CN101644047A - Method of adhering loose sands employing microbial mineralization - Google Patents
Method of adhering loose sands employing microbial mineralization Download PDFInfo
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- CN101644047A CN101644047A CN200910183240A CN200910183240A CN101644047A CN 101644047 A CN101644047 A CN 101644047A CN 200910183240 A CN200910183240 A CN 200910183240A CN 200910183240 A CN200910183240 A CN 200910183240A CN 101644047 A CN101644047 A CN 101644047A
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Abstract
The invention provides a method of adhering loose sands employing microbial mineralization, comprising the following steps: adopting peptone, beef extract and distilled water to prepare culture medium, adjusting pH value to 7.0, sterilizing and drying, inoculating Bacillus pasteurii strain to the prepared culture medium, culturing at 30 DEG C for 24-36h while keeping shaking in the oscillation frequency of 170r/min to ensure that the bacteria solution is completely muddy and the activity k of the bacteria solution is not less than 0.5mmol-min<-1>, mixing urea and calcium chloride solutions ofwhich volumes are equal, and injecting the obtained solution and the bacteria solution separately according the volume ratio of 2:1 in sands or mixing the obtained solution and the bacteria solution and injecting in sands. When the obtained solution and the bacteria solution are separately injected in sands, the bacteria solution is first injected, the mixed solution of urea and calcium chloride is then injected until the bacteria solution leaks out of the sands form top bottom, then the bacteria solution is injected until the solution leaks out and the above steps are repeated. When injectedin sands, the mixed solution is pressed in sands from bottom to top and is discharged from the top of sands. The adhered sand bodies have environmental friend property, simple process and low cost, and the compressive strength can be up to about 2MPa.
Description
Technical field
The present invention relates to a kind of method of glued discrete particles, especially a kind of method of utilizing the glued loose sand grains of microbial technique.
Background technology
Along with the continuous destruction of ecological environment, the desertification degree is serious day by day; In some areas, be soft state owing to local soil property water content is higher, so in some Important Project construction, then need soil property is reinforced such as building subway etc.; And many cities are based upon based near the sandy seabeach, entire city exist glide, subside, the danger of depression.In the control of desert, often adopt kind of the mode of grass planting tree, often adopt cementitious material such as pouring cement based in the reinforcement measure, but this class material is waterproof, heavy damage the ecological performance of soil property, be unfavorable for its sustainable development, just seem particularly important so search out a kind of environmentally friendly reinforcement means.
Biomineralization is a kind of very general natural phenomena, and almost each biology can both synthetic mineral.Biomineralization often can form natural composite organic-inorganic material orderly arrangement, that structure is very excellent, and is wherein nearly 2nd/3rd, the calcium ore deposit, and quite a few has glued function.They are by the vital movement of himself, and between the surrounding environment medium constantly circulation enzyme is taking place is turning usefulness into, mineralising forms a kind of binding material gradually---calcite (calcium carbonate), pass through the accumulation of very long time again, the hard rock of the glued formation of the most loose detrital material of occurring in nature deposition.And utilize the solid soil of microbial technique glue sand still to exist some problems: how to select the bacterial strain that suits and it is cultivated and tames; How to improve weatherability of microorganism or the like, and adopt the price of microbial technique also too expensive.
Summary of the invention
In order to adopt the method for watering cementitious material such as cement based in the reinforcement measure that solves the prior art existence, heavy damage the shortcoming of ecological performance of soil property, the invention provides a kind of method of utilizing the glued loose sand grains of microorganism mineralization, have environment friendly, fixed good sand body has certain compression strength.
The present invention adopts following technical scheme:
A kind of method of utilizing the glued loose sand grains of microorganism mineralization, step is:
The first step, preparation culture medium: with peptone, beef extract and distilled water preparation culture medium, regulating pH is 7.0, behind the sterilizing, drying, bacterial strain Pasteur bacillus Bacillus pasteurii is seeded in the prepared culture medium, under 30 ℃, carries out shaken cultivation, frequency of oscillation is 170r/min, cultivate 24～36h, muddy fully to bacterium liquid, bacteria solution active k 〉=0.5mmolmin
-1
Second step was 3molL with concentration
-1Urea liquid and concentration be 3molL
-1The calcium chloride solution equal-volume mix the back and inject sand grains respectively with 2: 1 by volume the ratio of bacterium liquid of first step preparation, inject sand grains together after perhaps mixing.
In second step, when the mixed solution of urea liquid and calcium chloride solution and bacterium liquid inject sand grains respectively, be to inject bacterium liquid earlier, after treating that bacterium liquid oozes out from top to bottom, the afenil mixed solution that reinjects, treat that solution oozes out after, bacterium liquid reinjects, alternate cycles, when the sand body upper epidermis did not have sedimentary deposit to hinder liquid to infiltrate, liquid 24h did not have and oozes out then glued success fully.
In second step, when the afenil mixed solution injects sand grains with bacterium liquid, be from lower to upper mixed liquor to be pressed in the sand grains and to discharge, when pressure is motionless, stop from the upper end.
The present invention is seeded to bacterial strain Bacillus pasteurii in the culture medium, is nutrient source with the substrate urea in the culture medium, and the urase of generation has the ability of decomposing substrate, shown in equation (1), along with the continuous decomposition of substrate, NH
3Constantly discharge, make the pH value of solution raise, shown in equation (2), pH value rising simultaneously impels following reversible reaction to carry out shown in equation (3) toward positive direction again, makes CO
3 2-Concentration improve constantly.Replenish proper C a this moment in solution
2+, the continuous immediately chelating Ca of SM (water soluble organic matter) that somatic cells membrane interface place is electronegative
2+, shown in equation (4), induce local crystal anion (CO
3 2-) concentration further increases, thereby attract more Ca
2+, increase to up to crystal predecessor concentration and to be beneficial to coring, thereby deposited the mineralized material CaCO of consolidation
3Particle is shown in equation (5).
NH
2-CO-NH
2+H
2O→2NH
3+CO
2 (1)
NH
3+H
2O→NH
4 ++OH
- (2)
CO
2+H
2O→H
2CO
3→HCO
3 -+H
+→CO
3 2-+2H
+ (3)
Cell+Ca
2+→Cell-Ca
2+ (4)
Cell-Ca
2++CO
3 2-→Cell-CaCO
3 (5)
Pack in the die trial loose sand grains and tamping, contain a large amount of space between the particle this moment.At first introduce bacterium liquid in sand grains, treat after the complete seepage flow of bacterium liquid, a large amount of thalline is just stayed in the sand body and is attached to sand grain surface.And then toward the mixed liquor that wherein injects urea and calcium chloride, under action of microorganisms, produce binding material CaCO
3Particle, binding material be also attached to the surface of sand, thereby and along with increasing of binding material the thalline parcel is replaced the former sand surface that occupies of thalline.Inject bacterium liquid and substrate mixed liquor by continuous circulation subsequently, make binding material in layer toward outgrowth up to the particle gap is filled up, thereby adjacent particle is coupled together, and make all sand grains form a integral body with certain intensity.
Principle according to the glued loose sand grains of mentioned microorganism has two kinds of methods to realize.The first has been taked bacterium liquid, urea liquid, calcium chloride solution equal-volume are mixed the concurrently injected mode in back, with speed slowly liquid is pressed in the sand grains, and it is slowly oozed out again; The 2nd, at first inject bacterium liquid, after treating fully to ooze out, thereby prevent that bacterium liquid and substrate from beginning reaction and stopping up upper surface on the sand body surface, inject the mixed liquor of urea and calcium chloride, alternate cycles.
Beneficial effect:
1. environmental friendliness because of it is different from cement-based gelling material or other types cementitious material, can keep sand body still can keep certain void content after fixed, the ecological functions of the soil property that adequately protected;
2. make full use of the nature microorganism resource, not only aboundresources, and technology is simple, and clean environment is with low cost;
3. microorganism or substrate solution all can penetrate in sandy soil or the soft soil property easily, and the grouting treatment of other cementitious material is very difficult;
4. fixed its compressive strength of successful sand body is the highest can be near 2MPa.
Description of drawings
Fig. 1 is the X-ray diffraction analysis collection of illustrative plates of fixed sand body, and wherein A is quartzy, and B is a calcite, and C is a calcium chloride.
Fig. 2 is fixed sand body observed surface topography under SEM, wherein (a) is that 200 times surface topography is amplified on the relatively poor sand body surface of fixed tight ness rating, (b) be that fixed tight ness rating is amplified 300 times surface topography in the sand body surface preferably, (c) amplify 1000 times surface topography (being irregularly shaped) for the calcarinate matter that is attached to fixed sand grain surface.
Fig. 3 is fixed sand body measured stress-strain diagram under the loading speed of electronic universal tester at 1N/s, and wherein (a) numbers the 1. stress-strain diagram of sand body, and its mode of failure is that the bottom is destroyed, shown in (c); (b) be to number the 2. stress-strain diagram of sand body, its mode of failure is for destroying fully, shown in (d).
The specific embodiment
The cultivation of bacterium liquid
Preparation culture medium (medium component is as shown in table 1), regulate pH=7.0,121 ℃ of high-temperature sterilization 25min, after in 60 ℃ of baking ovens, drying, bacterial strain Bacillus pasteurii is seeded to (sterile working) in the triangular flask that culture medium is housed, 30 ℃ of shaken cultivation of shaking table (frequency of oscillation 170r/min), 24～36h.
Table 1 medium component
The measurement of bacteria solution active
The activity of bacterium liquid is come secondary indication with the electrical conductivity of solution.
The urea liquid of preparation 3mol/L is got 25ml and is put in the beaker, and adds the laboratory tap water of 20ml, puts into water-bath constant temperature at 25 ℃, and the speed of adjusting agitator is 200r/min.Get 5ml through the cultured bacterium liquid of shaking table (guarantee that the operation to bacterium liquid is to finish in gnotobasis, avoid contaminated bacteria liquid), and be expelled in the beaker immediately.Numerical value every 30s record electrical conductivity.In the operation of measuring bacteria solution active, the ultimate density of substrate is 1.5mol/L, and bacterium liquid is diluted to 1/10th, so the activity of bacterium liquid should be 10 times of the activity value that records.As bacteria solution active k 〉=0.5mmolmin
-1The time, can think that this bacterium liquid has higher activity, possesses the ability of glued loose sand grains.
Glue sand technology
The calcium chloride solution equal-volume mixing of the urea liquid of bacterium liquid, 3mol/L, 3mol/L from bottom to top is pressed into simultaneously oozes out.With speed slowly mixed liquor is pressed in the sand grains, it is slowly oozed out again.The prerequisite that adopts the method can not add urea when being the cultivation of bacterium liquid, otherwise just can produce a large amount of precipitations after the three mixes, and has both reduced the composition of calcium chloride in the substrate solution, and can be to the sand body effect of resulting in blockage.The advantage of the method is that the time that produces glued matter is wanted much shorter, has effectively shortened the glued time, has improved the efficient of glue sand because bacterium liquid and substrate solution are mixed the back injection; But owing to be that manually-operated is pressed into mixed liquor, dynamics and speed all are difficult to control, so form a depression in the bottom of sand body easily.Though can in sand body, produce deposition very soon after mixing, but also be more prone to simultaneously produce precipitation on the surface of sand body, later stage is pressed into mixed liquor causes difficulty, this point can be that the method for " mix from bottom to top be pressed into simultaneously and from the upper end discharge " overcomes by changing " mix from bottom to top to be pressed into simultaneously and ooze out ".From practical engineering application, all there is the shortcoming of performance difficulty in technology from lower to upper in addition, because no matter be desert control or solid reinforcement, infiltration technology from top to bottom all has more operability than from the bottom up.But produce prefabrication and can adopt this process fully.This technology belongs to continuous process, stops when pressure is motionless.
The mixed liquor of urea, calcium chloride and bacterium liquid earlier are that 1: 1 ratio is mixed by volume with urea and calcium chloride solution, and urea, calcium chloride mixed solution and 2: 1 by volume ratio of bacterium liquid are separately alternately from top to bottom oozed out then.At first inject bacterium liquid, treat fully to ooze out the mixed liquor that urea and calcium chloride are injected in the back, thereby can prevent that like this bacterium liquid and substrate from beginning reaction and stopping up upper surface, alternate cycles on the sand body surface.This technological advantage is that technology is simplified as far as possible, and is simple to operate, and is difficult for forming the seepage flow that deposition hinders liquid on the sand body surface, but shortcoming is that originally flow velocity is very fast, needs testing crew constantly to repeat work.This technology belongs to batch process, and when the sand body upper epidermis did not have sedimentary deposit to hinder liquid to infiltrate, liquid 24h did not have and oozes out then glued success fully.
Fixed sand body is carried out X-ray diffraction analysis, the analysis collection of illustrative plates that obtains as shown in Figure 1, wherein A is quartzy, B is a calcite, C is a calcium chloride, atlas analysis can obtain, and the calcium carbonate that the microorganism mineralization obtains is the calcite of rhombohedron type, is not the aragonite of needle-like or the vaterite of polycrystalline sphere; And by in the collection of illustrative plates also as can be seen from the quartz of sand grains and from the CaCl of auxiliary material
2Phase.
Fixed sand body is carried out its surface topography of sem observation, as shown in Figure 2, wherein (a) is that 200 times surface topography is amplified on the relatively poor sand body surface of fixed tight ness rating, (b) be that fixed tight ness rating is amplified 300 times surface topography in the sand body surface preferably, can observe in the particle gap of (b) and be filled with a large amount of glued matter, when adjacent particle is connected, almost particle is wrapped up, and still have a large amount of spaces (a).(c) be that the calcarinate matter that is attached to fixed sand grain surface is amplified 1000 times surface topography (being irregularly shaped).
Compression strength value
Fixed sand body is carried out compression test.Before the test sand body is carried out surface rubbing and handle, and in 60 ℃ of baking oven 24h oven dry.Electronic universal tester applies pressure to the sand body surface with the loading speed of 1N/s, measured stress-strain diagram as shown in Figure 3, wherein (a) numbers the 1. stress-strain diagram of sand body, its mode of failure is that the bottom is destroyed, shown in (c); (b) be to number the 2. stress-strain diagram of sand body, its mode of failure is for destroying fully, shown in (d).Two each index of numbering sand body are as shown in table 3, wherein bacterium liquid consumption for the test at the beginning of to the used bacterium liquid of off-test total amount, diameter is the diameter after sample takes out, highly be the distance between the upper and lower surface of specimen surface after polishing, maximum pressure is the pressure maximum value that sample is reached in the load-bearing process, and revising compressive strength is that compressive strength multiply by the value that obtains after the coefficient.Relation between coefficient and the sample ratio of height to diameter is with reference to the correction factor of concrete cylindrical shape test specimen.The compressive strength of fixed sand body reaches as high as about 2MPa.
The relation of table 2 correction factor and ratio of height to diameter
Table 3 sand body sample resistance to compression result
Claims (3)
1. method of utilizing the glued loose sand grains of microorganism mineralization is characterized in that step is:
The first step, preparation culture medium: with peptone, beef extract and distilled water preparation culture medium, regulating pH is 7.0, behind the sterilizing, drying, bacterial strain Pasteur bacillus Bacillus pasteurii is seeded in the prepared culture medium, under 30 ℃, carries out shaken cultivation, frequency of oscillation is 170r/min, cultivate 24～36h, muddy fully to bacterium liquid, bacteria solution active k 〉=0.5mmolmin
-1
Second step was 3molL with concentration
-1Urea liquid and concentration be 3molL
-1The calcium chloride solution equal-volume mix the back and inject sand grains respectively with 2: 1 by volume the ratio of bacterium liquid of first step preparation, inject sand grains together after perhaps mixing.
2. the method for utilizing the glued loose sand grains of microorganism mineralization as claimed in claim 1, it is characterized in that, in second step, when the mixed solution of urea liquid and calcium chloride solution and bacterium liquid inject sand grains respectively, be to inject bacterium liquid earlier, after treating that bacterium liquid oozes out from top to bottom, the urea that reinjects, calcium chloride mixed solution, treat that solution oozes out after, bacterium liquid reinjects, alternate cycles, when the sand body upper epidermis did not have sedimentary deposit to hinder liquid to infiltrate, liquid 24h did not have and oozes out then glued success fully.
3. the method for utilizing the glued loose sand grains of microorganism mineralization as claimed in claim 1, it is characterized in that, in second step, when the afenil mixed solution injects sand grains with bacterium liquid, be from lower to upper mixed liquor to be pressed in the sand grains and to discharge, when pressure is motionless, stop from the upper end.
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