CN117230777A - Method for solidifying granite residual soil - Google Patents
Method for solidifying granite residual soil Download PDFInfo
- Publication number
- CN117230777A CN117230777A CN202311194871.7A CN202311194871A CN117230777A CN 117230777 A CN117230777 A CN 117230777A CN 202311194871 A CN202311194871 A CN 202311194871A CN 117230777 A CN117230777 A CN 117230777A
- Authority
- CN
- China
- Prior art keywords
- bacillus
- residual soil
- granite
- solidifying
- granite residual
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000002689 soil Substances 0.000 title claims abstract description 55
- 239000010438 granite Substances 0.000 title claims abstract description 47
- 238000000034 method Methods 0.000 title claims abstract description 31
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 73
- 239000007788 liquid Substances 0.000 claims abstract description 36
- 230000001580 bacterial effect Effects 0.000 claims abstract description 27
- 241000976738 Bacillus aryabhattai Species 0.000 claims abstract description 23
- 239000013535 sea water Substances 0.000 claims abstract description 20
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 11
- 238000011049 filling Methods 0.000 claims abstract description 8
- 239000002609 medium Substances 0.000 claims description 34
- 241000589158 Agrobacterium Species 0.000 claims description 14
- 150000003839 salts Chemical class 0.000 claims description 14
- 239000001963 growth medium Substances 0.000 claims description 12
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 11
- 239000004202 carbamide Substances 0.000 claims description 11
- 238000004321 preservation Methods 0.000 claims description 10
- 238000000855 fermentation Methods 0.000 claims description 8
- 230000004151 fermentation Effects 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 5
- 239000012530 fluid Substances 0.000 claims description 3
- 239000004576 sand Substances 0.000 abstract description 33
- 108010046334 Urease Proteins 0.000 abstract description 22
- 238000012216 screening Methods 0.000 abstract description 4
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 17
- 230000000694 effects Effects 0.000 description 17
- 241001401069 Terribacillus saccharophilus Species 0.000 description 12
- 241001143891 Bacillus aquimaris Species 0.000 description 11
- 244000005700 microbiome Species 0.000 description 10
- 239000000243 solution Substances 0.000 description 10
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 8
- 229910000019 calcium carbonate Inorganic materials 0.000 description 8
- 229940041514 candida albicans extract Drugs 0.000 description 8
- 239000012138 yeast extract Substances 0.000 description 8
- 108091028043 Nucleic acid sequence Proteins 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 238000009629 microbiological culture Methods 0.000 description 6
- 239000002245 particle Substances 0.000 description 6
- 239000000834 fixative Substances 0.000 description 5
- 230000010412 perfusion Effects 0.000 description 5
- 235000019270 ammonium chloride Nutrition 0.000 description 4
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 4
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 4
- 235000011130 ammonium sulphate Nutrition 0.000 description 4
- 238000009826 distribution Methods 0.000 description 4
- 238000001556 precipitation Methods 0.000 description 4
- 241000006384 Jeotgalibacillus marinus Species 0.000 description 3
- 238000001000 micrograph Methods 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 150000007523 nucleic acids Chemical class 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 230000002787 reinforcement Effects 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 238000002441 X-ray diffraction Methods 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 125000003275 alpha amino acid group Chemical group 0.000 description 2
- 239000001166 ammonium sulphate Substances 0.000 description 2
- 229910001424 calcium ion Inorganic materials 0.000 description 2
- 238000007596 consolidation process Methods 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 241000304886 Bacilli Species 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 229910021532 Calcite Inorganic materials 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000004927 clay Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000012669 compression test Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000003628 erosive effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 230000002572 peristaltic effect Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a method for solidifying granite residual soil. The method comprises the following steps: providing bacillus; sequentially filling bacillus, a fixing agent and cementing liquid into granite residual soil; wherein the bacillus is selected from one or more of bacillus seawater, bacillus aryabhattai and bacillus terrestris. According to the method for solidifying the granite residual soil, three bacterial liquids which are obtained by screening in nature and can produce urease are adopted to perfuse the granite residual soil, so that the solidified granite residual soil with high uniaxial compressive strength is formed. The method can quickly and efficiently adhere loose sand grains together, so that the strength of foundation sand is improved; in addition, the strains used in the invention are isolated from the natural world, and the strains cannot pollute the natural environment.
Description
Technical Field
The invention relates to the technical field of foundation reinforcement, in particular to a method for solidifying granite residual soil.
Background
The granite residual soil is a special geological material formed in arid and semiarid desert climatic environments, and is also a cheap material with the most extensive distribution, the most abundant reserves and the most convenient material acquisition. However, the granite residual soil has loose structure, less content of clay particles, no cohesive force, poor stability and low bearing capacity, and is subjected to frequent wind power transportation and wind erosion throughout the year, and has high fluidity. In addition, the content of the saline-alkali in the granite residual soil is high, and large-scale construction equipment cannot enter or the construction cost is too high; limited by various factors, the general foundation treatment technology is not applicable to granite residual soil areas.
The essence of the microorganism-induced calcium carbonate precipitation technique (Microbially Induced Calcite Precipitation, MICP) is that bacteria of a certain class in nature are metabolized to produce urease that breaks down urea, and carbonate ions produced after urea breaking down can combine with free metal cations in nature to produce gelled crystals.
At present, researchers often spray bacterial liquid of bacillus pasteurizer onto the surface of granite residual soil, and the strength of the granite residual soil is improved by utilizing the urease producing capability of the bacillus pasteurizer; however, it is not known whether other strains than Bacillus pasteuris can be used to consolidate granite residual soil.
Disclosure of Invention
Based on the above, the invention provides a method for solidifying granite residual soil by using bacterial liquid of one or more strains of bacillus sea, bacillus aryabhattai and bacillus terrestris.
The specific technical scheme is as follows:
according to one aspect of the present invention, there is provided a method of curing granite residual soil comprising the steps of:
providing bacillus; and
sequentially filling the bacillus, the fixing agent and the cementing liquid into granite residual soil;
wherein the bacillus is selected from one or more of bacillus seawater, bacillus aryabhattai and bacillus terrestris.
In one embodiment, the preservation number of the bacillus seawater is CGMCC No.18068.
In one embodiment, the bacillus aryabhattai has a preservation number of CGMCC No.18069.
In one embodiment, the preservation number of the agrobacterium is CGMCC No.18070.
In one embodiment, after the step of providing bacillus, before the step of filling the bacillus, the method further comprises:
inoculating single colony of bacillus into fermentation culture medium, and culturing at 25-37 deg.c and 150-250 rpm for 12-60 hr.
In one of the factsIn an embodiment, the fermentation medium is NH 4 -YE medium or salt medium.
In one embodiment, the fixative comprises 0.04M-0.06M CaCl 2 。
In one embodiment, the cementing fluid comprises 0.4M to 0.6M CaCl 2 And 0.4M to 0.6M urea.
In one embodiment, the ratio of the bacterial liquid of the bacillus, the fixing agent and the cementing liquid is (40 mL to 160 mL): (0.01M-0.1M): (140 mL-160 mL).
In one embodiment, the grouting speed of the bacterial liquid of the bacillus is 0.05 mL/min-1.00 mL/min; and/or
The grouting speed of the fixing agent is 0.05 mL/min-1.00 mL/min; and/or
The grouting speed of the cementing liquid is 0.05 mL/min-0.15 mL/min.
Compared with the prior art, the invention has the following beneficial effects:
according to the method for solidifying the granite residual soil, three bacterial liquids which are obtained by screening in nature and can produce urease are adopted to perfuse the granite residual soil, so that the solidified granite residual soil with high uniaxial compressive strength is formed. The method can quickly and efficiently adhere loose sand grains together, so that the strength of foundation sand is improved; in addition, the strains used in the invention are isolated from the natural world, and the strains cannot pollute the natural environment.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 is an electron microscope view of Bacillus seawater Bacillus aquimaris H216;
FIG. 2 is an electron microscopic view of Bacillus aryabhattai Bacillus aryabhattai H961;
FIG. 3 is an electron microscope view of Agrobacterium Terribacillus saccharophilus H986;
FIG. 4 shows NH of three strains selected according to the invention 4 -statistical graphs of bacterial liquid concentration in YE medium and salt medium;
FIG. 5 shows NH of three strains selected according to the invention 4 -a statistical map of urease activity in YE medium and salt medium;
FIG. 6 is a statistical chart of urease activity of three strains screened according to the present invention under different culture temperature conditions;
FIG. 7 is a statistical chart of urease activity of three strains screened according to the present invention under different pH media conditions;
FIG. 8 is a graph showing the particle size distribution of granite residual soil sample particles used in example 1;
FIG. 9 is a scanning electron microscope image of a sand column after the Bacillus seawater Bacillus aquimaris H is filled;
FIG. 10 is a scanning electron microscope image of a sand column after perfusion of Bacillus aryabhattai Bacillus aryabhattai H961;
FIG. 11 is a scanning electron microscope image of a sand column after perfusion of Agrobacterium Terribacillus saccharophilus H986;
FIG. 12 is a statistical plot of uniaxial compressive strength of a granite residual soil sand column filled with three strains screened in accordance with the present invention.
Detailed Description
The detailed description of the present invention will be provided to make the above objects, features and advantages of the present invention more obvious and understandable. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. The present invention may be embodied in many other forms than described herein and similarly modified by those skilled in the art without departing from the spirit of the invention, whereby the invention is not limited to the specific embodiments disclosed below.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Unless otherwise specifically indicated, the various raw materials, reagents, instruments, equipment and the like used in the present invention are commercially available or may be prepared by existing methods.
Terminology
Unless otherwise indicated or contradicted, terms or phrases used herein have the following meanings:
the term "and/or," "and/or," as used herein, includes any one of two or more of the listed items in relation to each other, as well as any and all combinations of the listed items in relation to each other, including any two of the listed items in relation to each other, any more of the listed items in relation to each other, or all combinations of the listed items in relation to each other. It should be noted that, when at least three items are connected by a combination of at least two conjunctions selected from the group consisting of "and/or", "and/or", it should be understood that, in the present invention, the technical solutions include technical solutions that all use "logical and" connection, and also include technical solutions that all use "logical or" connection. For example, "a and/or B" includes three parallel schemes A, B and a+b. For another example, the technical schemes of "a, and/or B, and/or C, and/or D" include any one of A, B, C, D (i.e., the technical scheme of "logical or" connection), and also include any and all combinations of A, B, C, D, i.e., any two or three of A, B, C, D, and also include four combinations of A, B, C, D (i.e., the technical scheme of "logical and" connection).
The terms "plurality", "plural", "multiple", and the like in the present invention refer to, unless otherwise specified, an index of 2 or more in number. For example, "one or more" means one kind or two or more kinds.
As used herein, "a combination thereof," "any combination thereof," and the like include all suitable combinations of any two or more of the listed items.
The "suitable" in the "suitable combination manner", "suitable manner", "any suitable manner" and the like herein refers to the fact that the technical scheme of the present invention can be implemented, the technical problem of the present invention is solved, and the technical effect expected by the present invention is achieved.
Herein, "preferred", "better", "preferred" are merely to describe better embodiments or examples, and it should be understood that they do not limit the scope of the invention.
In the present invention, "further", "still further", "particularly" and the like are used for descriptive purposes to indicate differences in content but should not be construed as limiting the scope of the invention.
In the present invention, "optional" means optional or not, that is, means any one selected from two parallel schemes of "with" or "without". If multiple "alternatives" occur in a technical solution, if no particular description exists and there is no contradiction or mutual constraint, then each "alternative" is independent.
In the invention, the technical characteristics described in an open mode comprise a closed technical scheme composed of the listed characteristics and also comprise an open technical scheme comprising the listed characteristics.
In the present invention, a numerical range (i.e., a numerical range) is referred to, and optional numerical distributions are considered to be continuous within the numerical range and include two numerical endpoints (i.e., a minimum value and a maximum value) of the numerical range and each numerical value between the two numerical endpoints unless otherwise specified. Where a numerical range merely refers to integers within the numerical range, including both end integers of the numerical range, and each integer between the two ends, unless otherwise indicated, each integer is recited herein as directly, such as where t is an integer selected from 1-10, and where t is any integer selected from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. Further, when a plurality of range description features or characteristics are provided, these ranges may be combined. In other words, unless otherwise indicated, the ranges disclosed herein are to be understood to include any and all subranges subsumed therein.
The temperature parameter in the present invention is not particularly limited, and may be a constant temperature treatment or may vary within a predetermined temperature range. It should be appreciated that the constant temperature process described allows the temperature to fluctuate within the accuracy of the instrument control. Allows for fluctuations within a range such as + -5 ℃, + -4 ℃, + -3 ℃, + -2 ℃, + -1 ℃.
Some embodiments of the present invention provide a method of curing granite residual soil.
In some of these embodiments, the method of curing granite residual soil includes steps S10 and S30.
S10: providing bacillus; and
s30: sequentially filling bacterial liquid, a fixing agent and cementing liquid of the bacillus into granite residual soil;
wherein in step S10, the bacillus is selected from one or more of Bacillus seawater, bacillus aryabhattai and Bacillus soil.
Understandably, the bacterial liquid can be selected from any bacterial liquid of bacillus seawater, bacillus aryabhattai and bacillus terrestris; or selecting any two mixed bacterial solutions; the mixed bacterial liquid of the three strains can also be selected.
In some specific examples, the bacillus seawater is selected from bacillus seawater Bacillus aquimaris H, and the strain has been preserved in China general microbiological culture Collection center (address: north Xielu No.1, 3, postal code: 100101, of the Korean-yang area of Beijing) at 7.4 days of 2019, and the preservation number is CGMCC No.18068.
In some specific examples, the 16s DNA sequence of the bacillus seawater is shown as SEQ ID NO. 1.
In addition, the Bacillus seawater may be selected from a sequence substantially similar to SEQ ID NO.1 in the 16s DNA sequence. By "substantially similar" is meant that a given nucleic acid or amino acid sequence shares at least 95% identity with a reference sequence, e.g., 96%, 97%, 98%, 98.5%, 99%, 99.5%. Alternatively, it is meant that a given nucleic acid or amino acid sequence has 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 nucleic acids or amino acids differences from a reference sequence.
In some specific examples, the Bacillus aryasis is selected from Bacillus aryasis Bacillus aryabhattai H961, which has been deposited in China general microbiological culture Collection center (address: north Xiyroad No.1, 3, postal code: 100101, of the area of Korea of Beijing) on day 7 and day 4, with a deposit number of CGMCC No.18069.
In some specific examples, the 16s DNA sequence of the Bacillus aryabhattai is shown in SEQ ID NO. 2.
In some specific examples, the agrobacterium is selected from agrobacterium Terribacillus saccharophilus H986, which has been deposited in China general microbiological culture Collection center (address: institute 1, 3, postal code: 100101, and accession number CGMCC No. 18070) by the China Committee for culture Collection of microorganisms, 7.4, 2019.
In some specific examples, the 16s DNA sequence of the above-mentioned Agrobacterium is shown in SEQ ID NO. 3.
In some embodiments, the method for solidifying residual soil of granite further includes step S20:
inoculating single colony of bacillus into fermentation culture medium, and culturing at 25-37 deg.c and 150-250 rpm for 12-60 hr.
In one preferred example, in step S20, individual colonies of Bacillus are inoculated into a fermentation medium and cultured at 30℃for 16 hours at 150rpm to 250 rpm; and (5) collecting bacterial liquid.
In some embodiments, in step S20, the fermentation medium is NH 4 -YE medium or high salt medium.
In some of these specific examples, NH 4 -YE medium comprising yeast extract and ammonium sulphate; the proportion of the yeast extract to the ammonium sulfate is (10-20) according to the mass volume ratio: 10.
understandably, NH 4 The ratio of yeast extract to ammonium sulphate in the YE medium is "(10-20): 10", i.e. including but not limited to the point values in the examples: specific ratios of 10:10, 11:10, 12:10, 13:10, 14:10, 15:10, 16:10, 17:10, 18:10, 19:10, 20:10, etc.
In some specific examples, the salt medium includes yeast extract and ammonium chloride; the mass volume ratio of the yeast extract to the ammonium chloride is (10-20): 10.
understandably, the ratio of yeast extract to ammonium chloride in the salt medium is "(10-20): 10", i.e. including but not limited to the point values in the examples: specific ratios of 10:10, 11:10, 12:10, 13:10, 14:10, 15:10, 16:10, 17:10, 18:10, 19:10, 20:10, etc.
Further, NH 4 The pH of the YE medium or the salt medium is 7.0-9.5. It is understood that controlling the pH of the fermentation medium within the above-described range may promote urease-producing ability of the strain at a relatively low cost.
In some embodiments, in step S30, the fixative includes 0.04M to 0.06M CaCl 2 。
In some embodiments, the cementing fluid comprises 0.4M to 0.6M CaCl 2 And 0.4M to 0.6M urea.
According to the microorganism-induced calcium carbonate precipitation technique (MICP), certain specific microorganisms can utilize organic matters such as urea in the surrounding environment, a calcium ion source and the like to generate calcium carbonate with gelling properties; because the generation speed and the strength of the calcium carbonate material with microbial causes are controllable, the calcium carbonate material can be used as a binder to bind loose sand grains into artificial sand gravel with controllable strength and permeability. Therefore, MICP is expected to find wide application in concrete, sandy soil, powder soil consolidation and the like.
The three bacillus strains can generate urease, so that urea in the sandy soil environment is decomposed into ammonium and carbon dioxide, and meanwhile, the pH value of the sandy soil environment around the bacillus strains is raised; and then pouring a fixing agent containing calcium ions and cementing liquid into the sand to generate water-insoluble calcium carbonate salt so as to bond the surrounding sand grains together.
In some embodiments, the ratio of the bacterial liquid of bacillus, the fixing agent and the cementing liquid is (40 mL-160 mL): (0.01M-0.1M): (140 mL-160 mL).
In some embodiments, the grouting speed of the bacterial liquid of the bacillus is 0.05 mL/min-1.00 mL/min. Preferably, the grouting speed of the bacterial liquid is 0.5mL/min.
In some embodiments, the fixative has a grouting rate of 0.05mL/min to 1.00mL/min. Preferably, the fixative has a grouting speed of 0.5mL/min.
In some embodiments, the grouting speed of the cementing liquid is 0.05mL/min to 0.15mL/min. Preferably, the grouting speed of the cementing liquid is 0.1mL/min.
The method applies a plurality of microorganism resources which are abundant in nature and have no toxicity to the foundation reinforcement engineering, can effectively change the mechanical properties of civil engineering and geotechnical engineering, and opens up a new idea for the foundation reinforcement field of the civil engineering and the geotechnical engineering.
It will be appreciated that the three bacilli of the present invention may be formulated to solidify granite residual soil.
In some embodiments, the formulation comprises one or more of bacillus seawater, bacillus aryabhattai, and bacillus terrestris.
One or more of the strains are prepared into a preparation, which is convenient to use, and the preparation with proper dosage is added according to the volume of the residual soil of granite during use.
The present invention will be further described with reference to specific examples and comparative examples, which should not be construed as limiting the scope of the invention.
Example 1:
(1) Screening and identification of strains
Enriching and culturing the collected sea sand test sample (offshore sea sand of south China sea) for 24 hours at 37 ℃ under the condition of 5M high-concentration urea, and killing various microbial vegetative cells which cannot tolerate and utilize the high-concentration urea; then carrying out gradient dilution on the treated culture solution, coating a urease screening culture plate, culturing at 37 ℃, picking out strains which turn the color of the culture medium red, streaking and separating single colonies to obtain three urease-producing microorganisms: bacillus seawater Bacillus aquimaris H216, bacillus albopictus Bacillus aryabhattai H961 and bacillus agrobacterium Terribacillus saccharophilus H986. The three isolated strains have urea-decomposing enzyme, can decompose urea to produce a large amount of ammonia, and make the culture medium alkaline and red.
The bacillus sea water, bacillus alnicosum and bacillus soil are all in rod shape, have spores, have no capsules and are positive in gram staining. FIG. 1 is an electron microscope view of Bacillus seawater Bacillus aquimaris H216; FIG. 2 is an electron microscopic view of Bacillus aryabhattai Bacillus aryabhattai H961; fig. 3 is an electron microscope view of agrobacterium Terribacillus saccharophilus H986.
At NH 4 On the YE culture medium, the bacterial colony is round, the surface is moist and smooth, the edge is neat, the size of the bacterial colony is 1 mm-2 mm, and the bacterial colony is light yellow; and the three isolated strains can grow at the temperature of the culture medium of 4-37 ℃ and the pH value of 7.0-9.5.
Three isolated bacteria are further identified by a 16S rDNA sequencing method, and the 16S DNA sequence of the bacillus marinus is shown as SEQ ID NO. 1; the 16s DNA sequence of the bacillus aryabhattai is shown as SEQ ID NO. 2; the 16s DNA sequence of the soil bacillus is shown as SEQ ID NO. 3.
SEQ ID NO.1：
ttgctcccatcagtcagcggcggacgggtgagtaacacgtgggtaacctgcctgtaagactgggataactccgggaaaccggggctaataccggataactcatttcctcgcatgaggaaatgttgaaaggtggcttttagctatcacttacagatggacccgcggcgcattagctagttggtgaggtaatggctcaccaaggcgacgatgcgtagccgacctgagagggtgatcggccacactgggactgagacacggcccagactcctacgggaggcagcagtagggaatcttccgcaatggacgaaagtctgacggagcaacgccgcgtgagtgatgaaggttttcggatcgtaaagctctgttgttagggaagaacaagtaccgttcgaatagggcggtaccttgacggtacctaaccagaaagccacggctaactacgtgccagcagccgcggtaatacgtaggtggcaagcgttgtccggaattattgggcgtaaagcgcgcgcaggtggttccttaagtctgatgtgaaagcccacggctcaaccgtggagggtcattggaaactggggaacttgagtgcagaagaggaaagtggaattccaagtgtagcggtgaaatgcgtagatatttggaggaacaccagtggcgaaggcgactttctggtctgtaactgacactgaggcgcgaaagcgtggggagcaaacaggattagataccctggtagtccacgccgtaaacgatgagtgctaagtgttagggggtttccgccccttagtgctgcagctaacgcattaagcactccgcctggggagtacggtcgcaagactgaaactcaaaggaattgacgggggcccgcacaagcggtggagcatgtggtttaattcgaagcaacgcgaagaaccttaccaggtcttgacatcctctgacaaccctagagatagggctttccccttcgggggacagagtgacaggtggtgcatggttgtcgtcagctcgtgtcgtgagatgttgggttaagtcccgcaacgagcgcaacccttgatcttagttgccagcattcagttgggcactctaagatgactgccggtgacaaaccggaggaaggtggggatgacgtcaaatcatcatgccccttatgacctgggctacacacgtgctacaatggacggtacaaagggcagcaagaccgcgaggtttagccaatcccataaaaccgttctcagttcggattgtaggctgcaactcgcctacatgaagctggaatcgctagtaatcgcggatcagcatgccgcggtgaatacgttcccgggccttgtacacaccgcccgtcacaccacgagagtttg
SEQ ID NO.2：
cgttagcggcggacgggtgagtaacacgtgggcaacctgcctgtaagactgggataacttcgggaaaccgaagctaataccggataggatcttctccttcatgggagatgattgaaagatggtttcggctatcacttacagatgggcccgcggtgcattagctagttggtgaggtaacggctcaccaaggcaacgatgcatagccgacctgagagggtgatcggccacactgggactgagacacggcccagactcctacgggaggcagcagtagggaatcttccgcaatggacgaaagtctgacggagcaacgccgcgtgagtgatgaaggctttcgggtcgtaaaactctgttgttagggaagaacaagtacgagagtaactgctcgtaccttgacggtacctaaccagaaagccacggctaactacgtgccagcagccgcggtaatacgtaggtggcaagcgttatccggaattattgggcgtaaagcgcgcgcaggcggtttcttaagtctgatgtgaaagcccacggctcaaccgtggagggtcattggaaactggggaacttgagtgcagaagagaaaagcggaattccacgtgtagcggtgaaatgcgtagagatgtggaggaacaccagtggcgaaggcggctttttggtctgtaactgacgctgaggcgcgaaagcgtggggagcaaacaggattagataccctggtagtccacgccgtaaacgatgagtgctaagtgttagagggtttccgccctttagtgctgcagctaacgcattaagcactccgcctggggagtacggtcgcaagactgaaactcaaaggaattgacgggggcccgcacaagcggtggagcatgtggtttaattcgaagcaacgcgaagaaccttaccaggtcttgacatcctctgacaactctagagatagagcgttccccttcgggggacagagtgacaggtggtgcatggttgtcgtcagctcgtgtcgtgagatgttgggttaagtcccgcaacgagcgcaacccttgatcttagttgccagcatttagttgggcactctaaggtgactgccggtgacaaaccggaggaaggtggggatgacgtcaaatcatcatgccccttatgacctgggctacacacgtgctacaatggatggtacaaagggctgcaagaccgcgaggtcaagccaatcccataaaaccattctcagttcggattgtaggctgcaactcgcctacatgaagctggaatcgctagtaatcgcggatcagcatgccgcggtgaatacgttcccgggccttgtacacaccgcccgtcacaccacgagagtttgta
SEQ ID NO.3：
tataatgcaagtcgagcgcaggaaccagatgatcccttcggggtgattctggtggaatgagcggcggacgggtgagtaacacgtgggcaacctgcctgtaagactgggataacttcgggaaaccggagctaataccggatagtatttcctttctcctgattggaaatggaaagacggtttcggctgtcacttacagatgggcccgcggtgcattagctagttggtggggtaatggcccaccaaggcgacgatgcatagccgacctgagagggtgatcggccacactgggactgagacacggcccagactcctacgggaggcagcagtagggaatcttccgcaatggacgaaagtctgacggagcaacgccgcgtgagcgatgaaggccttcgggtcgtaaagctctgttgttagggaagaacaagtacgagagtaactgctcgtaccttgacggtacctaaccagaaagccccggctaactacgtgccagcagccgcggtaatacgtagggggcaagcgttgtccggaattattgggcgtaaagggctcgtaggcggtttcttaagtctgatgtgaaagcccacagctcaactgtggagggtcattggaaactggggaacttgagtgcagaagaggagagtggaattccacgtgtagcggtgaaatgcgtagatatgtggaggaacaccagtggcgaaggcgactctctggtctgtaactgacgctgaggagcgaaagcgtggggagcaaacaggattagataccctggtagtccacgccgtaaacgatgagtgctaggtgttagggggtttccgccccttagtgctgaagttaacgcattaagcactccgcctggggagtacggccgcaaggctgaaactcaaaagaattgacgggggcccgcacaagcggtggagcatgtggtttaattcgaagcaacgcgaagaaccttaccaggtcttgacatccgctgacaatcttggagacaagacgttcccttcggggacagcgtgacaggtggtgcatggttgtcgtcagctcgtgtcgtgagatgttgggttaagtcccgcaacgagcgcaacccttgattctagttgccagcattaagttgggcactctagagtgactgccggtgacaaaccggaggaaggtggggatgacgtcaaatcatcatgccccttatgacctgggctacacacgtgctacaatggatggtacaaagggcagcaaagtcgcgaggctaagcaaatcccataaaaccattctcagttcggattgcaggctgcaactcgcctgcatgaagccggaatcgctagtaatcgcggatcagcatgccgcggtgaatacgttcccgggccttgtacacaccgcccgtcacaccacgagagttggtaacacccgaagtcggtgaggtaaccttttaggagccagccgccgaaggtgggatcaatga
The bacillus marinus Bacillus aquimaris H216 is preserved in China general microbiological culture Collection center (address: north Xila No.1, 3, postal code: 100101) of the China Committee for culture Collection of microorganisms, 7 months and 4 days of 2019, and the preservation number is CGMCC No.18068.
The bacillus aryasis Bacillus aryabhattai H961 is preserved in China general microbiological culture Collection center (address: north Chen Xila No.1, 3, postal code: 100101) of China Committee for culture Collection of microorganisms, 7.4.2019, and the preservation number is CGMCC No.18069.
The agrobacterium Terribacillus saccharophilus H and 986 is preserved in China general microbiological culture Collection center (address: north Xiyusa No.1, 3, postal code: 100101) of the China Committee for culture Collection of microorganisms, 7 and 4 of 2019, and the preservation number is CGMCC No.18070.
(2) Strain culture
NH was formulated separately according to the following formulation 4 -YE medium and high salt medium:
NH 4 -YE medium: yeast extract 20g/L, ammonium sulfate 10g/L, and pH is adjusted to 7.5-9.0.
High salt medium: yeast extract 20g/L, ammonium chloride 10g/L and pH adjusted to 7.5-9.0.
Taking 100mL of NH 4 -YE culture medium or high-salt culture medium, and filling into 500mL culture bottles, and sterilizing at high temperature for later use. Single colonies of bacillus seawater Bacillus aquimaris H, bacillus albopictus Bacillus aryabhattai H961 and bacillus agrobacteria Terribacillus saccharophilus H986 are respectively picked from a flat plate culture medium, inoculated into a 500mL culture bottle, cultured for 16 hours at 30 ℃ and 200rpm, and then bacterial liquid is collected for standby; and detecting the biomass (OD) of the bacterial liquid 600 ) And urease activity.
FIG. 4 shows three strains in NH respectively 4 OD after cultivation in YE medium and high salt medium 600 Values. As can be seen from the figure: compared with the other two strains, the bacillus aryabhattai Bacillus aryabhattai H961 has relatively higher bacterial liquid concentration; and the growth state of the three strains cultured in the high-salt culture medium is slightly better than that of NH 4 -YE medium.
FIG. 5 shows three strains in NH respectively 4 Urease activity after incubation in YE medium and high salt medium. As can be seen from the figure: whichever culture medium is used for the culture, the bacillus marinus Bacillus aquimaris H21Both the urease activity of 6 and the urease activity of agrobacterium Terribacillus saccharophilus H986 are high with Yu Ashi bacillus Bacillus aryabhattai H961; and, three strains are in NH 4 The urease activity after cultivation in the YE medium is higher than that in the high-salt medium.
FIG. 6 shows urease activity of three strains after cultivation in high salt medium at different temperatures. As can be seen from the figure: the three strains have extremely wide growth temperature range, can survive and grow at the temperature of between-4 ℃ and 80 ℃ and have higher urease activity; wherein, the urease activity of the strain cultured in the range of 28-50 ℃ is obviously higher.
FIG. 7 shows urease activity of three strains after cultivation in high salt medium at different pH. As shown in the figure, all three microorganisms have certain alkali resistance activity, can survive and grow in the pH range of 7.0-10.0, and have higher urease activity. In addition, the urease activity of agrobacterium Terribacillus saccharophilus H986 was higher than the other two strains at different pH.
(3) Preparation of solidified sand column
The granite residual soil is sampled to Zhaoqing city of Guangdong, the granularity distribution condition is shown in figure 8, and about 95 percent of the granite residual soil particles have the granularity between 0.1mm and 0.4 mm; XRD analysis shows that the main component of the granite residual soil sample is quartz.
In order to control the particle size, the granite residual soil is screened, and sand particles with the diameter of 0.1 mm-0.4 mm are collected and used as samples for the next step.
100 dry granite residual soil is filled into a sterile model column with the volume of 50mL, the height of 110mm and the inner diameter of 30mm, and the sand column to be solidified is prepared. In order to avoid the situation of blocking a grouting opening, two layers of gauze filter layers are respectively arranged at two ends of the sand column to be solidified.
(4) Grouting
Sequentially pouring bacterial liquid and fixative (0.05M CaCl) by adopting a distributed grouting method 2 ) And cementing liquid (0.5 MCaCl) 2 And 0.5M urea). The injector is adopted for filling, the upper end of the injector is tightly plugged by a rubber plug with a three-way pipe,two layers of coarse sand were used at both ends of the syringe for filtration to avoid clogging during calcium carbonate precipitation.
150mL of deionized water was first pumped using a peristaltic pump at 10rpm (about 1 mL/min) to saturate the sand column as much as possible. Respectively pouring three bacterial liquids collected in the step (2) into three groups of sand columns (each group is provided with 3 repetitions) at the speed of 0.5 mL/min; 50mL of 0.05M CaCl was then poured into each column at a rate of 0.5mL/min 2 A solution; finally, 150mL of cementing liquid is filled into each sand column at the speed of 0.1mL/min, so that microorganism-calcium carbonate-sand consolidation bodies are formed in a sand-solution system, and the bonding degree and strength of sand are enhanced.
(5) Analysis of results
XRD analysis was performed on the white precipitated material in each column, and the material formed was calcium carbonate. Further electron microscopic observation is carried out on the precipitate, and the results are shown in the figures 9-11 (the figure 9 is the perfusion of bacillus sea water Bacillus aquimaris H and the figure 10 is the perfusion of bacillus albopictus Bacillus aryabhattai H961 and the figure 11 is the perfusion of bacillus soil Terribacillus saccharophilus H986); after the bacterial solutions of the three strains are used for pouring, spherical and hexahedral calcium carbonate crystals are obviously generated between sand grains.
Further, the obtained sand column is subjected to demolding, and the sand column is bonded and molded and has certain strength; carrying out a single-shaft compression test on all sand columns, wherein a sand column sample adopts a cylinder with the diameter of 50mm and the height of 100mm, the parallelism deviation of two end faces of the sample is not more than 0.1m, placing the sample at the center of a bearing plate of a press, adjusting the bearing plate to uniformly stress the sample, and loading the sample at a loading speed of 0.5-0.8 MPa/s until the sample is destroyed; the number of samples in each group is not less than 3. The results are shown in FIG. 12: the uniaxial compressive strength of the sand column filled with bacillus aryabhattai Bacillus aryabhattai H961 was about 0.6Mpa; the uniaxial compressive strength of the sand column filled with the seawater bacillus Bacillus aquimaris H216 is about 0.4Mpa; the uniaxial compressive strength of the sand column perfused with the agrobacterium Terribacillus saccharophilus H986 was about 0.2Mpa.
The results show that the three bacillus of the invention can bond loose sand grains together, so that the compressive strength of granite residual soil is improved to a certain extent; among them, the effect of the bacillus aryabhattai Bacillus aryabhattai H961 to solidify granite residual soil is relatively more excellent than that of the bacillus maritima Bacillus aquimaris H and the bacillus agrobacterium Terribacillus saccharophilus H986.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The above examples illustrate only a few embodiments of the invention, which are described in detail and are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (10)
1. A method of curing granite residual soil, comprising the steps of:
providing bacillus; and
sequentially filling the bacillus, the fixing agent and the cementing liquid into granite residual soil;
wherein the bacillus is selected from one or more of bacillus seawater, bacillus aryabhattai and bacillus terrestris.
2. The method for solidifying residual soil of granite of claim 1, wherein the preservation number of the bacillus seawater is CGMCC No.18068.
3. The method of solidifying residual soil of granite of claim 1, wherein the bacillus aryabhattai has a preservation number of CGMCC No.18069.
4. The method for solidifying residual soil of granite of claim 1, wherein the preservation number of the agrobacterium is CGMCC No.18070.
5. The method of solidifying residual soil of granite of any one of claims 1-4, further comprising, after the step of providing bacillus, prior to the step of filling the bacillus:
inoculating single colony of bacillus into fermentation culture medium, and culturing at 25-37 deg.c and 150-250 rpm for 12-60 hr.
6. The method of solidifying residual soil of granite of claim 5, wherein the fermentation medium is NH 4 -YE medium or salt medium.
7. The method of solidifying residual soil of granite according to any one of claims 1-4 and 6, wherein the fixing agent comprises 0.04-0.06M CaCl 2 。
8. The method of curing residual soil of granite of any of claims 1-4 and 6, wherein the cementing fluid comprises 0.4M to 0.6M CaCl 2 And 0.4M to 0.6M urea.
9. The method of solidifying residual soil of granite according to any one of claims 1-4 and 6, wherein the ratio of the bacterial liquid of bacillus, the fixing agent and the cementing liquid is (40-160 mL): (0.01M-0.1M): (140 mL-160 mL).
10. The method for solidifying residual soil on granite according to any one of claims 1-4 and 6, wherein the grouting speed of the bacterial liquid of bacillus is 0.05-1.00 mL/min; and/or
The grouting speed of the fixing agent is 0.05 mL/min-1.00 mL/min; and/or
The grouting speed of the cementing liquid is 0.05 mL/min-0.15 mL/min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311194871.7A CN117230777A (en) | 2023-09-15 | 2023-09-15 | Method for solidifying granite residual soil |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311194871.7A CN117230777A (en) | 2023-09-15 | 2023-09-15 | Method for solidifying granite residual soil |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117230777A true CN117230777A (en) | 2023-12-15 |
Family
ID=89094253
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311194871.7A Pending CN117230777A (en) | 2023-09-15 | 2023-09-15 | Method for solidifying granite residual soil |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117230777A (en) |
-
2023
- 2023-09-15 CN CN202311194871.7A patent/CN117230777A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Tang et al. | Factors affecting the performance of microbial-induced carbonate precipitation (MICP) treated soil: a review | |
| Gebru et al. | Bio-cement production using microbially induced calcite precipitation (MICP) method: A review | |
| Mahawish et al. | Effect of particle size distribution on the bio-cementation of coarse aggregates | |
| Ivanov et al. | Basics of construction microbial biotechnology | |
| CN103173376B (en) | Method for preparing high-strength microbial mortar by using urease-producing microbes | |
| CN103266070A (en) | Bacillus lentus and method for generating by inducing bacillus lentus for reinforcing foundation | |
| KR101030761B1 (en) | Cememtation of soft ground using bacteria | |
| CN103773376B (en) | Method for solidifying loose coral sand by using microorganisms | |
| CN112322519B (en) | Microbial composite flora for biomineralization and preparation and application thereof | |
| CN110685266A (en) | Method for improving mechanical property of calcareous sand by solidifying fiber reinforced microorganisms | |
| Canakci et al. | Bacterail calcium carbonate precipitation in peat | |
| CN111827258A (en) | Method for reinforcing soil body by combining organic matters with plant urease | |
| CN103223278A (en) | Biocementation of particulate material in suspension | |
| DONG et al. | Evaluation of the effect of natural seawater solidifying calcareous sand based on MICP | |
| Chen et al. | Evaluating Mechanical Strength of Peat Soil Treated by Fiber Incorporated Bio-Cementation. | |
| CN103789242B (en) | Iron oxygen bacterium and utilize the method for its improvement Liquefaction of Light Loam characteristic | |
| XIAO et al. | Study on domestication of Sporosarcina pasteurii and cementation effect of calcareous sand in seawater environment | |
| Erdmann et al. | Influencing factors on ureolytic microbiologically induced calcium carbonate precipitation for biocementation | |
| CN117230777A (en) | Method for solidifying granite residual soil | |
| CN114894578A (en) | Method for preparing microbial mortar by using natural seawater and sea sand | |
| Li et al. | A genetically engineered composite biofilm for microbial induced calcium carbonate precipitation by synergic effect of urease, protein adhesive and xanthan gum | |
| Gao et al. | Staphylococcus derived from desert and its solidified ability on aeolian sandy soil | |
| CN112723958A (en) | Ecological fishpond organic fertilizer and preparation method thereof | |
| Shannoon et al. | Bio-Cementation of Sandy Soil through Bacterial Processing to Precipitate Carbonate | |
| CN116813226B (en) | Fly ash-bacteria complex biological curing agent and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |