本文選題：地下水污染　切入點(diǎn)：生物強(qiáng)化　出處：《吉林大學(xué)》2017年博士論文　論文類型：學(xué)位論文

【摘要】：隨著城市化和社會經(jīng)濟(jì)的高速發(fā)展,污染物的泄漏導(dǎo)致地下水的嚴(yán)重污染,特別是有機(jī)污染普遍存在,對人類飲用水安全構(gòu)成了嚴(yán)重的威脅。苯胺是現(xiàn)代農(nóng)業(yè)、醫(yī)藥和化工產(chǎn)品的常用原料,應(yīng)用范圍非常廣泛,對地下水的污染比較普遍。由于苯胺對人體具有毒性、致癌、致突變性和在環(huán)境中的持久性,苯胺污染地下水的防治受到了人們的普遍關(guān)注。在苯胺污染地下水修復(fù)方法中,物理修復(fù)技術(shù)能耗及處理成本高,化學(xué)修復(fù)技術(shù)則易對環(huán)境造成二次污染,而原位生物修復(fù)技術(shù)由于其相對經(jīng)濟(jì)、有效和環(huán)境友好且易和多種修復(fù)手段聯(lián)合應(yīng)用等特點(diǎn),具有明顯優(yōu)勢。其中,生物強(qiáng)化可通過向地下環(huán)境中注入外源降解菌,達(dá)到縮短修復(fù)時(shí)間、提高修復(fù)效率的目的。但是,實(shí)際修復(fù)中往往由于外源降解菌在投加到場地后活性和數(shù)量大幅度降低,導(dǎo)致生物強(qiáng)化失敗。所以,外源降解菌在地下環(huán)境中的存活和遷移對生物強(qiáng)化至關(guān)重要。本論文通過研究含水層中的降解菌存活、降解菌遷移和生物強(qiáng)化修復(fù)效率三者之間的關(guān)系,為實(shí)際生物強(qiáng)化原位修復(fù)苯胺污染地下水提供理論基礎(chǔ)。通過之前的研究表明,Pseudomonas migulae AN-1是一株適冷、高效的苯胺降解菌,其在低溫下的降解性能已經(jīng)清楚,降解條件也被優(yōu)化,但AN-1在地下環(huán)境中的存活、遷移和對苯胺污染含水層的修復(fù)效能尚未明確。本論文研究了AN-1在含水層中存活和遷移的相關(guān)性能;構(gòu)建了一種新的綠色熒光蛋白(Green fluorescent protein,GFP)標(biāo)記結(jié)合光投射技術(shù)的方法體系,直觀并實(shí)時(shí)監(jiān)測了降解菌在模擬含水層中的遷移,首次實(shí)現(xiàn)了無擾動(dòng)同時(shí)捕捉游離菌和附著菌的遷移動(dòng)態(tài),揭示了降解菌在含水層中的遷移機(jī)制,對明確修復(fù)范圍及強(qiáng)化調(diào)控生物修復(fù)具有重要的指導(dǎo)意義;運(yùn)用皂素對降解菌在含水層中的遷移進(jìn)行強(qiáng)化,通過研究AN-1—液體—固體介質(zhì)三者的表面熱力學(xué)性質(zhì)、界面表面能、吉布斯自由能和DLVO相互作用能,進(jìn)一步闡明了皂素強(qiáng)化遷移的機(jī)理,為強(qiáng)化生物修復(fù)和緩解生物堵塞在理論和技術(shù)上奠定了基礎(chǔ);考察并驗(yàn)證了AN-1在模擬苯胺污染含水層中的存活、遷移和修復(fù)效能,建立了一套較為完整的原位生物修復(fù)苯胺污染地下水技術(shù)體系,對實(shí)際修復(fù)具有重要參考價(jià)值。通過上述研究,本論文主要得出以下結(jié)論:⑴AN-1在含水層中存活和遷移的相關(guān)特性AN-1可適應(yīng)苯胺濃度的變化,自動(dòng)調(diào)節(jié)自身的細(xì)胞膜通透性、疏水性和自聚集性。AN-1具有附著成膜能力,且AN-1形成的生物膜也具有較好的苯胺降解及抗苯胺沖擊性能。地下水中的Ca2+和SO42-(≥5 m M)對AN-1成膜有促進(jìn)作用,Fe2+有抑制作用。⑵AN-1在含水層中的遷移機(jī)制采用GFP標(biāo)記結(jié)合光透射技術(shù),無擾動(dòng)同時(shí)捕捉了模擬含水層中游離菌和附著菌的遷移動(dòng)態(tài),直觀并定量的揭示了AN-1在含水層中的遷移機(jī)制,即降解菌在含水層中的遷移受自身生物特性和地下水對流、彌散的共同作用,為生物因素和水文地質(zhì)因素共同影響。(1)降解菌遷移速度:AN-1遷移速度比地下水流速快,且含水層介質(zhì)粒徑越小,對AN-1遷移速度促進(jìn)越大,AN-1遷移越快。(2)降解菌遷移能力:含水層介質(zhì)粒徑越大、地下水流速越快、地下水中離子強(qiáng)度越小、溶解性有機(jī)質(zhì)越多,AN-1在飽和多孔介質(zhì)中的質(zhì)量回收率就越大,沉積速率系數(shù)越小,越易在含水層中遷移。其中,相同離子強(qiáng)度、相同陰離子,二價(jià)陽離子比一價(jià)陽離子對AN-1遷移的抑制作用大;相同離子強(qiáng)度、相同陽離子,二價(jià)陰離子比一價(jià)陰離子對AN-1遷移的抑制作用小。(3)降解菌遷移動(dòng)態(tài):對于菌羽的初始形狀,菌羽在中砂中沿注入井形成一條窄帶,在粗砂中于注入井下半部形成一個(gè)橢球;對于菌羽的面積,與時(shí)間呈線性關(guān)系,而且在相同降解菌注入量、相同時(shí)間內(nèi)、相同地下水流速條件下,中砂中的面積大于粗砂;對于菌羽的范圍,從注入井至降解菌羽鋒面,可形成連續(xù)的微生物帶,包括游離菌羽區(qū)域和后方的附著菌區(qū)域,對明確修復(fù)范圍及強(qiáng)化調(diào)控生物修復(fù)具有重要的指導(dǎo)意義。⑶AN-1在含水層中遷移的強(qiáng)化皂素對AN-1無毒害作用。當(dāng)0.1%、0.3%或0.5%皂素存在時(shí),AN-1對苯胺的降解速率比無皂素時(shí)略高,生物量略大。確立了0.1%為皂素沖洗溶液的最佳濃度,該濃度下AN-1—液體—固體介質(zhì)三者構(gòu)成體系的界面表面能最小、吉布斯自由能最大、細(xì)菌截留量最小。采用0.1%皂素溶液沖洗的方法(不用皂素對AN-1進(jìn)行預(yù)處理,直接注入含水層,再用0.1%皂素溶液進(jìn)行沖洗),是強(qiáng)化AN-1在含水層中遷移最經(jīng)濟(jì)、最有效的方式。該技術(shù)方法可使AN-1在含水層中發(fā)生有效的二次遷移,進(jìn)一步擴(kuò)大潛在的生物強(qiáng)化修復(fù)范圍。離子不影響皂素對AN-1的促遷移作用。當(dāng)皂素與離子共存時(shí),與離子單獨(dú)存在相比,皂素顯著提高了AN-1與介質(zhì)顆粒之間的能量壁壘,基本消除了離子對AN-1遷移的抑制作用,促進(jìn)了AN-1在飽和多孔介質(zhì)中的遷移。⑷AN-1修復(fù)模擬苯胺污染含水層在模擬苯胺污染含水層中,AN-1能夠較好的存活,表現(xiàn)出良好的降解性能,并可維護(hù)土著微生物群落結(jié)構(gòu),使種群適應(yīng)苯胺的沖擊。從注入井至降解菌羽鋒面,可形成連續(xù)的微生物反應(yīng)帶,有效的修復(fù)范圍包括游離菌羽區(qū)域和后方的降解菌附著區(qū)域。當(dāng)降解菌在污染含水層中注入時(shí),降解菌會大量附著生長并聚集在注入井的附近,雖然該區(qū)域?qū)Ρ桨返慕到馑俾瘦^大,為主要降解區(qū)域,但可能會形成潛在的生物堵塞。降解菌聚集區(qū)域會隨水流緩慢向下游運(yùn)移、擴(kuò)大,并伴隨游離細(xì)胞的不斷剝落、隨水流向下游遷移。AN-1對模擬苯胺污染含水層具有良好的生物強(qiáng)化修復(fù)效能。本論文的主要?jiǎng)?chuàng)新體現(xiàn)在:構(gòu)建了一種新的GFP標(biāo)記結(jié)合光投射技術(shù)的方法體系,直觀并實(shí)時(shí)監(jiān)測了降解菌在模擬含水層中遷移,實(shí)現(xiàn)了無擾動(dòng)同時(shí)捕捉游離菌和附著菌的遷移動(dòng)態(tài),首次直觀的揭示了生物強(qiáng)化的有效修復(fù)范圍;建立了一種新的運(yùn)用皂素強(qiáng)化降解菌在含水層中遷移的技術(shù)方法,具有相對經(jīng)濟(jì)、有效且環(huán)境友好的特點(diǎn),并闡明了其強(qiáng)化機(jī)理;首次驗(yàn)證了高效苯胺降解菌Pseudomonas migulae AN-1在模擬苯胺污染含水層中的存活、遷移和修復(fù)效能,系統(tǒng)且全面的研究了含水層中降解菌存活、降解菌遷移和生物強(qiáng)化修復(fù)效率三者之間的內(nèi)在關(guān)系。綜上所述,本論文為實(shí)際生物強(qiáng)化原位修復(fù)苯胺污染地下水奠定了理論基礎(chǔ),對實(shí)際工程應(yīng)用具有重要的指導(dǎo)意義。
[Abstract]:With the rapid development of city economy and society, the serious pollution of groundwater due to the leakage of pollutants, especially widespread organic pollution, constitutes a serious threat to human safety of drinking water. Aniline is commonly used materials of modern agriculture, medicine and chemical products, a very wide range of applications, the pollution of groundwater due to cancer is relatively common. Aniline has on the human body, toxicity, mutagenicity and persistence in the environment, the prevention and control of groundwater pollution was affected by people's attention. In the remediation of groundwater contaminated by aniline method, physical repair technology of energy consumption and high cost of processing, chemical remediation technology is easy to cause two pollution on the environment, and in situ bioremediation due to its relative economic characteristics, effective and environmentally friendly and easy to repair and various means of combination, has obvious advantages. Among them, bioaugmentation through to the underground Exogenous degrading bacteria into the environment, to shorten the time to repair, improve the repair efficiency. However, the actual repair is often due to exogenous degrading activity in the added site and quantity is greatly reduced, lead to enhanced biological failure. Therefore, exogenous degradation bacteria in underground environment is vital to the survival and migration of bioaugmentation. Through the study on degrading bacteria in water containing survival, the relationship between migration and degradation bacteria bioaugmentation remediation efficiency of three, provide a theoretical basis for the actual biological remediation of groundwater pollution. Through the study of aniline ago showed that Pseudomonas migulae AN-1 is a cold, high efficient aniline degrading bacteria, in the low temperature the performance degradation is clear, the degradation conditions were also optimized, but the survival of AN-1 in the underground environment, and the migration of aniline contaminated aquifer on the repair efficiency is not yet clear. This paper studies the performance of AN-1 survival and migration in the aquifer; build a new green fluorescent protein (Green fluorescent protein, GFP) marker combination method system of optical projection technology, visual and real-time monitoring of the degrading bacteria in water in the simulation of migration, the first migration dynamic disturbance at the same time capture the free bacteria and attached bacteria, revealing the mechanism of migration in the aquifer degrading bacteria, has an important guiding significance to clear the scope and strengthen the regulation of repair of bioremediation; using saponin on degrading bacteria migration in the aquifer are strengthened by surface thermodynamic study of AN-1 liquid - solid medium three properties. The interface surface energy, Gibbs free energy and the DLVO interaction energy, and further clarified the mechanism for strengthening saponin migration, bioremediation and mitigation of Biological Clogging in theory and technology laid the foundation The basic investigation and verification of the AN-1; in the simulation of aniline polluted aquifer containing survival, migration and repair efficiency, establishes a relatively complete set of in situ bioremediation of aniline polluted groundwater system, has important reference value for the actual repair. Through the above research, this thesis mainly draws the following conclusions: the survival of AN-1 in aquifer in the migration and relevant characteristics of AN-1 can adapt to changes in the concentration of aniline, automatically adjust their cell membrane permeability, hydrophobicity and self aggregation of.AN-1 with the attached film forming ability of biofilm formation of AN-1, and also has good impact properties of aniline degradation and anti aniline. Groundwater in Ca2+ and SO42- (m M = 5) on AN-1 film role, the inhibitory effect of Fe2+. The migration mechanism of AN-1 in the aquifer by labeling with GFP optical transmission technology, without disturbance while capturing the simulated aquifer in the free and attached bacteria The dynamic migration of bacteria, intuitive and quantitative reveals the mechanism of AN-1 migration in the aquifer, the migration of degrading bacteria in the aquifer by its biological characteristics and groundwater convection, dispersion interaction, for the common effect of biological factors and hydrogeological factors. (1) degradation bacteria migration speed: AN-1 migration speed the ratio of groundwater flow fast and aquifer and the smaller the particle size of AN-1, promote the migration speed is bigger, faster and faster. The migration of AN-1 (2) degradation bacteria migration: the aquifer of larger particle size, groundwater flow is fast, the smaller the ionic strength in groundwater, dissolved organic matter more, the quality of AN-1 recovery in a saturated porous medium rate is greater, the coefficient of deposition rate is small, the more easy migration in the aquifer. The same ionic strength, the same anion, the two divalent cations than monovalent cations in inhibition of AN-1 migration; the same ionic strength, the same cation Two, anion than monovalent anions inhibit the migration of AN-1. (3) degradation bacteria dynamic migration: for the initial shape of the plume plume in bacteria, bacteria in the sand along the injection well to form a narrow band, in the sand to form a ellipsoid in underground injection part; for bacteria plume area. A linear relationship with time, and the injection amount in the same degrading bacteria, the same time, the same groundwater flow conditions in the sand in the area is larger than the range of bacteria for coarse sand; Yu, from the injection well to the degradation of feather fronts, which form a continuous microbial zone, including free plume region and the rear of the attachment of bacteria by region, has an important guiding significance to the clear repair scope and enhanced bioremediation. The regulation of AN-1 migration in the aquifer to strengthen saponin has no toxic effect on AN-1. When there are 0.3% or 0.1%, 0.5% saponin, AN-1 degradation rate of aniline is slightly higher than when no saponin, The biomass is slightly larger. The optimum concentration of 0.1% saponin washing solution, the concentration of AN-1 in liquid - solid medium three form interface surface energy of the system is minimum, Gibbs free energy, the minimum amount of bacteria retention method using 0.1% saponin flushing (without saponin pretreated for AN-1 direct injection containing water, then 0.1% saponin solution flushing), is to strengthen the AN-1 migration in the aquifer in the most economical, the most effective way. This method can make the AN-1 two effective migration in the aquifer, further expand the scope of strengthening the potential biological repair. Ion does not affect the function of promoting migration of saponin. When AN-1 saponin and ions, and ion alone compared to saponin significantly increased between AN-1 and medium particle energy barriers, the inhibition of AN-1 ion migration basically eliminated, promote AN-1 in saturated The migration in porous media. The AN-1 repair aniline contaminated aquifer in simulated aniline polluted aquifer, AN-1 can better survive, showed a good degradation performance, and can maintain the indigenous microbial community structure, make the population adapt to aniline impact. From injection well to degradation of plume fronts, can form a continuous microbial reaction with the effective range of the repair degrading bacteria free bacteria including the plume region and the rear of the attachment region. When injected in degrading bacteria in the aquifer polluted, degrading bacteria can adhesion growth and aggregation in the injection wells nearby, although the larger degradation rate of the area of aniline, the main degradation region, but may the formation of potential biological clogging. Degrading bacteria gathering area with water slowly moves downstream, and continue to expand, with peeling free cells, along with the water flow to the downstream migration of.AN-1 on aniline pollution water layer. Have good biological strengthening repair efficiency. The main innovation of this thesis lies in: to construct a new GFP marker combining method system of light projection technology, visual and real-time monitoring of the degrading bacteria in simulated migration in the aquifer, realizes dynamic migration without disturbance also capture free bacteria and bacterial attachment, the first intuitive to reveal the effective repair range bioaugmentation; establish a new use of saponin strengthen degrading bacteria migration in aquifer techniques, with relative economic, effective and environmentally friendly, and expounds its strengthening mechanism; the first to verify the efficient aniline degrading bacteria Pseudomonas migulae AN-1 in the simulation of aniline polluted with survival in the water, migration and repair efficiency, systematic and comprehensive research on water degradation bacteria survival layer, the intrinsic relationship between migration and degradation bacteria bioaugmentation remediation efficiency of three. To sum up This paper lays a theoretical foundation for the actual biological reinforcement of the remediation of groundwater contaminated by aniline in situ, and has important guiding significance for practical engineering applications.

【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：X172

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