Characterizing seasonal changes in physicochemistry and bacterial community composition in hyporheic sediments

The hyporheic zone of stream ecosystems is a critical habitat for microbial communities. However, the factors influencing hyporheic bacterial communities along spatial and seasonal gradients remain poorly understood. We sought to characterize patterns in bacterial community composition among the sediments of a small stream in southern Ontario, Canada. We used sampling cores to collect monthly hyporheic water and sediment microbial communities in 2006 and 2007. We described bacterial communities terminal-restriction fragment length polymorphism (TRFLP) and tested for spatial and seasonal relationships with physicochemical parameters using multivariate statistics. Overall, the hyporheic zone appears to be a DOC, oxygen, and nitrogen sink. Microbial communities were distinct from those at the streambed surface and from soil collected in the adjacent watershed. In the sediments, microbial communities were distinct between the fall, spring, and summer seasons, and bacterial communities were more diverse at streambed surface and near-surface sites compared with deeper sites. Moreover, bacterial communities were similar between consecutive fall seasons despite shifting throughout the year, suggesting recurring community assemblages associated with season and location in the hyporheic zone. Using canonical correspondence analysis, seasonal patterns in microbial community composition and environmental parameters were correlated in the following way: temperature was related to summer communities; DOC (likely from biofilm and allochthonous inputs) influenced most fall communities; and nitrogen associated strongly with winter and spring communities. Our results also suggest that labile DOC entering the hyporheic zone occurred in concert with shifts in the bacterial community. Generally, seasonal patterns in hyporheic physicochemistry and microbial biodiversity remain largely unexplored. Therefore, we highlight the importance of seasonal and spatial resolution when assessing surface- and groundwater interactions in stream ecosystems.     

Litter Breakdown and Microbial Succession on Two Submerged Leaf Species in a Small Forested Stream

Microbial succession during leaf breakdown was investigated in a small forested stream in west-central Georgia, USA, using multiple culture-independent techniques. Red maple (Acer rubrum) and water oak (Quercus nigra) leaf litter were incubated in situ for 128 days, and litter breakdown was quantified by ash-free dry mass (AFDM) method and microbial assemblage composition using phospholipid fatty acid analysis (PLFA), ribosomal intergenic spacer analysis (RISA), denaturing gradient gel electrophoresis (DGGE), and bar-coded next-generation sequencing of 16S rRNA gene amplicons. Leaf breakdown was faster for red maple than water oak. PLFA revealed a significant time effect on microbial lipid profiles for both leaf species. Microbial assemblages on maple contained a higher relative abundance of bacterial lipids than oak, and oak microbial assemblages contained higher relative abundance of fungal lipids than maple. RISA showed that incubation time was more important in structuring bacterial assemblages than leaf physicochemistry. DGGE profiles revealed high variability in bacterial assemblages over time, and sequencing of DGGE-resolved amplicons indicated several taxa present on degrading litter. Next-generation sequencing revealed temporal shifts in dominant taxa within the phylum Proteobacteria, whereas γ-Proteobacteria dominated pre-immersion and α- and β-Proteobacteria dominated after 1 month of instream incubation; the latter groups contain taxa that are predicted to be capable of using organic material to fuel further breakdown. Our results suggest that incubation time is more important than leaf species physicochemistry in influencing leaf litter microbial assemblage composition, and indicate the need for investigation into seasonal and temporal dynamics of leaf litter microbial assemblage succession.     

Patterns in the Composition of Microbial Communities from a Subtropical River: Effects of Environmental,Spatial and Temporal Factors

Microbes are key components of aquatic ecosystems and play crucial roles in global biogeochemical cycles. However, the spatiotemporal dynamics of planktonic microbial community composition in riverine ecosystems are still poorly understood. In this study, we used denaturing gradient gel electrophoresis of PCR-amplified 16S and 18S rRNA gene fragments and multivariate statistical methods to explore the spatiotemporal patterns and driving factors of planktonic bacterial and microbial eukaryotic communities in the subtropical Jiulong River, southeast China. Both bacterial and microbial eukaryotic communities varied significantly in time and were spatially structured according to upper stream, middle-lower stream and estuary. Among all the environmental factors measured, water temperature, conductivity, PO₄-P and TN/TP were best related to the spatiotemporal distribution of bacterial community, while water temperature, conductivity, NO_x-N and transparency were closest related to the variation of eukaryotic community. Variation partitioning, based on partial RDA, revealed that environmental factors played the most important roles in structuring the microbial assemblages by explaining 11.3% of bacterial variation and 17.5% of eukaryotic variation. However, pure spatial factors (6.5% for bacteria and 9.6% for eukaryotes) and temporal factors (3.3% for bacteria and 5.5% for eukaryotes) also explained some variation in microbial distribution, thus inherent spatial and temporal variation of microbial assemblages should be considered when assessing the impact of environmental factors on microbial communities.     

Spatial and temporal impacts of a diesel fuel spill on stream invertebrates

1. We assessed the effects of a 26 500 L diesel fuel spill on the macroinvertebrate fauna of a small trout stream in central New York, U.S.A. To determine the spatial extent of the spill we sampled three locations (0.7, 5.0 and 11.8 km downstream of the spill), each containing a reference site (unaffected tributary) and an impact site (downstream of spill). Sampling was repeated four times over a 15‐month period to assess temporal recovery. 2. Immediately after the spill, invertebrate density at all three locations below the spill was significantly lower than reference density. Three months after the spill, density up to 5 km below the spill was still far lower (<100 individuals per sample) than reference density (800–1200 individuals per sample). A year after the spill, density was similar between reference and impact sites, suggesting that invertebrates had recovered numerically. 3. Taxonomic richness up to 5.0 km below the spill was less than half the reference taxonomic richness and this difference persisted for at least 3 months. Some significant differences between reference and impact sites were observed after 15 months, but these differences could not be attributed to the oil spill. 4. For at least 3 months following the spill, the site immediately downstream of the spill was dominated by Optioservus, a petrochemical‐tolerant riffle beetle. Twelve to 15 months after the spill, both the reference and impact sites near the spill were dominated numerically by the mayfly Ephemerella, but the degree of dominance was twice as large at the impact site. 5. We concluded that the diesel fuel spill significantly reduced the density of invertebrates (by 90%) and taxonomic richness (by 50%) at least 5.0 km downstream, but density recovered within a year. Throughout the study, however, the fauna immediately below the spill was species poor and significantly over‐represented by a single dominant taxon, suggesting that 15 months was not sufficient for full community recovery from the oil spill.     

Bacterial communities in sediments of a drinking water reservoir

Sediment microbial communities play crucial roles in biogeochemical cycles of aquatic systems. Knowledge of microbial community structure could aid in our understanding of these roles. In this study, the sediment bacterial community structure of a drinking water reservoir in China was investigated. A large difference in major bacterial groups was observed at different sampling sites. Proteobacteria was the largest bacterial phylum, with a marked shift in the proportions of its major subdivisions. Microorganisms within phylum Proteobacteria might play important roles in various biogeochemical processes.     

Spatial and temporal response of stream bacteria to sources of dissolved organic carbon in a blackwater stream system

1. We hypothesized that changes in bacterial colony growth would be correlated to shifts in riparian vegetation (via leachate quality) along a river continuum of a south-eastern, blackwater stream (U.S.A.). Spatially, we expected bacterial assemblages from downstream reaches to utilize more sources of leachate and at higher concentrations than bacteria collected from headwater reaches. Temporally, we predicted higher colony growth on leachate from autumn-shed (senescent) leaves compared with leachate from fresh, green leaves.
2. We examined spatial differences in assemblage growth by culturing bacteria sampled along the stream continuum on gradient plates using leachates from four common riparian species ( Taxodium distichum , Carya spp., Acer rubrum and Decumaria barbara ). Bacteria from the lowest site were able to use all sources provided and at all concentrations, whereas bacteria from upper reaches could not. Colony density was correlated to relative leachate concentration at all sites along the continuum.
3. Leachates from fresh and senescent A. rubrum leaves were used to determine temporal differences. Winter assemblages of bacteria could not grow on fresh leaf leachate at any concentration but grew well on autumn leaf leachate at higher concentrations. Differential response of bacterial assemblages indicated local adaptation to potential sources of dissolved organic matter.
4. Growth response of stream bacterial colonies appeared to be dependent on the timing and source of leachate as well as on sources of dissolved organic carbon from further upstream. Growth of bacterial assemblages exhibited 'generalist' characteristics in headwater reaches and 'specialist' characteristics at the mouth of our study stream drainage. Thus, our findings lend support to the argument that variable resource habitats favour a small, generalist assemblage, while environments with stable resource supplies allow for highly diverse assemblages dominated by specialists.     

Spatial and temporal response of stream bacteria to sources of dissolved organic carbon in a blackwater stream system

1. We hypothesized that changes in bacterial colony growth would be correlated to shifts in riparian vegetation (via leachate quality) along a river continuum of a south-eastern, blackwater stream (U.S.A.). Spatially, we expected bacterial assemblages from downstream reaches to utilize more sources of leachate and at higher concentrations than bacteria collected from headwater reaches. Temporally, we predicted higher colony growth on leachate from autumn-shed (senescent) leaves compared with leachate from fresh, green leaves.
2. We examined spatial differences in assemblage growth by culturing bacteria sampled along the stream continuum on gradient plates using leachates from four common riparian species ( Taxodium distichum , Carya spp., Acer rubrum and Decumaria barbara ). Bacteria from the lowest site were able to use all sources provided and at all concentrations, whereas bacteria from upper reaches could not. Colony density was correlated to relative leachate concentration at all sites along the continuum.
3. Leachates from fresh and senescent A. rubrum leaves were used to determine temporal differences. Winter assemblages of bacteria could not grow on fresh leaf leachate at any concentration but grew well on autumn leaf leachate at higher concentrations. Differential response of bacterial assemblages indicated local adaptation to potential sources of dissolved organic matter.
4. Growth response of stream bacterial colonies appeared to be dependent on the timing and source of leachate as well as on sources of dissolved organic carbon from further upstream. Growth of bacterial assemblages exhibited 'generalist' characteristics in headwater reaches and 'specialist' characteristics at the mouth of our study stream drainage. Thus, our findings lend support to the argument that variable resource habitats favour a small, generalist assemblage, while environments with stable resource supplies allow for highly diverse assemblages dominated by specialists.     

Direct and indirect influence of parental bedrock on streambed microbial community structure in forested streams

A correlative study was performed to determine if variation in streambed microbial community structure in low-order forested streams can be directly or indirectly linked to the chemical nature of the parental bedrock of the environments through which the streams flow. Total microbial and photosynthetic biomass (phospholipid phosphate [PLP] and chlorophyll a), community structure (phospholipid fatty acid analysis), and physical and chemical parameters were measured in six streams, three located in sandstone and three in limestone regions of the Bankhead National Forest in northern Alabama. Although stream water flowing through the two different bedrock types differed significantly in chemical composition, there were no significant differences in total microbial and photosynthetic biomass in the sediments. In contrast, sedimentary microbial community structure differed between the bedrock types and was significantly correlated with stream water ion concentrations. A pattern of seasonal variation in microbial community structure was also observed. Further statistical analysis indicated dissolved organic matter (DOM) quality, which was previously shown to be influenced by geological variation, correlated with variation in bacterial community structure. These results indicate that the geology of underlying bedrock influences benthic microbial communities directly via changes in water chemistry and also indirectly via stream water DOM quality.     

Bacterial profiling of White Plague Disease in a comparative coral species framework

Coral reefs are threatened throughout the world. A major factor contributing to their decline is outbreaks and propagation of coral diseases. Due to the complexity of coral-associated microbe communities, little is understood in terms of disease agents, hosts and vectors. It is known that compromised health in corals is correlated with shifts in bacterial assemblages colonizing coral mucus and tissue. However, general disease patterns remain, to a large extent, ambiguous as comparative studies over species, regions, or diseases are scarce. Here, we compare bacterial assemblages of samples from healthy (HH) colonies and such displaying signs of White Plague Disease (WPD) of two different coral species (Pavona duerdeni and Porites lutea) from the same reef in Koh Tao, Thailand, using 16S rRNA gene microarrays. In line with other studies, we found an increase of bacterial diversity in diseased (DD) corals, and a higher abundance of taxa from the families that include known coral pathogens (Alteromonadaceae, Rhodobacteraceae, Vibrionaceae). In our comparative framework analysis, we found differences in microbial assemblages between coral species and coral health states. Notably, patterns of bacterial community structures from HH and DD corals were maintained over species boundaries. Moreover, microbes that differentiated the two coral species did not overlap with microbes that were indicative of HH and DD corals. This suggests that while corals harbor distinct species-specific microbial assemblages, disease-specific bacterial abundance patterns exist that are maintained over coral species boundaries.     

Profiling and monitoring of microbial populations by denaturing high-performance liquid chromatography

We describe a new molecular technique for the analysis of microbial species and complex microbial populations based on the separation of PCR-amplified 16S rDNA fragments by denaturing high-performance liquid chromatography (DHPLC). Using marine bacterial samples, we determined the optimum conditions for the analysis of bacterial species and the examination of complex bacterial assemblages obtained from different environments. The incorporation of a 40-bp GC clamp into the amplification primer was essential to effectively discriminate genetic differences in DHPLC-primers with a 20-, 10-, or 0-bp GC clamp length were less efficient. A 64.5 degrees C column temperature in DHPLC allowed optimal separation of species in a complex bacterial population. PCR-DHPLC analysis of bacterial assemblages demonstrated profiles with distinguishable peaks, which constituted the different populations and their degree of abundance. Fraction collection and DNA sequencing from profile peaks enabled bacterial identification. PCR-DHPLC analysis can also provide opportunities for describing bacterial communities, cloning bacteria, and monitoring bacterial populations in environments of interest.     

Microbial characterization of a JP-4 fuel-contaminated site using a combined lipid biomarker/polymerase chain reaction--denaturing gradient gel electrophoresis (PCR-DGGE)-based approach

The impact of pollution on soil microbial communities and subsequent bioremediation can be measured quantitatively in situ using direct, non-culture- dependent techniques. Such techniques have advantages over culture-based methods, which often account for less than 1% of the extant microbial community. In 1988, a JP-4 fuel spill contaminated the glacio-fluvial aquifer at Wurtsmith Air Force Base, Michigan, USA. In this study, lipid biomarker characterization of the bacterial and eukaryotic communities was combined with polymerase chain reaction– denaturing gradient gel electrophoresis (PCR–DGGE) analysis of the eubacterial community to evaluate correlation between contaminant (JP-4 fuel) concentration and community structure shifts. Vadose, capillary fringe and saturated zone samples were taken from cores within and up- and down-gradient from the contaminant plume. Lipid biomarker analysis indicated that samples from within the plume contained increased biomass, with large proportions of typically Gram-negative bacteria. Outside the plume, lipid profiles indicated low-biomass microbial communities compared with those within the initial spill site. 16S rDNA sequences derived from DGGE profiles from within the initial spill site suggested dominance of the eubacterial community by a limited number of phylogenetically diverse organisms. Used in tandem with pollutant quantification, these molecular techniques should facilitate significant improvements over current assessment procedures for the determination of remediation end-points.     

Temporal variability in the diversity and composition of stream bacterioplankton communities

Bacterioplankton in freshwater streams play a critical role in stream nutrient cycling. Despite their ecological importance, the temporal variability in the structure of stream bacterioplankton communities remains understudied. We investigated the composition and temporal variability of stream bacterial communities and the influence of physicochemical parameters on these communities. We used barcoded pyrosequencing to survey bacterial communities in 107 streamwater samples collected from four locations in the Colorado Rocky Mountains from September 2008 to November 2009. The four sampled locations harboured distinct communities yet, at each sampling location, there was pronounced temporal variability in both community composition and alpha diversity levels. These temporal shifts in bacterioplankton community structure were not seasonal; rather, their diversity and composition appeared to be driven by intermittent changes in various streamwater biogeochemical conditions. Bacterial communities varied independently of time, as indicated by the observation that communities in samples collected close together in time were no more similar than those collected months apart. The temporal turnover in community composition was higher than observed in most previously studied microbial, plant or animal communities, highlighting the importance of stochastic processes and disturbance events in structuring these communities over time. Detailed temporal sampling is important if the objective is to monitor microbial community dynamics in pulsed ecosystems like streams.     

Effects of pulsed and pressed disturbances on the benthic invertebrate community following a coal spill in a small stream in northeastern USA

In September 1999, a coal-carrying train derailed and spilled 180,000–270,000 kg of coal into the Cayuga Inlet near Ithaca, New York. This study determined the immediate effects of the coal spill and the clean up procedures on the aquatic invertebrate community, and whether the stream recovered from this event after 2 years. Benthic invertebrate samples were taken both upstream and downstream of the coal spill immediately following clean-up efforts and two years later. Just after the coal spill, the total abundance and species richness of aquatic invertebrates were significantly lower downstream of the spill, suggesting that the disturbance caused increased mortality and/or emigration compared to a reference site upstream. Taxa affected most were grazers and turbidity-susceptible invertebrates. Two years later the invertebrate communities were similar upstream and downstream of the spill, except for an increase in the percent of the dominant genus, Hydropsyche(Trichoptera: Hydropsychidae). We speculate that long-term effects of channelization of the stream that occurred during the clean-up prevented the invertebrate assemblage from returning to the conditions observed in a reference site upstream of the coal spill. We propose that large scale environmental clean-ups should be designed to avoid altering ecosystems permanently, and that streams should be allowed to recover naturally without destructive human intervention.     

Rapid biological assessment charts the recovery of stream macroinvertebrate assemblages after sewage discharges cease

Rapid biological assessment (RBA) methods involving non-quantitative sampling of aquatic macroinvertebrates, sub-sampling of only a proportion of the organisms collected, and identification only to genus-level were used to assess responses over up to 7 years following the cessation of discharges from five sewage treatment plants into small streams in the Blue Mountains, Australia, and an increased volume of discharge into another stream. Multivariate analysis (ordination and analysis of similarities) did not always show an obvious recovery after discharge cessation: assemblages sampled upstream and downstream of discharge points were not always clearly closer in ordination space after cessation than before and statistically significant differences between upstream and downstream assemblages generally remained after cessation, apparently because of natural medium-scale spatial variability. However, the tolerance-based SIGNAL-SG biotic index demonstrated some recovery in all cases of discharge cessation, and a lag in recovery of about a year. A lesser recovery in the more urbanised catchments was attributed to persistent pollution from sources other than treatment plant discharges. The biotic index also revealed subtle biological change that was not predictable from monitoring data on nutrient concentrations. The SIGNAL-SG biotic index derived from RBA sampling provides an easily communicated measure of the effectiveness of investment in abatement of sewage impacts on stream macroinvertebrates in the Blue Mountains.     

Impact of Oil-Spill Contamination on a Soil Bacterial Community: A 40-Year History of Rehabilitation in the Arava Valley

The present study evaluated the effect of crude oil contamination on a microbial community in hyper-arid soils. The Evrona Nature Reserve of Israel, situated in the Arava, was exposed twice in the last 40 years to petroleum-hydrocarbon-spill pollution. The first pollution event took place 40 years ago and was never treated, presenting a unique future time-point perspective to the second (2014) contamination event. Soil samples were collected after the second spill, and abiotic properties and bacterial diversity in the sampled soil were analyzed. The results showed that there is a significant decrease over time in the number of observed bacterial species in the contaminated samples, coupled together with bacterial species replacement toward species capable of using source oil as the sole carbon and energy source. The presence of petroleum in soil significantly changed the composition and functional diversity of a microbial community, and the Evrona Nature Reserve is still in the middle of a bioremediation process even 40 years after the crude oil contamination.     

Biome-level biogeography of streambed microbiota

A field study was conducted to determine the microbial community structures of streambed sediments across diverse geographic and climatic areas. Sediment samples were collected from three adjacent headwater forest streams within three biomes, eastern deciduous (Pennsylvania), southeastern coniferous (New Jersey), and tropical evergreen (Guanacaste, Costa Rica), to assess whether there is biome control of stream microbial community structure. Bacterial abundance, microbial biomass, and bacterial and microbial community structures were determined using classical, biochemical, and molecular methods. Microbial biomass, determined using phospholipid phosphate, was significantly greater in the southeastern coniferous biome, likely due to the smaller grain size, higher organic content, and lower levels of physical disturbance of these sediments. Microbial community structure was determined using phospholipid fatty acid (PLFA) profiles and bacterial community structure from terminal restriction fragment length polymorphism and edited (microeukaryotic PLFAs removed) PLFA profiles. Principal component analysis (PCA) was used to investigate patterns in total microbial community structure. The first principal component separated streams based on the importance of phototrophic microeukaryotes within the community, while the second separated southeastern coniferous streams from all others based on increased abundance of fungal PLFAs. PCA also indicated that within- and among-stream variations were small for tropical evergreen streams and large for southeastern coniferous streams. A similar analysis of bacterial community structure indicated that streams within biomes had similar community structures, while each biome possessed a unique streambed community, indicating strong within-biome control of stream bacterial community structure.     

Natural attenuation of diesel-oil contamination in a subantarctic soil (Crozet Island)

An accidental contamination occurred in subantarctic Crozet Island between July and November 1997 near the "Alfred Faure" scientific station (51°51'E-46°25'S). More than 20,000 l of diesel fuel was spilled in the soil in the vicinity of the power station. In order to evaluate the efficiency of the expected bioattenuation process, the contaminated area was sampled in December 1999 for bacterial and chemical analysis. All samples were analysed for total bacteria, heterotrophic viable assemblages and hydrocarbon-utilising microflora. The results of this first analytical survey clearly show a significant response of subantarctic microbial soil communities to the hydrocarbon contamination. Significant increases of total, heterotrophic and hydrocarbon-utilising micro-organisms occurred in the more contaminated zone (from less than 5᎒⁴ MPN g^-1 wet soil to more than 10⁸ MPN g^-1 wet soil for hydrocarbon-degrading micro-organisms). The very high numbers of hydrocarbon-degrading micro-organisms present in the more contaminated zone are clearly linked to early bioattenuation activities. Chemical results provided some clear indications that spilled fuel was still well preserved from chemical and biological weathering 1 year after the spill, as light aliphatics and aromatics were present in all oiled samples and little differences were usually observed between samples.     

Fate of glacier surface snow-originating bacteria in the glacier-fed hydrologic continuums

Glaciers represent important biomes of Earth and are recognized as key species pools for downstream aquatic environments. Worldwide, rapidly receding glaciers are driving shifts in hydrology, species distributions and threatening microbial diversity in glacier-fed aquatic ecosystems. However, the impact of glacier surface snow-originating taxa on the microbial diversity in downstream aquatic environments has been little explored. To elucidate the contribution of glacier surface snow-originating taxa to bacterial diversity in downstream aquatic environments, we collected samples from glacier surface snows, downstream streams and lakes along three glacier-fed hydrologic continuums on the Tibetan Plateau. Our results showed that glacier stream acts as recipients and vectors of bacteria originating from the glacier environments. The contributions of glacier surface snow-originating taxa to downstream bacterial communities decrease from the streams to lakes, which was consistently observed in three geographically separated glacier-fed ecosystems. Our results also revealed that some rare snow-originating bacteria can thrive along the hydrologic continuums and become dominant in downstream habitats. Finally, our results indicated that the dispersal patterns of bacterial communities are largely determined by mass effects and increasingly subjected to local sorting of species along the glacier-fed hydrologic continuums. Collectively, this study provides insights into the fate of bacterial assemblages in glacier surface snow following snow melt and how bacterial communities in aquatic environments are affected by the influx of glacier snow-originating bacteria.     

Short-Term Temporal Variability in Airborne Bacterial and Fungal Populations

Airborne microorganisms have been studied for centuries, but the majority of this research has relied on cultivation-dependent surveys that may not capture all of the microbial diversity in the atmosphere. As a result, our understanding of airborne microbial ecology is limited despite the relevance of airborne microbes to human health, various ecosystem functions, and environmental quality. Cultivation-independent surveys of small-subunit rRNA genes were conducted in order to identify the types of airborne bacteria and fungi found at a single site (Boulder, CO) and the temporal variability in the microbial assemblages over an 8-day period. We found that the air samples were dominated by ascomycete fungi of the Hypocreales order and a diverse array of bacteria, including members of the proteobacterial and Cytophaga-Flavobacterium-Bacteroides groups that are commonly found in comparable culture-independent surveys of airborne bacteria. Bacterium/fungus ratios varied by 2 orders of magnitude over the sampling period, and we observed large shifts in the phylogenetic diversity of bacteria present in the air samples collected on different dates, shifts that were not likely to be related to local meteorological conditions. We observed more phylogenetic similarity between bacteria collected from geographically distant sites than between bacteria collected from the same site on different days. These results suggest that outdoor air may harbor similar types of bacteria regardless of location and that the short-term temporal variability in airborne bacterial assemblages can be very large.