Name: sandy soil improvement through microbially induced calcite precipitation (micp) by immersion
Uploaded: 2019-09-12T14:30:00
Description: Watch this Scientific Journal Video about Sandy Soil Improvement through Microbially Induced Calcite Precipitation MICP by Immersion at JoVE.com

这项工作得到了国家科学基金会第1531382号赠款和MarTREC的支持。

注：以下程序中使用的所有相关材料均无危险。个人防护设备（安全眼镜、手套、实验室外套、全长裤子、闭趾鞋）仍需要。
1. 细菌溶液的制备
<ol><li>培养介质的准备（NH4-YE介质） 注：每升去离子水生长培养基的成分为：20克酵母提取物;10 g 的 （NH4）2SO4;和 0.13 M Tris 缓冲液 （pH 9.0）。<ol>
<li>高压灭菌成分分开。</li>
		<li>溶解20克酵母提取...

<ol>
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T., Brown, D. G., Kavazanjian, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+behavior+of+sands+treated+by+microbially+induced+carbonate+precipitation.">Mechanical behavior of sands treated by microbially induced carbonate precipitation.</a> Journal of Geotechnical and Geoenvironmental Engineering. 142 (2), 04015066 (2015).</li><li>Lauchnor, E. G., Topp, D. M., Parker, A. E., Gerlach, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole+cell+kinetics+of+ureolysis+by+sporosarcina+pasteurii.">Whole cell kinetics of ureolysis by sporosarcina pasteurii.</a> Journal of Applied Microbiology. 118 (6), 1321-1332 (2015).</li><li>Nafisi, A., Montoya, B. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+framework+for+identifying+cementation+level+of+MICP-treated+sands.">A new framework for identifying cementation level of MICP-treated sands.</a> IFCEE. , (2018).</li><li>Zhao, Q., Li, L., Li, C., Zhang, H., Amini, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+full+contact+flexible+mold+for+preparing+samples+based+on+microbial-induced+calcite+precipitation+technology.">A full contact flexible mold for preparing samples based on microbial-induced calcite precipitation technology.</a> Geotechnical Testing Journal. 37 (5), 917-921 (2014).</li><li>Bu, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+Rigid+Full-Contact+Mold+for+Preparing+Biobeams+through+Microbial-Induced+Calcite+Precipitation.">Development of a Rigid Full-Contact Mold for Preparing Biobeams through Microbial-Induced Calcite Precipitation.</a> Geotechnical Testing Journal. 42 (3), 656-669 (2018).</li><li>Li, M., Wen, K., Li, Y., Zhu, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+oxygen+availability+on+microbially+induced+calcite+precipitation+(MICP)+treatment.">Impact of oxygen availability on microbially induced calcite precipitation (MICP) treatment.</a> Geomicrobiology Journal. 35 (1), 15-22 (2018).</li><li>Martinez, B. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+optimization+of+microbial-induced+carbonate+precipitation+for+soil+improvement.">Experimental optimization of microbial-induced carbonate precipitation for soil improvement.</a> Journal of Geotechnical and Geoenvironmental Engineering. 139 (4), 587-598 (2013).</li></ol>

本文提出了浸没的MICP技术。土壤样品被浸入批次反应器中，在MICP过程中被凝固介质完全渗透。在该方法中，应用全接触柔性模具、刚性全接触模具和芯砖模具制备MICP处理样品。
可针对不同的几何形状设计不同的模具。土工布纤维结构增加了沙与固结介质的接触面积，有效地增加了固结介质对土壤样品的渗透。大量的土工毛孔还使模具内部发生更多的降水，以提高MICP处理样?...

微生物诱导方解石沉淀（MICP）作为一种生物地面改良技术，能够提高土壤的工程性能。它用于增强土壤的强度、刚度和渗透性。MICP技术在土壤改良方面得到了全世界1、2、3、4的高度重视。碳酸盐降水自然发生，并且可能由土壤环境原生的非致病生物诱发。MICP生物地球化学反应是由尿素细菌、尿素和富含钙的溶液5、6的驱动。孢子菌糊化是一种高度活跃的尿酶，能催化方解石7、8的沉淀反应网络。尿素水解工艺产生溶解铵（NH4+）和无机碳酸盐（CO32-）。碳酸盐离子与钙离子发生反应，作为碳酸钙晶体沉淀。尿素水解反应如下所示：
<img alt="Equation 1" src="/files/ftp_upload/60059/60059equ01.jpg">
<img alt="Equation 2" src="/files/ftp_upload/60059/60059equ02.jpg">
沉淀的CaCO3可以将沙粒粘合在一起，提高MICP处理土壤的工程性能。MICP技术已应用于各种应用，如提高土壤的强度和刚度，修复混凝土，以及环境修复9，10，11，12，13,14,15.
赵等人16日研制出一种浸没方法制备MICP处理的样品。该方法采用土工纺织品制成的全接触柔性模具。沉淀的CaCO3均匀地分布在其MICP处理的样品中。Bu等人17开发出一种刚性全接触模具，通过浸入法制备MICP处理的光束样品。使用刚性全接触模具制备的 MICP 处理样品可形成合适的梁形。将MICP处理的样品分为4个，并测量CaCO3含量。按重量计，CaCO3含量从 8.4 ± 1.5% 到 9.4 ± 1.2% 不等，这表明 CaCO3通过浸没方法均匀分布于 MICP 处理的样品中。这些MICP处理样品也实现了更好的机械性能。这些MICP处理的生物标本达到950 kPa的弯曲强度，类似于20-25%的水泥处理样品（600-1300 kPa）。Li等人10将随机分布的离散纤维加入沙土中，采用MICP浸入法对土壤进行处理。他们发现，通过添加适当的纤维，可明显增强MICP处理土壤的剪切强度、延展性和失效应变。
MICP的浸入法已经不断改进10，16，17。该方法可用于制备 MICP 处理的土壤样品和 MICP 处理的预制建筑材料，如砖块和横梁。开发了样品制备模具的不同几何尺寸。在MICP处理的样品中加入纤维，以提高其性能。此详细协议旨在记录 MICP 治疗的浸入式方法。

<table>
	<tbody>
		<tr>
			<td>Ammonium Chloride, &#62;99%</td>
			<td>Bio-world</td>
			<td>40100196-3 (705033)</td>
			<td></td>
		</tr>
		<tr>
			<td>Ammonium Sulfate</td>
			<td>Bio-world</td>
			<td>30635330-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Calcium Chloride Dihydrate, &#62;99%</td>
			<td>Bio-world</td>
			<td>40300016-3 (705111)</td>
			<td></td>
		</tr>
		<tr>
			<td>Nutrient Broth</td>
			<td>Bio-world</td>
			<td>30620056-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Bicarbonate, &#62;99%</td>
			<td>Bio-world</td>
			<td>41900068-3 (705727)</td>
			<td></td>
		</tr>
		<tr>
			<td>Sporosarcina pasteurii</td>
			<td>American Type Culture Collection</td>
			<td>ATCC 11859</td>
			<td></td>
		</tr>
		<tr>
			<td>Synthetic fiber</td>
			<td>FIBERMESH</td>
			<td>Fibermesh 150e3</td>
			<td></td>
		</tr>
		<tr>
			<td>Tris-Base, Biotechnology Grade, &#62;99.7%</td>
			<td>Bio-world</td>
			<td>42020309-2 (730205)</td>
			<td></td>
		</tr>
		<tr>
			<td>Urea, USP Grade, &#62;99%</td>
			<td>Bio-world</td>
			<td>42100008-2 (705986)</td>
			<td></td>
		</tr>
		<tr>
			<td>Yeast Extract</td>
			<td>Bio-world</td>
			<td>30620096-3 (760095)</td>
			<td></td>
		</tr>
	</tbody>
</table>

sandy soil improvement through microbially induced calcite precipitation (micp) by immersion

本文旨在开发一种浸入法，以改善微生物诱导方解石沉淀（MICP）处理样品。组装了一批反应器，将土壤样品浸入水泥介质中。凝固介质可以自由扩散到批次反应器中的土壤样品中，而不是注入水泥介质。使用全接触柔性模具、刚性全接触模具和芯砖模具来制备不同的土壤样品支架。选择合成纤维和天然纤维来强化MICP处理的土壤样品。测量了MICP处理样品不同区域的沉淀CaCO3。CaCO3分布结果表明，沉淀的CaCO3采用浸入法在土壤样品中均匀分布。

图7显示了沉淀的CaCO3在整个MICP处理样品中的分布情况。MICP处理的样本分为三个不同的区域。用酸洗法对每个区域的CaCO3含量进行了测试。为了溶解沉淀的碳酸盐，在HCl溶液（0.1 M）中清洗干MICP处理的样品，然后冲洗、排空和烘干48小时。酸洗前后样品质量的差异值被认为是MICP处理样品中沉淀的碳酸盐的质量。CaCO3内容以样本重量的百分比表示。浸没?...

Watch this Scientific Journal Video about Sandy Soil Improvement through Microbially Induced Calcite Precipitation MICP by Immersion at JoVE.com

Sandy Soil Improvement through Microbially Induced Calcite Precipitation MICP by Immersion

这里介绍了微生物诱导的方解石沉淀（MICP）技术，通过浸泡改善土壤性能。

通过浸没通过微生物诱导的钙化物沉淀（MICP）改善沙土

The goal of this article is to develop an immersion method to improve the microbially induced calcite precipitation (MICP) treated samples. A batch ...

该协议中演示的浸入方法可帮助研究人员通过不同的模具去除未经中断的 MICP 处理土壤样品进行机械测试。该技术有助于 MICP 处理土壤样本获得更均匀的碳酸钙沉淀。这些 MICP 处理的土壤样品也通过这种方法获得更好的机械性能。这项技术最重要的方面是确保细菌的活性足以使MICP过程避免耗时和材料浪费。对于首次选择这种技术的研究人员来说，为不同的应用设计合适的模具被认为是另一个重要方面。首先，将0.1毫升的细菌悬浮液转移到充满10毫升新鲜生长介质的离心管中。通过反转混合，然后松开管盖，以保持有氧条件。准备六管细菌悬浮液与生长介质和一个控制管与10毫升的新鲜生长介质单独。在200 RPM和30摄氏度的摇床中孵育所有管子48至72小时。如果生长介质在48小时后变得浑浊，则停止孵育。用细菌和生长介质将管子以4000倍g离心20分钟。取出上流水线，用25毫升的新鲜生长介质代替，使用涡流机混合。然后，为了增强细菌的培养，将0.25毫升悬浮液从管子转移到新管中，用25毫升的生长培养中培养接种。在200 RPM和30摄氏度的摇床中孵育所有管子48小时。用细菌和生长介质将管子以4000倍g离心20分钟。拆下上流水器，用新鲜生长介质代替，并使用涡流机很好地混合。在分光光度计上测量悬浮液在 600 纳米的光学密度。使用新鲜生长介质调整细菌浓度，使其消耗性为 0.6 的 OD600。通过空气吹水法将约 140 克干渥太华砂与 99.7% 的绳索加入模具中，以达到中位密度，相对密度在 42 至 55% 之间，砂干密度在 1.58 至 1.64 克/立方厘米范围内。在样品上盖上盖子，然后缝制。将细菌溶液通过可渗透的土工用膜倒入样品中。当在土壤样品底部观察到流动的细菌溶液时，它们被认为是饱和的。接下来，将样品放在架子上。将整个架子浸入一个装满凝固介质的批次反应器中。打开空气供应并调整空气输出，以保持 100% 空气饱和度。等待七天的 MICP 反应。从反应器中拿出样品。使用刀切割全接触柔性模具。然后，用水清洗样品，去除孔隙空间中的残留溶液。将样品放入105摄氏度的烤箱中48小时，直到其重量保持不变。对于合成纤维，用手以小增量混合2.7克纤维和900克干砂，以获得纤维含量为0.3%的均匀混合物。对于天然棕榈纤维，准备四部分沙子，每个部分190克。准备三层纤维，每层约1.8克。每隔一段时间在刚性全接触模具中加入砂的四个部分和三层纤维。接下来，重复MICP治疗，如以前一样。在此协议中，在整个 MICP 处理的样品中观察到沉淀的碳酸钙。MICP 处理的样品分为三个不同的区域。通过浸入法处理的MICP处理样品的碳酸钙含量在9.0至9.5%之间，表明沉淀的碳酸钙在整个土壤样品中均匀分布。用三层棕榈纤维和未强化生物砖加固的生物砖应力应变曲线显示，未强化生物砖的柔性强度为1，150千帕，而增强生物砖的弯曲强度为980千帕。通过添加棕榈纤维，弯曲应变显著改善。在 MICP 过程之前确保细菌溶液的活性非常重要。如果没有，那么在实验中能否成功还不确定。一旦实验失败，这应该是浪费时间和材料。按照此过程，可以通过 MICP 技术为不同的实验目的设计不同的模具。这些模具可以模拟MICP对不同结构或不同材料的影响。

Research

JoVE Journal

Engineering

Sandy Soil Improvement through Microbially Induced Calcite Precipitation (MICP) by Immersion

Investigation of Early Plasma Evolution Induced by Ultrashort Laser Pulses

超短激光脉冲诱导的等离子体演化早期的调查

Early plasma is generated owing to high intensity laser irradiation of target and the subsequent target material ionization. Its dynamics plays a ...

科研

工程学

Real-Time DC-dynamic Biasing Method for Switching Time Improvement in Severely Underdamped Fringing-field Electrostatic MEMS Actuators

实时直流偏置的动态法切换时间改善严重欠阻尼边缘化场静电MEMS致动器

Mechanically underdamped electrostatic fringing-field MEMS actuators are well known for their fast switching operation in response to a unit step input ...

Visible-light Induced Reduction of Graphene Oxide Using Plasmonic Nanoparticle

可见光诱导的还原氧化石墨烯的使用电浆纳米粒子

Present work demonstrates the simple, chemical free, fast, and energy efficient method to produce reduced graphene oxide (r-GO) solution at RT using ...

Visualizing Hyporheic Flow Through Bedforms Using Dye Experiments and Simulation

可视化潜流流通过底形用染料试验及仿真

Advective exchange between the pore space of sediments and the overlying water column, called hyporheic exchange in fluvial environments, drives solute ...

Automation of Mode Locking in a Nonlinear Polarization Rotation Fiber Laser through Output Polarization Measurements

锁模自动化的非线性偏振旋转光纤激光器通过输出偏振测量

When a laser is mode-locked, it emits a train of ultra-short pulses at a repetition rate determined by the laser cavity length. This article outlines a ...

Laser-induced Forward Transfer of Ag Nanopaste

银纳米糊膏的激光诱导正向传输

Over the past decade, there has been much development of non-lithographic methods1-3 for printing metallic inks or other functional materials. Many of ...

Magnetically Induced Rotating Rayleigh-Taylor Instability

磁感应旋转瑞利 - 泰勒不稳定性

Classical techniques for investigating the Rayleigh-Taylor instability include using compressed gasses1, rocketry2 or linear electric motors3 to reverse ...

Preparation of Liquid Crystal Networks for Macroscopic Oscillatory Motion Induced by Light

光诱导的宏观振荡运动液晶网络的制备

A strategy based on doped liquid crystalline networks is described to create mechanical self-sustained oscillations of plastic films under continuous ...

Measurements of Soil Carbon by Neutron-Gamma Analysis in Static and Scanning Modes

静态和扫描模式下中子伽马分析法测量土壤碳

The herein described application of the inelastic neutron scattering (INS) method for soil carbon analysis is based on the registration and analysis of ...

Quantitative Analysis of Vacuum Induction Melting by Laser-induced Breakdown Spectroscopy

激光击穿光谱对真空感应熔融的定量分析

Vacuum induction melting is a popular method for refining high purity metal and alloys. Traditionally, standard process control in metallurgy involves ...