本文選題：黑臭河道 + ATU�。� 參考：《華東師范大學(xué)》2017年碩士論文

【摘要】：為了盡快遏制我國城市水污染日益加重的趨勢、改善城市的水環(huán)境質(zhì)量,近年來許多城市根據(jù)其自身特點(diǎn),開展了相應(yīng)的黑臭河道治理工程。然而,隨著相應(yīng)的黑臭河道治理工程的實(shí)施,污染負(fù)荷得到了大幅度削減,氮去除難以達(dá)標(biāo)的問題卻逐漸突顯出來。其主要原因是目前黑臭河道治理過程中側(cè)重于對氨氮的硝化轉(zhuǎn)化,而忽視氮的脫除過程,導(dǎo)致總氮去除率偏低的現(xiàn)象普遍存在。為了解決黑臭河道"氮超標(biāo)"難題,本研究基于曝氣、調(diào)控C/N比和底泥投加硝酸鈣三種治理措施,借助同位素示蹤技術(shù)和物料平衡分析,從氨氧化、反硝化、厭氧氨氧化和硝酸鹽異化還原成銨4個方面對相應(yīng)措施下的氮轉(zhuǎn)化途徑進(jìn)行分型表征,探討在黑臭河道氮轉(zhuǎn)化途徑中的地位和作用,并通過高通量測序和熒光定量PCR技術(shù)解析對應(yīng)的微生物作用機(jī)制,以期為底泥內(nèi)源氮的有效控釋和黑臭河道治理工程提供基礎(chǔ)的理論指導(dǎo)和借鑒。主要研究結(jié)果如下:(1)在對氨芐青霉素、雙氰胺(DCD)、烯丙基硫脲(ATU)、辛炔、氯仿和乙炔抑制氨氧化細(xì)菌(AOB)性能評定的基礎(chǔ)上,篩選出適用于黑臭河道相對較優(yōu)的硝化抑制劑為ATU,適宜濃度為25μmol·L-1,其主要通過降低AOB(主要是亞硝化螺菌屬)種群豐度且抑制AOB菌群活性的方式實(shí)現(xiàn)其良好的硝化抑制效果。(2)基于曝氣、調(diào)控C/N比和底泥投加硝酸鈣三種治理措施的研究發(fā)現(xiàn),曝氣(工況3)對于氨氮的削減最為有效,在第18 d濃度便降低為2.07±0.24 mg·L-1,曝氣條件下增加C/N比(工況5和工況7)不利于氨氮的削減,使得氨氮降為2 mg·L-1的時間推遲10～15d;但對于TN來說,增大C/N比有利于TN的快速削減,TN均在34～38d后降為2mg·L-1以下,相比曝氣組達(dá)標(biāo)的時間平均縮短22～26 d,C/N比為10(工況5)或者20(工況7)無顯著影響差別,表明當(dāng)C/N比為10條件下,碳源不是影響TN去除的限制因素;投加硝酸鈣(工況9)不利于氨氮和TN的去除;所有添加抑制劑組均使氨氮和TN達(dá)標(biāo)時間顯著延遲。(3)曝氣調(diào)控條件下,AOB在黑臭河道氨氧化過程中起主導(dǎo)作用,其貢獻(xiàn)率為62.63～78.28%,而AOA占21.72～37.37%;同時發(fā)現(xiàn)穩(wěn)定時氮素的削減以反硝化作用為主導(dǎo),其速率高達(dá)3.61μmol N·kg-1·h-1,對氮素的削減貢獻(xiàn)率約為83.26～92.46%;各工況厭氧氨氧化速率均在0.27～0.48μmol N·kg-1·h-1,貢獻(xiàn)率為7.54～16.74%;曝氣過程中DNRA速率低于檢測限。(4)不同C/N比調(diào)控條件下,AOB均在氨氧化過程中起主導(dǎo)作用,其貢獻(xiàn)率約為72.74～83.74%,AOA占16.26～27.26%,且受C/N比影響不大;除C/N比為20(工況7)條件下,厭氧氨氧化速率與反硝化速率相近外,各工況氮素削減均以反硝化過程為主,貢獻(xiàn)率分別為64.58～89.17%,厭氧氨氧化貢獻(xiàn)約為10.83～35.42%。特別地,在C/N比為10(工況5)中檢測到DNRA過程,但其對氮素削減的貢獻(xiàn)率較低,約為5.18～7.45%。(5)投加硝酸鈣能夠增大AOA對氨氧化過程的貢獻(xiàn)(24.37～50.26%),且能夠促進(jìn)反硝化速率和厭氧氨氧化速率,但仍然以反硝化過程為主,其貢獻(xiàn)率約為79.56～85.52%,厭氧氨氧化作用的貢獻(xiàn)率為14.48～20.44%;且整個過程中發(fā)現(xiàn)DNRA過程速率低于檢測限。(6)黑臭河道中細(xì)菌以變形菌門、厚壁菌門和綠彎菌門為優(yōu)勢菌群,不同調(diào)控措施均能顯著降低厚壁菌門豐度,而對變形菌門來說,投加硝酸鈣調(diào)控對其豐度的影響最大;古菌中優(yōu)勢菌群主要為深古菌門、廣古菌門、泉古菌門和奇古菌門,與起始工況相比,各工況運(yùn)行穩(wěn)定時優(yōu)勢菌群豐度均顯著增加,但受不同調(diào)控措施影響較小。(7)基于amoA基因?qū)Π毖趸⑸锏姆肿由飳W(xué)分析表明,各工況AOB數(shù)量明顯高于AOA數(shù)量(4～9倍),約為3.71×105～1.13×106拷貝數(shù)/(克·干重),添加抑制劑主要抑制AOB中的Nitrosomonas和Nitsomospira菌屬,而對AOA無明顯影響,同時發(fā)現(xiàn)溶解氧是影響AOA菌群結(jié)構(gòu)差異性的主要環(huán)境因子。(8)對反硝化微生物來說,各工況nirS型反硝化菌的數(shù)量明顯高于narG、norB和nosZ型反硝化菌數(shù)量,約為5.39×107～5.47×108拷貝數(shù)/(克·干重),其優(yōu)勢菌屬為Pseudowonas、Sulfuritalea和Dechloromonas;在不同調(diào)控因子中,硝態(tài)氮是影響反硝化菌菌群結(jié)構(gòu)的主要因素,投加硝酸鈣后Thiobacillus菌屬的豐度劇烈增加,成為優(yōu)勢菌屬,推測可能促進(jìn)了硫自養(yǎng)反硝化過程的發(fā)生,其中抑制劑的添加影響了部分narG型反硝化菌,對其他反硝化菌無顯著影響。(9)對于厭氧氨氧化微生物來說,各工況優(yōu)勢菌屬為Candidatus_Jettenia和Candidatus_Kuenenia,在不同調(diào)控措施中,底泥投加硝酸鈣能夠顯著改變厭氧氨氧化菌的豐度和菌群組成,其數(shù)量由2.06×104～8.79×104拷貝數(shù)/(克·干重)增加到2.66×105～1.95×106拷貝數(shù)/(克·干重),其中Candidatus_Jettenia菌屬的相對豐度顯著增加,由19.53%增加為73.30～73.83%,而其他調(diào)控措施相對來說影響不大。(10)對于DNRA微生物來說,發(fā)現(xiàn)增加C/N比能夠促進(jìn)其生長,各工況數(shù)量為3.45×106～9.19×106拷貝數(shù)/(克·干重),其優(yōu)勢菌屬均為Desulfovibrio屬,豐度約為 0.7%～1.4%,主要以 Desulfovibrio_vulgaris 和 Desulfovibrio_termitidis種為主,但可能其活性較低或者相關(guān)功能酶未表達(dá)。
[Abstract]:In order to restrain the increasing trend of urban water pollution and improve the quality of urban water environment as soon as possible, in recent years, many cities have carried out corresponding control projects of black and smelly river channels according to their own characteristics. However, with the implementation of the corresponding control projects of the black and smelly river, the negative pollution is greatly reduced and the nitrogen removal is difficult to reach the standard. The main reason is that the main reason is that at present, the process of nitrification and conversion of ammonia nitrogen is emphasized in the process of controlling the black and smelly river, but the removal process of nitrogen is ignored and the removal rate of total nitrogen is low. In order to solve the problem of "nitrogen exceeding standard" in the black and smelly channel, this study is based on aeration, regulating the C/N ratio and adding calcium nitrate to the sediment. With the help of isotopic tracer and material balance analysis, the nitrogen conversion pathway under the corresponding measures was characterized by 4 aspects of ammonia oxidation, denitrification, anaerobic ammonia oxidation and nitrate dissimilation reduction into ammonium. The status and application of the nitrogen conversion in the black and smelly channel were discussed, and the high throughput sequencing and fluorescence quantitative PCR technique were used. The corresponding mechanism of microbial action is analyzed in order to provide theoretical guidance and reference for effective controlled release of endogenous nitrogen in sediment and treatment of black odor river. The main results are as follows: (1) on the basis of the performance evaluation of ampicillin, Dicyandiamide (DCD), allyl thiourea (ATU), octyyne, chloroform and acetylene inhibition ammonia oxidizing bacteria (AOB) A relatively excellent nitrification inhibitor suitable for black and smelly rivers was selected as ATU, with a suitable concentration of 25 mu mol. L-1, which achieved good nitrification inhibition effect mainly by reducing the abundance of AOB (mainly the genus snnail) and inhibiting the activity of AOB bacteria. (2) three control measures based on aeration, control C/N ratio and sediment cast calcium nitrate. The study found that aeration (working condition 3) is the most effective for ammonia nitrogen reduction, and the concentration of eighteenth D is reduced to 2.07 + 0.24 mg L-1. The increase of C/N ratio under the aeration condition (working condition 5 and working condition 7) is not conducive to the reduction of ammonia nitrogen, and the time of ammonia nitrogen reduction to 2 mg. L-1 is delayed 10 to 15d; but for TN, the increase of C/N ratio is beneficial to the rapid reduction of TN, TN is both in After 34 ~ 38d to below 2mg L-1, the time of the aeration group was reduced by 22~26 D, the C/N ratio was 10 (working condition 5) or 20 (working condition 7). It showed that when the C/N ratio was 10, the carbon source was not the limiting factor for the removal of TN; the addition of calcium nitrate (working condition 9) was not conducive to the removal of ammonia nitrogen and TN; all addition inhibitor groups were all made The time of ammonia nitrogen and TN standard was delayed significantly. (3) under the condition of aeration, AOB played a leading role in the process of ammoxidation of black and smelly rivers, and its contribution rate was 62.63 to 78.28%, while AOA accounted for 21.72 ~ 37.37%. At the same time, the reduction of nitrogen was dominated by denitrification, and its rate was up to 3.61 mol N. Kg-1. H-1, and the contribution rate of nitrogen reduction was about The rate of anammox is 83.26 to 92.46%; the rate of anammox is 0.27 to 0.48 Mu mol N kg-1. H-1, and the contribution rate is 7.54 to 16.74%, and the DNRA rate is lower than the detection limit during aeration. (4) under the condition of different C/N ratio, AOB plays the leading role in the process of ammonia oxidation, and its contribution rate is about 72.74 to 83.74%, AOA is 16.26 ~ 27.26%, and influenced by C/N ratio. In addition to the C/N ratio of 20 (working condition 7), the anaerobic ammonia oxidation rate is similar to the denitrification rate, and the nitrogen reduction is mainly denitrification, the contribution rate is 64.58 to 89.17%, the anammox contributes about 10.83 to 35.42%., and the DNRA process is detected in the C/N ratio 10 (working condition 5), but its contribution to nitrogen reduction The rate is low. The addition of 5.18 ~ 7.45%. (5) calcium nitrate can increase the contribution of AOA to the process of ammonia oxidation (24.37 ~ 50.26%), and can promote the denitrification rate and the anaerobic ammonia oxidation rate, but still mainly denitrification process, the contribution rate is about 79.56 to 85.52%, the contribution rate of anaerobic ammonia oxidation is 14.48 to 20.44%; and the whole process is in the whole process. It was found that the DNRA process rate was lower than the detection limit. (6) the bacteria in the black and smelly rivers were dominated by the deformable bacteria gate, the thick wall bacteria gate and the green flexural bacteria gate, and the different control measures could significantly reduce the abundances of the thick wall bacteria, while the control of calcium nitrate on the abundance of the bacteria was the most important for the deformable bacteria. Bacteria gate, spring Archaea and archaea door, compared with the starting condition, the abundance of dominant flora increased significantly when the operating conditions were stable, but affected by different control measures. (7) the molecular biological analysis of ammoxidation microorganism based on amoA gene showed that the number of AOB in each working condition was obviously higher than that of AOA (4~9 times), about 3.71 * 105 ~ 1.13 x 10 6 copy number / (g dry weight), adding inhibitor mainly inhibited Nitrosomonas and Nitsomospira in AOB, but had no obvious effect on AOA. At the same time, it was found that dissolved oxygen was the main environmental factor affecting the structural difference of AOA bacteria. (8) for denitrifying microorganisms, the number of nirS denitrifying bacteria in each condition was significantly higher than that of narG, norB and nosZ type. The number of nitrifying bacteria is about 5.39 x 107 ~ 5.47 * 108 copies / (k dry weight). The dominant bacteria are Pseudowonas, Sulfuritalea and Dechloromonas. In different regulatory factors, nitrate nitrogen is the main factor affecting the structure of denitrifying bacteria. After adding calcium nitrate, the abundance of Thiobacillus genera is greatly increased, and it is presumed possible. The process of sulfur autotrophic denitrification was promoted, in which the addition of inhibitors affected partial narG denitrifying bacteria and had no significant effect on other denitrifying bacteria. (9) for anaerobic ammonia oxidizing microorganisms, the dominant bacteria were Candidatus_Jettenia and Candidatus_Kuenenia in various operating conditions. In different control measures, calcium nitrate could be added to the sediment. The abundances and flora composition of anammox were significantly changed from 2.06 x 104 to 8.79 x 104 copies / (k dry weight) to 2.66 x 105 to 1.95 * 106 copies / (k dry weight). The relative abundance of Candidatus_Jettenia bacteria increased significantly from 19.53% to 73.30 ~ 73.83%, and the other regulation measures were relatively not affected. (10) for DNRA microbes, it was found that the increase of C/N ratio could promote its growth. The number of working conditions was 3.45 x 106 ~ 9.19 x 106 copies / (k dry weight). The dominant genus were all Desulfovibrio genera, and the abundance was about 0.7% to 1.4%, mainly with Desulfovibrio_vulgaris and Desulfovibrio_termitidis species, but their activity was lower or lower. The related functional enzymes were not expressed.

【學(xué)位授予單位】：華東師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：X522

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