基于微生物完整性指数的苏州市湿地健康评价

为建立在不同区域和不同湿地等大尺度范围内M-IBI（微生物生物完整性指数）评价方法体系,以苏州市15个湿地公园作为研究对象,构建符合区域性特征的生物完整性评价体系并进行应用研究.分别于2015年、2016年和2017年每年的7月对苏州市15个湿地公园的入流口和出流口水质进行采样调查,将采样点分为对照点、受损点和修复点,基于湿地水环境中微生物群落Illumina高通量测序技术,依据候选指标分布范围检验、敏感性分析和相关性检验,筛选出香农指数、前2优势分类单元相对丰度（相对丰度最多的2个属）总和、高温耐受属相对丰度、污染耐受属相对丰度、TN（总氮）敏感属相对丰度5个指标,提出了参数标准化公式和健康评价标准,依此将苏州市湿地健康状态划分为健康、亚健康、一般、较差、差5个等级.结果表明:①在10个受损点3 a所采集的30个样品中,有2个处于较差水平,有5个处于一般水平,有18个处于亚健康水平,有5个处于健康水平;而对于修复点,有23个样点处于健康水平,7个样点处于亚健康水平.②在农业面源污染较多的湿地入流口健康状况最差,工业用水汇入较多的湿地入流口次之,而城市景观水汇入的湿地入流口的健康状况最好.研究显示,M-IBI可以较合理地评价苏州市湿地生态系统健康状况,湿地对水环境的修复作用明显,经修复后的水体均处于健康或亚健康水平.      

植物微生物燃料电池修复Cr(Ⅵ)污染湿地土壤及机理研究

本研究通过引入沼生植物香蒲构建植物微生物燃料电池系统(P-MFC)修复Cr(VI)污染湿地土壤,考察了植物、不同初始Cr(VI)浓度对系统产电及去除效率的影响.结果显示,香蒲种植能显著提高P-MFC运行性能,系统最大功率密度与Cr(VI)去除率分别提高至23.83 mW·m~(-2)、33.01%,随着Cr(VI)暴露浓度的升高,系统运行性能降低.利用P-MFC修复Cr(VI)污染土壤过程中,电化学还原作用是Cr(VI)去除的主要机制,近90%的Cr(VI)通过电化学还原去除,系统中0.3%～1.86%的Cr(VI)被香蒲吸收富集,3.5%～9.5%的Cr(VI)被微生物与还原性有机物直接还原.通过高通量测序技术分析发现,香蒲种植与低浓度Cr(VI)暴露下阳极微生物群落多样性较大,优势门类Proteobacteria相对丰度最高为63.9%,较未种植香蒲与高浓度Cr(VI)暴露条件下提高了3.4%～19.0%,电化学活性微生物Geobacter相对丰度最高为12.4%,较未种植香蒲与高浓度Cr(VI)暴露条件下提高了4.4%～6.8%.系统中对Cr(VI)具有较强耐受性与还原能力的Acinetobacter、Bacillus占有较大比例,且相对丰度随暴露浓度升高而增大,最高分别为19.0%、14.4%,进一步说明微生物群落在Cr(VI)去除上发挥了一定作用.上述结果表明,P-MFC在去除湿地土壤Cr(VI)污染方面具有良好的潜力.     

Dynamic membrane bioreactor performance enhancement by powdered activated carbon addition: Evaluation of sludge morphological, aggregative and microbial properties

The effects of powdered activated carbon(PAC) addition on sludge morphological, aggregative and microbial properties in a dynamic membrane bioreactor(DMBR) were investigated to explore the enhancement mechanism of pollutants removal and filtration performance. Sludge properties were analyzed through various analytical measurements. The results showed that the improved sludge aggregation ability and the evolution of microbial communities affected sludge morphology in PAC-DMBR, as evidenced by the formation of large, regularly shaped and strengthened sludge flocs. The modifications of sludge characteristics promoted the formation process and filtration flux of the dynamic membrane(DM) layer. Additionally, PAC addition did not exert very significant influence on the propagation of eukaryotes(protists and metazoans)and microbial metabolic activity. High-throughput pyrosequencing results indicated that adding PAC improved the bacterial diversity in activated sludge, as PAC addition brought about additional microenvironment in the form of biological PAC(BPAC), which promoted the enrichment of Acinetobacter(13.9%), Comamonas(2.9%), Flavobacterium(0.31%) and Pseudomonas(0.62%), all contributing to sludge flocs formation and several(such as Acinetobacter) capable of biodegrading relatively complex organics. Therefore, PAC addition could favorably modify sludge properties from various aspects and thus enhance the DMBR performance.     

PCR-DGGE技术解析固体碳源表面生物膜的微生物群落结构

以聚乳酸/聚羟基丁酸戊酸共聚酯(PLA/PHBV)作为填充床反应器的碳源和生物膜载体,对受硝酸盐污染的水进行生物反硝化脱氮.采用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术分析了PLA/PHBV表面生物膜中微生物群落的结构和动态变化.结果表明,反应器运行初期,生物膜中微生物多样性下降.当反应器稳定运行时,DGGE图谱特征条带的香农威尔指数和辛普森指数均变化不大,微生物群落结构保持相对稳定.DGGE图谱特征条带的16S rDNA序列分析及扫描电镜分析的结果表明,生物膜中的主要微生物均为革兰氏阴性杆菌,包括Diaphorobacter、Acidovorax、Rubrivivax、Azospira、Thermomonas和Devosia,它们分别属于变形菌门(Proteobacteria)的α-,β-和γ-变形菌纲,其中Diaphorobacter为反应器稳定运行期生物膜中丰度最高的菌群.     

太子河着生藻类群落结构空间分布特征

以辽宁省太子河流域为研究范例,调查了全流域内69个采样点的着生藻类群落和水环境理化特征,并且以着生藻类的群落结构为基础,对太子河流域的水生态系统类型区进行了划分. 结果表明,太子河流域着生藻类群落结构具有明显的空间异质性. CCA(典范对应分析)结果显示,驱动全流域着生藻类群落结构形成的水环境因子为电导率、ρ(TDS)(TDS为总溶解固体)和ρ(TN). 基于着生藻类的群落结构特征,将太子河流域划分为3个着生藻类类型区:第1分区为太子河上游的森林多水区,第2分区为中游低山丘陵森林区,第3分区为下游平原农业区. CCA结果还显示,显著影响第1分区、第2分区及第3分区的着生藻类群落结构的环境因子分别为ρ(NO₃--N)、ρ(TN)及ρ(总溶解固体)和硬度.      

不同植被群落表层土壤中细菌群落多样性

对3种不同植被群落(海桐、雪松、桂花)表层土壤中的pH、w(TOC)、w(TN)和w(TP)进行了测定,采用DGGE(变性梯度凝胶电泳)方法对各植被群落表层土壤细菌的16S rDNA V3~V6可变区扩增片段进行了分析,利用分析得到的图谱数据与表层土壤中的pH、w(TOC)、w(TN)和w(TP)进行了群落-理化因子相关性分析以及CCA(典范对应分析). 结果表明,不同植被群落表层土壤的pH和养分含量不同,其中w(TOC)和w(TN)的差异显著(P<0.05),而pH和w(TP)的差异不显著(P>0.05). DGGE图谱分析结果揭示,不同植被群落表层土壤的细菌群落结构明显不同. 聚类分析显示,海桐和雪松的细菌群落组成和结构最为相似(82%),而它们与桂花样地的相似性(55%)相对较低,桂花样地的Shannon-Wiener指数最高且与其他2个样地差异显著,雪松样地的均匀度指数最高. 相关性分析和CCA结果进一步表明,w(TOC)和w(TN)是表层土壤细菌群落结构和多样性的显著影响因素,并且植被群落可能通过其根系分泌物产生间接影响.      

Absolute dominance of hydrogenotrophic methanogens in full-scale anaerobic sewage sludge digesters

Anaerobic digestion （AD） is gaining increasing attention due to the ability to covert organic pollutants into energy-rich biogas and, accordingly, growing interest is paid to the microbial ecology of AD systems. Despite extensive efforts, AD microbial ecology is still limitedly understood, especially due to the lack of quantitative information on the structures and dynamics of AD microbial communities. Such knowledge gap is particularly pronounced in sewage sludge AD processes although treating sewage sludge is among the major practical applications of AD. Therefore, we examined the microbial communities in three full-scale sewage sludge digesters using qualitative and quantitative molecular techniques in combination： denaturing gradient gel electrophoresis （DGGE） and real-time polymerase chain reaction （PCR）. Eight out of eleven bacterial sequences retrieved from the DGGE analysis were not affiliated to any known species while all eleven archaeal sequences were assigned to known methanogen species. Quantitative real-time PCR analysis revealed that, based on the 16S rRNA gene abundance, the hydrogenotrophic order Methanomicrobiales is the most dominant methanogen group （〉 94% of the total methanogen population） in all digesters. This corresponds well to the prevailing occurrence of the DGGE bands related to Methanolinea and Methanospirillum, both belonging to the order Methanomicrobiales, in all sludge samples. It is therefore suggested that hydrogenotrophic methanogens, especially Methanomicrobiales strains, are likely the major players responsible for biogas production in the digesters studied. Our observation is contrary to the conventional understanding that aceticlastic methanogens generally dominate methanogen communities in stable AD environments, suggesting the need for further studies on the dominance relationship in various AD systems.     

Influence of fuel moisture, charge size, feeding rate and air ventilation conditions on the emissions of PM, OC, EC, parent PAHs, and their derivatives from residential wood combustion

Controlled combustion experiments were conducted to investigate the influence of fuel charge size, moisture, air ventilation and feeding rate on the emission factors (EFs) of carbonaceous particulate matter, parent polycyclic aromatic hydrocarbons (pPAHs) and their derivatives from residential wood combustion in a typical brick cooking stove. Measured EFs were found to be independent of fuel charge size, but increased with increasing fuel moisture. Pollution emissions from the normal burning under an adequate air supply condition were the lowest for most pollutants, while more pollutants were emitted when an oxygen deficient atmosphere was formed in the stove chamber during fast burning. The impacts of these factors on the size distribution of emitted particles was also studied. Modified combustion efficiency and the four investigated factors explained 68%, 72%, and 64% of total variations in EFs of PM, organic carbon, and oxygenated PAHs, respectively, but only 36%, 38% and 42% of the total variations in EFs of elemental carbon, pPAHs and nitro-PAHs, respectively.     

Abundance and diversity of ammonia-oxidizing archaea in response to various habitats in Pearl River Delta of China, a subtropical maritime zone

Ammonia-oxidizing archaea (AOA) are widely considered key to ammonia oxidation in various environments. However, little work has been conducted to simultaneously investigate the abundance and diversity of AOA as well as correlations between archaeal amoA genotypes and environmental parameters of different ecosystems at one district. To understand the abundance, diversity, and distribution of AOA in Pearl River Delta of China in response to various habitats, the archaeal amoA genes in soil, marine, river, lake, hot spring and wastewater treatment plant (WWTP) samples were investigated using real-time fluorescent quantitative PCR and clone libraries. Our analyses indicated that the diversity of AOA in various habitats was different and could be clustered into five major clades, i.e., estuary sediment, marine water/sediment, soil, hot spring and Cluster 1. Phylogenetic analyses revealed that the structure of AOA communities in similar ecological habitats exhibited strong relation. The canonical correspondence method indicated that the AOA community structure was strongly correlated to temperature, pH, total organic carbon, total nitrogen and dissolved oxygen variables. Assessing AOA amoA gene copy numbers, ranging from 6.84×10⁶ to 9.45×10⁷ copies/g in dry soil/sediment, and 6.06×10⁶ to 2.41×10⁷ copies/L in water samples, were higher than ammonia-oxidizing bacteria (AOB) by 1-2 orders of magnitude. However, AOA amoA copy numbers were much lower than AOB in WWTP activated sludge samples. Overall, these studies suggested that AOA may be a major contributor to ammonia oxidation in natural habitats but play a minor role in highly aerated activated sludge. The result also showed the ratio of AOA to AOB amoA gene abundance was positively correlated with temperature and less correlated with other environmental parameters. New data from our study provide increasing evidence for the relative abundance and diversity of ammonia-oxidizing archaea in the global nitrogen cycle.     

长期秸秆还田对潮土真菌群落、酶活性和小麦产量的影响

为了研究秸秆还田对黄淮海平原潮土真菌群落的影响,开展连续10 a田间定位试验,设置不施肥（NF）、单施化肥（NPK）和秸秆还田配施化肥（NPKS）这3个处理,通过网络分析和结构方程模型,揭示真菌群落对土壤肥力、酶活性和小麦产量的影响机制.结果发现,NPKS处理土壤有机质（SOM）较NPK处理和NF处理分别升高9.20%和34.75%,碱解氮（AN）升高12.03%和39.17%,脱氢酶（DHA）升高37.21%和50.91%,β-葡糖苷酶（β-GC）升高17.29%和73.48%,小麦增产16.22%和125.53%.不同施肥处理真菌α-多样性并未发生显著变化,却导致β-多样性出现明显分异.冗余分析表明,速效磷（AP）、SOM和AN是潮土真菌群落组成变化的主要调控因素.差异物种分析表明,NF处理富集了Mortierella、Aspergillus、Ceriporia和Acremonium等具有溶磷/解钾功能的物种,以及Leohumicola和Hyalodendriella等植物共生菌;NPK处理中Sarocladium、Fusarium和Fusicolla等植物病原菌丰度显著升高;NPKS处理则激发了Pseudogymnoascus和Schizothecium等抑病菌生长,并提高Trichocladium和Lobulomyces等秸秆降解物种丰度.网络分析发现整个网络由4个主要模块组成,其中模块2物种累积丰度在NPKS处理显著升高,且与DHA和β-GC呈线性正相关关系.结构方程模型结果进一步表明小麦产量主要受SOM直接正向调控,而模块2物种可通过正向调节DHA和β-GC,间接影响SOM和小麦产量.综上所述,黄淮海平原潮土区秸秆还田可通过调节真菌种间互作关系,刺激特定物种集群生长,抑制病原菌活性,提高土壤酶活性,促进SOM累积,最终获得作物高产.