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Application of biological enzyme Used in Sludge Solidification
|關鍵字:||Sludge;淤泥;Bacillus pasteurii;巴氏桿菌||出版社:||土木工程學系所||引用:|| Bang, S. S., Galinat, J. K., and Ramakrishnan, V. (2001) Calcite precipitation induced by polyurethane-immobilized Bacillus pasteurii. Enzyme and Microb. Technol. 28, 404-409.  Bachmeier, K. L., Williams, A. E., Warmington, J. R., and Bang, S. S. (2002) Urease activity in microbiologically-induced calcite precipitation. J. Biotech. 93, 171-181.  Day, J. L., Ramakrishnan, V., and Bang, S. S. (2003) Microbiologically induced sealant for concrete crack remediation. Proceedings of the 16th Engineering Mechanics Conference, Seattle, WA.  DeJong, J. T., Fritzges, M. B., and Nusslein, K. (2006) Microbially induced cementation to control sand response to undrained shear. J. Geotech. Geoenv. Eng. 132, 1381-1392.  Fernandes, P. (2006) Applied microbiology and biotechnology in the conservation of stone cultural heritage materials. Appl Microbiol. Biotechnol. 73, 291–296.  Bennett, R. H., Bryant, W. R., and Hulbert, M. H. (1991). Microstructure of fine-grained sediments, Springer, New York.  Tiano, P. (1995) Stone reinforcement by calcite crystals precipitation induced by organic mat rix macromolecules [J]. Studies in Conservation, 40 (3):171~176.  Ruixing, W. Chunxiang, Q. Jianyun, W. Study on Microbiological precipitation of CaCO3. 東南大學學報(自然科學版)  Lochhead M J, Letellier S R, Vogel V. (1997) Assessing the role of interfacial electrostatics in oriented mineral nucleation at charged monolayers[J]. J Phys Chem, 101B: 10821-10827.  Kitamura, M., Konno, H. (2002) Controlling factors and mechanism of reactive crystallization of calcium carbonate polymorphs from calcium hydroxide suspensions [J]. Journal of Crystal Growth, (236): 323-332.  Zeshan, H., Yulin, D. (2003) Supersaturation control in aragonite synthesis using sparingly soluble calcium sulfate as reactants [J]. Journal of Colloid and Interface Science, (266): 359-365.  Ramakrishnan, V., Ramesh, K. P., Bang. S. S. (2001) Bacterial Concrete [C]. Proceedings of SPIE, 4234, Smart Material s, Alan R. Wilson, Hiroshi Asanuma, Editors, 168~176.  Lirong Zhong, M.R. Islam. 一種新型微生物封堵裂縫的試驗研究. 國外油田工程.  Zeshan, H., Yulin, D. (2004) Synthesis of needle-like aragonite from calcium chloride and sparingly soluble magnesium carbonate [J]. Powder Technology, (140):10-16.  Liang, C., Chun-Xiang, Q., Rui-Xing, W. and Jian-Yun, W. Study on the Mechanism of Calcium Carbonate Formation Inducedby Carbonate-mineralization Microbe. 東南大學學報(化學)  Ruixing, W., Chunxiang, Q. Restoration of Defects on the Surface of Cement-Based Materials by Microbiologically precipitated CaCO3. 東南大學學報(矽酸鹽).  Hill, D. D. (2002) Sleep B. E. Effects of biofilm growth on flow and transport through a glass parallel plate f racture[J]. Journal of Contaminant Hydrology, 56, 227~246.  Mobley, H.L.T., Island, M.D., and Hausinger, R.P. (1995) Molecular biology of microbial ureases. Microbiol. Rev. 59, 451–480.  Blakeley, R. L., Webb, E. C. and Zerner, B. (1969) Jack bean urease (EC 188.8.131.52). A new purification and reliable rate essay. Biochemistry 8, 19841990.  Weatherburn, M. W. (1967) Phenol-hypochlorite reaction for determination of ammonia. Anal. Chem. 39, 971–974.  McGee, D. J., May, C. A., Garner, R. M., Himpsl, J. M., and Mobley, H. L. T. (1999) Isolation of Helicobacter pylori genes that modulate urease activity. J. Bacteriol. 181, 2477–2484.  Kaltwasser, H., and Schlegel, H. G. (1966) NADH-dependent coupled enzyme assay for urease and other ammonia-producing systems. Anal. Biochem. 16, 132–138.  American Public Health Association (APHA). (1989) Standard methods for the examination of water and wastewater, 17th ed. American public health association, Washington, DC.  Ramachandran, S. K., Ramakrishnan, V., and Bang, S. S. (2001). “Remediation of concrete using micro-organisms.” ACI Material Journal, 98(1), 3-9.  Hammes, F., Boon, N., Villiers, J. D., Verstraete, W. and Siciliano, S. D. (2003). “Strain-specific ureolytic microbial calcium carbonate precipitation.” Applied and Environmental Microbiology Journal, 69(b), 4901-4909.  Chen, J. W., and Shiue, J. H. (1995). "Liquefaction and deformation of a sandy seabed: A Model Test Study." Proceedings of the National Science Council Journal, 19(5), 430-439.  LI Pei-hao,QU Wen-jun. State of Art in Application of Bioremedying and Bioreinforceing Materials in Civil Engineering. Vol.26, No5. 材料科學與工程學報.  許員豪、王明光、王一雄，2000，蒙特石及至蛭石吸持十六烷基三甲銨形成有機黏粒機制之探討，土壤與環境，3(2),113-120。  林季融，2009年，黏土礦物對土壤細菌生長影響之研究，國立中興大學土壤環境科學系碩士論文。  蔡攀鰲，民國89年，公路工程學。  陳銘鴻，土壤液化成因、災害與復健。  Nakano MM, Zuber P (1998). "Anaerobic growth of a "strict aerobe" (Bacillus subtilis)". Annu Rev Microbiol 52: 165–90. doi:10.1146/annurev.micro.52.1.165. PMID 9891797  中興工程顧問股份有限公司，2000，石門水庫淤泥元化處置方案評估規劃綜合報告。||摘要:||
研究結果顯示，應用B. pasteurii菌液於淤泥固化時，菌液的濃度越高所能誘導的碳酸鈣沉澱量越多，在濃度70%的Urea-CaCl2 medium條件下，所固化的淤泥試體抗壓強度以菌液濃度100%的最高，相較於對照組(菌液濃度0%者)，其抗壓強度約高13.48%，可知利用細菌固化淤泥，確實可提升其抗壓強度。Urea-CaCl2 medium的濃度越高所能誘導的碳酸鈣沉澱量越多，濃度70%的Urea-CaCl2 medium所固化的淤泥試體，其抗壓強度較濃度30%與10%高。
根據利用菌體共拌副產石灰固化純淤泥之試驗結果，固化後純淤泥試體28天之抗壓強度約介於11 kgf/cm2 ~17 kgf/cm2。無論在何種淤泥與副產石灰共拌比例與菌液濃度下，各齡期抗壓強度值均大於對照組(即菌液濃度為0%者)，可知利用細菌與副產石灰確實可提升淤泥之強度。綜合試驗結果，發現以副產石灰及細菌進行淤泥固化之工作，可充分發揮副產石灰與細菌之功效，達到良好之淤泥固化成效；另一方面，使用副產石灰亦可算是資源再利用，不但可減少資源浪費，還可降低淤泥固化之成本，可說是一舉兩得。
本研究在牧場與堆肥場所取樣的尿素含量高之土壤中，確實可分離出能產生尿素酶的菌株。由分離本土尿素酶菌的結果，我們獲得了Sporosarcina sp. CN3及Sporosarcina sp. CN6。Sporosarcina sp. CN3及Sporosarcina sp. CN6與Sporosarcina pasteurii(B. pasteurii)的外型皆為桿菌，而且與16S rDNA的相似度高達99%。可以說我們很幸運的找到與B. pasteurii相似的本土菌株。
In recent years, due to the climate change, the reservoir sedimentation has always been a disturbance to Taiwan government. And it has been an urgent topic for the authority to dispose reservoir sludge effectively. Researchers of civil engineering materials and environmental engineering, cooperating with microbiologists and geochemists, have recently attempted the solidification of sludge granules into sandstone-like material employing microorganisms. The aim of the present study is to investigate how to solidify sludge or soil particle rapidly to attain strength during the growth of bacteria, and apply the result in engineering to gain benefits.
In this research we tried to use Bacillus pasteurii in the solidification of reservoir sludge. The sludge used in the experiment has a moisture content of 40%. The change of bacteria broth culture concentration(0%, 25%, 50%, 75% and 100%) and CaCl2 ratio in the culturing Urea-CaCl2 medium(10%, 30% and 70%) were used as the experimental variables. The culture broth was mixed directly with reservoir sludge to form a cubic specimen (50 mm x 50 mm x 50 mm), which was then used in compressive strength, XRD and SEM test. The test result showed that when cultured in 70%-CaCl2 medium, the specimen was blended with 100% bacteria broth had the highest compressive strength, which, compared with the control group(0% bacteria broth), was elevated approximately 13.48%. the result suggest that the solidification by bacteria can indeed enhance the compressive strength of the cube. The result also showed that the higher the content of CaCl2 we use in the culturing medium, the more CaCO3 deposit can be induced by the bacteria. The sludge solidified with bacteria cultured in 70%-CaCl2 medium had the highest compressive strength.
Furthermore, when substituted CaCl2 with By-product Lime of circulating fluidized bed(CFBC), the compressive strength of the specimen at the 28th day was 15-41% higher than control group. The result showed that the sludge solidification utilizing CFBC and bacteria can bring the two component into effect and attain excellent solidification result. On the other hand, the reuse of CFBC can also reduce the consumption of resource and diminish the cost of sludge solidification.
According to literature, Sporosarcina pasteurii (also known as Bacillus pasteurii) could hydrolyze urea to carbon dioxide and ammonium. Carbon dioxide turns to carbonate in the resulting alkaline environment and forms calcium carbonate in the presence of calcium. The calcium carbonate precipitates could fill the gaps between sand grains. In this study we tried to test the idea of whether the sizes of silt from water reservoir can be increased by adding S. pasteurii, urea, and calcium into the slurry. If it works, this new concept may facilitate the rate of removing silt from water reservoir. In this study, we optimized the culture medium and condition for growing S. pasteurii in scales up to 300 liters. Then we incubated S. pasteurii with silt for various periods of time and observed the formation of calcium carbonate with scanning electron microscope and X-ray powder diffractometer. We found that the average size of sediment was increased. Indigenous strains of S. pasteurii (Sporosarcina sp. CN3 and Sporosarcina sp. CN6) were isolated from compost. They can be used for further application study to avoid potential conflicts of intellectual rights.
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