Evaluation of Shear Strength Parameters of Sandy Soils upon Microbial Treatment

A new method for soil stabilization known as microbial-induced calcite precipitation (MICP) has been the focus of research in this area during the last decade. In this method, the reaction of microorganisms in the presence of urea and calcium chloride is used to produce calcite. Despite the large numbers of bacteria in soil, Sporobacillus pasteurii (previously known as Bacillus pasteurii) has the most capability to create cementation between soil particles in the MICP method due to its high urease activity. In this paper, the effect of MICP treatment on the shear strength characteristics of a sandy soil was studied. The change in the shear strength of sandy soil upon MICP treatment was measured using a strain-controlled direct shear test before and after treatment of the soil samples. The results showed an increase of 44–86% in the shear strength of the sandy soil after 15 days of MICP treatment compared to the untreated soil. The enhanced shear strength was the result of an increase in both the cohesion intercept and angle of internal friction. The increase in the cohesion intercept was more significant than the increase in the angle of internal friction.     

Biocalcification using B. pasteurii for strengthening brick masonry civil engineering structures

Microbiologically induced calcite precipitation in bricks by bacterium Bacillus pasteurii (NCIM 2477) using a media especially optimized for urease production (OptU) was demonstrated in this study. Effect of biocalcification activity on compressive strength and water absorption capacity of bricks was investigated. Various other parameters such as pH, growth profile, urease activity, urea breakdown and calcite precipitated were monitored during the 28 days curing period. Efficiency of B. pasteurii to form microbial aided calcite precipitate in OptU media resulted into 83.9 % increase in strength of the bricks as compared to only 24.9 % with standard media, nutrient broth (NB). In addition to significant increase in the compressive strength, bricks treated with B. pasteurii grown in OptU media resulted in 48.9 % reduction in water absorption capacity as compared to control bricks immersed in tap water. Thus it was successfully demonstrated that microbial calcification in optimized media by Bacillus pasteurii has good potential for commercial application to improve the life span of structures constructed with bricks, particularly structures of heritage importance.     

State-of-the-Art Review of Biocementation by Microbially Induced Calcite Precipitation (MICP) for Soil Stabilization

Biocementation is a recently developed new branch in geotechnical engineering that deals with the application of microbiological activity to improve the engineering properties of soils. One of the most commonly adopted processes to achieve soil biocementation is through microbially induced calcite precipitation (MICP). This technique utilizes the metabolic pathways of bacteria to form calcite (CaCO₃) that binds the soil particles together, leading to increased soil strength and stiffness. This paper presents a review of the use of MICP for soil improvement and discusses the treatment process including the primary components involved and major affecting factors. Envisioned applications, potential advantages and limitations of MICP for soil improvement are also presented and discussed. Finally, the primary challenges that lay ahead for the future research (i.e. treatment optimization, upscaling for in situ implementation and self-healing of biotreated soils) are briefly discussed.     

Stimulation of Native Microorganisms for Improving Loose Salty Sand

Prolonged droughts and excessive water harvesting in western Asia has accelerated desertification and caused longer dry seasons of salt lakes. The Aral Sea experience has proven that dust from saline soil is a serious health issue. Various stabilization techniques to reduce wind erosion have been used in the past. However, in recent years, a potentially viable method has been developed; microbial induced calcite precipitation (MICP) has been introduced as a method of soil stabilization, though its effectiveness in saline soils remains to be examined. The effect of salt content in loose sandy soil on calcite precipitation of calcite through stimulation of native bacteria is investigated in this article. Samples with salinity up to 30% salt content were prepared and treated with different culture medium compounds. A number of tests were used to evaluate the effect of the mentioned parameters. The results show that improvement increases with increasing salinity up to 5% salt, and further increase in salinity reduces the effectiveness of improvement. It is also shown that the addition of urea in the culture medium has a significant effect on the urea hydrolysis which resulted in a five-fold increase in compressive strength. Four native strains of halotolerant urease-positive bacteria were also identified.     

Reducing Soil Permeability Using Microbial Induced Carbonate Precipitation (MICP) Method: A Case Study of Shiraz Landfill Soil

Abstract

Improvement of engineering properties of soils to meet project requirements has long been subject of interest to civil engineers. One of the environment-friendly methods that have recently been used for this purpose is the biological method. These methods that actually benefit from various sciences such as biology, biochemistry, and civil engineering, use biological products or organisms such as bacteria that are commonly found in soils. In this study, the reduction of permeability or hydraulic conductivity of Shiraz landfill base soil using microbial induced calcite precipitation (MICP) has been explored. B. sphaericus was used to treat the soil. Falling head permeability tests are conducted to measure soil samples’ permeability before and after biological treatment. The target variables were the curing time, bacterial density, optimal nutrient content, and soil unit weight. The test results demonstrated that the permeability of the samples treated with Bacillus sphaericus decreases by increasing curing time, the density of calcium chloride solution and bacterial density of samples. This study showed that the MICP can be utilized as a new environment-friendly method for reducing the soil permeability at the base and walls of the landfill to form a barrier between the waste and the groundwater and substrata.     

Remediation of copper-contaminated soil by Kocuria flava CR1, based on microbially induced calcite precipitation

An indigenous calcifying bacterial strain CR1, identified as Kocuria flava, was isolated from soil of a mining area, Urumqi, China. An extensive copper bioremediation capacity of this isolate was studied based on microbially induced calcite precipitation (MICP). K. flava CR1 removed 97% of copper when initial Cu concentration was 1000 mg L⁻¹. The isolate produced significant amount of urease (472 U mL⁻¹), an enzyme that leads to calcite precipitation. The isolate removed 95% of copper from contaminated soil. The MICP process in bioremediation was further confirmed by FTIR and XRD analyses. FTIR analysis showed two different forms of calcium carbonate, i.e., calcite and aragonite, and the results were well supported by XRD. For the first time, the ability of K. flava has been documented in the bioremediation of polluted soil. This study showed that MICP-based bioremediation by K. flava is a viable, environmental friendly technology for cleaning-up the copper-contaminated site.     

Assessment of the Parameters Influencing Microbial Calcite Precipitation in Injection Experiments Using Taguchi Methodology

Soil improvement is one of the major concerns in civil engineering. Therefore, a variety of approaches have been employed for different soil types. The loose granular soils and sediments have always imposed challenges due to their low strength and bearing capacity as well as presenting difficulties in drilling and excavation. Biomediated soil improvement, i.e., utilizing some bacteria to precipitate calcite on soil particles, has recently been introduced as a novel link of biotechnology and civil engineering to improve the problematic soils. Biogrout as a branch of biomediated soil improvement is based upon microbial calcium carbonate precipitation (MICP). In the present study, the Taguchi method with the aim of optimizing the process was utilized to design the experiments (DOE). A standard L9 orthogonal array with four parameters comprising bacterial cell concentration, molar concentration ratio of nutrient solution, curing time, and flow rate, each assigned to three levels, was selected. In this regard, soil samples were stabilized in sandy soil columns. Two-phase injections were conducted by injecting the bacterium Sporosarcina pasteurii PTCC 1642 in the first phase and nutrient in the second phase. Specimens were subjected to an unconfined compressive strength (UCS) test. ANOVA pointed out how effectual each parameter was. The most effective parameter was curing time, which accounted for 45.97% of the overall variance of the experimental data followed by bacterial cell concentration (22.01%), nutrient strength (19.98%), and flow rate (12.04%). Predicted UCS values for the optimum condition were validated in a confirmation test. Indeed, the UCS of the soil increased from 85 kPa in the control sample to 930 kPa for the optimally treated specimen. It was concluded that rather than curing time, the other parameters are almost equally influential in the applied injection procedure.     

Immobilization of cadmium in soil by microbially induced carbonate precipitation with Exiguobacterium undae at low temperature

Microbially induced carbonate precipitation (MICP) is an advanced biological treatment technology to immobilize heavy metals in form of carbonate salts. In this MICP study, ureolytic Exiguobacterium undae was employed for immobilization of cadmium in contaminated soil at low temperature (10 °C). The sequential extraction test revealed conversion of more than 90% of cadmium in the tested soil from the soluble-exchangeable fraction to carbonate-bound fraction in 14 days of treatment. The cadmium may be precipitated in a separate CdCO₃ phase or be co-precipitated in calcite crystals. Activities of urease and dehydrogenase were enhanced during MICP, which were not affected by the testing temperatures. MICP with E. undae is a biological approach that may be worth investigating further to immobilize cadmium in soils of cold regimes.     

产脲酶芽胞杆菌的微波诱变育种

巴氏芽胞杆菌是目前微生物诱导碳酸钙沉淀(MICP)方法中应用最为热门的一种细菌。为提高巴氏芽胞杆菌尿素分解以及矿化能力,以巴氏芽胞杆菌YB-B为出发菌株,采用微波诱变育种技术,通过诱变菌株特性筛选及其遗传稳定性检测,成功选育出2株突变菌株YB-3和YB-4。与出发菌株相比,诱变菌株尿素分解能力较原菌株提高1.5倍左右,矿化能力提高114%。诱变菌株具有生长速度快,环境适应性强,矿化能力高等优点,这为MICP更广层次的应用奠定了坚实的基础。     

Strain improvement of <Emphasis Type="Italic">Sporosarcina pasteurii</Emphasis> for enhanced urease and calcite production

Phenotypic mutants of Sporosarcina pasteurii (previously known as Bacillus pasteurii) (MTCC 1761) were developed by UV irradiation to test their ability to enhance urease activity and calcite production. Among the mutants, Bp M-3 was found to be more efficient compared to other mutants and wild-type strain. It produced the highest urease activity and calcite production compared to other isolates. The production of extracellular polymeric substances and biofilm was also higher in this mutant than other isolates. Microbial sand plugging results showed the highest calcite precipitation by Bp M-3 mutant. Scanning electron micrography, energy-dispersive X-ray and X-ray diffraction analyses evidenced the direct involvement of bacteria in CaCO₃ precipitation. This study suggests that calcite production by the mutant through biomineralization processes is highly effective and may provide a useful strategy as a sealing agent for filling the gaps or cracks and fissures in any construction structures.     

Efficacy of Fe3O4/Starch Nanoparticles on Sporosarcina pasteurii Performance in MICP Process

The microbial induced calcite precipitation (MICP) has been explored using well-known urease producer bacterium Sporosarcina pasteurii for many applications including soil stabilization. Urease enzyme hydrolyzes urea and in the presence of calcium chloride causes calcium carbonate precipitation between sand particles increasing sand stiffness and strength. In this study, the liquefied soil samples from Anzali coast were positioned inside injection columns by standard positioning technique. The columns were treated by injecting S. pasteurii suspension and cementation solution (CaCl₂ and urea). The effect of different conditions consisting of number of injections, injection intervals, flow rate, and ratio of injection solution on unconfined compression strength (USC) of sands formed inside the columns were evaluated. The results indicated that soil strength was increased when ratio of reactant solutions and injection time were elevated. Moreover, the maximum Ca-precipitation in MICP reaction in liquid medium was obtained while Fe₃O₄/starch concentration and time of addition of nanoparticle to culture medium were 10.8?mg/L and 1.4?h, respectively. The USC results showed that the columns injected by bacterial suspension treated by Fe₃O₄/starch under optimized conditions improved the soil strength up to 1200?kPa in comparison to the control column as 220?kPa.     

生物炭添加对不同水氮条件下芦苇生长和氮素吸收的影响

生物炭能改良土壤从而促进植物生长和氮素吸收，但其作用效果是否受水氮条件的影响尚不清楚。以湿地植物芦苇为研究对象，在3种氮添加水平（无添加，30 kg hm^-2 a^-1和60 kg hm^-2 a^-1）和两种水分（淹水和非淹水）条件下分别进行生物炭添加和不添加处理，结果表明：（1）生物炭添加能促进芦苇根系生长，在非淹水条件下根系生物量增加了40.5%，在淹水条件下根系生物量增加了20.1%。（2）生物炭添加能促进非淹水条件下芦苇的氮素吸收，能提高淹水条件下芦苇的氮素生产力。（3）生物炭添加加剧了土壤氮素损失，且在非淹水高氮条件下作用最强，可能是由于生物炭促进了芦苇的氮素吸收。芦苇氮素吸收速率与土壤氮损失之间存在显著的正相关关系。因此，在添加生物炭时，需要考虑土壤水分状况和氮素富集程度以及植物的氮素吸收偏好。该研究结果可为生物炭在湿地生态系统中的应用提供参考。     

Impact of Oxygen Availability on Microbially Induced Calcite Precipitation (MICP) Treatment

Most microbially induced calcite precipitation (MICP) processes are induced by aerobic bacteria; thus, oxygen availability plays an important role in MICP treatment. To determine the effects of oxygen supply on MICP treatment catalyzed by Sporosarcina pasteurii, contrast tests under an aerated condition, air-restricted condition, and open air condition were conducted. The results showed that dissolved oxygen (DO) in the air-restricted reactor decreased with time and was almost exhausted within 7 days; DO in the open box decreased by 50% after 7 days of treatment because of the superficial air supply; and DO in the aerated box maintained an initial high level because the consumed oxygen was supplied immediately by adequate air bubbles in the treatment solution. Unconfined compressive strength (UCS) and CaCO₃ content are high under the aerated condition, moderate under the open condition, and poor under the air-restricted condition. The UCS can be 100 times different depending on the different oxygen supply conditions. The overall influence process is as follows: oxygen is dissolved to supply DO for life and activity of the aerobic urea hydrolysis bacteria; then, urea is hydrolyzed to carbonate anions for CaCO₃ precipitation in the presence of Ca²⁺; and finally, CaCO₃ precipitation results in the strengthening of sand. The results indicate that a sufficient air supply is essential to improve MICP processes catalyzed by aerobic bacteria.     

Microbially Induced Calcite Precipitation for Seepage Control in Sandy Soil

Microbially induced calcite precipitation (MICP) can reduce the permeability of soil by reducing the pore volumes. A MICP-based soil improvement method to control water leakage in irrigation channels and reservoirs built on sandy soil grounds is presented in this article. Using this method, a low-permeable hard crust can be formed at the soil surfaces. An experimental study was carried out to evaluate the effect of this method. Sandy soil samples were treated using four different schemes, namely, (1) surface spray, (2) surface spray with the addition of fibers, (3) surface spray and bulk stabilization, and (4) immersion stabilization. By applying around 2.6?L treatment liquid (consisting of ureolytic bacteria, 0.5?mol/L calcium chloride and 0.5?mol/L urea) to the top 2-cm thick soil, the seepage rates of the samples treated by the four different schemes could be reduced by up to 379 times. The conversion rates of calcium source in the tests were up to 89.7%. The results showed that a method of treating the soil in bulk before the formation of a crust on top of the soil layer was effective in reducing the seepage rates. After the bio-treatment, the formed low-permeable hard crust layer was 10 to 20?mm thick with a calcite content higher than 5%. Below the hard crusts, the calcite content was less than 5% and the soil was not properly cemented. Using the mercury intrusion test, it was found that both pore volumes and pore sizes of the bio-treated soil reduced significantly as compared with the untreated soil. Penetration tests using a flat-bottom penetrometer were used to assess the mechanical behavior of the bio-treated soil. The results indicated that the penetration resistance of the bio-treated soil layer was much higher than that of the untreated soil.     

Comparison of conditions for mycelial growth of <Emphasis Type="Italic">Lepista sordida</Emphasis> causing fairy rings on <Emphasis Type="Italic">Zoysia matrella</Emphasis> turf to those on <Emphasis Type="Italic">Agrostis palustris</Emphasis> turf

Symptoms of fairy rings caused by Lepista sordida have been reported on Zoysiagrass (Zoysia spp.) turf maintained at fairway height (2 cm), but not on bentgrass (Agrostis spp.) maintained at putting green height (0.5 cm). The mycelia of this fungus inhabit primarily the upper 0–2 cm layer of the soil extending into the thatch. To compare conditions for the mycelial growth in Z. matrella turf to those in A. palustris turf, we examined the effects of nutrients, temperature, water potential, and pH in the field as well as in the laboratory. Greater growth of the mycelia was observed in medium that included hot water extracts from soil of the 0–1 cm zone in Z. matrella turf compared to that from A. palustris. The upper soil layer in Z. matrella turf contained more organic matter from clippings than that in A. palustris. The temperature and water potential of the 0–2 cm soil zone in Z. matrella turf were also more favorable for the mycelial growth. The soil pH values of this zone in Z. matrella turf were less favorable compared to A. palustris but within the range for accelerating mycelial growth. Part of this study was presented orally at the 46th meeting of the Mycological Society of Japan in 2002     

Study on Strength and Leaching Behavior of Biogeochemical Cemented Sand

Abstract

Conventional ground improvement techniques involve densification of soil either by mechanical compaction or chemical grouting while others involve inclusion of reinforcements, etc. Many conventional grout materials were found posing a threat to environment due to their toxic nature and release of greenhouse gases. In this regard, research is initiated in developing more environmentally sustainable additives for soil improvement in which biological based alternatives are gaining momentum. In the present study, a noble technique Microbial Induced Calcite Precipitation (MICP) using Sporosarcina pasteurii was adopted to modify the properties of sand and improve its efficiency by supplementing with Cellulomonas flavigena. The mineralogy of treated specimens was studied using X-Ray Diffraction and Scanning Electron Microscope analyses. The leachability of precipitated calcite was studied under constant flowing conditions and the material was found to be stable. The Unconfined Compressive Strength (UCS) and elastic modulii of the treated specimens were found to be in the range of 266–343?kPa and 14–35?MPa respectively. The angle of internal friction found from Direct Shear Test on treated specimens was observed to be a little lower compared to virgin specimens. The permeability of treated specimen showed a reduction in magnitude by one order approximately.     

Sugarcane Molasses: A Cheap Carbon Source for Calcite Production in Different Class of Soils using Stimulation of Indigenous Urease-producing Bacteria

Abstract

In this research, we investigated the abilities of three different concentration of sugarcane molasses as a carbon source to stimulate indigenous bacterial growth in different classes of soil, namely poorly graded sand (SP), silty sand (SM), and clayey sand (SC) (according to the Unified classification system). A total of 7, 10, and 15 days after the treatment, direct shear tests were performed on the untreated and treated samples. The calcite content on all direct shear samples was determined to further correlate it with the strength gains in the treated samples. The scanning electron microscopy (SEM) images, EDX analysis, and X-ray diffraction (XRD) patterns were taken before and after treatment for all samples to analyze the microbial-induced calcite precipitation (MICP) process. The SP soil samples showed the highest strength gains and also highest calcite content as compared with other two soil type. The peak cohesion intercept for SP-treated samples increased by 2.7–5.5 times as compared to the untreated samples for molasses concentration of 1–3?g/L, respectively. The treated samples became more dilative with the increase in molasses concentration. The sample with highest molasses concentration showed stiffer behavior in shear than the samples with lower concentration.     

Carbonate Mineral Precipitation for Soil Improvement Through Microbial Denitrification

Microbially induced carbonate precipitation (MICP) and associated biogas production may provide sustainable means of mitigating a number of geotechnical challenges associated with granular soils. MICP can induce interparticle soil cementation, mineral precipitation in soil pore space and/or biogas production to address geotechnical problems such as slope instability, soil erosion and scour, seepage of levees and cutoff walls, low bearing capacity of shallow foundations, and earthquake-induced liquefaction and settlement. Microbial denitrification has potential for improving the mechanical and hydraulic properties of soils because it promotes precipitation of calcium carbonate (CaCO₃) and produces nitrogen (N₂) gas without generating toxic by-products. We evaluated the potential for inducing carbonate precipitation in soil via bacterial denitrification using bench-scale experiments with the facultative anaerobe Pseudomonas denitrificans. Bench-scale experiments were conducted (1) without calcium in an N-rich bacterial growth medium in 2.0 L glass batch reactors and (2) with a source of calcium in sand-filled acrylic columns. Changes of pH, alkalinity, NO₃^? and NO₂^? in the batch reactors and columns, quantification of biogas production and observations of calcium-carbonate precipitation in the sand-filled columns indicate that denitrification led to carbonate precipitation and particle cementation in the pore water as well as a substantial amount of biogas production in both systems. These results document that bacterial denitrification has potential as a soil improvement mechanism.     

Effects of environmental factors on microbial induced calcium carbonate precipitation

Aims: To gain an understanding of the environmental factors that affect the growth of the bacterium Sporosarcina pasteurii, the metabolism of the bacterium and the calcium carbonate precipitation induced by this bacterium to optimally implement the biological treatment process, microbial induced calcium carbonate precipitation (MICP), in situ. Methods and Results: Soil column and batch tests were used to assess the effect of likely subsurface environmental factors on the MICP treatment process. Microbial growth and mineral precipitation were evaluated in freshwater and seawater. Environmental conditions that may influence the ureolytic activity of the bacteria, such as ammonium concentration and oxygen availability, as well as the ureolytic activities of viable and lysed cells were assessed. Treatment formulation and injection rate, as well as soil particle characteristics are other factors that were evaluated for impact on uniform induction of cementation within the soils. Conclusions: The results of the study presented herein indicate that the biological treatment process is equally robust over a wide range of soil types, concentrations of ammonium chloride and salinities ranging from distilled water to full seawater; on the time scale of an hour, it is not diminished by the absence of oxygen or lysis of cells containing the urease enzyme. Significance and Impact of Study: This study advances the biological treatment process MICP towards field implementation by addressing key environmental hurdles faced with during the upscaling process.     

Lactose mother liquor as an alternative nutrient source for microbial concrete production by <Emphasis Type="Italic">Sporosarcina pasteurii</Emphasis>

Microbiologically induced calcite precipitation by the bacterium Sporosarcina pasteurii (NCIM 2477) using the industrial effluent of the dairy industry, lactose mother liquor (LML) as growth medium was demonstrated for the first time in this study. The urease activity and the calcite precipitation by the bacterium was tested in LML and compared with the standard media like nutrient media and yeast extract media. Calcite constituted 24.0% of the total weight of the sand samples plugged by S. pasteurii and urease production was found to be 353 U/ml in LML medium. The compressive strength of cement mortar was increased by S. pasteurii in all the media used compared to control. No significant difference in the growth, urease production and compressive strength of mortar among the media suggesting LML as an alternative source for standard media. This study demonstrates that microbial calcite acts as a sealing agent for filling the gaps or cracks and fissures in constructed facilities and natural formations alike.