Alterations in Membrane Phospholipid Fatty Acids of Gram-Positive Piezotolerant Bacterium Sporosarcina sp. DSK25 in Response to Growth Pressure

Pressure is an important thermodynamic property of the ocean and the deep biosphere that affects microbial physiology and biochemistry. Here, we report on our investigation of the response of Gram-positive piezotolerant bacterium Sporosarcina sp. DSK25 to hydrostatic pressure. Strain DSK25 responded in an adaptive manner to upshifts of growth pressure and showed systematic changes in phospholipid fatty acids. As the pressure increased from 0.1 to 10 MPa (Megapascal), unsaturated fatty acids in DSK25 increased from 21.7 to 31.1 % of total fatty acids, while the level of iso- and anteiso-branched fatty acids remained unchanged. At higher pressures (30, 50, and 60 MPa), the amount of unsaturated fatty acids decreased, and that of anteiso-branched fatty acids increased from 34.4 to 49.9 % at the expense of iso-branched fatty acids. For the first time, two polyunsaturated fatty acids (PUFA), 18:2n-6 and 18:2n-x, with the latter having much higher abundance than the former, were identified in DSK25. The concentration of the PUFA increased with growth pressure. These results indicate the involvement of unsaturated and methyl-branched fatty acids in the modulation of bacteria membrane fluidity and function over environmentally relevant parameter (pressure). Piezotolerant bacterium Sporosarcina sp. DSK25 appears to utilize two regulatory mechanisms for adaptation to high pressure, a rapid-responding mechanism on transient scale, expressed as increased biosynthesis of monounsaturated fatty acids, and a long-term adaptation mechanism in increased synthesis of anteiso-branched and polyunsaturated fatty acids. Our results further suggest that Gram-positive piezophilic bacteria respond differently than Gram-negative bacteria in adaptation to high pressure.     

砂土微生物固化过程中尿素的影响研究

砂土固化技术可改善砂土力学特性,被广泛应用于工程中,而在菌种培养液中添加尿素可大大加快固化反应提高砂土固化效率。开展了在菌种培养液中添加尿素分析其对砂土固化影响的试验研究。首先将尿素添加方式分为灭菌前加尿素、灭菌后加尿素和不加尿素3种,研究不同方式对菌种生长和脲酶活性的影响;再控制不同尿素添加量对比分析得到适合的尿素添加量以便后续研究;然后在砂土固化试验中采用灭菌后加尿素的方式研究其对砂土固化的影响;最后通过控制砂柱长度、胶凝液灌注速度和砂土颗粒粒径等因素,得到各因素对砂柱固化效果的影响。结果表明培养液中添加尿素可提高脲酶活性,但稍微抑制菌种生长;适合的尿素添加量为5~20 g/L;灭菌后添加尿素能显著提高灌注部位的强度,但长砂柱却因强度不均而整体强度较低;灌注速度越快,整体强度越高;添加20 g/L尿素时,整体强度随颗粒粒径增大而增大,因此,该方法适用于粗砂。研究成果对后期砂土固化技术的应用具有重要指导意义。     

微生物诱导碳酸镁沉淀试验研究

碳酸镁和碳酸钙一样也具有胶结作用,且镁矿强度远大于钙矿,因此研究碳酸镁固化技术具有重要意义。本文测量了巴氏芽孢杆菌在培养过程中的吸光度和脲酶活性,并计算得到单位脲酶活性;研究了尿素、氯化钠、醋酸根、Ca2+和Mg2+浓度对脲酶活性的影响;控制温度、pH值和离子浓度对比了钙和镁沉淀效率和不同温度不同尿素浓度下碳酸镁沉淀产率;对比研究了碳酸钙和碳酸镁沉淀下砂土固化效果。结果表明,48 h培养过程中,吸光度和脲酶活性的增长都先缓慢后迅速增长再减小最后停止,单位脲酶活性则是先增加后减小。适当增加尿素或Mg2+浓度可增强细菌脲酶活性,氯化钠和醋酸根浓度对酶活性无明显影响,Ca2+浓度对脲酶活性有明显抑制作用。同一温度、pH值和离子浓度条件下,碳酸镁产率明显小于碳酸钙。而在菌液中添加尿素可促进碳酸镁沉淀生成,且温度相同,尿素浓度越高,碳酸镁产率越大。菌液中添加尿素可使得碳酸镁固化砂土成型并具有一定强度,因此,该方法可解决砂土固化碳酸镁沉淀不足的问题,为后续碳酸镁固化试验奠定基础。     

微生物改性水泥土的制备与力学特性研究

花岗岩残积土在我国华南地区广泛分布,由于其具有明显的遇水崩解特性,往往带来山体滑坡等地质灾害。基于微生物诱导碳酸钙沉淀技术(MICP)采用巴氏芽孢杆菌对水泥加固花岗岩残积土的工程性质、工程技术进行改良研究,探究以水泥掺入量、钙离子浓度和钙源等因素为变量时,MICP技术对水泥土强度、应力应变关系等力学性能的影响。通过无侧限抗压强度(UCS)试验,分析了经MICP加固后水泥土的力学强度,得到以下结论:①MICP技术可以明显增强花岗岩残积土为基材的水泥土的强度、刚度和韧性等工程性质;②与对照组相比,试验组强度最大增长率为87.5%,最经济的水泥掺入量为15%;③氯化钙和乙酸钙为钙源都可以改善试样的韧性,而乙酸钙的效果更好,与对照组相比,试验组应变最大增长率为69.67%,此时所对应的钙离子浓度为0.5 mol/L。     

Sporosarcina pasteurii诱导碳酸盐-铀共沉淀修复低浓度铀废水的试验研究

对有钙源条件下Sporosarcina pasteurii诱导碳酸盐-铀共沉淀修复低浓度铀废水的性能开展了试验研究。试验结果表明,该细菌能分泌脲酶水解尿素诱导产生方解石并促使其与铀发生共沉淀,使废水中铀的去除率达到95.38%;该细菌能够耐受的铀浓度高达500 mg/L;铀的去除率随Ca²⁺浓度的升高而增加;温度在30℃左右和碱性环境有利于方解石与铀的共沉淀。对沉淀产物进行XRD、SEM-EDS表征分析表明,其主要成分为方解石,且包含铀元素;消解分析表明,在方解石形成过程中铀以共沉淀的方式被固定到了沉淀产物中。本研究表明,Sporosarcina pasteurii诱导碳酸盐-铀共沉淀对修复低浓度铀废水有潜在应用前景。     

Compressibility of biocemented loose sands under constant rate of strain,loading, and pseudo K0-triaxial conditions

The compressibility behavior of loose sands treated with Microbially Induced Carbonate Precipitation (MICP) is presented in this paper. The paper discusses the strain rate effects and evolution of at-rest earth pressure coefficient and elastic shear modulus during $K_0$-loading. The soil samples were prepared in a triaxial cell in which a biological solution containing the ureolytic bacteria Sporosarcina pasteurii was injected and held under a small back pressure. Cementation treatments were injected following an alternated top and bottom sequence. The constant rate of strain, constant rate of loading, and pseudo $K_0$-triaxial tests were performed at different strain and stress rates. On-specimen internal instrumentation consisting of a submersible load cell, three Hall Effect transducers, and vertical Bender Elements were used to control radial strains during $K_0$-loading and measure small-strain shear modulus changes. Based on shear wave velocity measurements, the MICP-treated sand was lightly cemented and displayed soil-like behavior. The experimental results demonstrated a significant reduction in soil compressibility after MICP treatment. The material response was remarkably similar for every tested strain rate. The very small values of axial strains measured for the biotreated samples in relation to untreated control specimens for vertical effective stress levels below 200 kPa is evidence of the suitability of this treatment and shows its potential for use in field applications at relatively shallow depths.     

微生物矿化技术固化石墨尾矿的影响因素研究

研究了不同因素对石墨尾矿固化效果的影响,包括菌液浓度、胶结液浓度、胶结液pH和环境温度。采用正交试验设计,以试样无侧限抗压强度（UCS）为指标,评价试样的固化效果,并给出各因素的推荐值及对固化效果影响的优先顺序。     

微生物在石英砂中诱导方解石沉积的实验研究

利用巴斯德芽孢杆菌和培养基来处理石英砂,与不含菌的对照样品相比,其固化效果比较明显。通过X-射线衍射分析处理后的样品,发现其中有方解石晶体生成。通过扫描电子显微镜可观察到石英砂颗粒的缝隙之间紧密地填充有大量的方解石晶体,由于方解石晶体本身具有胶结作用,从而使得石英砂颗粒能够胶结在一起。放大至10000倍可看到这些晶体表面附着有杆状菌体。能谱分析的结果也证明巴斯德芽孢杆菌在石英砂中诱导沉积出了方解石晶体。通过测算得出在本试验条件下（500mL含活菌5×10^9个的巴斯德芽孢杆菌菌液用urea—CaCl2重悬后与50g石英砂混合装入60mL注射器中,urea—CaCl2以重力作用流过注射器）方解石的产量平均为2．178g。     

微生物诱导方解石沉积加固的影响因素

通过比较用巴斯德芽孢杆菌处理介质前后的重量,应用单因素方法研究各种条件和环境对微生物诱导方解石沉积加固的影响。结果表明：①当培养基中ca^2＋浓度为25．2mM时,巴斯德芽孢杆菌诱导沉积的方解石产量最高;②培养基为碱性时,巴斯德芽孢杆菌所诱导沉积出的方解石的产量则较为接近;③巴斯德芽孢杆菌诱导沉积出的方解石的终重量与活菌数呈反比;④在30℃时,巴斯德芽孢杆菌诱导沉积的方解石产量最高;⑤巴斯德芽孢杆菌所诱导沉积出的方解石的产量随石英砂和海砂混合物中海砂所占比例的增大而减小;⑥随着培养基中Ni^2＋浓度增大,巴斯德芽孢杆菌诱导沉积的方解石产量显著降低。     

环境变化对微生物活性及矿化的影响

微生物诱导碳酸钙沉积（MICP）技术因具有低能耗、低污染等环境友好性优势成为文物保护、水处理、混凝土修补加固等领域的研究热点。MICP的矿化效果受多因素的制约与影响。本文开展了环境温度、pH值等因素对细菌生长和微生物矿化的影响研究。结果表明,环境温度不超过40℃时对菌液脲酶活性影响不大,42.5~45℃时6h后脲酶活性急剧降低,超过45℃时30min后菌液浓度及脲酶活性均大幅下降,直至失活;pH值越大,脲酶活性衰减越明显,pH≥11时脲酶活性急剧下降,大量的细菌不能承受高碱性环境进一步分解死亡。MICP技术在30℃时碳酸钙产量最高,在pH值为11~13的环境中细菌很难生长繁殖,碳酸钙生成量较低。