低DO下AGS-SBR处理低COD/N生活污水长期运行特征及种群分析

本研究在序批式活性污泥反应器(SBR)中接种好氧颗粒污泥(AGS),构成AGS-SBR系统,研究其在低DO(0.5~1.0mg·L~(-1))条件下,处理低COD/N比(4.0)生活污水同步脱氮除磷的长期稳定运行特性,并解析反应器的主要菌群构成.结果表明,在反应器运行的180d里,AGS-SBR系统表现出了良好且稳定的除污能力,反应器对水体中COD、NH~+_4-N、TN和TP平均去除率分别达到87.17%、95.21%、77.05%和91.11%.好氧颗粒污泥沉降性能一直很好,污泥始终保持着完整的颗粒外观和密实紧凑的结构,并没有出现明显的颗粒污泥解体的现象.同时,高通量测序结果表明,变形菌门、厚壁菌门、绿菌门、绿弯菌门和拟杆菌门为SBR-AGS反应器中主要优势菌群.Denitratisoma、Planctomycetaceae、Thauera、Comamonas、Nitrosomonas和Nitrospira是反应器中与脱氮有关菌群;Clostridium和Anaerolinea是除磷相关细菌.     

A fluorescence-based bioassay for aquatic macrophytes and its suitability for effect analysis of non-photosystem II inhibitors

Background, Goals and Scope During the last years the miniaturization of toxicity test systems for rapid and parallel measurements of large quantities of samples has often been discussed. For unicellular algae as well as for aquatic macrophytes, fluorescence-based miniaturized test systems have been introduced to analyze photosystem II (PSII) inhibitors. Nevertheless, high-throughput screening should also guarantee the effect detection of a broad range of toxicants in order to ensure routinely applicable, high-throughput measuring device experiments which can cover a broad range of toxicants and modes of action others than PSII inhibition. Thus, the aim of this study was to establish a fast and reproducible measuring system for non-PSII inhibitors for aquatic macrophyte species to overcome major limitations for use. Methods A newly developed imaging pulse-amplitude-modulated chlorophyll fluorometer (I-PAM) was applied as an effect detector in short-term bioassays with the aquatic macrophyte species Lemna minor. This multiwell-plate based measuring device enabled the incubation and measurement of up to 24 samples in parallel. The chemicals paraquat-dichloride, alizarine and triclosan were chosen as representatives for the toxicant groups of non-PSII herbicides, polycyclic aromatic hydrocarbons (PAHs) and pharmaceuticals and personal care products (PPCPs), which are often detected in the aquatic environment. The I-PAM was used (i) to establish and validate the sensitivity of the test system to the three non-PSII inhibitors, (ii) to compare the test systems with standardized and established biotests for aquatic macrophytes, and (iii) to define necessary time scales in aquatic macrophyte testing. For validation of the fluorescence-based assay, the standard growth test with L. minor (ISO/DIS 20079) was performed in parallel for each chemical. Results The results revealed that fluorescence-based measurements with the I-PAM allow rapid and parallel analysis of large amounts of aquatic macrophyte samples. The I-PAM enabled the recording of concentration-effect-curves with L. minor samples on a 24-well plate with single measurements. Fluorescence-based concentration-effect-curves could be detected for all three chemicals after only 1 h of incubation. After 4–5 h incubation time, the maximum inhibition of fluorescence showed an 80–100% effect for the chemicals tested. The EC50 after 24 h incubation were estimated to be 0.06 mg/L, 0.84 mg/L and 1.69 mg/L for paraquatdichloride, alizarine and triclosan, respectively. Discussion The results obtained with the I-PAM after 24 h for the herbicide paraquat-dichloride and the polycyclic aromatic hydrocarbon alizarine were in good accordance with median effective concentrations (EC50s) obtained by the standardized growth test for L. minor after 7 d incubation (0.09 mg/L and 0.79 mg/L for paraquat-dichloride and alizarine, respectively). Those results were in accordance with literature findings for the two chemicals. In contrast, fluorescence-based EC50 of the antimicrobial agent triclosan proved to be two orders of magnitude greater when compared to the standard growth test with 7 d incubation time (0.026 mg/L) as well as with literature findings. Conclusion Typically, aquatic macrophyte testing is very time consuming and relies on laborious experimental set-ups. The I-PAM measuring device enabled fast effect screening for the three chemicals tested. While established test systems for aquatic macrophytes need incubation times of ≥ 7 d, the I-PAM can detect inhibitory effects much earlier (24 h), even if inhibition of chemicals is not specifically associated with PSII. Thus, the fluorescence-based bioassay with the I-PAM offers a promising approach for the miniaturization and high-throughput testing of chemicals with aquatic macrophytes. For the chemical triclosan, however, the short-term effect prediction with the I-PAM has been shown to be less sensitive than with long-term bioassays, which might be due to physicochemical substance properties such as lipophilicity. Recommendations and Perspectives The results of this study show that the I-PAM represents a promising tool for decreasing the incubation times of aquatic macrophyte toxicity testing to about 24 h as a supplement to existing test batteries. The applicability of this I-PAM bioassay on emergent and submerged aquatic macrophyte species should be investigated in further studies. Regarding considerations that physicochemical properties of the tested substances might play an important role in microplate bioassays, the I-PAM bioassay should either be accompanied by evaluating physicochemical properties modeled from structural information prior to an experimental investigation, or by intensified chemical analyses to identify and determine nominal concentrations of the toxicants tested. The chemicals paraquat-dichloride, alizarine and triclosan were chosen as representatives for the toxicant groups of non-PSII herbicides, PAHs and PPCPs which are often detected in the aquatic environment. Nevertheless, in order to ensure a routinely applicable measuring device, experiments with a broader range of toxicants and samples of surface and/or waste waters are necessary. ESS-Submission Editor: Dr. Markus Hecker (MHecker@Entrix.com)     

北京市4种不同污水处理系统中病原菌变化研究

采用定量PCR技术,对北京市4种不同污水处理系统中大肠杆菌、军团菌和沙门氏菌随工艺及四季的数量变化进行了追踪研究,以评估病原菌的去除效果及污水回用的健康风险.结果发现,大肠杆菌在夏季进水和出水中的浓度分别在107~108copies·m L-1和105copies·m L-1左右;G-A/O系统对大肠杆菌的去除率最高,平均去除率达99.88%.军团菌在4个污水处理系统中进水浓度为104~105copies·m L-1,出水浓度约为104copies·m L-1,其在剩余污泥样品中浓度较高,达到105~106copies·m L-1.沙门氏菌进水浓度较低,为102~103copies·m L-1,且其在多个工艺段样品中未检出.季节变化对于病原菌的去除具有较大影响.研究表明,大肠杆菌在各污水处理系统中均可检出,且其分布具有一定的季节性,夏季的进出水中浓度相对较高;而军团菌和沙门氏菌浓度在各工艺中则并未呈现出明显的季节性变化.G-A/O系统对3种细菌的整体去除效果最为稳定,去除率较高.大肠杆菌在污水处理厂的出水及剩余污泥中浓度仍然较高,此外,冬季出水中也能检测到沙门氏菌的存在,因此,污水处理厂的出水和污泥排放仍存在一定的生态和健康风险.     

贵州喀斯特地区土壤细菌群落结构特征及变化

为探明贵州喀斯特不同植被演替群落下的土壤细菌群落结构及变化特征,本文利用高通量测序技术对5个主要植被演替群落(稀灌草丛、藤刺灌丛、灌木林、乔灌过渡林、乔木林)的根际(竹叶椒)、非根际土壤细菌群落结构及环境因子进行了分析研究。结果表明:贵州喀斯特高原土壤细菌类群主要为变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)、疣微菌门(Verrucomicrobia)、放线菌门(Actinobacteria)、厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)和浮霉菌门(Planctomycetes),其相对丰富度分别为43.35%、12.97%、7.53%、7.12%、6.19%、5.35%、5.05%,未分类类群占7.36%。样品中检测到了较低丰度的广古菌门和泉古菌门。随植被群落演替,根际土壤中变形菌门、厚壁菌门和浮霉菌门丰度逐渐增加;非根际土壤中酸杆菌门和疣微菌门丰度随植被演替逐渐减小。贵州喀斯特高原土壤细菌的影响因子大小为土壤有机碳、土壤总氮、含水量、电导率等,其中土壤有机碳和土壤总氮有显著性影响。     

闽江河口互花米草海向入侵对湿地土壤nirS型反硝化微生物群落结构及多样性的影响

2020年秋季,在闽江河口鳝鱼滩东部的互花米草分布区,由陆向海方向选择互花米草海向入侵前的光滩(MF)、入侵1～2年的互花米草湿地(SAN)和入侵6～7年的互花米草湿地(SA)为研究对象.基于高通量测序技术,探讨了互花米草海向入侵对土壤nir S型反硝化微生物群落结构及多样性的影响.结果表明,变形菌门(Proteobacteria)均是互花米草不同入侵阶段土壤中nir S型反硝化微生物的优势门(90.41%～97.36%),其次是厚壁菌门(Firmicutes)和放线菌门(Actinobacteria),三者在SAN和MF土壤中的丰度均与SA存在显著差异(p<0.05).不同入侵阶段土壤中共有的nir S型反硝化细菌属为假单胞菌属(Pseudomonas)、芽孢杆菌属(Bacillus)、偶氮弧菌属(Azoarcus)、陶厄氏菌属(Thauera)、Sulfurifustis属、副球菌属(Paracoccus)、鲁杰氏菌属(Ruegeria)、红肠命菌属(Rubrivivax)、趋磁螺菌属(Magnetospirillum)和福格斯氏菌属(Vogesella),其中,假单胞菌属是...     

三峡库区消落带落羽杉人工林土壤细菌群落结构多样性及动态变化

消落带是连接陆地与水域的交错地带,具有重要的生态功能,其适生木本植物的种植对消落带土壤的生物地球化学循环起着重要的作用.为探究适生植物对三峡库区消落带环境的适应机制,采用高通量测序技术对消落带退水后适生植物落羽杉(Taxodium distichum)不同生长时期(T1:5月、 T2:7月和T3:9月)的土壤细菌群落组成及多样性进行研究,同时采用PICRUSt2对细菌功能进行预测.结果表明,土壤pH值、硝态氮、铵态氮、土壤蔗糖酶、磷酸酶和脲酶等理化指标随时间变化显著(P<0.05);土壤细菌多样性、丰富度和结构也随时间变化而变化,除Chao1以外,根际土壤细菌的α多样性均表现为：T1>T2>T3,非根际土壤细菌α多样性则表现为：T3>T1>T2. RDA分析表明,影响细菌群落的理化指标主要有土壤pH值、脲酶、铵态氮和硝态氮.所有土壤样本中共检测到细菌60门,其中以变形菌门和酸杆菌门为优势细菌门.根据PICRUSt2预测,代谢是落羽杉土壤细菌群落中普遍存在的基本功能,与C、 N和P有关的各代谢途径存在一定的时间差异.以上研究结果有助于加深对三峡消落带植被修复...     

长期覆膜条件下农田土壤微生物群落的响应特征

地膜覆盖是农业生产中保障粮食增产增收的重要措施.为明确长期地膜覆盖对农田土壤微生物群落结构特征的影响,采集4个不同覆膜年限的农田土壤,利用高通量测序技术分析土壤细菌和真菌群落结构变化,探讨长期覆膜农田土壤中微生物群落的变化及其对微生物生态环境效应的影响.结果表明,长期覆膜对土壤细菌多样性无显著影响,但降低真菌多样性;长期覆膜使土壤细菌酸杆菌（Acidobacteriota）和真菌被孢霉菌（Mortierellomycetes）物种丰度降低,增加土壤放线菌（Actinobacteriota）物种丰度.长期覆膜可以使土壤富集细菌中的芽孢杆菌（Bacillus）和类诺卡氏菌（Nocardioidaceae）,及真菌中的肉座菌目（Hypocreales）和曲霉菌（Aspergillus）等有益微生物菌群.然而长期覆膜使土壤真菌共生网络变得简单而脆弱,其关键物种仅有子囊菌门中的粪壳菌目（Sordariales）中的未知菌属一种,因此对农田土壤生态环境带来潜在风险.本研究为深化了解长期覆膜对农田微生物生态环境效应的影响提供理论依据.     

丹江口水库真核浮游植物群落分布特征及其与环境因子的关系

浮游植物是湖库生态系统物质循环和能量流动的主要参与者,了解其群落组成对保护水生生态系统平衡具有重要意义.2021年6月在南水北调中线工程水源地丹江口水库淅川库区设置7个采样点,分4层分别在0.5、5、10和20 m处共采取28个样品.采用高通量测序测定真核浮游植物群落结构,分析真核浮游植物群落在水平和垂向上的分布特征.采用冗余分析探讨了浮游植物主要类群与环境因子的关系.结果表明：①库区真核浮游植物共12门68属,物种为甲藻-硅藻-绿藻型结构.水平方向上浮游植物群落差异较大,各采样点垂直方向上浮游植物丰度和多样性存在较大差异.宋岗、土门和党子口浮游植物群落多样性和丰度随水深增加呈下降趋势,其他采样点的浮游植物群落与水深关系趋势不明显.②硝氮（NO₃^--N）、水深、溶解氧（DO）、pH和水温（WT）是影响库区浮游植物垂直分布的重要环境因子.环境因子对浮游植物门类影响各异,甲藻门与NO₃^--N和水深呈负相关,与其他环境因子呈正相关;硅藻门与NO₃^--N和水深呈正相关,与其他环境因子呈负相关;绿藻门与WT、pH和DO呈负相关,与水深无明显相关性.     

嘉陵江滨岸带不同土地利用类型土壤真菌群落结构与功能多样性

通过明确嘉陵江滨岸带不同土地利用类型土壤真菌群落多样性的差异,为嘉陵江生态环境保护修复提供理论依据.选择嘉陵江中下游滨岸带的人工湿地、天然湿地、林地和农田这4种典型土地利用类型作为研究样地,利用高通量测序技术对土壤真菌群落进行测序,分析不同土地利用类型对土壤真菌群落多样性、结构和功能的影响.结果表明,林地与天然湿地土壤真菌的Chao1指数显著高于其他两种土地利用类型（P<0.05）,林地土壤真菌的Shannon指数显著高于农田和人工湿地两种土地利用类型（P<0.05）;嘉陵江流域滨岸带土壤真菌被划分为15个菌门,在门分类水平上的优势真菌群落（相对丰度>0.01）分别为：子囊菌门（Ascomycota）、担子菌门（Basidiomycota）、油壶菌门（Olpidiomycota）、罗兹菌门（Rozellomycota）、被孢霉门（Mortierellomycota）和壶菌门（Chytridiomycota）;相比之下,罗兹菌门偏好选择林地生境,而油壶菌门和被孢霉门偏好选择农田,担子菌门在人工湿地具有显著优势;天然湿地中的优势功能类群为植物病原菌,人工湿地中的优势功能类群为粪腐菌,农田中优势功能类群为动物病原菌-真菌寄生菌;冗余分析表明,土壤含水率（MC）、全氮（TN）、有机碳（TOC）和碱解氮（AN）是影响真菌群落变化的主要环境因子.由此可见,林地是嘉陵江流域真菌多样性最高和功能类群最均衡的土地利用类型,其次为天然湿地,人为干扰导致嘉陵江滨岸带土壤真菌群落多样性水平降低.     

氮添加对不同坡度退化高寒草甸土壤真菌多样性的影响

为明确不同坡度退化高寒草甸土壤真菌多样性对外源氮添加的响应规律,选取三江源区果洛州不同坡度退化程度相近的高寒草甸,进行外源氮添加试验,探讨不同氮添加水平：低等量氮添加（LN,2 g ·m^-2）、中等量氮添加（MN,5 g ·m^-2）和高等量氮添加（HN,10 g ·m^-2）对不同坡度退化草地土壤真菌多样性的影响.结果表明：①土壤中真菌分布类群集中于子囊菌门（Ascomycota）、担子菌门（Basidiomycota）、被孢霉门（Mortierellomycota）和球囊菌门（Glomeromycota）,其中优势菌门为子囊菌门和担子菌门.子囊菌门、根肿黑粉门和油壶菌门等相对丰度在不同坡度间差异显著（P<0.05）.而同一坡度不同氮添加处理下真菌各门丰度变化差异均不显著.②优势属为被孢霉属和Archaeorhizomyces,它们对不同坡度和不同氮添加水平的响应无差异性.③已鉴定95个菌属在不同坡度间存在显著差异（P<0.05）.④一些相对丰度<1%的属在同一坡度不同水平氮添加间存在显著差异（P<0.05）.5 α和β多样性分析整体表现为在不同坡度和不同氮添加水平下土壤真菌群落无差异性.表明退化高寒草甸土壤真菌对外源氮添加的响应不敏感.