CN109900880A - A kind of MICP test method using immobilized microorganism technique - Google Patents
A kind of MICP test method using immobilized microorganism technique Download PDFInfo
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- CN109900880A CN109900880A CN201910226179.5A CN201910226179A CN109900880A CN 109900880 A CN109900880 A CN 109900880A CN 201910226179 A CN201910226179 A CN 201910226179A CN 109900880 A CN109900880 A CN 109900880A
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Abstract
The invention discloses a kind of MICP test methods using immobilized microorganism technique, comprising the following steps: one: the activation culture of microorganism fungus kind；Two: carrier, microbial culture medium are uniformly mixed and (are uniformly mixed soil sample and carrier in proportion) by selected carrier；Three: bacterium solution, consolidating fluid being proportionally mixed into fine grained soil at a temperature of being placed in 30 ° and conserved 5-10 days；Four: the particulate soil sample conserved being made into three axis samples, then carries out stress-strain tester.Immobilized microorganism technology is applied to MICP technology by this kind of method innovation, so that microorganism it is more be attached to fixation support, good living environment is provided for microorganism, enhance the process that microorganism induction calcium carbonate generates precipitating (MICP), adhesive bond is formed by core of carrier, significantly improves the intensity of fine grained soil.The method in two different fields is combined, novel, environmental protection, production carbonic acid ca efficiency greatly improve.
Description
Technical field
The present invention relates to a kind of MICP technology more particularly to a kind of test sides MICP using immobilized microorganism technique
Method belongs to microorganism ground, environmental wastewater processing technology field.
Background technique
The reinforcing of geotechnical engineering is always one of most important project of engineering field, since the 21th century, microbiology
It is combined with each other between geotechnical engineering science, promote mutually, biotechnology gradually penetrates into geotechnical engineering field.MICP technology
It is currently the technology of microorganism geotechnical engineering aspect comparative maturity.MICP technology is heavy by producing gland enzyme bacteria-induction carbonate
Product, gelling ambient substance particle play the role of reinforcing.Immobilized microorganism technique is that selected microorganism is fixed on choosing
On fixed carrier, makes its highly dense and keep bioactivity, the biological skill that can quickly, be largely proliferated under optimum conditions
Art.The technology of this " bio-synergistic " substantially increases the efficiency of wastewater treatment.But existing there is presently no by both technology knots
Application example altogether, there are no the introductions of the specific implementation effect in engineering.
Summary of the invention
The technical problem to be solved by the present invention is a kind of MICP test method using immobilized microorganism technique is provided,
By the way that immobilized microorganism technology is applied in MICP technology so that microorganism it is more be attached to fixation support, to be micro-
Biology provides good living environment, enhances the process that microorganism induction calcium carbonate generates precipitating (MICP), is with carrier
Core forms adhesive bond, significantly improves the intensity of fine grained soil, and the method in two different fields is combined, novel,
Environmental protection, production carbonic acid ca efficiency greatly improve.
The technical solution of the present invention is as follows: a kind of MICP test method using immobilized microorganism technique, the method packet
Include following steps:
Step 1: microorganism fungus kind needed for purchase microorganism induction precipitation of calcium carbonate technology (MICP) carries out activation training
It supports；
Step 2: absorption method, cross-linking method, investment, root are had according to the process for fixation that immobilized microorganism technique uses
Different carriers is selected according to different methods, and carrier is put into manipulated soil and is evenly mixed；
Step 3: bacterium solution, consolidating fluid are uniformly blended into the soil body conserve in proportion；
Step 4: the soil sample conserved is fabricated to three axis samples, then carries out soil test.
In the step 1, strain is Bacillus pasteurii, and the culture medium of activation is NH4-YE culture medium, at subpackage
Include: Bacillus pasteurii is put into culture solution, will activated by yeast extract, ammonium sulfate, 0.13M Tris buffer (PH=9)
Culture medium is put into 190 turns of 30 ° of isothermal vibration water tanks, places 40-42h.
In the step 2, carrier is active carbon, and active carbon is crossed 2 ㎜ sieve, weighs the active carbon of soil body quality 7%, then
It is put into manipulated soil and is evenly mixed.
In the step 3, the soil body is red clay, is mixed using bacterium solution, consolidating fluid as liquid proportional uniform, remodeling
Red clay is configured according to original state soil's water content, is placed under 30 ° of constant temperature water tanks and is conserved 5-10 days.
The bacterium solution, the mixed method of consolidating fluid are as follows: bacterium solution, consolidating fluid are proportionally uniformly blended into soil, according to
The unit of 1mol/L configures volume 1:1, and consolidating fluid is configured to MICP process and provides urea and calcium ion, and CaCl is selected in test2Make
For calcium source, match 1mol CaCl respectively2With the consolidating fluid of urea, every liter of CaCl containing 1mol2With 1mol CO (NH2)2。
In the step 4, test controls formula triaxial machining permeameter using SLB-1 type ess-strain, carries out under K0 consolidation
Conventional triaxial compression test.
The beneficial effects of the present invention are: compared with prior art, using technical solution of the present invention, by immobilized microorganism
Technology is applied to a kind of new method of the method for microorganism induction calcium carbonate technology (MICP) as microorganism geotechnical engineering, compares
Microorganism induction precipitation of calcium carbonate technology had not only greatly enhanced the effect that microorganism solidifies ground, but also environment-friendly high-efficiency.This kind of method wound
It is new that immobilized microorganism technology is applied to MICP technology so that microorganism it is more be attached to fixation support, be micro- life
Object provides good living environment, the process that microorganism induction calcium carbonate generates precipitating (MICP) is enhanced, using carrier as core
It is formed centrally adhesive bond, significantly improves the intensity of fine grained soil.The method in two different fields is combined, novel, ring
Guarantor, production carbonic acid ca efficiency greatly improve, and generate better effect, extremely agree with requirement of engineering.
Detailed description of the invention
Fig. 1 is stress strain diagram of the present invention under 400 confining pressures；
Fig. 2 is stress strain diagram of the present invention under 300 confining pressures；
Fig. 3 is stress strain diagram of the present invention under 250 confining pressures；
Fig. 4 is Bacillus pasteurii growth curve chart.
Specific embodiment
To make the object, technical solutions and advantages of the present invention clearer, below with reference to this specification attached drawing to this hair
It is bright to be described in further detail.
Embodiment 1: as shown in attached drawing 1~4, a kind of MICP test method using immobilized microorganism technique, the side
Method the following steps are included:
Step 1: microorganism fungus kind needed for purchase microorganism induction precipitation of calcium carbonate technology (MICP) carries out activation training
It supports；
Step 2: absorption method, cross-linking method, investment, root are had according to the process for fixation that immobilized microorganism technique uses
Different carriers is selected according to different methods, and carrier is put into manipulated soil and is evenly mixed；
Step 3: bacterium solution, consolidating fluid are uniformly blended into the soil body conserve in proportion；
Step 4: the soil sample conserved is fabricated to three axis samples, then carries out soil test.
In the step 1, strain is Bacillus pasteurii, and the culture medium of activation is NH4-YE culture medium, at subpackage
Include: Bacillus pasteurii is put into culture solution, will activated by yeast extract, ammonium sulfate, 0.13M Tris buffer (PH=9)
Culture medium is put into 190 turns of 30 ° of isothermal vibration water tanks, places 40-42h.
In the step 2, carrier is active carbon, and active carbon is crossed 2 ㎜ sieve, weighs the active carbon of soil body quality 7%, then
It is put into manipulated soil and is evenly mixed.
In the step 3, the soil body is red clay, is mixed using bacterium solution, consolidating fluid as liquid proportional uniform, remodeling
Red clay is configured according to original state soil's water content, is placed under 30 ° of constant temperature water tanks and is conserved 5 days.
The bacterium solution, the mixed method of consolidating fluid are as follows: bacterium solution, consolidating fluid are proportionally uniformly blended into soil, according to
The unit of 1mol/L configures volume 1:1, and consolidating fluid is configured to MICP process and provides urea and calcium ion, and CaCl is selected in test2Make
For calcium source, match 1mol CaCl respectively2With the consolidating fluid of urea, every liter of CaCl containing 1mol2With 1mol CO (NH2)2。
In the step 4, test controls formula triaxial machining permeameter using SLB-1 type ess-strain, carries out under K0 consolidation
Conventional triaxial compression test.
Here is as follows according to done test of illustrating:
One, the soil body used is Kweiyang red clay in the present embodiment, takes native position: Guiyang City, Guizhou Province Huaxi District highway base
Hole, soil body type: remodeling red clay.Take native depth: 1-10m.Its physico-mechanical properties is as shown in table 1.Active carbon is chosen in test
For the cocoanut active charcoal that Immohilized microorganism carrier is bought, active carbon particle size Wei≤2mm.
The physical mechanical property index of 1 red clay of table
Two, Bacillus pasteurii (Sporosarcina pasteurii) is selected in the present embodiment, belongs to Gram-positive
Bacterium, the strain are widely used one kind in MICP at present, and the ability of the urase of secretion is stronger.The present embodiment is from ATCC
Bacillus pasteurii freeze-dried powder is bought, using the NH4-YE culture medium that it is recommended, ingredient is yeast extract 20g, ammonium sulfate
10g, 0.13M Tris buffer (PH=9) 15.782g.The strain of activation is placed in 30 DEG C, the constant-temperature shaking culture of 180rpm
40-42h in case.Seminar's early period carries out Bacillus pasteurii concentration mensuration, and in oscillation 40-42h, bacterium solution increases to enter and stablize
Phase, OD600It is worth highest, the growth curve chart of Bacillus pasteurii is as shown in Figure 4.
Three, urea and calcium ion are provided for MICP process, CaCl is selected in test2As calcium source, match 1MCaCl respectively2With urine
The consolidating fluid of element, every liter contains 1MCaCl2 and 1MCO (NH2)2。
Four, the preparation of soil sample: red clay natural air drying is crossed into 2mm sieve sterilizing remodeling, preparation remodeling red clay is 65% aqueous
Rate (original state red clay natural moisture content is 65%), active carbon is crossed 2mm sieve and is added in soil sample.4 groups of tests are set, respectively without adding
It aggravates modeling red clay plus microorganism remodeling red clay plus 7% active carbon red clay plus microorganism and the remodeling of 7% active carbon is red glutinous
Soil.Specification is carried out in the form of soil sample, bacterium solution and consolidating fluid, the uniform mix of active carbon as three axis of diameter 3.9cm high 8.1cm
Sample preparation.
Five, testing program: soil sample is placed in 30 DEG C of insulating boxs and is conserved 5 days, three axis soil samples are then made, pass through SLB-1
Type ess-strain controls formula triaxial machining permeameter, carries out the conventional triaxial compression test under K0 consolidation, chooses consolidation and does not drain
It shears, does one group of Duplicate Samples under every kind of consolidation pressure, totally 24 three axis soil samples, specific testing program data are as shown in table 2.
2 testing program of table
Note: CU indicates that consolidation does not drain scissor test, and CTC indicates that mechanical property, subscript indicate consolidation stresses final value, under
Mark indicates consolidation stress ratio σ3/σ1。
Six, test result analysis
1, the stress-strain variation of four groups of comparative tests
In the case where K0 consolidates und rained condition, the mechanical property of progress is tested, it can be deduced that following analysis: 1. four groups not
Shear stress peak value with addition remodeling red clay is addition active carbon and microorganism highest, and with the increase of confining pressure, is cut
Peak stress also increases with it, and shear stress peak value averagely increases 16.8% under three kinds of confining pressures.2. the remodeling of four groups of differences addition
The shear stress peak value that microorganism, addition active carbon and microorganism are added in red clay is above no added remodeling under three kinds of confining pressures
Red clay, and add microorganism remodeling red clay comparison it is no added, averagely increased under 3 kinds of confining pressures of shear stress peak value
5.1%.3. and add the remodeling red clay of active carbon and no added remodeling red clay peak stress remains basically stable, under 250 confining pressures
The remodeling red clay peak stress of addition active carbon is slightly above no added remodeling red clay.
It is undrained shear according to four groups of tests, it is as shown in table 3 that done data calculate C, φ value.It is effectively anti-by comparing
Intensity index C, φ value is cut, following analysis conclusion is obtained: 1. by comparison addition microorganism and active carbon, no added two groups of tests
Effective Shear Strength Index, it can be seen that effective C value of addition microorganism and active carbon increases, and effective φ value reduces.2. right
Than addition microorganism, effective C, φ value of no added two groups of tests, the effective C value for dosing microorganism also has enhancing, effective φ value
Become smaller.3. and comparison addition active carbon, no added effective Shear Strength Index C, the φ value of two groups of tests, effective C of two groups of tests
Value difference is smaller, and the addition effective φ value of activated-carbon test group becomes smaller.4. comparing effective C, φ value of four groups of tests, it can be seen that effectively
The height of C value enhancing is successively: the test group highest of addition microorganism and active carbon is added taking second place for microorganism, addition activity
Charcoal experimental group C value variation it is smaller, effective φ value of four groups of tests is successively from big to small: addition it is no added, addition active carbon,
Add microorganism, addition microbial activity charcoal.
3 four groups of proof stress intensity indexs of table
2, Analysis on Mechanism
It is analyzed by above-mentioned data, it is believed that active carbon combines microorganism in the soil body, so that microorganism is largely proliferated,
Urase yield is improved, MICP process is promoted, and then generate relatively large number of calcium carbonate, plays better cementation, increase
The strong solidification effect to the soil body.
Active carbon has the characteristics that huge specific surface area, flourishing pore structure and excellent absorption property, with active carbon
Good living environment as vector construction microorganism.It is analyzed in terms of the technical method in test due to the micro- life of immobilization
" fixation " of the carrier to microorganism in object technology, i.e. suction-operated of the active carbon of this paper to Bacillus pasteurii, so that micro- life
Object can be in its huge specific surface area and quick, mass propagation in flourishing hole, and then quickly generates more urase, is accelerating
During MICP, a large amount of urase combination urea elements and Ca ion of output, with active carbon be " core " so that surrounding Ca from
Son generates more CaCO3, plays the role of to soil body particle preferably cementing, plays better " synergy " to MICP.Therefore number
It is obvious according to the enhancing of middle shearing strength.
Secondly MICP belongs to the induced biomineralization (BIM) in developing stages, and induced biomineralization (BIM) controls journey
Lower, the not limitation in space is spent, is guided without special cell tissue or large biological molecule.Induced biomineralization (BIM) shape
At crystal without specific modality, orientation, forming process and inorganic chemistry precipitating mineral are closely similar, and identical living matter is not
With different crystal can be formed in environment.And active carbon is added, microorganism has the carrier that can be depended on, and the process of biomineralization is just
It focuses mostly in the carrier or carrier surface, there has been the setting in " region ", and then make with the biomineralization that active carbon is " core "
With formed calcium carbonate crystal, due to biomineralization environment optimization or make calcium carbonate crystallization when to crystal structure, form generate
Certain control action, so that the calcium carbonate that the remodeling red clay that active carbon and microorganism is added generates is after the cementing soil body,
The growth effect of its Shear Strength Index is not 1+1=2, that is, the remodeling red clay shearing stress peak of active carbon and microorganism is added
Value, Shear Strength Index enhancing be greater than only be added active carbon with only be added microorganism remodeling red clay and.
3, conclusion
The present embodiment is tested by mechanical property, and it is special to carry out the mechanics that active carbon solidifies Kweiyang red clay to microorganism
Property experimental study is concluded that
(1) conventional triaxial compression test is not drained by K0 consolidation, four groups of check experiment peak stress sizes are successively: adding
Remodeling red clay > addition microorganism remodeling red clay > the additions active carbon for entering active carbon and microorganism remolds red clay, no added
Remold red clay；And the remodeling red clay peak stress that active carbon and microorganism is added enhances maximum, and is greater than and microorganism is added
It remolds red clay and active carbon is added and remold the increased summation of red clay peak stress.
(2) conventional triaxial compression test consolidated by K0 under 400,300,250 this 3 kinds of confining pressures, obtains its shearing strength
Index C, φ value compares no added remodeling red clay, adds the remodeling red clay of microorganism and active carbon, adds the weight of microorganism
Moulding red clay is mainly C value, and φ value does not change substantially, and the remodeling red clay C value for adding microorganism and active carbon enhances most
Obviously；The φ value for adding active carbon reduces, and C value is essentially unchanged.
(3) variation of the peak stress variation and Shear Strength Index C, φ value of four groups of tests is compared, analysis obtains: activity
Charcoal plays an important role during microorganism induction precipitation of calcium carbonate (MICP), and microorganism is a large amount of in active carbon, quick
Proliferation, so that generating the cementing soil body of more calcium carbonate minerals, plays the role of " synergy " to MICP；It is served as into during MICP
Mine " core ", due to biomineralization environment optimization or make calcium carbonate crystallization when crystal structure, form are produced it is certain
Control action, and then change the mechanical characteristic of red clay.
Immobilized microorganism technique is applied to the method for microorganism induction calcium carbonate technology (MICP) as Microbialites
A kind of new method of geotechnological journey had both greatly enhanced the effect that microorganism solidifies ground compared to microorganism induction precipitation of calcium carbonate technology
Fruit, and environment-friendly high-efficiency.Immobilized microorganism technology is applied to MICP technology by this kind of method innovation, so that microorganism is more
It is attached to fixation support, provides good living environment for microorganism, microorganism induction calcium carbonate is enhanced and generates precipitating
(MICP) process forms adhesive bond by core of carrier, significantly improves the intensity of fine grained soil.It is different by two
The method in field combines, and novel, environmental protection, production carbonic acid ca efficiency greatly improve, and generate better effect, and extremely agreeing with engineering needs
It wants.
Place is not described in detail by the present invention, is the well-known technique of those skilled in the art of the present technique.Finally, it is stated that the above reality
It applies example to be only used to illustrate the technical scheme of the present invention and not to limit it, although having carried out specifically the present invention referring to preferred embodiment
It is bright, those skilled in the art should understand that, can with modification or equivalent replacement of the technical solution of the present invention are made, and
The objective and range for not departing from technical solution of the present invention, are intended to be within the scope of the claims of the invention.
Claims (5)
1. a kind of MICP test method using immobilized microorganism technique, it is characterised in that: the described method comprises the following steps:
Step 1: microorganism fungus kind needed for purchase microorganism induction precipitation of calcium carbonate technology (MICP) carries out activation culture；
Step 2: absorption method, cross-linking method, investment are had according to the process for fixation that immobilized microorganism technique uses, according to not
Same method selectes different carriers, and carrier is put into manipulated soil and is evenly mixed；
Step 3: bacterium solution, consolidating fluid are uniformly blended into the soil body conserve in proportion；
Step 4: the soil sample conserved is fabricated to three axis samples, then carries out soil test.
2. the MICP test method according to claim 1 using immobilized microorganism technique, it is characterised in that: the step
In rapid one, strain is Bacillus pasteurii, and the culture medium of activation is NH4-YE culture medium, and ingredient includes: yeast extract, sulphur
Sour ammonium, buffer(PH=9 0.13M Tris), Bacillus pasteurii is put into culture solution, activation medium is put into 30 ° of constant temperature
40-42h is placed in 190 turns of water tank of concussion.
3. the MICP test method according to claim 1 using immobilized microorganism technique, it is characterised in that: the step
In rapid two, carrier is active carbon, and active carbon is crossed 2 ㎜ sieve, the active carbon of soil body quality 7% is weighed, places into manipulated soil and carry out
It evenly mixes.
4. the MICP test method according to claim 1 using immobilized microorganism technique, it is characterised in that: the step
In rapid three, the soil body is red clay, is mixed using bacterium solution, consolidating fluid as liquid proportional uniform, remodeling red clay according to original state
Soil's water content is configured, and is placed under 30 ° of constant temperature water tanks and is conserved 5-10 days.
5. the MICP test method according to claim 4 using immobilized microorganism technique, it is characterised in that: the bacterium
The mixed method of liquid, consolidating fluid are as follows: bacterium solution, consolidating fluid are proportionally uniformly blended into soil, configured according to the unit of 1mol/L
Volume 1:1, consolidating fluid are configured to MICP process and provide urea and calcium ion, and CaCl is selected in test2As calcium source, match 1mol respectively
CaCl2With the consolidating fluid of urea, every liter of CaCl containing 1mol2With 1mol CO(NH2)2。
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| CN110240439A (en) * | 2019-06-28 | 2019-09-17 | 湖北工业大学 | A kind of preparation method of the luminous ecological substrate of the cured high-strength light of microorganism |
| CN111672900A (en) * | 2020-06-01 | 2020-09-18 | 合肥工业大学 | Method for restoring heavy metal pollution of soil by microorganism-induced calcium carbonate precipitation |
| CN111875306A (en) * | 2020-08-10 | 2020-11-03 | 中国科学院武汉岩土力学研究所 | Microorganism enhancement method for three-dimensional printing of solid cement-based solid model |
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| CN113231457A (en) * | 2021-04-30 | 2021-08-10 | 合肥工业大学 | Method for restoring heavy metal polluted soil by magnesium oxide and microorganisms |
| CN113373139A (en) * | 2021-06-15 | 2021-09-10 | 佛山市南海区苏科大环境研究院 | Immobilized microbial material for treating wastewater containing heavy metal ions and preparation method thereof |
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| CN114262066A (en) * | 2021-12-23 | 2022-04-01 | 西安建筑科技大学 | Method and reactor for synchronously removing organic matters by microorganism-induced calcium precipitation |
| CN114396038A (en) * | 2022-01-21 | 2022-04-26 | 中煤航测遥感集团有限公司 | Method for reinforcing soil strength by using microorganism induced calcium carbonate deposition |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110240439A (en) * | 2019-06-28 | 2019-09-17 | 湖北工业大学 | A kind of preparation method of the luminous ecological substrate of the cured high-strength light of microorganism |
| CN110240439B (en) * | 2019-06-28 | 2021-06-04 | 湖北工业大学 | Preparation method of light high-strength luminous ecological base material solidified by microorganisms |
| CN111672900A (en) * | 2020-06-01 | 2020-09-18 | 合肥工业大学 | Method for restoring heavy metal pollution of soil by microorganism-induced calcium carbonate precipitation |
| CN111672900B (en) * | 2020-06-01 | 2021-08-10 | 合肥工业大学 | Method for restoring heavy metal pollution of soil by microorganism-induced calcium carbonate precipitation |
| CN111875306A (en) * | 2020-08-10 | 2020-11-03 | 中国科学院武汉岩土力学研究所 | Microorganism enhancement method for three-dimensional printing of solid cement-based solid model |
| CN112974512A (en) * | 2021-02-03 | 2021-06-18 | 宁夏大学 | Application of bacillus fusiformis with MICP function in tailing greening repair |
| CN113231457A (en) * | 2021-04-30 | 2021-08-10 | 合肥工业大学 | Method for restoring heavy metal polluted soil by magnesium oxide and microorganisms |
| CN113373139A (en) * | 2021-06-15 | 2021-09-10 | 佛山市南海区苏科大环境研究院 | Immobilized microbial material for treating wastewater containing heavy metal ions and preparation method thereof |
| CN113445490A (en) * | 2021-06-29 | 2021-09-28 | 南京林业大学 | Method for uniformly solidifying soft clay by microorganisms |
| CN114262066A (en) * | 2021-12-23 | 2022-04-01 | 西安建筑科技大学 | Method and reactor for synchronously removing organic matters by microorganism-induced calcium precipitation |
| CN114396038A (en) * | 2022-01-21 | 2022-04-26 | 中煤航测遥感集团有限公司 | Method for reinforcing soil strength by using microorganism induced calcium carbonate deposition |
| CN114751685A (en) * | 2022-05-11 | 2022-07-15 | 辽宁工程技术大学 | MICP principle-based ecological solid carbon filling material and preparation method thereof |
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Application publication date: 20190618 |