized microbial inoculum produced by the present invention have greatly improved, and degradation property is excellent.
Embodiment 4
Mushroom bran immobilized microbial inoculum prepared by embodiment 2 is applied to the reparation of polycyclic aromatic hydrocarbon pollution.
Experiment is Shengli Oil Field oil-polluted soils with soil, and soil is added with the polycyclic aromatic hydrocarbon solution of 50mg/kg mass concentration
In earth, soil sample crushing is stirred to loose, is sieved after air-drying, takes 80 mesh to the soil between 40 mesh, that is, remove larger impurity and
It is used for flowerpot reparative experiment after the too small fine earth of diameter, and adjusting soil C, N, P ratio to be 100:10:1, it uses in repair process
Weight reduction adjusts the moisture content in each flowerpot, is allowed to be maintained at 15-30%.
With natural reparation（CK）For blank control group, it is divided into target contaminant with four groups in petroleum hydrocarbon, is separately added into free
Bacterium group（JK1,10% free bacterium）, mushroom bran group（JK2,5% mushroom bran）, mushroom bran and free bacterium group（JK3 ,+10% free bacterium of 5% mushroom bran）,
Mushroom bran immobilized microbial inoculum group（JK4,10% immobilized microbial inoculum）.Every 2-7d is stirred once in repair process, is placed in 20-30 DEG C, humidity
For 50d in the constant temperature and humidity incubator of 50-70%.
Polycyclic aromatic hydrocarbon China and Philippines, pyrene, anthracene content are measured using Soxhlet extraction-gas chromatography.
After measured, after 50d is repaired, CK group is respectively 10.33%, 11.09%, 8.02% to phenanthrene, pyrene, anthracene removal rate,
JK1 group is respectively 15.89%, 16.33% to phenanthrene, pyrene, anthracene removal rate, and 13.99%, JK2 group is respectively to phenanthrene, pyrene, anthracene removal rate
20.45%, 22.98%, 19.53%, JK3 group are respectively 36.89%, 38.9% to phenanthrene, pyrene, anthracene removal rate, 34.87%, JK4 group pair
Phenanthrene, pyrene, anthracene removal rate are respectively 59.90%, 53.78%, 56.01%.Illustrate mushroom bran immobilized microbial inoculum to polycyclic aromatic hydrocarbon pollution
Repairing effect is best.
And in conjunction with attached drawing 5 as can be seen that compared to reparation naturally, free bacterium is individually repaired, mushroom bran is individually repaired, mushroom bran
The reparation of+free bacterium, immobilized microbial inoculum produced by the present invention have greatly improved to luxuriant and rich with fragrance, pyrene and anthracene degradation effect, degradability
It can be excellent.
Each embodiment in this specification is described in a progressive manner, the highlights of each of the examples are with other
The difference of embodiment, the same or similar parts in each embodiment may refer to each other.For device disclosed in embodiment
For, since it is corresponded to the methods disclosed in the examples, so being described relatively simple, related place is said referring to method part
It is bright.
The foregoing description of the disclosed embodiments enables those skilled in the art to implement or use the present invention.
Various modifications to these embodiments will be readily apparent to those skilled in the art, as defined herein
General Principle can be realized in other embodiments without departing from the spirit or scope of the present invention.Therefore, of the invention
It is not intended to be limited to the embodiments shown herein, and is to fit to and the principles and novel features disclosed herein phase one
The widest scope of cause.
Claims (10)
1. a kind of preparation method of mushroom bran immobilized microbial inoculum, which is characterized in that include the following steps：
（1）The preparation of bacteria suspension：Petroleum hydrocarbon degradation bacterium addition physiological saline is prepared into bacteria suspension；
（2）The preparation of carrier：It is air-dried after mushroom bran is cleaned up, it is spare to cross 20-80 mesh；
（3）Absorption：By the step（2）In mushroom bran and the step（1）In bacteria suspension with 1:15-25（v/v）Ratio is mixed
It closes, is subsequently placed in 30-37 DEG C, adsorbs 16-24h in 120-160rpm shaking table, discard upper solution；
（4）Centrifugation：The mushroom bran after absorption is transferred in centrifuge tube with physiological saline, is then centrifuged for discarding supernatant liquid；In repetition
State centrifugally operated it is multiple after, centrifugation gained precipitating be immobilized microbial inoculum.
2. the preparation method of mushroom bran immobilized microbial inoculum according to claim 1, which is characterized in that the step（1）In
Petroleum hydrocarbon degradation bacterium is two plants, respectively anthropi（Ochrobactrum sp.）And microbacterium（Microbacterium
sp.）；The petroleum hydrocarbon degradation bacterium acquisition modes are：By anthropi（Ochrobactrum sp.）And microbacterium
（Microbacterium sp.）The bacterium solution of the two is with 2:The mixing of 3 volume ratios, abandons supernatant after centrifugation, then use sterile physiological salt
Water rinses to obtain；Bacterium solution absorption values in the case where ultraviolet specrophotometer measures its 600nm wavelength are denoted as OD600, measurement
For 0.4-0.8.
3. the preparation method of mushroom bran immobilized microbial inoculum according to claim 1, which is characterized in that the step（1）Middle bacterium
Suspension absorption values in the case where ultraviolet specrophotometer measures its 600nm wavelength are denoted as OD600, are measured as 0.4-0.8.
4. the preparation method of mushroom bran immobilized microbial inoculum according to claim 1, which is characterized in that the step（3）Middle bacterium
Chaff and bacteria suspension mixed proportion are 1:20（v/v）.
5. the preparation method of mushroom bran immobilized microbial inoculum according to claim 1, which is characterized in that the step（4）In from
The heart is to be centrifuged 10min at 1000rpm；The centrifugally operated number is 3-5 times.
6. the preparation method of mushroom bran immobilized microbial inoculum according to claim 1 or 2, which is characterized in that the step（1）In
Petroleum hydrocarbon degradation bacterium acquisition modes be：Supernatant will be abandoned after the petroleum hydrocarbon degradation bacterium bacterium solution cultivated in liquid medium centrifugation
Then liquid rinses to obtain with sterile saline；The petroleum hydrocarbon degradation bacterium bacterium solution is bacterium solution of the culture to late log phase, described
Bacterium solution absorption values in the case where ultraviolet specrophotometer measures its 600nm wavelength are denoted as OD600, are measured as 0.4-0.8.
7. the preparation method of mushroom bran immobilized microbial inoculum according to claim 6, which is characterized in that the fluid nutrient medium is
Beef extract-peptone fluid nutrient medium, the beef extract-peptone Liquid Culture based component are：Beef extract 5g, peptone 10g,
NaCl 5g, 1000 mL of distilled water；The preparation of the fluid nutrient medium be by beef extract, peptone, NaCl, distilled water in proportion
After mixing adjust pH 7. 0~7.5, then 121 DEG C of 20 min of sterilizing to obtain the final product.
8. the preparation method of mushroom bran immobilized microbial inoculum according to claim 1, which is characterized in that the step（2）Middle bacterium
Chaff is residual waste after planting edible mushroom harvest；The edible bacterium includes Pleurotus eryngii, oyster mushroom and needle mushroom.
9. the application method for the mushroom bran immobilized microbial inoculum that any one of the claim 1-8 preparation method obtains, which is characterized in that
Applied to the biological prosthetic of petroleum hydrocarbon contaminated soil.
10. the application method of mushroom bran immobilized microbial inoculum according to claim 9, which is characterized in that by the mushroom bran immobilization
Microbial inoculum is added in oil-polluted soils with 10% mass ratio and is repaired 35-60 days.
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| WO2021077888A1 (en) * | 2019-10-22 | 2021-04-29 | 中国石油大学(华东) | Preparation method and use method for heavy oil viscosity reduction and degradation mixed microbial inoculum |
| CN114105714A (en) * | 2021-12-15 | 2022-03-01 | 桂润环境科技股份有限公司 | Soil remediation agent, preparation method and application thereof, and soil remediation method |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109679665A (en) * | 2019-02-25 | 2019-04-26 | 大连润鸣材料技术有限公司 | A kind of soil conditioner, preparation method and applications |
| WO2021077888A1 (en) * | 2019-10-22 | 2021-04-29 | 中国石油大学(华东) | Preparation method and use method for heavy oil viscosity reduction and degradation mixed microbial inoculum |
| CN114105714A (en) * | 2021-12-15 | 2022-03-01 | 桂润环境科技股份有限公司 | Soil remediation agent, preparation method and application thereof, and soil remediation method |
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