成果

2018
苏志国, 张衍, 代天娇, 陈嘉瑜, 张永明, 温东辉. 环境中抗生素抗性基因与I型整合子的研究进展. 微生物学通报 [Internet]. 2018;45(10):2217~2233. 访问链接AbstractPKU 
 抗生素抗性基因(Antibiotic resistance genes,ARGs)作为一种新型污染物在不同环境中广泛分布、来源复杂,对生态环境和人类健康造成了很大的潜在风险。同时,I型整合子(Int I)介导的ARGs水平转移是环境中微生物产生耐药性的重要途径,I型整合子整合酶基因(intI1)与ARGs丰度在环境中表现出了较高的正相关性,Int I可以作为标记物在一定程度上反映ARGs在环境中的迁移转化规律和人类活动影响程度。本文介绍ARGs与Int I在环境中的来源与分布,总结Int I介导的ARGs迁移转化机制以及相关研究方法,并展望未来的研究发展趋势。
2017
何静林, 温东辉, 高波. 杭州萧山区饮用水源地铁污染评价及控制对策研究. 中国环境监测 [Internet]. 2017;33(1):97-105. 访问链接AbstractPKU 
以杭州市萧山区2008—2014年饮用水源地铁含量监测数据为分析对象,研究饮用水源地的铁含量时空分布并评价铁污染状况,结果表明,铁含量单次测值历年超标,超标倍数0.01~8.80,超标率16.7%~86.1%,7年间超标率呈现阶段性的螺旋式上升,铁污染日趋严重,到2014年有所减轻。通过对饮用水源地周边环境和污染源调查,结合文献资料,分析铁污染成因,提出防治对策:制定地表水体中铁的分类标准,完善废水的铁排放标准;建立跨行政区域的水环境管理机制,全面实施河长制;加强水土保持和酸雨控制,进行河流综合整治;加强饮用水源管理,扩建备用水源和另找水源。
Zhang Y, Li A, Dai T, Li F, Xie H, Chen L, Wen D. Cell-free DNA: a neglected source for antibiotic resistance genes spreading from WWTPs. Environmental Science & Technology [Internet]. 2017;52(1):248-257. 访问链接Abstract
Cell-associated ARGs in wastewater treatment plants (WWTPs) has been concerned, however, cell-free ARGs in WWTPs was rarely studied. In this study, the abundances of four representative ARGs, sulII, tetC, blaPSE‑1,and ermB, in a large municipal WWTP were investigated in both cell-associated and cell-free fractions. Cell-associated ARGs was the dominant ARGs fraction in the raw wastewater. After biological treatment, sludge settling, membrane filtration, and disinfection, cell-associated ARGs were substantially reduced, though the ratios of ARG/16S rRNA gene were increased with disinfection. Cell-free ARGs persisted in the WWTP with a removal of 0.36 log to 2.68 logs, which was much lower than the removal of cell-associated ARGs (3.21 logs to 4.14 logs). Therefore, the abundance ratio of cell-free ARGs to cell-associated ARGs increased from 0.04−1.59% to 2.00−1895.08% along the treatment processes. After 25-day-storage, cell-free ARGs in both biological effluent and disinfection effluent increased by 0.14 log to 1.99 logs and 0.12 log to 1.77 logs respectively, reflecting the persistence and low decay rate of cell-free ARGs in the discharge water. Therefore, cell-free ARGs might be a kind of important but previously neglected pollutant from WWTPs, which added potential risks to the effluent receiving environments.
张衍, 陈吕军, 谢辉, 李奥林, 代天娇. 两座污水处理系统中细胞态和游离态抗生素抗性基因的丰度特征. 环境科学 [Internet]. 2017;38(9):3823-3830. 访问链接AbstractPKU 
为探究污水处理系统中抗生素抗性基因(ARGs)、特别是胞外游离态ARGs的赋存特征,本研究选取生活污水处理系统和工业废水处理系统各一座,采用荧光定量PCR对细胞态及游离态ARGs丰度变化开展研究.在生活污水处理系统M中,进水sulⅡ、tet C、bla PSE-1erm B这4种ARGs细胞态的绝对丰度均大幅高于游离态的绝对丰度,生物处理未对抗生素抗性菌(ARBs)产生富集效应;MBR的超滤膜有效削减了水中细胞态和游离态DNA,最终ARGs的总去除率为2.54~4.95 logs.在焦化废水处理系统C中,生物处理对携带sulⅡ的ARBs产生了富集效应,但游离态sulⅡ的相对丰度和绝对丰度均有所降低;其后混凝-砂滤工艺使水中细胞态和游离态sulⅡ的绝对丰度分别出现了下降和上升,游离态sulⅡ在总sulⅡ中的比例从生物处理出水中的0.05%,上升到混凝-砂滤出水中的1.33%,并在25℃恒温避光静置5d后进一步上升至9.31%.ARBs深度去除及残留细胞裂解,使污水处理系统出水中游离ARGs在总ARGs中的比例有所上升.游离态ARGs介导ARGs在污水处理系统出水受纳环境中的传播扩散风险有待后续研究进行深入评估.
陶怡乐, 张晨枫, 徐琳, 温东辉. 北京大学未名湖水质与底泥菌群结构的时空变化. 北京大学学报(自然科学版) [Internet]. 2017;53(6):1150-1160. 访问链接Abstract
通过测定不同季节北京大学未名湖不同区域的水质指标,发现在无显著人为外源影响的条件下,水体氮相关指标较好,四季全部采样点的氨氮都优于Ⅱ类水指标,总氮都优于Ⅳ类水指标,但COD,TP和DO等指标在有的采样点仅达到、甚至劣于Ⅴ类。未名湖的平均综合营养状态指数(TLI)为56.1,属于轻度富营养化。利用PCR-DGGE分析未名湖的底泥微生物群落,结果表明其多样性随季节变化较明显,春、秋季多样性指数都较高,夏、秋季多样性指数与地理位置及水体营养盐含量呈显著正相关关系。线性回归结果提示未名湖底泥微生物群落可能受藻类固氮作用的影响。
方元狄, 张静, 郑中原, 温东辉. 焦化废水处理试验系统出水的生物毒性变化. 生态毒理学报 [Internet]. 2017;12(3):317-326. 访问链接AbstractPKU 
焦化废水是一种典型的难降解工业废水,组分复杂,生物毒性高,大多采用生物处理联合物化深度处理的工艺,以满足炼焦化学工业的污染排放标准,但其排水安全性仍然令人担忧。为研究工艺排水安全性,选择发光细菌青海弧菌Q67、稀有鮈鲫(Gobiocypris rasus)血红细胞、活性污泥微生物群落为测试生物,研究了焦化废水及各处理阶段出水的急性毒性和遗传毒性变化,进而识别影响生物毒性的水质因子。焦化废水经过序批式生物膜反应器处理后,出水急性毒性比进水下降71%,遗传毒性下降为90%以上的轻度以下损伤,显示生物强化处理对焦化废水生物毒性有良好的去除作用。生物处理出水再经过深度处理后,则表现出不同的毒性变化:活性炭吸附法对生物急性毒性的消除最佳,但遗传毒性较生物处理出水有所升高;臭氧氧化法不仅水质改善效率差,且最终出水的生物急性毒性与遗传毒性均升高;臭氧催化氧化法对水中残留有机物去除效率较高,但也造成出水急性毒性与遗传毒性的升高。各水样对青海弧菌Q67的急性毒性与有机物、氮等水质指标表现出较强相关性,而遗传毒性与水质指标之间的相关性不显著。研究结果可为评价和改进处理工艺、保障水体生态安全提供参考。
2016
Dai T, Zhang Y, Tang Y, Bai Y, Tao Y, Huang B, Wen D. Identifying the key taxonomic categories that characterize microbial community diversity using full-scale classification: a case study of microbial communities in the sediments of Hangzhou Bay. FEMS Microbiology Ecology [Internet]. 2016;92(10):fiw150. 访问链接Abstract
Coastal areas are land–sea transitional zones with complex natural and anthropogenic disturbances. Microorganisms in coastal sediments adapt to such disturbances both individually and as a community. The microbial community structure changes spatially and temporally under environmental stress. In this study, we investigated the microbial community structure in the sediments of Hangzhou Bay, a seriously polluted bay in China. In order to identify the roles and contribution of all microbial taxa, we set thresholds as 0.1% for rare taxa and 1% for abundant taxa, and classified all operational taxonomic units into six exclusive categories based on their abundance. The results showed that the key taxa in differentiating the communities are abundant taxa (AT), conditionally abundant taxa (CAT), and conditionally rare or abundant taxa (CRAT). A large population in conditionally rare taxa (CRT) made this category collectively significant in differentiating the communities. Both bacteria and archaea demonstrated a distance decay pattern of community similarity in the bay, and this pattern was strengthened by rare taxa, CRT and CRAT, but weakened by AT and CAT. This implied that the low abundance taxa were more deterministically distributed, while the high abundance taxa were more ubiquitously distributed.
Yu C, Li X, Zhang N, Wen D, Liu C, Li Q. Inhibition of biofilm formation by D-tyrosine: Effect of bacterial type and D-tyrosine concentration. Water research [Internet]. 2016;92:173-9. 访问链接Abstract
D-Tyrosine inhibits formation and triggers disassembly of bacterial biofilm and has been proposed for biofouling control applications. This study probes the impact of D-tyrosine in different biofilm formation stages in both G+ and G- bacteria, and reveals a non-monotonic correlation between D-tyrosine concentration and biofilm inhibition effect. In the attachment stage, cell adhesion was studied in a flow chamber, where D-tyrosine caused significant reduction in cell attachment. Biofilms formed by Pseudomonas aeruginosa and Bacillus subtilis were characterized by confocal laser scanning microscopy as well as quantitative analysis of cellular biomass and extracellular polymeric substances. D-Tyrosine exhibited strong inhibitive effects on both biofilms with  an effective concentration as low as 5 nM; the biofilms responded to D-tyrosine concentration change in a non-monotonic, bi-modal pattern. In addition, D-tyrosine showed notable and different impact on EPS production by G+ and G- bacteria. Extracellular protein was decreased in P. aeruginosa biofilms, but increased in those of B. subtilis. Exopolysaccharides production by P. aeruginosa was increased at low concentrations and reduced at high concentrations while no impact was found in B. subtilis. These results suggest that distinct mechanisms are at play at different D-tyrosine concentrations and they may be species specific. Dosage of D-tyrosine must be carefully controlled for biofouling control applications.
Zhang Y, Chen L, Sun R, Dai T, Tian J, Zheng W, Wen D. Population and diversity of ammonia-oxidizing archaea and bacteria in a pollutants’ receiving area in Hangzhou Bay. Applied Microbiology and Biotechnology [Internet]. 2016;100(13):6035-6045. 访问链接Abstract
The community structure of ammonia-oxidizing microorganisms is sensitive to various environmental factors, including pollutions. In this study, real-time PCR and 454 pyrosequencing were adopted to investigate the population and diversity of ammonia-oxidizing archaea (AOA) and bacteria (AOB) temporally and spatially in the sediments of an industrial effluent receiving area in the Qiantang River’s estuary, Hangzhou Bay. The abundances of AOA and AOB amoA genes fluctuated in 105–107 gene copies per gram of sediment; the ratio of AOA amoA/AOB amoA ranged in 0.39–5.52. The AOA amoA/archaeal 16S rRNA, AOB amoA/bacterial 16S rRNA, and AOA amoA/AOB amoA were found to positively correlate with NH4+-N concentration of the seawater. Nitrosopumilus cluster and Nitrosomonas-like cluster were the dominant AOA and AOB, respectively. The community structures of both AOA and AOB in the sediments exhibited significant seasonal differences rather than spatial changes in the effluent receiving area. The phylogenetic distribution of AOB in this area was consistent with the wastewater treatment plants (WWTPs) discharging the effluent but differed from the Qiantang River and other estuaries, which might be an outcome of long-term effluent discharge.
Zhang Y, Chen L, Sun R, Dai T, Tian J, Zheng W, Wen D. Temporal and spatial changes of microbial community in an industrial effluent receiving area in Hangzhou Bay. Journal of Environmental Sciences [Internet]. 2016;44:57-68. 访问链接Abstract
Anthropogenic activities usually contaminate water environments, and have led to the eutrophication of many estuaries and shifts in microbial communities. In this study, the temporal and spatial changes of the microbial community in an industrial effluent receiving area in Hangzhou Bay were investigated by 454 pyrosequencing. The bacterial community showed higher richness and biodiversity than the archaeal community in all sediments. Proteobacteria dominated in the bacterial communities of all the samples; Marine_Group_I and Methanomicrobia were the two dominant archaeal classes in the effluent receiving area. PCoA and ANOVA revealed strong seasonal but minor spatial changes in both bacterial and archaeal communities in the sediments. The seasonal changes of the bacterial community were less significant than those of the archaeal community, which mainly consisted of fluctuations inabundance of a large proportion of longstanding species rather than the appearance and disappearance ofmajor archaeal species. Temperaturewas found to positively correlatewith the dominant bacteria, Betaproteobacteria, and negatively correlate with the dominant archaea,Marine_Group_I; and might be the primary driving force for the seasonal variation of the microbial community.
张楠, 熊富忠, 温东辉, 于聪, 李琪琳. 环境因素对降解型生物膜形成的影响. 北京大学学报(自然科学版) [Internet]. 2016;52(2):345-353. 访问链接Abstract
采用改良微孔板法,考察p H、温度、培养时间和目标污染物浓度4个环境因子对3株氮杂环芳烃降解菌成膜的影响。结果表明,p H、温度、培养时间对生物膜的形成影响显著,且各降解菌的最佳成膜条件分别为:BC026成膜的最适p H为7,最适温度为35℃,培养时间为36小时;BW001成膜的最适p H为8,最适温度为35℃,培养时间为48小时;BW004成膜的最适p H为7~9,最适温度为40℃,培养时间为36小时。在0~1600 mg/L的目标污染物浓度内,目标污染物对生物膜形成的影响不显著。
陶怡乐, 温东辉. 细菌硝酸盐异化还原成铵过程及其在河口生态系统中的潜在地位与影响. 微生物学通报 [Internet]. 2016;43(1):172-181. 访问链接AbstractPKU 
细菌硝酸盐异化还原成铵(DNRA)过程能够将河口沉积物中的硝氮转化为氨氮,是河口生态系统中潜在的重要氮循环过程之一。本文介绍DNRA机理与分类,综述河口生态系统中DNRA的地位与影响,并总结河口生态系统中几种重要生态因子对DNRA过程的调控与影响。目前DNRA的机理还有待完善。深入研究各类河口生态系统中环境因子对DNRA的调控与影响机制,并研发新的研究方法,将为我国河口地区的水资源保护和生态治理提供科学依据。
2015
Zhang Y, Chen L, Dai T, Sun R, Wen D. Ammonia manipulates the ammonia-oxidizing archaea and bacteria in the coastal sediment-water microcosms. Applied Microbiology and Biotechnology [Internet]. 2015;99(15):6481-6491. 访问链接
Zhang Y, Chen L, Sun R, Dai T, Tian J, Wen D. Ammonia-oxidizing bacteria and archaea in wastewater treatment plant sludge and nearby coastal sediment in an industrial area in China. Applied Microbiology and Biotechnology [Internet]. 2015;99(10):4495-4507. 访问链接
Xing R, Zheng Z, Wen D. Comparison between UV and VUV photolysis for the pre-and post-treatment of coking wastewater. Journal of Environmental Sciences [Internet]. 2015;29:45-50. 访问链接
Zhang Y, Chen L, Dai T, Tian J, Wen D. The influence of salinity on the abundance, transcriptional activity, and diversity of AOA and AOB in an estuarine sediment: a microcosm study. Applied Microbiology and Biotechnology [Internet]. 2015;99(22):9825-9833. 访问链接
2014
Zhang J, Wen D, Zhao C, Tang X. Bioaugmentation accelerates the shift of bacterial community structure against shock load: a case study of coking wastewater treatment by zeolite-sequencing batch reactor. Applied Microbiology and Biotechnology [Internet]. 2014;98(2):863-873. 访问链接
Chen C, Wen D, Wang J. Cellular surface characteristics of Saccharomyces cerevisiae before and after Ag (I) biosorption. Bioresource Technology [Internet]. 2014;156:380-383. 访问链接
Zhang Y, Chen L, Sun R, Dai T, Tian J, Liu R, Wen D. Effect of wastewater disposal on the bacterial and archaeal community of sea sediment in an industrial area in China. FEMS Microbiology Ecology [Internet]. 2014;88(2):320-332. 访问链接Abstract
Human activities usually leave footprints in the environment. By using 454 pyrosequencing, the impact of effluent from an industrial park on the coastal microecology in Hangzhou Bay, China, was interpreted by analysing the microbial communities of the activated sludge from three wastewater treatment plants and the sediment from the effluent receiving area. Based on richness and biodiversity, the sediments were more diversified than the activated sludge, although the seawater environment was highly contaminated. Both bacterial and archaeal communities were niche-determined. The bacterial phylum Proteobacteria dominated in all samples; and certain pollutant-resistant genera, such as Thauera and Truepera, were found in all samples. Archaeal phyla Euryarchaeota and Thaumarchaeota dominated the activated sludge and sediment samples, respectively. According to the analysis of shared operational taxonomic units (OTUs) and reads among different samples, more bacterial OTUs and reads were shared between two samples from sites with a direct effluent connection, showing a clear correlation between the wastewater treatment plants and the effluent receiving bay area. The impact of second-hand pollution can be evaluated by comparing the bacterial community in different eco-environments with a direct effluent connection, especially when pristine samples are not available.
Yu C, Ma J, Zhang J, Lou J, Wen D, Li Q. Modulating particle adhesion with micro-patterned surfaces. ACS Applied Materials & Interfaces [Internet]. 2014;6(11):8199-8207. 访问链接

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