CN101368384A - Method for curing soil by using carbonate mineralized bacterium - Google Patents
Method for curing soil by using carbonate mineralized bacterium Download PDFInfo
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- CN101368384A CN101368384A CNA2008101569419A CN200810156941A CN101368384A CN 101368384 A CN101368384 A CN 101368384A CN A2008101569419 A CNA2008101569419 A CN A2008101569419A CN 200810156941 A CN200810156941 A CN 200810156941A CN 101368384 A CN101368384 A CN 101368384A
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Abstract
The invention discloses a soil curing method through carbonate-mineralization microbes, including the following steps: first, preparing high concentration microbe liquid: inoculating Bacillus pasteurii strain into the culture fluid of beef extract and peptone; wherein each litre of culture fluid contains 4-6g peptone and 2-4g beef extract; cultivating for16-24h under 30DEG C with the pH value controlled at 6-8, then taking out the mixture and centrifuging for 5-8min under the speed of 5000-8000rpm, thus obtaining high-concentration microbe liquid at concentration of 2 X 10<9>-2 X 10<11>cell/ml; second, mixing the soil: mixing the high-concentration microbe liquid prepared in the first step and the newly prepared culture fluid into the dried soil sample through lime(3%-7% the weight of the dry soil) and carbamide(2%-6% the weight of the dry soil) according the proportioning of 100g dry soil per 2-3mL high-concentration microbe liquid and 0-50mL newly prepared culture fluid, blending to be even and then cultivating for 0.5-8days; third; molding through a die: molding the soil mixture cultivated in second step in a die when the mixture reaches the optimal water ratio, then demoulding and conserving in constant temperature of 20 DEG C. Seven days later, the soil is cured, with the compressive resistance more than 1.0MPa, the pure oil improved by 38% relatively.
Description
Technical field
The present invention relates to a kind of method of utilizing the carbonate mineralized bacterium curing soil, mainly utilize fiber filled effect and the biomineralization mineralising consolidated soil of microorganism.
Background technology
The occurring in nature certain micro-organisms can be by the vital movement of himself, and between the surrounding environment medium constantly circulation enzyme is taking place is turning usefulness into, mineralising forms calcite gradually.Pass through the accumulation in very long period again, the hard rock of the glued formation of the most loose detrital material of occurring in nature deposition.
At present, in the defect repair of cement-based material, the original appearance reparation of building historical relic, aspects such as the fixed and desert curing of heavy metal in soil ion have obtained broad research and application to this technology.At present this technology aspect soil solidification still in the experimental study stage.
Summary of the invention
The present invention relates to a kind of method of utilizing the carbonate mineralized bacterium curing soil, can investigate the application of this bacterium on soil solidification as leading indicator with compressive strength and water stability.
The present invention adopts following technical scheme: a kind of method of utilizing the carbonate mineralized bacterium curing soil, and step is:
The first step, prepare highly enriched bacterium liquid: bacterial strain Pasteur bacillus Bacillus pasteurii is inoculated in the beef extract-peptone nutrient solution, every liter of nutrient solution contains peptone 4～6g, beef extract 2～4g, and control pH is 6～8, cultivate 16～24h down in 30 ℃, taking-up obtains highly enriched bacterium liquid with centrifugal 5～8min under 5000～8000rpm, and bacterial strain concentration is 2 * 10
9～2 * 10
11Cell/mL
Second step, mixing with soil: add highly enriched bacterium liquid and the new system nutrient solution of the amount of the highly enriched bacterium liquid of 2～3mL and 0～100mL new system nutrient solution by every 100g dry ground with first step preparation, 3%～7% the lime of used dry ground quality and the urea of 2%～6% mass fraction are admixed in the soil sample of oven dry, and cultivated 0.5～8 day the back that stirs; When the amount of fresh medium was 0mL, the incubation time of soil was preferably 0.5 day.The amount that adds the new system nutrient solution in the soil of drying is 20～100mL, and incubation time is preferably 3～8 days.
In the 3rd step, moulded section: in the moulding of mixing soil materials optimum moisture content dip mold, the demoulding is also put to 20 ℃ of thermostatic curings of thermostatic chamber with second soil that goes on foot the process cultivation.Test specimen is the cylinder of Φ 50 * 50mm.
Beneficial effect: earth consolidation after 7 days, compressive strength reach more than the 1.0MPa, and plain relatively soil improves 38%.The part group reaches more than the 1.2MPa, and plain relatively soil improves 67%.Bacterium is after carrying out 5～8 days cultivation in the soil, because its increase active and quantity, be significantly improved than the test specimen intensity of straight forming.
Description of drawings
Fig. 1 is urea content and the linear fitted figure of OD value relation.
Fig. 2 is each group test specimen 7d compressive strength column diagram.
Fig. 3 (a) and (b) be earth consolidation sample SEM image and energy spectrum analysis figure.
The specific embodiment
Definite method of optimum moisture content
Adopt compaction test to measure the maximum dry density and the optimum moisture content of used soil sample, the density that obtains used plain soil in this experiment is 1.880g/cm
3, optimum moisture content is 14.9%wt, the density that adds the soil sample behind the lime that accounts for plain soil property amount 3% is 1.795g/cm
3, optimum moisture content is 15.6%wt, the density that adds the soil sample behind the lime that accounts for plain soil property amount 5% is 1.788g/cm
3, optimum moisture content is 15.9%wt, the shaping test piece of pressing Φ 50 * 50mm size calculates, degree of compaction 98%, the desired substance proportioning sees Table 1.
The plain soil of table 1, added each component content of rendzinas compressive strength test specimen of lime
Adopt light-duty compaction cylinder in the compaction test, JTJ051-93 experimentizes according to " highway earthwork test rule ", measures the relation curve of dry density and water content (ratio of the quality of water and dry ground quality).Estimate five water content 12%wt, 14%wt, 16%wt, 18%wt and 20%wt in the experiment, wet soil and dry ground quality in the compaction cylinder under each water content of weighing, and calculate dry density under this water content, draw the curve of dry density and water content according to the gained result, curve is parabolic shape, the corresponding maximum dry density of peak, its corresponding water content is an optimum moisture content.Rendzinas compaction test method roughly the same adds the optimum moisture content moulding of lime group by mixed soil sample under this volume.
According to the native compaction test gained of element result, take by weighing the plain native 180.9g of required oven dry, measure 27mL water and be sprayed to soil surface and stir, one night of sealing shelving.Moulding on pressure testing machine, compression leg and die trial maintain an equal level about being depressed into, and keep pressure about 1 minute, and its demoulding is taken out, and weighing test specimen quality is 208.5g, with vernier caliper measurement test specimen height 50.0mm.Test specimen is put in the thermostatic chamber maintenance, takes out measured intensity after reaching the length of time.Test specimen after the maintenance is taken out, adopt the tensile testing machine of specification 10KN to measure its intensity, during measurement, keeping its rate of deformation is 5mm/min.Maximum pressure P when the record test specimen destroys, compressive strength R
c=P/A, A are the sectional area of test specimen.Obtaining 7 days average compressive strength of plain soil at last is 0.72MPa.Plain native test specimen water stability is very poor, meets water and promptly destroys.
Among the present invention, strength measurement method thereafter is with herein.
The preparation of highly enriched bacterium liquid:
Bacterial strain Pasteur bacillus Bacillus pasteurii is inoculated in the beef extract-peptone nutrient solution, every liter of nutrient solution contains peptone 4～6g, beef extract 2～4g, and control pH is 6～8, cultivate 16～24h down in 30 ℃, taking-up is taken off layer with centrifugal 5～8min under 5000～8000rpm and is obtained highly enriched bacterium liquid, and bacterial strain concentration is 2 * 10
9～2 * 10
11Cell/mL.
Embodiment 1
Is 2 * 10 with soil with the lime, the bacterial strain concentration that account for native quality 3%～7%
11Highly enriched bacterium liquid 6mL, the urea 4.8g of cell/mL, water, supernatant make up respectively, the centrifugal supernatant liquor that obtains when wherein supernatant is for the highly enriched bacterium liquid of preparation, here replace water to test shown in table 2 design, evenly compacting behind the mix, the demoulding and in 20 ℃ of maintenances of thermostatic chamber, 3 test specimens of every composing type, taking-up test compressive strength is as shown in table 3 after 7 days.
Table 2 microbial technique solidified earth schematic design
Table 3 is respectively organized 7 days compressive strength of soil test specimen
Test record 7 days compressive strength the highest be T6 (soil+bacterium+urea liquid) group, what intensity was minimum is T3 (soil+urea liquid) group.The group of admixture lime is generally good than what do not mix, but T12 intensity then less than T6, this is because the existence of calcium ion causes urea to decompose slowly, decomposes 20% back and just is difficult to continue to decompose; Supernatant place of water test specimen compressive strength is slightly improved, but amplification is very little; The adding of bacterium makes the soil test specimen that raising in various degree all be arranged.Wherein 7 days compressive strength of T4 group (soil+highly enriched bacterium liquid+water) group test specimen reaches 1.20MPa, and plain relatively soil raising surpasses 60%, compares plain its compressive strength of native test specimen and obviously improves, and object bacteria plays the effect of fiber filled in the soil the inside.And T3 and T6 group, bacterium can also generate urase by enzymolysis urea except fiber filled effect and catalytic action, thereby improves resistance of soil; Record soil+highly enriched bacterium liquid+urea+compressive strength of 7 days of water group test specimen by the T6 prescription and reach 1.67MPa, plain relatively soil doubles many, T6 group is because the existence of bacterium, and urea decomposes very fast, impels test specimen intensity to be improved and produce material such as biology enzyme in the enzymolysis process.And in T3 group soil+urea+water prescription, 7 days intensity of test specimen is lower, and 0.32MPa is only arranged, and the very big test specimen intensity that reduced of urea existence be described, and this is to be caused by the suction of urea when the maintenance of soil test specimen.
Embodiment 2
Making one group of test specimen T12 ＇ in addition at the prescription of T12 group contrasts: with bacterial strain concentration is 2 * 10
11The highly enriched bacterium liquid 6mL of cell/mL joins in the dry ground, and adding 50mL new system nutrient solution, every liter contains peptone 5g, beef extract 3g in the nutrient solution, left standstill respectively 4 days, 8 days, mix accounts for lime 5.2g and the urea 4.8g and the moulding of dry ground quality 3% then, and it is as shown in table 4 to test two groups of test specimen intensity.
7 days compressive strength of two groups of test specimens of table 4
Press the prescription of T12 ＇, by with the quadrat method moulding, the compressive strength of measuring 4 days and 8 days reaches 1.22 respectively, 1.16MPa, and the intensity of comparing straight forming in the T12 group has improved about 27%, than original plain native raising about 70%.This is because bacterium descends in centrifugal back activity, and culture medium can improve activity and the quantity of bacterium after adding, thereby strengthens its effect to soil, improves the compressive strength of test specimen.
Embodiment 3
Investigate the water stability of part group test specimen: with the maintenance test specimen taking-up curing in water 24h of last day, the water surface is higher than test specimen upper surface 2.5cm, measures immersion and does not soak test specimen intensity separately, and both ratios are the steady coefficient of water, and test result is as shown in table 5.
The fixed back of table 5 part group test specimen water stability energy
At T4, T5, T6, T10, T11, the T12 group is carried out the water stability test, although we find that former three intensity is higher, but water stability is very poor, and this is inapplicable in actual engineering, then behind three's admixture 3% lime, soak and to keep test specimen intact in 1 day, but its unconfined compression strength is lower, and water stability is not good, can consider that the volume that improves lime improves the water stability energy.
Embodiment 4
At microorganism enzymolysis urea group, carry out the measurement of urea resolution ratio.In the presence of highly enriched bacterium liquid, urea can decompose, and according to the developer principle, measures the urea content in the different time sections soil sample, calculates the urea resolution ratio.Utilizing normal concentration (40g/L) urea liquid reading in ultraviolet-uisible spectrophotometer under the developer effect is that the OD value is measured the urea resolution ratio with the volume Changing Pattern.Therefrom take out 0.125mL respectively with pipettor, 0.25mL, 0.5mL, 1mL, 2mL, 4mL standard urea solution, and drop in 6 50mL volumetric flasks, add the deionized water of 25mL, vibrate a little, and then in each volumetric flask, drip 2mLH
2SO
4, vibration then drips the 2mL paradime thylaminobenzaldehyde more respectively, uses the deionized water constant volume after the vibration, and fully rocks evenly.Open spectrophotometer, the adjustment wavelength is 430nm, measures solution O D value in each volumetric flask, and the relation curve that obtains a substantially linear is seen shown in Figure 1.
Do not consider the decomposition of soil to urea, T6, the T12 group is can be ureaclastic.The urea resolution ratio data of measuring loose soil sample and closely knit soil sample are shown in table 6,7.
The urea resolution ratio of bacterial strain in the loose sample of table 6
The urea resolution ratio of bacterial strain in the closely knit sample of table 7
As can be seen, T6 group urea all decomposes very fast in loose soil sample and closely knit soil sample from table 6,7, just can decompose more than 90% less than 40h, and the T12 group then slowly and after decomposing 20% is decomposed with regard to being difficult to continuation owing to there is decomposition in calcium ion.So T12 intensity is adding the height that does not have T6 not add on the contrary behind the lime, the T6 group can very fast decomposing urea, not only basic neutralisation the negative effect of urea to test specimen, and its enzymolysis process and self-acting make intensity improve greatly on the contrary.
Embodiment 5
Admixture accounts for the lime of plain soil property amount 3% in soil sample, add highly enriched bacterium liquid and water (supernatant) again, just T7 and T8 the group prescription, stir, one night of shelving, shaping and demoulding, record its 7 days compressive strength and reach 1.26 (1.35) MPa, plain relatively soil has improved more than 70%, simultaneously because the adding of lime, test specimen immersion 24h remains intact, but the steady coefficient of water is not high, have only about 0.4, and lime intermingled quantity is less in this test specimen, water stability depends on the adding of lime, strengthens lime intermingled quantity and can improve the steady coefficient of water.
Embodiment 6
Admixture accounts for 3% lime of plain soil property amount in soil sample, add highly enriched bacterium liquid, urea and water again, the prescription of T12 group just, stir one night of shelving, shaping and demoulding, record its 7 days compressive strength and reach 0.96MPa, plain relatively soil has improved 1/3, and the steady coefficient of its water reaches 0.50, needs to strengthen lime intermingled quantity equally and improves the steady coefficient of water.T12 group urea also can be decomposed by bacterium, measures through the urea resolution ratio, and this component is separated slower, and this is because the existence of calcium ion has suppressed the decomposition of urea.
The urea of the lime of admixture 3%, water and equivalent in soil sample, just the prescription of T9 group is 0.59MPa by record its 7 days compressive strength with the quadrat method moulding, illustrates that also the existence of urea can cause test specimen intensity to reduce, and both contrasts illustrate that the adding of bacterium still has no small raising to intensity.
From SEM image (Fig. 3), can see the existence of some oblique side's particles, (a) in the corresponding power spectrum of black surround part show that wherein Ca, O, C constituent content are higher, be CaCO
3, (b) the middle corresponding power spectrum of black surround part shows that then constituent contents such as O, Si, Al are higher, this also conforms to the contained element of soil, illustrates that promptly this place is soil particle.
Embodiment 7
A kind of method of utilizing the carbonate mineralized bacterium curing soil, step is:
The first step, prepare highly enriched bacterium liquid: bacterial strain Pasteur bacillus Bacillus pasteurii is inoculated in the beef extract-peptone nutrient solution, every liter of nutrient solution contains peptone 4g, beef extract 2g, and control pH is 6～8, cultivate 16h down in 30 ℃, taking-up obtains highly enriched bacterium liquid with centrifugal 5min under the 5000rpm, and bacterial strain concentration is 2 * 10
9Cell/mL;
In second step, mixing with soil: add the highly enriched bacterium liquid of the highly enriched bacterium liquid of 2mL with first step preparation by every 100g dry ground, 3% the lime of used dry ground quality and the urea of 2% mass fraction are admixed in the soil sample of oven dry, and cultivated 0.5 day the back that stirs;
In the 3rd step, moulded section: in the moulding of mixing soil materials optimum moisture content dip mold, the demoulding is also put to 20 ℃ of thermostatic curings of thermostatic chamber with second soil that goes on foot the process cultivation.Test specimen is the cylinder of Φ 50 * 50mm.
Embodiment 8
A kind of method of utilizing the carbonate mineralized bacterium curing soil, step is:
The first step, prepare highly enriched bacterium liquid: bacterial strain Pasteur bacillus Bacillus pasteurii is inoculated in the beef extract-peptone nutrient solution, every liter of nutrient solution contains peptone 6g, beef extract 4g, and control pH is 6～8, cultivate 24h down in 30 ℃, taking-up obtains highly enriched bacterium liquid with centrifugal 8min under the 8000rpm, and bacterial strain concentration is 2 * 10
11Cell/mL;
Second step, mixing with soil: add highly enriched bacterium liquid and the new system nutrient solution of the amount of highly enriched bacterium liquid of 3mL and 100mL new system nutrient solution by every 100g dry ground with first step preparation, 7% the lime of used dry ground quality and the urea of 6% mass fraction are admixed in the soil sample of oven dry, and cultivated 8 days the back that stirs;
In the 3rd step, moulded section: in the moulding of mixing soil materials optimum moisture content dip mold, the demoulding is also put to 20 ℃ of thermostatic curings of thermostatic chamber with second soil that goes on foot the process cultivation.Test specimen is the cylinder of Φ 50 * 50mm.
Embodiment 9
A kind of method of utilizing the carbonate mineralized bacterium curing soil, step is:
The first step, prepare highly enriched bacterium liquid: bacterial strain Pasteur bacillus Bacillus pasteurii is inoculated in the beef extract-peptone nutrient solution, every liter of nutrient solution contains peptone 5g, beef extract 3g, and control pH is 6～8, cultivate 20h down in 30 ℃, taking-up obtains highly enriched bacterium liquid with centrifugal 7min under the 6500rpm, and bacterial strain concentration is 2 * 10
10Cell/mL;
Second step, mixing with soil: add highly enriched bacterium liquid and the new system nutrient solution of the amount of highly enriched bacterium liquid of 2.5mL and 20mL new system nutrient solution by every 100g dry ground with first step preparation, 5% the lime of used dry ground quality and the urea of 4% mass fraction are admixed in the soil sample of oven dry, and cultivated 3 days the back that stirs;
In the 3rd step, moulded section: in the moulding of mixing soil materials optimum moisture content dip mold, the demoulding is also put to 20 ℃ of thermostatic curings of thermostatic chamber with second soil that goes on foot the process cultivation.Test specimen is the cylinder of Φ 50 * 50mm.
Embodiment 10
A kind of method of utilizing the carbonate mineralized bacterium curing soil, step is:
The first step, prepare highly enriched bacterium liquid: bacterial strain Pasteur bacillus Bacillus pasteurii is inoculated in the beef extract-peptone nutrient solution, every liter of nutrient solution contains peptone 5g, beef extract 3g, and control pH is 6～8, cultivate 20h down in 30 ℃, taking-up obtains highly enriched bacterium liquid with centrifugal 7min under the 6500rpm, and bacterial strain concentration is 2 * 10
10Cell/mL;
Second step, mixing with soil: add highly enriched bacterium liquid and the new system nutrient solution of the amount of highly enriched bacterium liquid of 2.5mL and 60mL new system nutrient solution by every 100g dry ground with first step preparation, 5% the lime of used dry ground quality and the urea of 4% mass fraction are admixed in the soil sample of oven dry, and cultivated 5 days the back that stirs;
In the 3rd step, moulded section: in the moulding of mixing soil materials optimum moisture content dip mold, the demoulding is also put to 20 ℃ of thermostatic curings of thermostatic chamber with second soil that goes on foot the process cultivation.Test specimen is the cylinder of Φ 50 * 50mm.
Claims (3)
1. a method of utilizing the carbonate mineralized bacterium curing soil is characterized in that, step is:
The first step, prepare highly enriched bacterium liquid: bacterial strain Pasteur bacillus Bacillus pasteurii is inoculated in the beef extract-peptone nutrient solution, every liter of nutrient solution contains peptone 4～6g, beef extract 2～4g, and control pH is 6～8, cultivate 16～24h down in 30 ℃, taking-up obtains highly enriched bacterium liquid with centrifugal 5～8min under 5000～8000rpm, and bacterial strain concentration is 2 * 10
9～2 * 10
11Cell/mL;
Second step, mixing with soil: add highly enriched bacterium liquid and the new system nutrient solution of the amount of the highly enriched bacterium liquid of 2～3mL and 0～100mL new system nutrient solution by every 100g dry ground with first step preparation, 3%～7% the lime of used dry ground quality and the urea of 2%～6% mass fraction are admixed in the soil sample of oven dry, and cultivated 0.5～8 day the back that stirs;
In the 3rd step, moulded section: in the moulding of mixing soil materials optimum moisture content dip mold, the demoulding is also put to 20 ℃ of thermostatic curings of thermostatic chamber with second soil that goes on foot the process cultivation.
2. the method for utilizing the carbonate mineralized bacterium curing soil according to claim 1 is characterized in that, when the amount of fresh medium was 0mL, the incubation time of soil was 0.5 day in second step.
3. the method for utilizing the carbonate mineralized bacterium curing soil according to claim 1 is characterized in that, the amount that adds the new system nutrient solution in second step in the soil of oven dry is 20～100mL, and incubation time is 3～8 days.
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