CN103266070A - Bacillus lentus and method for generating by inducing bacillus lentus for reinforcing foundation - Google Patents
Bacillus lentus and method for generating by inducing bacillus lentus for reinforcing foundation Download PDFInfo
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Abstract
The invention provides bacillus lentus and a method for generating by inducing bacillus lentus for reinforcing foundation, the bacillus lentus URIC, the bacillus lentus UR9D, and the bacillus lentus UR52 are preserved in China General Microbiological Culture Collection Center (CGMCC) on August 29, 2012; the method comprises: firstly, respectively fermenting and culturing a single bacterium colony of bacillus lentus in a fermentation medium, then grouting in two steps, filling bacteria liquid into a sand column pumped with deionized water, and filling a mixed solution of reaction solution calcium chloride and urea, to form a microbe-calcium carbonate-sand grains consolidating body in a sand grain-solution system, and furthermore to enhance bonding degree and intensity of the sand grains. The bacillus lentus and the method of the invention has advantages of short solidification time, good effect and low cost, and will not cause pollution on environment.
Description
Technical field
The present invention relates to the foundation consolidating technology field, be specifically related to bacillus lentus and induce the method that calcium carbonate is used for foundation stabilization that produces.
Background technology
Foundation stabilization is that one extensively and the important engineering practical activity in building and the geotechnical engineering.The bearing capacity deficiency of foundation soil can cause roadbed settlement, dykes and dams, sand dune and the side slope unstability of highway and railway and the excessive erosion on seashore or riverbank etc.Earthquake or traffic vibration can cause the liquefaction of weak loose foundation soil.And the oil gas drilling engineering in loose rock-soil layer or foundation ditch tunnel excavation also often are subjected to having a strong impact on of quicksand stream soil.In economically developed area, delta, carry out marine reclamation land, strengthening soft foundation improves ground degree of compactness and stability, also is an important problem.
Traditional chemical grouting technique and geotextile reinforcement technique can be described as the major project measure that the ground original position is reinforced, in being usually used in above-mentioned all kinds of foundation stabilization and handling.But there is the problem of weather resistance deficiency in traditional geotextile reinforcement technique; Chemical grouting environmental influence based on cement or lime or organic gel is big, and has greatly reduced the water-permeable of ground, has hindered seepage action of ground water, and the effective grouting radius of single is little, economic performance is lower, and then causes large-scale foundation stabilization very difficult; And physics reinforcement means technical and economic performances such as ground displacement, pile foundation and strong rammer are low.Therefore, seek the novel engineering method means of big area ground consolidation process, be worth constantly studying, developing and put into practice.
Along with the fast development of microbial technique in recent years, the effect in building and geotechnical engineering has had more deep understanding to people to microbial technique.The latest developments of microorganism metallogeny show, some specific microorganism, organism and calcium ion sources such as urea around can utilizing, the very fast calcium carbonate with gelating property that generates.Different with the calcium carbonate that general chemical action generates, this microbiogenic calcium carbonate material formation speed and controllable intensity, and can be used as binding agent, loose sand grains is bonded to intensity and the controlled artificial sandy gravel of perviousness.The newest research results of this basic biomaterial science is for a brand-new field has been opened up in the research of building and geotechnical engineering.In Base Consolidation Engineering, in conjunction with growing microbial engineering, effectively utilize abundant and this natural resources of microorganism of toxicological harmless, change the engineering mechanics property of building and geotechnical engineering, will bring technological revolution for building and geotechnical engineering foundation stabilization field.
Summary of the invention
In order to overcome the deficiencies in the prior art, the object of the present invention is to provide bacillus lentus and induce the method that calcium carbonate is used for foundation stabilization that produces, adopt microorganism bacillus lentus foundation stabilization treatment process of the present invention, have set time short, effective, cost is low, and can not pollute environment.
For achieving the above object, the technical solution adopted in the present invention is:
Bacillus lentus, it is characterized in that: described bacillus lentus Bacillus lentus UR1C, bacillus lentus Bacillus lentus UR9D and bacillus lentus Bacillus lentus UR52 have been preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on August 29th, 2012, deposit number is respectively CGMCC No.6485, CGMCC No.6483, CGMCC No.6484, CGMCC is called for short at this preservation center, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101.
It is oval shaft-like that described bacillus lentus Bacillus lentus UR1C, bacillus lentus Bacillus lentus UR9D and bacillus lentus Bacillus lentus UR52 all are, and gemma arranged, no pod membrane, Gram-positive; At NH
4-YE is yeast extract 20g/L, and on the ammonium sulfate 10g/L flat board, bacterium colony is rounded, and surface wettability is smooth, neat in edge, and the bacterium colony size is 1-2mm, and it is faint yellow that bacterium colony is, and this bacterium all can grow under the scope of 4 ℃-37 ℃ substratum temperature and pH7-9.5.
Bacillus lentus described above is induced and is produced the method that calcium carbonate is used for foundation stabilization, comprises the steps:
Step 1: bacillus lentus Bacillus lentus UR1C, bacillus lentus Bacillus lentus UR9D and the single bacterium colony of bacillus lentus Bacillus lentus UR52 were obtained three kinds of bacterium liquid respectively in fermentation culture 12-60 hour in fermention medium under 25 ℃ of-37 ℃ of conditions;
Step 2: adopt the substep injection process, be in the milk in two steps, pour into bacterium liquid earlier, pour into reaction solution calcium chloride and urea mixing solutions then, concrete grammar is: form three sand posts in three the sand grains moulds of at first sand grains being packed into, pump into the deionized water of 3 times of sand-grain volumes then respectively in three sand posts, make the sand post saturated, in three sand posts, pour into one of described three kinds of bacterium liquid of step 1 respectively subsequently, the volume that pours into bacterium liquid is 1-3 times of sand-grain volume, at last divide 2-4 to criticize the mixing solutions that pours into reaction solution calcium chloride and urea respectively in three sand posts, original position generates MCP in the sand post; The mixing solutions of described reaction solution calcium chloride and urea contains 0.5M CaCl for every liter
2With 0.5M urea, every batch of mixed liquor volume that pours into is 3 times that bacteria liquid amasss, and makes and forms microorganism-calcium carbonate-sand grains induration in sand grains-solution system, and then strengthen bonding degree and the intensity of sand grains.
The described fermention medium of step 1 comprises yeast extract 10-20g/L, and ammonium sulfate or ammonium chloride 10g/L, pH are 7-9.5.
The described speed that pumps into deionized water in three sand posts of step 2 is 0.5-1.5ml/min.
The described speed that pours into the mixing solutions of reaction solution calcium chloride and urea in the sand post of step 2 is 0.05-0.15ml/min.
The present invention compares with prior art, has following advantage:
1, screen the microorganism that can produce calcium carbonate from existing urease-producing bacterial strain, soil sample and water sample, and unknown bacterium is carried out Molecular Identification, the microorganism strains of consolidated subsoil sand provides the candidate resource in order to obtain effectively.
2, urea is decomposed into ammonium root and carbonic acid gas under the effect of the urase that Institute of Micro-biology produces, and makes bacterium pH rising on every side simultaneously, is providing under the situation of calcium ion, generates water-fast carbonate salt, and sand grains is bonded together on every side; Has the ground sand intensity time of raising weak point, good effectiveness.
3, the cost of used nutritive medium is low.
4, among the present invention the microorganism that utilizes be microorganism itself that exist in the ground (soil) or the culture of certain microorganism wherein, nutritive substance also is crude substance, can not pollute environment.
Description of drawings
Fig. 1 is grouting experimental installation and percolation path synoptic diagram, wherein:
Fig. 1 a is grouting experimental installation synoptic diagram;
Fig. 1 b is the percolation path synoptic diagram.
Fig. 2 (uses OD for the biomass of cultivating bacterial strain
600Value representation) and urease activity, X-coordinate is different types of urease-producing bacterial strain, and ordinate zou is biomass and urease activity.
Fig. 3 is the stress-strain(ed) curve of bonding gained sand post, wherein:
Fig. 3 a is the stress-strain(ed) curve of bacterial strain UR9D institute bonded sand post;
Fig. 3 b is the stress-strain(ed) curve of bacterial strain UR1C institute bonded sand post;
Fig. 3 c is the stress-strain(ed) curve of bacterial strain UR52 institute bonded sand post.
Fig. 4 is the stereoscan photograph of gained sand post, wherein:
Fig. 4 a is the stereoscan photograph of bacterial strain UR9D institute bonded sand post;
Fig. 4 b is the stereoscan photograph of bacterial strain UR1C institute bonded sand post;
Fig. 4 c is the stereoscan photograph of bacterial strain UR52 institute bonded sand post.
Embodiment
Below in conjunction with the drawings and specific embodiments the present invention is described in further detail.
Urease-producing induces calcium carbonate to produce the separation screening of bacterial strain
The urease-producing strains separation is gathered pedotheque from the Tsing-Hua University flower nursery.
Bacterium has urea decomposing enzyme, can produce a large amount of ammonia by decomposing urea, makes substratum be alkalescence, shows red.This experiment utilizes this characteristic, test sample is first at 37 ° of C, under the high concentration urea condition of 5mol/L behind the enrichment culture 24h, kill the various microbial nutrition somatocyte that can not tolerate and utilize high concentration urea, nutrient solution after will handling again carries out gradient dilution, coating urase screening and culturing flat board, cultivate under 37 ° of C, the bacterial strain that picking reddens the substratum color, line separates single bacterium colony, the microorganism that obtains is the urease-producing microorganism, and utilize 16S rDNA method to identify, difference called after bacillus lentus Bacillus lentus UR1C, bacillus lentus Bacillus lentus UR9D and bacillus lentus Bacillus lentus UR52.
The cultivation of urease-producing bacterial strain
Preparation fermention medium: yeast extract 10-20g/L, ammonium sulfate or ammonium chloride 10g/L, pH is 7-9.5, the 100ml fermention medium packed into sterilize in the 500ml culturing bottle, the single bacterium colony bacillus lentus of picking Bacillus lentus UR1C, bacillus lentus Bacillus lentus UR9D and bacillus lentus Bacillus lentus UR52 are inoculated in the fermention medium respectively from flat board respectively, cultivate under 30 ℃ of temperature, rotating speed is 150rpm-250rpm.Cultivate and collect bacterium liquid after 16 hours, detect the biomass of bacterium liquid and (use OD
600Value representation) and urease activity, detected result as shown in Figure 2.
The bonding of sand grains
The substep injection process is adopted in this experiment, is in the milk in two steps, and perfusion bacterium liquid pours into reaction solution calcium chloride and urea mixing solutions then earlier.The grouting system as shown in Figure 1.The sand post moulding compound shell of this use, volume is 50ml, internal diameter 30mm, long 110mm.Be the long sand post to be bonded of about 100mm in the middle of the sand post mould.The situation that the grouting mouth that occurs for fear of last joint stops up has the double gauze filtering layer respectively at the two ends of sand post to be bonded.The upper end of the syringe rubber plug jam-pack of band Y-tube.Used sand is Chinese iso standard sand, manually sieves into different-grain diameter, and the sand grains particle diameter of screening is 0.16-0.315mm.Each syringe shell about 100g of dress sand grains to be bonded.
Before the experiment at first earlier with peristaltic pump with the about 1ml/min of 10rpm() speed pump into the deionized water 150ml of 3 times of sand-grain volumes, make the sand post saturated as far as possible.Pour into sand-grain volume 1-3 bacterium liquid 50-150ml doubly then in each sand post, divide 3 batches in the individual sand post afterwards and pour into reaction solution calcium chloride and urea mixing solutions, original position generates MCP in the sand post.Used reaction solution calcium chloride and urea mixing solutions contain 0.5M CaCl for every liter
2With 0.5M urea.Pour into reaction solution calcium chloride and urea mixed liquor volume is 3 times that bacteria liquid amasss at every turn, 150-450ml, the about 0.1ml/min of filling speed.Make and form microorganism-calcium carbonate-sand grains induration in sand grains-solution system, and then strengthen bonding degree and the intensity of sand grains.
Analysis to formation sand post
1, the uniaxial compressive strength of sand post detects
The sand post that obtains is carried out form removal, the equal molding bonded of sand post, observation has certain intensity.Carry out uniaxial compression test, the result as shown in Figure 3.The uniaxial compressive strength of gained sand post is between 0.5-2.4Mpa, and wherein the uniaxial compressive strength of UR9D bacterial strain is the highest, is 2.4Mpa.
2, sand grain surface generates the analysis of material
White mass to precipitation carries out XRD analysis, and formed material is calcium carbonate.Throw out is carried out electron microscopic observation, the result as shown in Figure 4, can obviously see has spherical between sand grains and hexahedron shape calcium carbonate crystal generates.
3, the contained calcium carbonate quality of sand post is analyzed
The sand post is soaked in the dilute hydrochloric acid solution, by detecting the reduction of soaking front and back sand post quality, calculates the content of calcium carbonate in every gram sand post.The result is as shown in table 1:
Table 1
Claims (6)
1. bacillus lentus, it is characterized in that: described bacillus lentus Bacillus lentus UR1C, bacillus lentus Bacillus lentus UR9D and bacillus lentus Bacillus lentus UR52 have been preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on August 29th, 2012, deposit number is respectively CGMCC No.6485, CGMCC No.6483, CGMCC No.6484, CGMCC is called for short at this preservation center, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101.
2. a kind of bacillus lentus according to claim 1, it is characterized in that: described bacillus lentus Bacillus lentus UR1C, bacillus lentus Bacillus lentus UR9D and bacillus lentus Bacillus lentus UR52 all are oval shaft-like, gemma is arranged, no pod membrane, Gram-positive; At NH
4-YE is yeast extract 20g/L, and on the ammonium sulfate 10g/L flat board, bacterium colony is rounded, and surface wettability is smooth, neat in edge, and the bacterium colony size is 1-2mm, and it is faint yellow that bacterium colony is, and this bacterium all can grow under the scope of 4 ℃-37 ℃ substratum temperature and pH7-9.5.
3. claim 1 or 2 each described bacillus lentuses are induced and are produced the method that calcium carbonate is used for foundation stabilization, it is characterized in that: comprise the steps:
Step 1: bacillus lentus Bacillus lentus UR1C, Bacillus lentus UR9D and the single bacterium colony of Bacillus lentus UR52 were obtained three kinds of bacterium liquid respectively in fermentation culture 12-60 hour in fermention medium under 25 ℃ of-37 ℃ of conditions;
Step 2: adopt the substep injection process, be in the milk in two steps, pour into bacterium liquid earlier, pour into reaction solution calcium chloride and urea mixing solutions then, concrete grammar is: form three sand posts in three the sand grains moulds of at first sand grains being packed into, pump into the deionized water of 3 times of sand-grain volumes then respectively in three sand posts, make the sand post saturated, in three sand posts, pour into one of described three kinds of bacterium liquid of step 1 respectively subsequently, the volume that pours into bacterium liquid is 1-3 times of sand-grain volume, at last divide 2-4 to criticize the mixing solutions that pours into reaction solution calcium chloride and urea respectively in three sand posts, original position generates MCP in the sand post; The mixing solutions of described reaction solution calcium chloride and urea contains 0.5M CaCl for every liter
2With 0.5M urea, every batch of mixed liquor volume that pours into is 3 times that bacteria liquid amasss, and makes and forms microorganism-calcium carbonate-sand grains induration in sand grains-solution system, and then strengthen bonding degree and the intensity of sand grains.
4. method according to claim 3, it is characterized in that: the described fermention medium of step 1 comprises yeast extract 10-20g/L, ammonium sulfate or ammonium chloride 10g/L, pH are 7-9.5.
5. method according to claim 3 is characterized in that: the described speed that pumps into deionized water in three sand posts of step 2 is 0.5-1.5ml/min.
6. method according to claim 3 is characterized in that: the described speed that pours into the mixing solutions of reaction solution calcium chloride and urea in the sand post of step 2 is 0.05-0.15ml/min.
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Application publication date: 20130828 |