Freshwater Bacterioplankton Communities Research Articles

Resistance, resilience, and functional redundancy of freshwater bacterioplankton communities facing a gradient of agricultural stressors in a mesocosm experiment.

Agricultural pollution with fertilizers and pesticides is a common disturbance to freshwater biodiversity. Bacterioplankton communities are at the base of aquatic food webs, but their responses to these potentially interacting stressors are rarely explored. To test the extent of resistance and resilience in bacterioplankton communities faced with agricultural stressors, we exposed freshwater mesocosms to single and combined gradients of two commonly used pesticides: the herbicide glyphosate (0-15mg/L) and the neonicotinoid insecticide imidacloprid (0-60μg/L), in high or low nutrient backgrounds. Over the 43-day experiment, we tracked variation in bacterial density with flow cytometry, carbon substrate use with Biolog EcoPlates, and taxonomic diversity and composition with environmental 16S rRNA gene amplicon sequencing. We show that only glyphosate (at the highest dose, 15mg/L), but not imidacloprid, nutrients, or their interactions measurably changed community structure, favouring members of the Proteobacteria including the genus Agrobacterium. However, no change in carbon substrate use was detected throughout, suggesting functional redundancy despite taxonomic changes. We further show that communities are resilient at broad, but not fine taxonomic levels: 24days after glyphosate application the precise amplicon sequence variants do not return, and tend to be replaced by phylogenetically close taxa. We conclude that high doses of glyphosate - but still within commonly acceptable regulatory guidelines - alter freshwater bacterioplankton by favouring a subset of higher taxonomic units (i.e., genus to phylum) that transiently thrive in the presence of glyphosate. Longer-term impacts of glyphosate at finer taxonomic resolution merit further investigation.

Genetic diversity of Actinobacteria inhabiting water and sponges of Lake Baikal

Actinobacteria is a ubiquitous major group in freshwater bacterioplankton and sponge microbial communities. We were first to analyse the diversity of Actinobacteria inhabiting Lake Baikal water and endemic sponges Lubomirskia baikalensis (healthy and diseased) using highthroughput 16S rRNA sequencing. In total, we have assigned of three classes, fourteen orders and twenty-six families of Actinobacteria. The diversity and composition of actinobacteria were common in bacterioplankton and healthy sponges. The environmental crisis in the coastal zone of Lake Baikal led to a colonization of the L. baikalensis by benthic and soil-derived representatives of Actinobacteria. Some amplicon sequences of opportunistic genera were detected in the coastal water. The example of Actinobacteria has dramatically demonstrated the response of bacterial populations caused by negative environmental impact.

Cyanobacterial blooms alter the relative importance of neutral and selective processes in assembling freshwater bacterioplankton community

CyanoHABs have substantial impacts on the functioning and sustainability of freshwater ecosystems by restricting light penetration, depleting dissolved oxygen, and producing various toxins. This study combined physicochemical variable measurements, 16S rRNA gene sequencing and microscopy observations to examine mechanisms that govern the assembly of bacterioplankton communities following the progress of cyanobacterial blooms in a freshwater reservoir. Throughout the sampling season, bacterioplankton distribution patterns were well predicted by a neutral model, which assumes passive dispersal and ecological drift as the predominate mechanisms for community assembly. The neutral model consistently explained the distribution of over 67% of bacterioplankton OTUs and its fit was weaker during the cyanobacterial blooms (R2 = 0.322) than the before- (R2 = 0.549) and after-bloom stages (R2 = 0.535). Variations of environmental factors, acting as selective pressures, explained shifts of non-neutral OTUs (above/under neutral prediction) (63.9%) better than neutral OTUs (34.5%). Co-occurrence network analysis organized microbial communities into modules and revealed strong positive correlations between bacterioplankton and cyanobacteria than with planktonic algae and zooplankton. Overall, our results suggest that neutral processes play significant roles in assembling bacterioplankton communities over a cyanobacterial bloom succession and its relative importance may be weakened by biotic pressures (interspecific interactions) during the bloom period. Our results also indicate that among biotic factors, cyanobacteria had greater impacts on bacterioplankton community assembly than planktonic algae and zooplankton.

Soils associated to different tree communities do not elicit predictable responses in lake bacterial community structure and function.

Freshwater bacterioplankton communities are influenced by the inputs of material and bacteria from the surrounding landscape, yet few studies have investigated how different terrestrial inputs affect bacterioplankton. We examined whether the addition of soils collected under various tree species combinations differentially influences lake bacterial communities. Lake water was incubated for 6 days following addition of five different soils. We assessed the taxonomic composition (16S rRNA gene sequencing) and metabolic activity (Biolog Ecoplates) of lake bacteria with and without soil addition, and compared these to initial soil communities. Soil bacterial assemblages showed a strong influence of tree composition, but such community differences were not reflected in the structure of lake communities that developed during the experiment. Bacterial taxa showing the largest abundance increases during incubation were initially present in both lake water and across most soils, and were related to Cytophagales, Burkholderiales and Rhizobiales. No clear metabolic profiles based on inoculum source were found, yet soil-amended communities used 60% more substrate than non-inoculated communities. Overall, we show that terrestrial inputs influence aquatic communities by stimulating the growth and activity of certain ubiquitous taxa distributed across the terrestrial-aquatic continuum, yet different forest soils did not cause predictable changes in lake bacterioplankton assemblages.

Inoculation history affects community composition in experimental freshwater bacterioplankton communities.

Priority effects occur when the arrival order of species or genotypes has a lasting effect on community or population structure. For freshwater bacteria, priority effects have been shown experimentally among individual species, but no experiments have been performed using complex natural communities. We investigated experimentally whether a foreign bacterioplankton community influences the community assembly trajectory when inoculated prior to the local community, whether inoculation time lag affects priority effects, and how the individual OTUs responded to time lag. Two bacterioplankton communities from dissimilar ponds were inoculated into one of the natural media with a time lag of 0, 12, 36 or 60 h, giving advantage in time to the foreign community. All three time lags resulted in priority effects, as the final community composition of these treatments differed significantly from that of the treatment with no time lag, but compositional shifts were not linear to inoculation time lag. The responses of individual OTUs to time lag were highly diverse and not predictable based on their immigration history or relative abundance in the inocula or control. The observed impact and complexity of priority effects in multispecies systems emphasize the importance of this process in structuring both natural and industrial bacterial communities.

Contrasting dynamics and environmental controls of dispersed bacteria along a hydrologic gradient

Freshwater bacterioplankton communities are influenced by the transport of bacteria from the surrounding terrestrial environments. It has been shown that, although most of these dispersed bacteria gradually disappear along the hydrologic continuum, some can thrive in aquatic systems and become dominant, leading to a gradual succession of communities. Here we aimed at exploring the environmental factors driving the structure of such contrasting bacterial populations as well as their functional properties. Using Illumina sequencing of the 16S rRNA gene, we characterized the taxonomic composition of bacterioplankton communities from 10 streams and rivers in Québec spanning the whole hydrologic continuum (river Strahler order 0 to 7), which were sampled in two occasions. With the aim to understand the fate and controls of the transported bacteria, among the taxa present at the origin of the hydrologic gradient (i.e., in the smallest headwater streams) we identified two types of dynamics: i) ‘Tourist’ taxa, which were those that decreased in abundance from the headwaters towards the largest rivers, and ii) ‘Seed’ taxa, those that increased their abundances along the hydrologic continuum. Communities changed gradually from the fast-flowing headwater streams dominated by ‘Tourist’ taxa (ca. 95% of the sequences) towards the largest rivers (Strahler order 4-7) where ‘Seed’ taxa comprised up to 80% of community sequences. Variation in taxonomic composition of the communities dominated by ‘Tourist’ taxa in streams seemed related to different degree of terrestrial inputs, whereas compositional changes in ‘Seed’ communities in the large rivers were linked to differences in autochthonous processes. Finally, the two types of communities differed significantly in their metabolic potential assessed through Biolog Ecoplates. All this suggests that hydrologic transport modulates the gradual replacement of two contrasting population types subjected to different environmental controls and with different metabolic potentials. Moreover, we show that the separate exploration of the two pools of taxa allows unveiling environmental drivers and processes operating on them that remain hidden if explored at the whole community level.

Differential impact of lytic viruses on the taxonomical resolution of freshwater bacterioplankton community structure

The significance of lytic viral lysis in shaping bacterial communities in temperate freshwater systems is less documented. Here we used Illumina sequencing of 16S rRNA genes to examine bacterial community structure and diversity in relation to variable viral lysis in the euphotic zone of 25 temperate freshwater lakes (French Massif Central). We captured a rich bacterial community that was dominated by a few bacterial classes and operational taxonomic units (OTUs) frequently detected in other freshwater ecosystems. In the investigated lakes with contrasting physico-chemical characteristics, the dominant bacterioplankton community was represented by major taxonomical orders, namely Actinomycetales, Burkholderiales, Sphingobacteriales, Acidimicrobiales, Flavobacteriales and Cytophagales covering about 70% of all sequences. Viral lysis was significantly correlated with the bacterial diversity indices (Chao, Shannon, OTUs) which explained about 33% and 45% of the variation in species diversity and observed richness respectively. Anosim and UniFrac analyses indicated a clear distinction of bacterial community structure among the lakes that exhibited high and low lytic viral infection (FIC) rates. Based on our findings, high FIC (>10%) supported higher species richness, whereas low FIC (<10%) resulted in less diverse community. Our study strongly suggests that lytic activity prevailed over the type of lake ecosystems in shaping bacterioplankton diversity.

Vertical partitioning of freshwater bacterioplankton community in a deep mesotrophic lake with a fully oxygenated hypolimnion (Lake Biwa, Japan).

In freshwater microbial ecology, extensive studies are attempting to characterize the vast majority of uncultivated bacterioplankton taxa. However, these studies mainly focus on the epilimnion and little is known regarding the bacterioplankton inhabiting the hypolimnion of deep holomictic lakes, despite its biogeochemical importance. In this study, we investigated the bacterioplankton community composition in a deep freshwater lake with a fully oxygenated hypolimnion (Lake Biwa, Japan) using high-throughput 16S rRNA gene amplicon sequencing. Sampling at a pelagic site over 15 months throughout the water column revealed that the community composition in the hypolimnion was significantly different from that in the epilimnion. The bacterial community in the hypolimnion was composed of groups dominating in the whole water layer (e.g., bacI-A1 and acI-B1) and groups that were hypolimnion habitat specialists. Among the hypolimnion specialists, members of Chloroflexi and Planctomycetes were highly represented (e.g., CL500-11, CL500-15 and CL500-37), followed by members of Acidobacteria, Chlorobi and nitrifiers (e.g., Ca. Nitrosoarchaeum, Nitrosospira and Nitrospira). This study identified the number of previously understudied taxa dominating the deep aerobic freshwater habitat, suggesting that the biogeochemical cycling there is driven by the microbial community that are different from that in the epilimnion.

Effects of Dispersal and Initial Diversity on the Composition and Functional Performance of Bacterial Communities.

Natural communities are open systems and consequently dispersal can play an important role for the diversity, composition and functioning of communities at the local scale. It is, however, still unclear how effects of dispersal differ depending on the initial diversity of local communities. Here we implemented an experiment where we manipulated the initial diversity of natural freshwater bacterioplankton communities using a dilution-to-extinction approach as well as dispersal from a regional species pool. The aim was further to test whether dispersal effects on bacterial abundance and functional parameters (average community growth rates, respiration rates, substrate utilisation ability) differ in dependence of the initial diversity of the communities. First of all, we found that both initial diversity and dispersal rates had an effect on the recruitment of taxa from a regional source, which was higher in communities with low initial diversity and at higher rates of dispersal. Higher initial diversity and dispersal also promoted higher levels of richness and evenness in local communities and affected, both, separately or interactively, the functional performance of communities. Our study therefore suggests that dispersal can influence the diversity, composition and functioning of bacterial communities and that this effect may be enhanced if the initial diversity of communities is depleted.

When, where and how does microbial community composition matter?

When, where and how does microbial community composition matter?

The Diversity of the Limnohabitans Genus, an Important Group of Freshwater Bacterioplankton, by Characterization of 35 Isolated Strains

Bacteria of the genus Limnohabitans, more precisely the R-BT lineage, have a prominent role in freshwater bacterioplankton communities due to their high rates of substrate uptake and growth, growth on algal-derived substrates and high mortality rates from bacterivory. Moreover, due to their generally larger mean cell volume, compared to typical bacterioplankton cells, they contribute over-proportionally to total bacterioplankton biomass. Here we present genetic, morphological and ecophysiological properties of 35 bacterial strains affiliated with the Limnohabitans genus newly isolated from 11 non-acidic European freshwater habitats. The low genetic diversity indicated by the previous studies using the ribosomal SSU gene highly contrasted with the surprisingly rich morphologies and different patterns in substrate utilization of isolated strains. Therefore, the intergenic spacer between 16S and 23S rRNA genes was successfully tested as a fine-scale marker to delineate individual lineages and even genotypes. For further studies, we propose the division of the Limnohabitans genus into five lineages (provisionally named as LimA, LimB, LimC, LimD and LimE) and also additional sublineages within the most diversified lineage LimC. Such a delineation is supported by the morphology of isolated strains which predetermine large differences in their ecology.

Links between metabolic plasticity and functional redundancy in freshwater bacterioplankton communities

Metabolic plasticity and functional redundancy are fundamental properties of microbial communities, which shape their response to environmental forcing, and also mediate the relationship between community composition and function. Yet, the actual quantification of these emergent community properties has been elusive, and we thus do not know how they vary across bacterial communities, and their relationship to environmental gradients and to each other. Here we present an experimental framework that allows us to simultaneously quantify metabolic plasticity and functional redundancy in freshwater bacterioplankton communities, and to explore connections that may exists between them. We define metabolic plasticity as the rate of change in single-cell properties (cell wall integrity, cell size, single-cell activity) relative to changes in community composition. Likewise, we define functional redundancy as the rate of change in carbon substrate uptake capacities relative to changes in community composition. We assessed these two key community attributes in transplant experiments where bacterioplankton from various aquatic habitats within the same watershed were transplanted from their original water to waters from other systems that differ in their main resources. Our results show that metabolic plasticity is an intrinsic property of bacterial communities, whereas the expression of functional redundancy appears to be more dependent on environmental factors. Furthermore, there was an overall strong positive relationship between the level of functional redundancy and of metabolic plasticity, suggesting no trade-offs between these community attributes but rather a possible co-selection. The apparent continuum in the expression of both functional redundancy and plasticity among bacterial communities and the link between them, in turn suggest that the link between community diversity and function may also vary along a continuum, from being very tight, to being weak, or absent.

A decade of seasonal dynamics and co-occurrences within freshwater bacterioplankton communities from eutrophic Lake Mendota, WI, USA

With an unprecedented decade-long time series from a temperate eutrophic lake, we analyzed bacterial and environmental co-occurrence networks to gain insight into seasonal dynamics at the community level. We found that (1) bacterial co-occurrence networks were non-random, (2) season explained the network complexity and (3) co-occurrence network complexity was negatively correlated with the underlying community diversity across different seasons. Network complexity was not related to the variance of associated environmental factors. Temperature and productivity may drive changes in diversity across seasons in temperate aquatic systems, much as they control diversity across latitude. While the implications of bacterioplankton network structure on ecosystem function are still largely unknown, network analysis, in conjunction with traditional multivariate techniques, continues to increase our understanding of bacterioplankton temporal dynamics.

Resistance and resilience of microbial communities – temporal and spatial insurance against perturbations

Bacteria play fundamental roles for many ecosystem processes; however, little empirical evidence is available on how environmental perturbations affect their composition and function. We investigated how spatial and temporal refuges affect the resistance and resilience of a freshwater bacterioplankton community upon a salinity pulse perturbation in continuous cultures. Attachment to a surface avoided the flushing out of cells and enabled re-colonization of the liquid phase after the perturbation, hence serving as a temporal refuge. A spatial refuge was established by introduction of bacteria from an undisturbed reservoir upstream of the continuous culture vessel, acting analogous to a regional species pool in a metacommunity. The salinity pulse affected bacterial community composition and the rates of respiration and the pattern of potential substrate utilization as well as the correlation between composition and function. Compared with the no-refuge treatment, the temporal refuge shortened return to pre-perturbation conditions, indicating enhanced community resilience. Composition and function were less disturbed in the treatment providing a spatial refuge, suggesting higher resistance. Our results highlight that spatial and temporal dynamics in general and refuges in particular need to be considered for conceptual progress in how microbial metacommunities are shaped by perturbations.

Freshwater bacterioplankton richness in oligotrophic lakes depends on nutrient availability rather than on species–area relationships

A central goal in ecology is to grasp the mechanisms that underlie and maintain biodiversity and patterns in its spatial distribution can provide clues about those mechanisms. Here, we investigated what might determine the bacterioplankton richness (BR) in lakes by means of 454 pyrosequencing of the 16S rRNA gene. We further provide a BR estimate based upon a sampling depth and accuracy, which, to our knowledge, are unsurpassed for freshwater bacterioplankton communities. Our examination of 22,669 sequences per lake showed that freshwater BR in fourteen nutrient-poor lakes was positively influenced by nutrient availability. Our study is, thus, consistent with the finding that the supply of available nutrients is a major driver of species richness; a pattern that may well be universally valid to the world of both micro- and macro-organisms. We, furthermore, observed that BR increased with elevated landscape position, most likely as a consequence of differences in nutrient availability. Finally, BR decreased with increasing lake and catchment area that is negative species-area relationships (SARs) were recorded; a finding that re-opens the debate about whether positive SARs can indeed be found in the microbial world and whether positive SARs can in fact be pronounced as one of the few 'laws' in ecology.

Interactive effect of temperature and resources on carbon cycling by freshwater bacterioplankton communities

Planktonic heterotrophic prokaryotes play an essential role in all aquatic ecosystems due to their short generation times and access to dissolved organic carbon and nutrients. In order to understand how rising temperatures and nutrient loading affect the biogeochemistry of whole lake ecosystems, it is essential to understand the interactive effects that temperature and resources have on the metabolism of natural bacterioplankton communities. To address this question we sampled bacterial communities from 1 mesotrophic and 1 oligotrophic lake in Clearwater County, Minnesota, USA, in winter and summer. Each community was exposed to a combination of carbon, nitrogen and phosphorus additions at 4 different levels (ambient, 2 ×, 5 ×, and 10 × ambient) and 4 temperatures (4, 14, 24 and 34°C). Community metabolic response to temperature depended on the resource treat- ment, and the season when the community was sampled. Bacterial respiration increased more with temperature than bacterial growth, resulting in decreased bacterial growth efficiencies at higher temperatures. This result was most pronounced in the high resource treatments, while at lower resource levels the results were more ambiguous. In addition, differences between seasons and lakes suggested that the bacterial communities had adapted or acclimated to in situ temperature such that communities sampled from cold temperature environments had higher respiration at high tempera- tures than those sampled from warmer environments. These results suggest that the effect of temper- ature on carbon cycling mediated by the bacterial community depends on both the in situ resource pool and the extent to which the bacterial community is acclimated to a specific temperature regime.

Microbial community dynamics in a humic lake: differential persistence of common freshwater phylotypes

In an effort to better understand the factors contributing to patterns in freshwater bacterioplankton community composition and diversity, we coupled automated ribosomal intergenic spacer analysis (ARISA) to analysis of 16S ribosomal RNA (rRNA) gene sequences to follow the persistence patterns of 46 individual phylotypes over 3 years in Crystal Bog Lake. Additionally, we sought to identify linkages between the observed phylotype variations and known chemical and biological drivers. Sequencing of 16S rRNA genes obtained from the water column indicated the presence of phylotypes associated with the Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria, TM7 and Verrucomicrobia phyla, as well as phylotypes with unknown affiliation. Employment of the 16S rRNA gene/ARISA method revealed that specific phylotypes varied independently of the entire bacterial community dynamics. Actinobacteria, which were present on greater than 95% of sampling dates, did not share the large temporal variability of the other identified phyla. Examination of phylotype relative abundance patterns (inferred using ARISA fragment relative fluorescence) revealed a strong correlation between the dominant phytoplankton succession and the relative abundance patterns of the majority of individual phylotypes. Further analysis revealed covariation among unique phylotypes, which formed several distinct bacterial assemblages correlated with particular phytoplankton communities. These data indicate the existence of unique persistence patterns for different common freshwater phylotypes, which may be linked to the presence of dominant phytoplankton species.

Specific Detection, Isolation, and Characterization of Selected, Previously Uncultured Members of the Freshwater Bacterioplankton Community

High-throughput cultivation was combined with rapid and group-specific phylogenetic fingerprinting in order to recover representatives of three freshwater bacterioplankton communities. A total of 570 bacterial cultures were obtained by employing the most probable number and MicroDrop techniques. The majority of the cultured bacteria were closely related to previously uncultured bacteria and grouped with the alpha-Proteobacteria, beta-Proteobacteria, Actinobacteria, Firmicutes, or Flavobacteria-Cytophaga lineage. Correspondingly, the natural bacterioplankton community was analyzed by high-resolution phylogenetic fingerprinting of these five bacterial lineages. 16S rRNA gene fragments were generated for each lineage and subsequently separated by denaturing gradient gel electrophoresis. By the combination of five group-specific PCR protocols, the total number of 16S rRNA gene fingerprints generated from the natural communities was increased sixfold compared to conventional (eubacterial) fingerprinting. Four of the environmental alpha-Proteobacteria 16S rRNA gene sequences obtained from the natural community were found to be identical to those of bacterial isolates. One of these phylotypes was detected in 14 different cultures and hence represented the most frequently cultured bacterium. Three of these 14 strains were characterized in detail. Their complete 16S rRNA gene sequences showed only 93% similarity to that of Sandaracinobacter sibiricus, the closest relative described so far. The novel phylotype of bacterium is a strict aerobe capable of using numerous organic carbon substrates and contains bacteriochlorophyll a bound to two different photosynthetic light-harvesting complexes. Dot blot hybridization revealed that the strains occur in lakes of different trophic status and constitute up to 2% of the microbial community.

Analysis of Microbial Gene Transcripts in Environmental Samples

We analyzed gene expression in marine and freshwater bacterioplankton communities by the direct retrieval and analysis of microbial transcripts. Environmental mRNA, obtained from total RNA by subtractive hybridization of rRNA, was reverse transcribed, amplified with random primers, and cloned. Approximately 400 clones were analyzed, of which approximately 80% were unambiguously mRNA derived. mRNAs appeared to be from diverse taxonomic groups, including both Bacteria (mainly alpha- and gamma-Proteobacteria) and Archaea (mainly Euryarchaeota). Many transcripts could be linked to environmentally important processes such as sulfur oxidation (soxA), assimilation of C1 compounds (fdh1B), and acquisition of nitrogen via polyamine degradation (aphA). Environmental transcriptomics is a means of exploring functional gene expression within natural microbial communities without bias toward known sequences, and provides a new approach for obtaining community-specific variants of key functional genes.

An introduction to the biogeography of aquatic microbes

Aquatic microbes, like all organisms, have biogeographies, but this subject has attracted relatively little attention. In this review, recent results exploiting techniques of molecular biology are summarized to place in perspective the studies of this Theme Section. The studies con- sidered concern large-scale patterns of spatial distribution among heterotrophic planktonic prokary- otic and eukaryotic microbes. For freshwater bacterioplankton communities, reported patterns are inconsistent. Taxonomic richness may increase with system size, and composition may be related among neighboring bodies of water. However, inconsistencies in patterns may be due to differences in the temporal and spatial scales considered. Among planktonic marine prokaryotes, biogeographic patterns are known only in terms of high level groups, e.g. Archea are perhaps dominant in deep oceanic waters. However, studies of large-scale patterns have just begun and they suggest that some ribotypes or species may be restricted to certain oceanic areas. Eukaryotic microbes appear to char- acterised by high capacities for both dispersal and gene flow. Recent studies appear to conclude that we can form morphological, genetic and physiological groupings but their inter-relationships are obscure at this point in time.