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题名: 猪场废水生物处理与土地利用过程抗生素抗性基因的转归研究
作者: 隋倩雯1
学位类别: 博士后
答辩日期: 2017-07
授予单位: 中国科学院大学
授予地点: 北京
导师: 魏源送
关键词: 抗生素抗性基因,猪场废水,生物处理,序批式膜生物反应器,土地利用 ; Antibiotic resistance genes, Swine wastewater, Biologicaltreatment, Sequencing batch membrane bioreactor, Land application
学位专业: 环境科学与工程
中文摘要: 抗生素抗性污染是二十一世纪公共卫生面临的巨大的威胁,畜禽养殖过程 抗生素用量大,而针对畜禽养殖过程抗生素抗性污染的数据却非常缺乏。抗生 素抗性基因(Antibiotic resistance genes, ARGs)作为一种新型污染物,通过畜 禽养殖废弃物的排放或还田利用等途径向环境中传播,引起环境抗性水平提高, 并对公共健康安全造成威胁。因此,本研究以猪场废水中典型的 ARGs为研究 对象,以现场调研和实验室模拟为研究手段,针对猪场废水的主要处理工艺和 利用过程开展研究,探讨猪场废水 ARGs的时空分布特征、生物处理过程抗性 基因去除及其参数优化、以及猪场废水土地利用过程抗性基因的归趋。以期为 畜禽养殖废水 ARGs消减提供科学与技术支持。 通过对我国南北方典型规模化猪场冬、夏季猪场废水处理与土地利用过程 ARGs的调研,结果表明 tetM、tetX、ermB、ermF、mefA、sul1和 sul2是猪场 废水中的优势 ARGs,冬季 ARGs的拷贝数大于夏季,冬、夏季拷贝数差异在 0.26-3.16 log。猪场废水处理过程对 ARGs拷贝数具有较好去除,冬季去除效果 优于夏季。但 ARGs丰度在不同处理工艺下,去除效果不一。tetG、ereA、sul1 和 sul2在猪场废水处理过程中丰度不断升高,其丰度与 intI1具有显著正相关 关系(p<0.01),这表明 ARGs可能在猪场废水处理过程中发生水平转移,其风 险需要特别关注。在猪场废水土地利用过程,北方冬季土地休耕停止施用沼液 可有效降低土壤中 ARGs的含量,消减了 1.66 log。而南方猪场在全年施用沼 液的情况下,冬、夏季土壤中 ARGs的拷贝数总数相似(p>0.05)。在猪粪水厌 氧消化与土地利用过程中持久性 ARGs(sul1和 sul2)和逐渐升高的 ARGs(tetG 和 ereA)需要特别关注。 序批式膜生物反应器(Sequencing batch membrane bioreactor, SMBR)小试 装置处理猪场废水的结果表明,SRT是影响 ARGs转归最大的单一因子,对 ARGs变化贡献了 17.4%。而环境因子(SRT、SS、温度、COD、NH4+-N、NO2--N 和 NO3--N)较菌群结构在影响 ARGs转归方面占主导地位,可以解释 ARGs变 化的 45.8%。较短的 SRT(12天)时,SMBR对 ARGs的拷贝数去除效率最高 为 2.91 log。通过 SMBR试验验证了现场调研结果,即猪场废水生物处理后 tetG、 ereA、sul1和 sul2丰度提高,且 SRT越长,反应器内上清液 tetG和 ereA丰度 提高幅度越大。提高 SRT,生物处理(进水与上清液比较)ARGs拷贝数和丰 度的去除率均呈现降低的趋势,反应器中可能发生了较高频率的 ARGs的增殖 以及水平转移。反应器内样品(混合液、微滤膜表面附着的生物膜、上清液) 受 SRT的影响较大,而出水受 SRT影响较小。 猪粪水、沼液土地利用的盆栽试验结果表明,猪粪水还田比沼液还田对土 壤的菌群结构、ARGs和 ARB影响大,化肥添加组和 CK组变化规律相似;经 过猪粪水、沼液还田处理,红土受影响最大,其次是黄土,黑土受粪源细菌、 ARGs和 ARB污染最小。tetG、tetM和 ermF是猪粪水土地利用土壤中的优势 ARGs,在土壤中衰减符合一级反应动力学反应,它们的消减速率黑土 >黄土> 红土。盆栽试验一定程度上验证了现场调研的结果,即 tetG和 ereA在粪水还 田的土壤中丰度升高,尤其在红土和黄土中,说明 tetG和 ereA在土壤中发生 了增长与传播,其影响不容忽视。通过偏 RDA,土壤菌群对 ARGs变化贡献 63.7%,土壤理化特性贡献 39.8%,对 ARGs变化的贡献总计 81.4%。
英文摘要: Antibiotic resistance is an increasing threat to global public health. Large amount of antibiotics was consumed by animal production. However, little information was on the antibiotic resistance from animal wastes. Antibiotic resistance genes (ARGs) as emerging contaminants were disseminated through discharge and land application of animal wastes, which evaluated the antibiotic resistance level in environment and also pose threat to public health. Therefore, in this study ARGs in swine wastewater during the biological treatment and application of swine wastewater was investigated through the methods of on-site survey and laboratary experiment. The temporal and spatial distribution of ARGs in swine wastewater, the ARGs removal during the biological treatment as well as the parameter optimization, and the occurrence of ARGs during the land application of swine wastewater were explored, in order to provide the theoretical and technological support for the control and elimination of ARGs in swine waster. On-site survey was conducted on biological treatment and land application of swine wastewater in typical large-scale swine farms located at sourth and north of China during winter and summer, respectively. The results indicated that tetM, tetX, ermB, ermF, mefA, sul1 and sul2 were the dominant ARGs in swine wastewater. The copy number of ARGs detected was higher in winter samples than those in summer, where there was a 0.26-3.16 log variation in ARGs between the two seasons. ARGs copy number was well reduced by wastewater treatment processes, and the removal efficiency was relatively high in winter compared to summer. But the removal effect of ARGs abundance varied among the treatment processes. The abundance of tetG, ereA, sul1 and sul2 was raised during the wastewater treatment process, and was positively correlated with intI1 (p<0.01), which suggested that horizontal gene transfer may occurred and the risk required more attention. Through the on-site survey on the land application of swine wastewater, the total ARGs quantity in soil fell down by 1.66 logs in idle period of winter compared to application period of summer in the northern region, whereas the total amount was steady with whole-year application in south (p>0.05). The persistent (sul1 and sul2) and elevated ARGs (tetG and ereA) in anaerobic digestion and land application of swine wastewater need more attention. A lab-scale sequencing batch membrane bioreactor (SMBR) was established to treat swine wastewater. The results showed that SRT was the biggest individual impacting factor, who contributed to ARGs variation on 17.4%. Environmental factors (SRT, SS, temperture, COD, NH4+-N, NO2--N, and NO3--N) were the leading contributor on the ARGs variation accounting for 45.8%. At the short SRT of 12, the SMBR showed the highest removal of ARGs copy number at 2.91 logs. The SMBR experiment verified the results of the on-site survey that the abundance of tetG, ereA, sul1 and sul2 increased through biological treatment. More tetG and ereA was increased in the supernant of the SMBR when prolonging the SRT. The ARGs increment and horizontal gene transfer was occurred at the relatively long SRT. The samples in the SMBR including the mixed liquid, the biofilm attached to the membrane, and the supernant was greatly influenced by SRT, while the effluent was little influcenced by SRT. Pot experiment was conducted on the land application of swine wastewater and digestate. The results showed that the disturbance of soil microbial community, ARGs and antibiotic resistance bacteria (ARB) was much greater for slurry than the digestate, while the NPK (chemical fertilizer) and control groups showed the similar variation trends. Soil-R was the most vulnerable, followed by soil-Y, and soil-B had the lowest contamination of ARGs, ARB and fecal bacteria from swine slurry application. tetG, tetM and ermF was the predominant ARGs in slurry amended soils, whose decay fitted to the first-order dynamic reaction, and the decay rates were the order as soil-B > soil-Y > soil-R. The pot experiment verified the results of the on-site survey to some extent that the abundance of tetG and ereA were raised in the soils amended with swine wastewater, especially in soil-Y and soil-R, indicating that tetG and ereA increased and horizontal gene transfer may occur. By partial RDA, soil microbial community (63.7%) was a major contributor compared with the physiochemical parameters (39.8%), with a total explanation of ARGs’ variance at 81.4%.
内容类型: 学位论文
URI标识: http://ir.rcees.ac.cn/handle/311016/38695
Appears in Collections:水污染控制技术研究室_学位论文

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作者单位: 1.中国科学院生态环境研究中心

Recommended Citation:
隋倩雯. 猪场废水生物处理与土地利用过程抗生素抗性基因的转归研究[D]. 北京. 中国科学院大学. 2017.
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