Freshwater Bacterial Communities Research Articles

Differential Impacts of Road De-icers on Freshwater Bacterial Communities

Concern about salt levels in freshwater habitats receiving road de-icer runoff has inspired the development of “eco-friendly” formulations that are intended to be less toxic to aquatic organisms, but few experiments have determined that these products are environmentally benign. Mesocosms containing lake water were established for 6 weeks to compare traditional road salt with two newer de-icers, one an inorganic mixture of chloride salts and the second of beet extract and brine. Amplicon sequencing and algal blocking sequences facilitated the identification of differentially impacted bacterial taxa. Ironically, although there was only a minor effect on bacterial structure at high road-salt concentrations, there was an increased relative abundance of salt-resistant genera in the mixed-salt formulation. After amendment with the beet brine de-icer, there was a turnover of taxa coincident with a 68-fold decrease in dissolved oxygen, with decreased diversity and displacement by anaerobic genera indicating a shift across a threshold to a new, apparently stable state, suggesting mesocosm recovery was unlikely. Overall, although we applaud the sentiment behind the formulation of less-damaging “eco-friendly” de-icers, they appear to have more negative environmental impacts than the traditional road salt that they were made to replace.

Top-down structuring of freshwater bacterial communities by mixotrophic flagellates

Mixotrophic and heterotrophic protists hold a key position in aquatic microbial food webs. Whereas they can account for the bulk of bacterivory in pelagic systems, the potential structuring effect of these consumers on bacterial communities is far from clear. We conducted short-term grazing experiments to test for the overall impact on bacterial community structure and possible prey preferences of phagotrophic protists. The protist taxa selected for this study include three mixotrophic flagellates, comprising two obligate- and one facultative mixotroph, and one phagoheterotrophic flagellate lacking phototrophic capacity. Bacterioplankton from seven different lakes were enriched and used to represent semi-natural prey communities. Our study demonstrated protist strain specific impacts on bacterial community composition linked to grazing. The three mixotrophs had variable impacts on bacterial communities where the two obligate mixotrophs exhibited lower grazing rates, while showing a tendency to promote higher bacterial diversity. The phagoheterotroph displayed the highest grazing rates and structured the bacterial communities via apparent selective grazing. Consistent selectivity trends were observed throughout the experiments, such as the apparent avoidance of all flagellates of Actinobacteria, and high grazing on dominant Burkholderiales taxa. However, there was no consistent “fingerprint” of mixotrophic grazing on prey communities, but the structuring impact rather seemed to depend on the trophic mode of the individual protist taxa, i.e. their dependence on phototrophy vs. phagotrophy. Our findings highlight the differential structuring impact of protist taxa on bacterial communities which may have important ecological implications, for example during periodic dominance of obligate mixotrophic bacterivores in changing lake ecosystems.

Nutrient availability and grazing influence the strength of priority effects during freshwater bacterial community coalescence.

When bacterial communities mix, immigration history can fundamentally affect the community composition as a result of priority effects. Priority effects arise when an early immigrant exhausts resources and/or alters habitat conditions, thereby influencing the establishment success of the late arriver. The strength of priority effects is context-dependent and expected to be stronger if environmental conditions favour the growth of the first arriver. In this study, we conducted a two-factorial experiment testing the importance of nutrient availability and grazing on the strength of priority effects in complex aquatic bacterial communities. We did so by mixing two dissimilar communities, simultaneously, and with a 38 h time-delay. Priority effects were measured as the invasion resistance of the first community to the invading second community. We found stronger priority effects in treatments with high nutrient availability and absence of grazing, but in general, the arrival timing was less important than the selection by nutrients and grazing. At the population level, the results were complex, but priority effects may have been driven by bacteria belonging to for example, the genera Rhodoferax and Herbaspirillum. Our study highlights the importance of arrival timing in complex bacterial communities, especially if environmental conditions favour rapid community growth.

Habitat-specific regulation of bacterial community dynamics during phytoplankton bloom succession in a subtropical eutrophic lake

Phytoplankton blooms, an important indicator of severe eutrophication, are a globally significant consequence of anthropogenic activities and climate change on freshwater lakes. Shifts in microbial communities during phytoplankton blooms have been extensively investigated, yet we have a limited understanding of how distinct assembly processes underlying the temporal dynamics of freshwater bacterial communities within different habitats respond to the succession of phytoplankton blooms. To address this knowledge gap, we collected both water and sediment samples in a subtropical eutrophic lake over a complete period of phytoplankton blooms to assess the dynamics of bacterial communities and the temporal shifts in assembly processes. Our results showed that phytoplankton blooms strongly altered the diversity, composition, and coexistence patterns of both planktonic and sediment bacterial communities (PBC and SBC), but the successional patterns differed between PBC and SBC. PBC were less temporally stable under bloom-induce disturbances, with higher variations in temporal dynamics and greater sensitivity to environmental fluctuations. Furthermore, the temporal assembly patterns of bacterial communities in both habitats were mainly driven by homogeneous selection and ecological drift. In the PBC, the role of selection decreased over time, while ecological drift became increasingly important. Conversely, in the SBC, the relative impact of selection and ecological drift on community assemblages fluctuated less over time, with selection remaining the dominant process throughout the bloom.

Functional gene transcription variation in bacterial metatranscriptomes in large freshwater Lake Ecosystems: Implications for ecosystem and human health

Little is known regarding the temporal and spatial functional variation of freshwater bacterial community (BC) under non-bloom conditions, especially in winter. To address this, we used metatranscriptomics to assess bacterial gene transcription variation among three sites across three seasons. Our metatranscriptome data for freshwater BCs at three public beaches (Ontario, Canada) sampled in the winter (no ice), summer and fall (2019) showed relatively little spatial, but a strong temporal variation. Our data showed high transcriptional activity in summer and fall but surprisingly, 89% of the KEGG pathway genes and 60% of the selected candidate genes (52 genes) associated with physiological and ecological activity were still active in freezing temperatures (winter). Our data also supported the possibility of an adaptively flexible gene expression response of the freshwater BC to low temperature conditions (winter). Only 32% of the bacterial genera detected in the samples were active, indicating that the majority of detected taxa were non-active (dormant). We also identified high seasonal variation in the abundance and activity of taxa associated with health risks (i.e., Cyanobacteria and waterborne bacterial pathogens). This study provides a baseline for further characterization of freshwater BCs, health-related microbial activity/dormancy and the main drivers of their functional variation (such as rapid human-induced environmental change and climate change).

A resistome survey across hundreds of freshwater bacterial communities reveals the impacts of veterinary and human antibiotics use.

Our decreasing ability to fight bacterial infections is a major health concern. It is arising due to the evolution of antimicrobial resistance (AMR) in response to the mis- and overuse of antibiotics in both human and veterinary medicine. Lakes integrate watershed processes and thus may act as receptors and reservoirs of antibiotic resistance genes (ARGs) introduced into the watershed by human activities. The resistome - the diversity of ARGs - under varying anthropogenic watershed pressures has been previously studied either focused on few select genes or few lakes. Here, we link the resistome of ~350 lakes sampled across Canada to human watershed activity, trophic status, as well as point sources of ARG pollution including wastewater treatment plants and hospitals in the watershed. A high percentage of the resistance genes detected was either unimpacted by human activity or highly prevalent in pristine lakes, highlighting the role of AMR in microbial ecology in aquatic systems, as well as a pool of genes available for potential horizontal gene transfer to pathogenic species. Nonetheless, watershed agricultural and pasture area significantly impacted the resistome. Moreover, the number of hospitals and the population density in a watershed, the volume of wastewater entering the lake, as well as the fraction of manure applied in the watershed as fertilizer significantly impacted ARG diversity. Together, these findings indicate that lake resistomes are regularly stocked with resistance genes evolved in the context of both veterinary and human antibiotics use and represent reservoirs of ARGs that require further monitoring.

Tracking the upstream history of aquatic microbes in a boreal lake yields new insights on microbial community assembly.

Bacterial community structure can change rapidly across short spatial and temporal scales as environmental conditions vary, but the mechanisms underlying those changes are still poorly understood. Here, we assessed how a lake microbial community assembles by following its reorganization from the main tributary, which, when flowing into the lake, first traverses an extensive macrophyte-dominated vegetated habitat, before reaching the open water. Environmental conditions in the vegetated habitat changed drastically compared to both river and lake waters and represented a strong environmental gradient for the incoming bacteria. We used amplicon sequencing of the 16S rRNA gene and transcript to reconstruct the shifts in relative abundance of individual taxa and link this to their pattern in activity (here assessed with RNA:DNA ratios). Our results indicate that major shifts in relative abundance were restricted mostly to rare taxa (<0.1% of relative abundance), which seemed more responsive to environmental changes. Dominant taxa (>1% of relative abundance), on the other hand, traversed the gradient mostly unchanged with relatively low and stable RNA:DNA ratios. We also identified a high level of local recruitment and a seedbank of taxa capable of activating/inactivating, but these were almost exclusively associated with the rare biosphere. Our results suggest a scenario where the lake community results from a reshuffling of the rank abundance structure within the incoming rare biosphere, driven by selection and growth, and that numerical dominance is not a synonym of activity, growth rate, or environmental selection, but rather reflect mass effects structuring these freshwater bacterial communities.

Time after time: detecting annual patterns in stream bacterial biofilm communities.

SummaryTo quantify the major environmental drivers of stream bacterial population dynamics, we modelled temporal differences in stream bacterial communities to quantify community shifts, including those relating to cyclical seasonal variation and more sporadic bloom events. We applied Illumina MiSeq 16S rRNA bacterial gene sequencing of 892 stream biofilm samples, collected monthly for 36‐months from six streams. The streams were located a maximum of 118 km apart and drained three different catchment types (forest, urban and rural land uses). We identified repeatable seasonal patterns among bacterial taxa, allowing their separation into three ecological groupings, those following linear, bloom/trough and repeated, seasonal trends. Various physicochemical parameters (light, water and air temperature, pH, dissolved oxygen, nutrients) were linked to temporal community changes. Our models indicate that bloom events and seasonal episodes modify biofilm bacterial populations, suggesting that distinct microbial taxa thrive during these events including non‐cyanobacterial community members. These models could aid in determining how temporal environmental changes affect community assembly and guide the selection of appropriate statistical models to capture future community responses to environmental change.

Comparative Analysis of Selective Bacterial Colonization by Polyethylene and Polyethylene Terephthalate Microplastics.

In this study, we report the biodiversity and functional characteristics of microplastic-attached biofilms originating from two freshwater bacterial communities. Even though the microplastic-biofilm (MPB) diversities are mostly determined by original bacteria instead of microplastic types, the results from 16S rRNA amplicon sequencing still showed that the dynamic biofilm successions on different microplastics were highly dissimilar. Furthermore, the analysis of biomarkers indicated distinct bacterial species with significant dissimilarities between different MPBs, which further determined the associated functions. The co-occurrence networks showed distinct interconnective characteristics in different MPBs: The structure of MPB incubated in the lake water sample was more robust under environmental stresses, and bacteria in the tap water MPB interacted more cooperatively. Regarding this cooperative interaction, the analysis of functional prediction, in this study, also showed that more symbionts and parasites colonized on microplastics in the tap water than in the lake water. Moreover, it was suggested that MPBs were more easily formed in the tap water sample. The overall results revealed significant dissimilarities in bacterial diversity, succession, and associated functions between MPBs, in which bacterial species with specific functions should be taken seriously.

Spatio-temporal dynamics of bacterial communities in the shoreline of Laurentian great Lake Erie and Lake St. Clair\u2019s large freshwater ecosystems

BackgroundLong-term trends in freshwater bacterial community composition (BCC) and dynamics are not yet well characterized, particularly in large lake ecosystems. We addressed this gap by temporally (15 months) and spatially (6 sampling locations) characterizing BCC variation in lakes Erie and St. Clair; two connected ecosystems in the Laurentian Great Lakes.ResultsWe found a spatial variation of the BCC between the two lakes and among the sampling locations (significant changes in the relative abundance of 16% of the identified OTUs at the sampling location level). We observed five distinct temporal clusters (UPGMA broad-scale temporal variation) corresponding to seasonal variation over the 15 months of sampling. Temporal variation among months was high, with significant variation in the relative abundance of 69% of the OTUs. We identified significant differences in taxonomic composition between summer months of 2016 and 2017, with a corresponding significant reduction in the diversity of BCC in summer 2017.ConclusionsAs bacteria play a key role in biogeochemical cycling, and hence in healthy ecosystem function our study defines the scope for temporal and spatial variation in large lake ecosystems. Our data also show that freshwater BCC could serve as an effective proxy and monitoring tool to access large lake health.

Human Fecal Contamination Corresponds to Changes in the Freshwater Bacterial Communities of a Large River Basin.

ABSTRACTMicrobial water quality is generally monitored by culturable fecal indicator bacteria (FIB), which are intended to signal human health risk due to fecal pollution. However, FIB have limited utility in most urbanized watersheds as they do not discriminate among fecal pollution sources, tend to make up a small fraction of the total microbial community, and do not inform on pollution impacts on the native ecosystem. To move beyond these limitations, we assessed entire bacterial communities and investigated how bacterial diversity relates to traditional ecological and human health-relevant water quality indicators throughout the Milwaukee River Basin. Samples were collected from 16 sites on 5 days during the summer, including both wet and dry weather events, and were processed by 16S rRNA gene amplicon sequencing. Historical water quality at each sampling location, as opposed to upstream land use, was associated significantly with bacterial community alpha diversity. Source partitioning the sequence data was important for determining water quality relationships. Sewage-associated bacterial sequences were detected in all samples, and the relative abundance of sewage sequences was strongly associated with the human Bacteroides fecal marker. From this relationship, we developed a preliminary threshold for human sewage pollution when using bacterial community sequence data. Certain abundant freshwater bacterial sequences were also associated with human fecal pollution, suggesting their possible utility in water quality monitoring. This study sheds light on how bacterial community analysis can be used to supplement current water quality monitoring techniques to better understand interactions between ecological water quality and human health indicators.IMPORTANCE Surface waters in highly developed mixed-use watersheds are frequently impacted by a wide variety of pollutants, leading to a range of impairments that must be monitored and remediated. With advancing technologies, microbial community sequencing may soon become a feasible method for routine evaluation of the ecological quality and human health risk of a water body. In this study, we partnered with a local citizen science organization to evaluate the utility of microbial community sequencing for identifying pollution sources and ecological impairments in a large mixed-use watershed. We show that changes in microbial community diversity and composition are indicative of both long-term ecological impairments and short-term fecal pollution impacts. By source partitioning the sequence data, we also estimate a threshold target for human sewage pollution, which may be useful as a starting point for future development of sequencing-based water quality monitoring techniques.

Increasing Freshwater Salinity Impacts Aerosolized Bacteria.

Increases in the salt concentration of freshwater result in detrimental impacts on water quality and ecosystem biodiversity. Biodiversity effects include freshwater microbiota, as increasing salinity can induce shifts in the structure of native freshwater bacterial communities, which could disturb their role in mediating basal ecosystem services. Moreover, salinity affects the wave breaking and bubble-bursting mechanisms via which water-to-air dispersal of bacteria occurs. Given this dual effect of freshwater salinity on waterborne bacterial communities and their aerosolization mechanism, further effects on aerosolized bacterial diversity and abundance are anticipated. Cumulative salt additions in the freshwater-euhaline continuum (0-35 g/kg) were administered to a freshwater sample aerosolized inside a breaking wave analogue tank. Waterborne and corresponding airborne bacteria were sampled at each salinity treatment and later analyzed for diversity and abundance. Results demonstrated that the airborne bacterial community was significantly different (PERMANOVA; F1,22 = 155.1, r2 = 0.38, p < 0.001) from the waterborne community. The relative aerosolization factor (r-AF), defined as the air-to-water relative abundance ratio, revealed that different bacterial families exhibited either an enhanced (r-AF ≫ 1), neutral (r-AF ∼ 1), or diminished (r-AF ≪ 1) transfer to the aerosol phase throughout the salinization gradient. Going from freshwater to euhaline conditions, aerosolized bacterial abundance exhibited a nonmonotonic response with a maximum peak at lower oligohaline conditions (0.5-1 g/kg), a decline at higher oligohaline conditions (5 g/kg), and a moderate increase at polyhaline-euhaline conditions (15-35 g/kg). Our results demonstrate that increases in freshwater salinity are likely to influence the abundance and diversity of aerosolized bacteria. These shifts in aerosolized bacterial communities might have broader implications on public health by increasing exposure to airborne pathogens via inhalation. Impacts on regional climate, related to changes in biological ice-nucleating particles (INPs) emission from freshwater, are also expected.

Impact of Zero-Valent Iron on Freshwater Bacterioplankton Metabolism as Predicted from 16S rRNA Gene Sequence Libraries.

The application of zero-valent iron particles (ZVI) for the treatment of heavily polluted environment and its biological effects have been studied for at least two decades. Still, information on the impact on bacterial metabolic pathways is lacking. This study describes the effect of microscale and nanoscale ZVI (mZVI and nZVI) on the abundance of different metabolic pathways in freshwater bacterial communities. The metabolic pathways were inferred from metabolism modelling based on 16S rRNA gene sequence data using paprica pipeline. The nZVI changed the abundance of numerous metabolic pathways compared to a less influencing mZVI. We identified the 50 most affected pathways, where 31 were related to degradation, 17 to biosynthesis, and 2 to detoxification. The linkage between pathways was two times higher in nZVI samples compared to mZVI, and was specifically higher considering the arsenate detoxification II pathway. Limnohabitans and Roseiflexus were linked to the same pathways in both nZVI and mZVI. The prediction of metabolic pathways increases our knowledge of the impacts of nZVI and mZVI on freshwater bacterioplankton.

Nano-Sized Polystyrene at 1mg/L Concentrations Does Not Show Strong Disturbance on the Freshwater Microbial Community.

In recent years, microplastics and nanoplastics have gained public attention, but their impacts on the freshwater microbial communities is rarely evaluated. In this study, the effects of 1mg/L nano-sized polystyrene (nPS) and its modified forms (carboxyl-modified and amino-modified nPS) on the structures and functions of freshwater microbial community were determined. The nPS were found to slightly reduce the chlorophyll-a and increase the phycocyanin contents of freshwater microbial communities. Moreover, the richness of the microbial communities temporarily decreased during this process, while their diversity remained uninfluenced by treatment with nPS. Although the three tested nPS types were found to disturb the compositions of both the prokaryotic and eukaryotic communities to some degree, they did not affect the functions of freshwater bacterial communities significantly due to functional redundancy. Our study demonstrated that the ecotoxicities of the nPS itself were found to be lower than what is generally expected.

Diel Dynamics of Freshwater Bacterial Communities at Beaches in Lake Erie and Lake St. Clair, Windsor, Ontario.

Bacteria play a key role in freshwater biogeochemical cycling as well as water safety, but short-term trends in freshwater bacterial community composition and dynamics are not yet well characterized. We sampled four public beaches in southern Ontario, Canada; in June, July, and August (2016) over a 24-h (diel) cycle at 2-h intervals. Using high-throughput sequencing of 16S rRNA gene, we found substantial bi-hourly and day/night variation in the bacterial communities with considerable fluctuation in the relative abundance of Actinobacteria and Proteobacteria phyla. Moreover, relative abundance of Enterobacteriaceae (associated with potential health risk) was significantly high at night in some dial cycles. Diversity was significantly high at night across most of the diel sampling events. qPCR assays showed a substantial bi-hourly variation of Escherichia coli levels with a significant high level of E. coli at night hours in comparison with day hours and the lowest levels at noon and during the afternoon hours. Taken together, these findings highlighted a considerable short-term temporal variation of bacterial communities which helps better understanding of freshwater bacterial dynamics and their ecology. E. coli monitoring showed that multiple samples in different hours will provide more accurate picture of freshwater safety and human health risk. Graphical abstract.

Microhabitats are associated with diversity-productivity relationships in freshwater bacterial communities.

Eukaryotic communities commonly display a positive relationship between biodiversity and ecosystem function (BEF) but the results have been mixed when assessed in bacterial communities. Habitat heterogeneity, a factor in eukaryotic BEFs, may explain these variable observations but it has not been thoroughly evaluated in bacterial communities. Here, we examined the impact of habitat on the relationship between diversity assessed based on the (phylogenetic) Hill diversity metrics and heterotrophic productivity. We sampled co-occurring free-living (more homogenous) and particle-associated (more heterogeneous) bacterial habitats in a freshwater, estuarine lake over three seasons: spring, summer and fall. There was a strong, positive, linear relationship between particle-associated bacterial richness and heterotrophic productivity that strengthened when considering dominant taxa. There were no observable BEF trends in free-living bacterial communities for any diversity metric. Biodiversity, richness and Inverse Simpson's index, were the best predictors of particle-associated production whereas pH was the best predictor of free-living production. Our findings show that heterotrophic productivity is positively correlated with the effective number of taxa and that BEF relationships are associated with microhabitats. These results add to the understanding of the highly distinct contributions to diversity and functioning contributed by bacteria in free-living and particle-associated habitats.

Differential impact of top-down and bottom-up forces in structuring freshwater bacterial communities.

Limited data exist on the simultaneous impact of bottom-up (nutrients) and top-down (viruses and heterotrophic nanoflagellates) forces in shaping freshwater bacterial communities. In our laboratory microcosms, nutrient additions (organic and inorganic) and viral reduction approach led to the proliferation of high nucleic acid (HNA) bacterial subpopulation without an increase in phage abundance. High viral-mediated bacterial lysis in the presence of nanoflagellates yielded high proportion of low nucleic acid bacterial subpopulation. 16S rRNA gene sequence analysis indicated that members of classes Proteobacteria and Bacteroidetes evoked differential responses to nutrients and mortality forces, thereby resulting in differences (P <0.001) in bacterial community composition and diversity, as observed from analysis of similarities and UniFrac analysis. Bacterial species richness (Chao) and diversity (Shannon) index was significantly higher (P <0.001) in the presence of both the top-down factors and viruses alone, whereas lower host diversity was observed under nutrient relaxation of growth-limiting substrates due to the explosive growth of opportunistic HNA bacterial subpopulation. Our results are in agreement with the theoretical model of 'killing the winner', where the availability of growth-limiting substrates can act as a stimulating factor for host community composition while top-down forces can operate in the control of host diversity.

Turnover is replaced by nestedness with increasing geographical distance in bacterial communities of coastal shallow lakes

In this study we measured the relative contribution of two components of β-diversity, turnover and nestedness, of bacterioplankton among 25 shallow lakes in southern Brazil and tested their relationship with local (environment, chlorophyll-a and biomass of phytoplanktonic classes) and landscape variables, as well as geographical distance. We predicted that turnover would be the largest share of total β-diversity due to the variation of local characteristics among lakes. Further, we expected nestedness to increase at the expense of turnover with increasing geographical distance among lakes due to dispersal limitation. The results indicated a higher contribution of turnover than nestedness to total β-diversity, which was driven by local factors. When the relationship between β-diversity components and the spatial extent between each lake and all other lakes was considered, turnover was replaced by nestedness with increasing geographical distance for 8 (the furthermost lakes) of the 25 lakes likely because of a combination of decreasing dispersal due to distance and richness differences due to wind-driven mass effects. The results of this study suggest a role for nestedness as an indicator of dispersal limitation owing to geographical distance and wind dispersal, and for turnover as an indicator of species sorting because of environmental filters for these freshwater bacterial communities.

Annual bacterial community cycle in a seasonally ice‐covered river reflects environmental and climatic conditions

AbstractBacteria are key players in biogeochemical cycles and control water quality in freshwater ecosystems. Nevertheless, little is known about the identity and ecology of riverine bacteria, especially during ice‐covered periods that are often mistakenly perceived as periods with negligible biological activities. Here, we analyzed in detail the effects of environmental and climatic conditions on freshwater bacterial community structure and diversity over a 2‐yr sampling campaign, targeting a seasonally ice‐covered river of the Quebec City (Canada) area, the Saint‐Charles River. Quantitative polymerase chain reaction and 16S ribosomal RNA gene high‐throughput sequencing demonstrated a strong seasonal cycle of the bacterial community composition with rapid successions of bacterial lineages, reflecting the harsh climatic condition of the region. During the summer, the bacterial community was dominated by typical freshwater microorganisms such as Limnohabitans, Sporichthyaceae hgcI/acI clade, and Pseudarcicella. In contrast, the results suggest that during the cold season, the low water temperatures, combined with other prevailing conditions such as reduced light availability and minimal particulate inputs from the catchment, created various environmental niches for potential methanotrophic Gammaproteobacteria, such as Crenothrix and Methylobacter, other Betaproteobacteria, such as Candidatus Methylopumilus, Candidatus Nitrotoga, and Rhodoferax, as well as Verrucomicrobia and Parcubacteria. The presence of these taxa in the winter suggests active carbon, iron, and nitrogen cycling under ice, whereas summer lineages are dormant or in a phase of reduced activity. These results increase our understanding of bacterial dynamic and potential metabolic processes occurring in seasonally ice‐covered inland waters, providing evidence that winter can be an especially important period for freshwater ecological processes.

Biogeography and Diversity of Freshwater Bacteria on a River Catchment Scale.

To illustrate how freshwater bacterial community changes with geographic gradient, we investigated the spatial changes of bacterial abundance and community structures from over 200 samples on a catchment scale in the Songhua River using heterotrophic plate counts, flow cytometry, denaturing gradient gel electrophoresis, and pyrosequencing analysis. The results showed that the mainstream had higher cultivable bacteria and total bacterial concentration than tributaries in the Songhua River catchment. Response model analysis demonstrated that the bacterial community exhibits a biogeographical signature even in an interconnected river network system, and the total bacterial concentration and biodiversity were significantly correlated to latitude (p< 0.001) and longitude (p< 0.001). Multivariate redundancy analysis indicated that temperature was the most important factor driving bacterial community structure in the Songhua River, which accounts for 35.30% variance of communities, then dissolved oxygen (17.60%), latitude (17.60%), longitude (11.80%), and pH (5.88%). High-throughput pyrosequencing revealed that at the phylum level, Proteobacteria was numerically dominant (89.6%) in river catchment, followed by Bacteroidetes (8.1%) and Cyanobacteria (1.2%). The overall results revealed that the bacterial community was driven by geographical distance regardless of the continuum of the river on a catchment scale.