Application of Ion Torrent Sequencing to the Assessment of the Effect of Alkali Ballast Water Treatment on Microbial Community Diversity

The impact of NaOH as a ballast water treatment (BWT) on microbial community diversity was assessed using the 16S rRNA gene based Ion Torrent sequencing with its new 400 base chemistry. Ballast water samples from a Great Lakes ship were collected from the intake and discharge of both control and NaOH (pH 12) treated tanks and were analyzed in duplicates. One set of duplicates was treated with the membrane-impermeable DNA cross-linking reagent propidium mono-azide (PMA) prior to PCR amplification to differentiate between live and dead microorganisms. Ion Torrent sequencing generated nearly 580,000 reads for 31 bar-coded samples and revealed alterations of the microbial community structure in ballast water that had been treated with NaOH. Rarefaction analysis of the Ion Torrent sequencing data showed that BWT using NaOH significantly decreased microbial community diversity relative to control discharge (p<0.001). UniFrac distance based principal coordinate analysis (PCoA) plots and UPGMA tree analysis revealed that NaOH-treated ballast water microbial communities differed from both intake communities and control discharge communities. After NaOH treatment, bacteria from the genus Alishewanella became dominant in the NaOH-treated samples, accounting for <0.5% of the total reads in intake samples but more than 50% of the reads in the treated discharge samples. The only apparent difference in microbial community structure between PMA-processed and non-PMA samples occurred in intake water samples, which exhibited a significantly higher amount of PMA-sensitive cyanobacteria/chloroplast 16S rRNA than their corresponding non-PMA total DNA samples. The community assembly obtained using Ion Torrent sequencing was comparable to that obtained from a subset of samples that were also subjected to 454 pyrosequencing. This study showed the efficacy of alkali ballast water treatment in reducing ballast water microbial diversity and demonstrated the application of new Ion Torrent sequencing techniques to microbial community studies.     

再生水补水对河道底泥细菌群落结构的影响

以北京市永定河麻峪湿地再生水补水口附近河道底泥为研究对象,应用末端限制性片段长度多态性(Terminal Restriction Fragment Length Polymorphism,T-RFLP)技术探究再生水补水口附近底泥细菌群落结构的空间差异特征以及环境因子所产生的影响,并借助典范对应分析(Canonical Correspondence Analysis,CCA)方法分析麻峪湿地底泥细菌群落结构空间差异特征的形成原因。分析结果表明:河道底泥微生物群落对再生水的净化主要发生在距补水口1200m的范围内,随着再生水与上游来水径向混合净化渐变过程,在补水口2000m处河道底泥微生物群落结构与河道再生水补水口上游趋于相似。基于样点聚类结果的群落结构多样性分析表明随着再生水净化过程的实现微生物综合多样性指数呈现下降的趋势,从均匀度指数的变化趋势看,再生水补水口具有最高均匀度指数,补水口下游1200米处由于非优势菌群所占相对丰度最低以及偶见菌群缺失,导致群落结构比较单一、群落多样性和均匀度指数都最低。CCA方面分析结果表明再生水补水口上游细菌群落与因累积效应形成的重金属有密切关系,补水口出口底泥细菌群落则主要受到总磷和总有机碳的影响较大,而再生水补水与上游来水汇水径向渐变过程可能与氨氮净化具密切关系。研究区的主要优势菌种为假单胞菌属、贪食菌属和芽孢杆菌属,是与再生水中有机碳、氮降解具有密切关系的菌属。     

城市河流改造工程对河流生态系统野生维管植物的影响

随着城市化的发展,许多城市对流经市区的河流进行了大规模工程改造,在获得了多种效益的同时,也引起了诸如生物多样性破坏和物种组成改变等一系列生态问题,因此对城市河流改造工程的生态影响进行研究非常重要,将有助于在未来城市河流改造中更有效地保护河流生态系统。该文以汾河太原段的河道改造工程为研究对象,用Shannon-Weiner 多样性指数、Sørensen群落相似性指数和单因素方差分析方法,研究了河流改造工程对河流生态系统中的野生维管植物物种多样性、频度和群落物种组成相似性的影响。结果表明:太原汾河段改造后比改造前的野生维管植物物种数和Shannon-Weiner 多样性指数显著降低;低频度的物种数明显增加而高频度的物种数明显减少;河流改造前后的群落相似性明显降低。由此可见,城市河流改造工程对河流生态系统中的野生维管植物物种多样性、频度和群落相似性有明显影响。     

Effect of land use on water quality and phytoplankton community in the tropical Khami River in semi-arid southwest Zimbabwe

The water quality and phytoplankton community assemblage of the Khami River, a tropical river sub catchment in semi-arid southwest Zimbabwe impacted by agriculture and urban land use, were examined in March 2015. Conductivity, sulphates, total dissolved solids and salinity were higher at urban sampling points than at agricultural sampling points. In contrast, agricultural sampling points were more turbid, and had a greater content of nitrates than urban sampling points. The phytoplankton community was dominated by Cyanobacteria, mainly Microcystis aeruginosa, with densities of up to 20 times higher at urban than at agricultural sampling points. There was an increasing dominance of Cyanobacteria (M. aerugionosa and Merismopedia glauca) and Chlorophyta (Eudorina elegans, Spirogyra sp. and Pediastrum duplex) and a decreasing importance of Bacillariophyta along the agriculture-to-urban gradient. Given the increasing scarcity of water in southern Africa, not only do our findings confirm the importance of land use types as drivers of water quality and phytoplankton community composition and structure. However, they also show that Cyanobacteria, Chlorophyta and Bacillariophyta can be indicators of changes in water quality, factors that will prove pertinent to management for effective water quality management using phytoplankton composition as bioindicators.     

Phylogenetic Diversity and Ecological Pattern of Ammonia-oxidizing Archaea in the Surface Sediments of the Western Pacific

The phylogenetic diversity of ammonia-oxidizing archaea (AOA) was surveyed in the surface sediments from the northern part of the South China Sea (SCS). The distribution pattern of AOA in the western Pacific was discussed through comparing the SCS with other areas in the western Pacific including Changjiang Estuary and the adjacent East China Sea where high input of anthropogenic nitrogen was evident, the tropical West Pacific Continental Margins close to the Philippines, the deep-sea methane seep sediments in the Okhotsk Sea, the cold deep sea of Northeastern Japan Sea, and the hydrothermal field in the Southern Okinawa Trough. These various environments provide a wide spectrum of physical and chemical conditions for a better understanding of the distribution pattern and diversities of AOA in the western Pacific. Under these different conditions, the distinct community composition between shallow and deep-sea sediments was clearly delineated based on the UniFrac PCoA and Jackknife Environmental Cluster analyses. Phylogenetic analyses showed that a few ammonia-oxidizing archaeal subclades in the marine water column/sediment clade and endemic lineages were indicative phylotypes for some environments. Higher phylogenetic diversity was observed in the Philippines while lower diversity in the hydrothermal vent habitat. Water depth and possibly with other environmental factors could be the main driving forces to shape the phylogenetic diversity of AOA observed, not only in the SCS but also in the whole western Pacific. The multivariate regression tree analysis also supported this observation consistently. Moreover, the functions of current and other climate factors were also discussed in comparison of phylogenetic diversity. The information collectively provides important insights into the ecophysiological requirements of uncultured ammonia-oxidizing archaeal lineages in the western Pacific Ocean.     

人工遮光和营养添加对河流反硝化活性和反硝化细菌群落结构的影响

河流生态系统受到人类活动例如河岸带森林植被毁损和农业活动施肥等的干扰日益加剧,而这些活动使河流接收的光照增多、河流的氮磷营养盐浓度增加。微生物的反硝化作用是河流去除氮的有效途径。在汉江的一级支流金水河上游核心保护区内选取6条溪流开展野外控制实验,利用营养添加模拟河流中营养的增加,遮盖河面来模拟源头溪流的隐蔽状态,来研究河流沉积物中微生物的反硝化作用对光照和营养改变的响应,并利用高通量测序(Mi Seq)技术研究在两种处理下河流沉积物中nir S型反硝化细菌的群落结构变化。结果显示:营养元素添加促进了沉积物的反硝化活性,河面遮盖抑制了沉积物的反硝化活性。营养添加和遮盖两种处理均降低了控制实验区域内脱氯单胞菌属(优势菌属)的相对丰度,同时也降低了该区域nir S型反硝化菌群落的Chao多样性。本研究初步证实了光照增加和河流的营养增加提高了河流沉积物反硝化活性,并为提高河流的脱氮能力提供科学依据。     

Quantitative and qualitative beta diversity measures lead to different insights into factors that structure microbial communities

The assessment of microbial diversity and distribution is a major concern in environmental microbiology. There are two general approaches for measuring community diversity: quantitative measures, which use the abundance of each taxon, and qualitative measures, which use only the presence/absence of data. Quantitative measures are ideally suited to revealing community differences that are due to changes in relative taxon abundance (e.g., when a particular set of taxa flourish because a limiting nutrient source becomes abundant). Qualitative measures are most informative when communities differ primarily by what can live in them (e.g., at high temperatures), in part because abundance information can obscure significant patterns of variation in which taxa are present. We illustrate these principles using two 16S rRNA-based surveys of microbial populations and two phylogenetic measures of community beta diversity: unweighted UniFrac, a qualitative measure, and weighted UniFrac, a new quantitative measure, which we have added to the UniFrac website (http://bmf.colorado.edu/unifrac). These studies considered the relative influences of mineral chemistry, temperature, and geography on microbial community composition in acidic thermal springs in Yellowstone National Park and the influences of obesity and kinship on microbial community composition in the mouse gut. We show that applying qualitative and quantitative measures to the same data set can lead to dramatically different conclusions about the main factors that structure microbial diversity and can provide insight into the nature of community differences. We also demonstrate that both weighted and unweighted UniFrac measurements are robust to the methods used to build the underlying phylogeny.     

城市河流的景观生态学研究:概念框架

城市河流作为城市景观中一种重要的生态廊道,其功能的正常实现与否关系到整个城市的可持续发展。通过分析当前城市河流的研究概况,发现应用景观生态学原理对城市河流展开多尺度、多学科的综合研究是实现“自然-人类-水体”可持续发展的必然趋势。从景观生态学的角度出发,结合城市河流的特点,提出了更为综合的、景观水平上的城市河流研究的概念框架。特别针对景观生态学研究的核心问题,对城市河流的研究尺度、格局分析、干扰程度等重要方面进行了详细论述,以期在景观水平上构建城市河流的可持续发展预案。     

基于eDNA metabarcoding探究北京市主要河流鱼类分布及影响因素

使用eDNA宏条形码(eDNA metabarcoding)和地笼法检测了北京市3条水系在夏季和秋季两个季节的鱼类多样性, 旨在研究北京市鱼类群落的空间格局特征, 探索适用于北京鱼类生物多样性监测及保护的新方法。结果表明: 在北京市的34个采样点中, 利用eDNA metabarcoding共检测出鱼类55种, 显著高于传统方法所捕获的鱼类种数(35种), 鱼类组成以鲤形目和鲈形目为主。山区河流清水鱼的多样性要显著高于城区河流, 城区河流(北运河水系)群落结构较为单一, 以鲫(Carassius auratus)、麦穗鱼(Pseudorasbora parva)、泥鳅(Misgurnus anguillicaudatus)等耐污种为优势种; 山区河流(潮白河水系及大清河水系)以宽鳍鱲(Zacco platypus)、拉氏鱥(Rhynchocypris lagowskii)、马口鱼(Opsariichthys uncirostris)等为优势种。不同季节影响清水鱼群落结构的环境因子不同, 夏季主要是总溶解固体和电导率, 秋季主要是海拔和温度。清水鱼丰富度与环境因子及人类活动的相关性表明, 清水鱼的丰富度随着总溶解固体及灯光指数增加而显著降低, 且均与海拔、温度等存在显著相关性。本研究证明了eDNA metabarcoding方法用于监测北京市鱼类多样性及其时空分布的可行性。     

北京市妫水河浮游动物群落结构与水质评价

由于浮游动物对水体环境变化敏感,可表征水体污染程度,因此在2017年对妫水河浮游动物群落结构进行调查研究,分析了浮游动物群落结构时空变化特征及其与环境因子的关系,并利用生物学评价方法对水质进行评价。结果表明:妫水河浮游动物有4门22属88种,其中原生动物种类最多,为42种,主要以轮虫和原生动物为主,浮游动物平均细胞密度和生物量分别为5041.58个/L和2.88 mg/L。浮游动物群落结构与环境因子的CCA分析显示,水温、pH、DO和氨氮是影响妫水河浮游动物群落结构变化的重要因素,其中裂痕龟纹轮虫、冠饰异尾轮虫和螺形龟甲轮虫等对水体中氮磷的相关性极为显著,具有富营养化指示作用,可作为监测水质的指示生物。妫水河浮游动物多样性指数H、均匀度指数J和丰富度指数D全年平均值分别为0.43、0.31和0.41,整体评价结果显示,妫水河水体处于中到富营养型水平,尤其是下段城区段污染严重,表明妫水河水体生态功能遭到破坏,水质还需进一步改善和治理。本研究结果可为妫水河水质评价、水环境监测及水污染治理提供基础数据资料和理论依据。     

洪潮江水库浮游细菌群落空间分布及其与环境因子的关系

【目的】为了解洪潮江水库浮游细菌群落组成、空间分布及其与环境因子的相互作用关系。【方法】基于16S rRNA基因高通量测序技术,对洪潮江水库浮游细菌群落结构与多样性进行了分析,同步对水体的理化指标进行检测。【结果】洪潮江水库共注释浮游细菌28门79纲168目243科325属85种。优势门为变形菌门、放线菌门、蓝细菌门、疣微菌门、拟杆菌门、浮霉菌门,分别占比21.95%、21.30%、17.98%、12.27%、11.72%、9.51%。基于Bray-Curtis距离的PCoA分析表明,洪潮江水库9个采样点可以被分为3组,细菌群落呈现沿上下游梯度变化的趋势。perMANOVA检验显示,各组差异显著,但是各组的多样性指数没有显著性差异。Mantel分析表明,透明度、浊度、总磷、亚硝酸盐氮、硝酸盐氮、叶绿素a、pH、溶解氧、氧化还原电位、比电导、总溶解固体以及营养状态会显著影响浮游细菌群落结构。【结论】洪潮江水库浮游细菌空间分布特征是自然环境理化因子和农业活动综合作用的结果。研究结果对了解浮游细菌群落空间格局及其对人类活动的响应,以及水库管理具有科学参考价值。     

Toolbox Approaches Using Molecular Markers and 16S rRNA Gene Amplicon Data Sets for Identification of Fecal Pollution in Surface Water

In this study, host-associated molecular markers and bacterial 16S rRNA gene community analysis using high-throughput sequencing were used to identify the sources of fecal pollution in environmental waters in Brisbane, Australia. A total of 92 fecal and composite wastewater samples were collected from different host groups (cat, cattle, dog, horse, human, and kangaroo), and 18 water samples were collected from six sites (BR1 to BR6) along the Brisbane River in Queensland, Australia. Bacterial communities in the fecal, wastewater, and river water samples were sequenced. Water samples were also tested for the presence of bird-associated (GFD), cattle-associated (CowM3), horse-associated, and human-associated (HF183) molecular markers, to provide multiple lines of evidence regarding the possible presence of fecal pollution associated with specific hosts. Among the 18 water samples tested, 83%, 33%, 17%, and 17% were real-time PCR positive for the GFD, HF183, CowM3, and horse markers, respectively. Among the potential sources of fecal pollution in water samples from the river, DNA sequencing tended to show relatively small contributions from wastewater treatment plants (up to 13% of sequence reads). Contributions from other animal sources were rarely detected and were very small (<3% of sequence reads). Source contributions determined via sequence analysis versus detection of molecular markers showed variable agreement. A lack of relationships among fecal indicator bacteria, host-associated molecular markers, and 16S rRNA gene community analysis data was also observed. Nonetheless, we show that bacterial community and host-associated molecular marker analyses can be combined to identify potential sources of fecal pollution in an urban river. This study is a proof of concept, and based on the results, we recommend using bacterial community analysis (where possible) along with PCR detection or quantification of host-associated molecular markers to provide information on the sources of fecal pollution in waterways.     

Aquatic diversity in a former floodplain: Remediation in an urban context

The Lobau, a former floodplain area of the Danube River situated within the city limits of Vienna (Austria), was strongly affected by the river regulation in 1875. The reduced hydrological connectivity led to an increasing loss of aquatic habitats. A water enhancement scheme with a maximum water input of 0.5 m³ s^?1 was initiated in 2001. The present study assesses the effect of this scheme on biodiversity using three target species groups – aquatic molluscs, dragonflies and fish – following a common Before–After Control-Impact design (BACI). Dragonflies and molluscs were positively affected, reflecting the habitat alterations in the system. For fish, no significant impact was observed. The aim of the scheme has been achieved: increased habitat diversity and improved habitat conditions for the system's initial community and further rheophilic/rheotolerant species. Water enhancement schemes can be effective remediation measures and deserve further attention in the management of urban wetlands.     

柴达木盆地荒漠灌丛群落谱系结构研究

谱系信息是群落生态学和保育生物学研究的主要内容之一。为探究柴达木盆地荒漠灌丛群落谱系结构及其与环境因子的关系,该研究以柴达木盆地荒漠灌丛为对象,基于群落中物种存在与否的物种组成数据,使用R语言中picante软件包计算了灌丛群落谱系多样性指数和谱系结构指数,并且分析了谱系结构指数与年均温度、年均降水以及土壤含水量之间的关系,以揭示柴达木盆地灌丛群落物种之间的亲缘关系和群落生物多样性维持机制。结果表明:(1)柴达木盆地灌丛群落谱系结构与土壤含水量之间存在极显著相关性(P=2.77×10-6),随着土壤含水量的增加,群落谱系结构聚集程度逐渐降低,表现出生境过滤作用逐步减弱,生物间相互作用逐渐加强的变化趋势。(2)群落谱系结构与年均温度、年均降水之间无显著相关性。(3)典型荒漠生境灌丛和河谷(河漫滩)生境灌丛群落的谱系结构差异显著(P0.05),整体上分别表现为谱系聚集状态和谱系发散状态;河谷(河漫滩)生境灌丛群落的谱系多样性显著高于典型荒漠生境灌丛(P0.05)。(4)将群落谱系信息应用在生物多样性保护实践中,发现河谷(河漫滩)生境灌丛群落较典型荒漠生境的灌丛群落可能具有更高的保护价值。研究认为,将群落谱系研究与保育生物学理论结合将会使生物多样性保护策略更加科学有效。     

Genetic structure of the marsh frog (Pelophylax ridibundus) populations in urban landscape

Urbanization is a pervasive process causing habitat fragmentation, spatial isolation of populations, and reduction of biological diversity. In this study, we applied 11 microsatellite loci and Bayesian analyses to investigate genetic diversity and population structure in marsh frogs (Pelophylax ridibundus) living in two types of environment—highly fragmented urban landscapes, and landscapes characterized by the presence of a river and artificial canals. Our results show reduced genetic diversity, lower effective population sizes, and higher genetic differentiation for spatially isolated urban populations in comparison with populations outside intensely urbanized areas. Reduction of allelic diversity in urban localities isolated for 13–37 generations is more conspicuous than reduction of expected heterozygosity. Populations living close to the River Danube, its branches, and artificial canals are genetically more homogenous. Our results also suggest that the Danube in Bratislava is not a natural barrier to gene flow. In contrast, it acts as a natural corridor for water frog dispersal. Population structure of P. ridibundus also shows higher genetic connectivity within water paths than between them, suggesting limited overland dispersal, and reflects the historical landscape structure associated with the distribution of the lost river branches.     

武夷山国家自然保护区不同植被类型土壤微生物群落特征

运用BIOLOG微平板技术,对武夷山国家自然保护区不同植被类型土壤微生物群落结构和代谢功能多样性进行研究,探讨不同植被类型对土壤微生物群落的影响.结果表明: 不论是土壤理化性质、酶活性,还是反映土壤微生物代谢功能多样性的平均颜色变化率(AWCD),青冈林和米槠林代表的天然林均明显优于马尾松林和杉木林代表的人工林,荒地最差.AWCD随着培养时间的延长而逐渐增加,但不同植被类型土壤AWCD值具有较大差异.碳水化合物和羧酸类碳源是各植被类型土壤微生物的主要碳源,其次为氨基酸类、酚酸类和聚合物类,胺类碳源的利用率最小.土壤微生物Simpson指数、Shannon指数、丰富度指数和McIntosh指数也呈现天然林高于人工林的趋势.主成分分析表明,从31个因素中提取的与碳源利用相关的主成分1、主成分2分别能解释变量方差的56.3%和30.2%,不同植被类型土壤微生物碳源利用特征出现分异,在主成分分离中起主要贡献作用的是胺类和氨基酸类碳源.研究结果可为进一步探讨植被多样性与土壤微生物多样性之间的关系奠定基础.     

Hydrological seasonality dictates fish fauna of the lower Araguaia River,Tocantins-Araguaia basin

Hydrological seasonality of flow dictates diversity in tropical aquatic ecosystems. Large tropical rivers and tributaries are typically more depth and shows increased flow velocity and area in flood than in dry season. A thorough analysis of seasonality effect on fish community structure can provide valuable information of major forces driving tropical communities. However, these types of analyses have been limited by poor knowledge of tropical diversity, human impacts on environment, and biased sampling methods. We used a pristine large tropical river (and tributaries) and five types (12 gill nets, beach seine nets, longlines, branch baited hooks, and cast nets) of fishing gears to present an evaluation of flow seasonality effect on fish community. We sampled fourteen sites in river channel and tributaries in the lower Araguaia River. Sampling was conducted in flood (March and May 2009) and dry (July and September 2009) seasons of a typical seasonal year. Species richness, Shannon Diversity and Evenness, mean similarity of binary data, and abundance (CPUE) were analyzed by PERMANOVA and PCoA. We also analyzed the species most correlated (by Spearman correlation rank) with flood and dry season. Features differed significantly between flood and dry seasons, but not between channel and tributaries. The composition and abundance of community, and the representative species of flood and dry changed quite along the year. Flood showed lotic-related species, and dry presented lentic-related species. Thus, we achieved an alternation of fish community between season, both for channel and tributaries.     

沙颍河流域人为输入对水体水化学组成的影响

人类活动输入影响河流水体化学组成,增加经河流体系向海洋输出物质的通量,影响全球物质循环过程.有效识别人为输入的影响途径和范围对于量化人类活动对全球物质循环的影响具有重要作用.沙颍河是淮河上游最大支流,流域水体受人为输入影响严重,通过研究沙颍河流域强人为输入对河水水化学组成的影响过程,有利于弄清楚强烈人为活动干扰下河流输...     

Managing water-use trade-offs in a semi-arid river delta to sustain multiple ecosystem services: a modeling approach

Managing trade-offs among water uses in a river basin to sustain multiple ecosystem services is crucial for adaptation to changing river flow regimes. Here we analyze the trade-off between irrigation and fisheries in the Amudarya, a semi-arid river basin in Central Asia, using an optimal control and an agent-based modeling approach. With the optimal control approach (OCA), we identify the economic and ecological conditions for water sharing in a regime where a social manager controls water withdrawals and fish harvesting. With the agent-based model (ABM), we relax some of the assumptions of the OCA to investigate how localized, individual agents with varied water use histories adapt their water use activities to local resource conditions. Variation in the farmers’ initial labor allocations to the two activities results in regimes with only one activity or both. Global returns and income equality are highest in a mixed regime. The mixed regimes also are more robust to water variability because fishing activities can compensate for decreased agricultural performance in the midstream regions. Thus, allowing for multiple uses can improve the coupled social-ecological system’s performance and its resilience. We also observe a lock-in effect similar to the current situation in the Amudarya, where agriculture is the dominant water use and transition to a more balanced allocation has proven to be extremely difficult. As in the ABM, this can to some extent be attributed to the difficulties of achieving sufficient revenues from fishing when agricultural activities upstream are high. Regulations or incentives are needed to overcome those barriers, and to facilitate progress towards integrated water management.     

Ecological effects of urban stormwater runoff on benthic macroinvertebrates inhabiting the Green River,Massachusetts

Although it has been demonstrated that urban stormwater can alter the quality of receiving waters, the corresponding impact on aquatic biota remains essentially undocumented. A year-long intensive study, therefore, was implemented to monitor and describe the ecological effects exerted by urban runoff on benthic macroinvertebrates.Rock-filled, basket-type artificial substrates deployed periodically in nonurban and urban river reaches yielded collections of macroinvertebrates that furnished data for: (1) species diversity (the Brillouin index, H), (2) hierarchical diversity, (3) major taxa composition, and (4) collection dissimilarity at the species level.The overall results from these four analytical procedures strongly indicate that the macrobenthic community became progressively disrupted downstream in the urban reach. The high degree of correspondence between the known sources of urban runoff and the observed effects on the benthic community are forceful arguments that urban runoff is the causal agent of disruption. The impact is not confined to periods following heavy rains. Instead the pollutants appear to remain in the system. The stress imposed by them was most acute during the summer low flow and was probably localized in or near the stream bed.To assess the impact of urban runoff on an aquatic ecosystem, physical, chemical, and biological monitoring should routinely consider the stream bed microzone. Urban runoff pollutant loading standards must take into account the apparent long-term residence of pollutants in the substrate and the associated stress of summer low flows.