Modern metagenomic environmental DNA studies are almost completely reliant on next-generation sequencing, making evaluations of these methods critical. We compare two next-generation sequencing techniques – amplicon and shotgun – on water samples across four of Brazil’s major river floodplain systems (Amazon, Araguaia, Paraná, and Pantanal). Less than 50% of phyla identified via amplicon sequencing were recovered from shotgun sequencing, clearly challenging the dogma that mid-depth shotgun recovers more diversity than amplicon-based approaches. Amplicon sequencing also revealed ~27% more families. Overall the amplicon data were more robust across both biodiversity and community ecology analyses at different taxonomic scales. Our work doubles the sampling size in similar environmental studies, and novelly integrates environmental data (e.g., pH, temperature, nutrients) from each site, revealing divergent correlations depending on which data are used. While myriad variants on NGS techniques and bioinformatic pipelines are available, our results point to core differences that have not been highlighted in any studies to date. Given the low number of taxa identified when coupling shotgun data with clade-based taxonomic algorithms, previous studies that quantified biodiversity using such bioinformatic tools should be viewed cautiously or re-analyzed. Nonetheless, shotgun has complementary advantages that should be weighed when designing projects.

Aim Evaluating how groups of organisms vary in dispersal capability and how environmental, spatial and temporal signals vary across multiple scales is critical to elucidating metacommunity theory. We examined whether the relative contributions of environmental, spatial and hydrological factors have different effects on organismal groups with different dispersal abilities at three spatial scales, and how this knowledge contributes to our understanding of metacommunity dynamics. Location Four major Brazilian floodplains, with the largest distance among them of 2,300 Km. Taxon 10 aquatic organismal groups, ranging from bacterioplankton to fish. Methods We sampled lakes connected to the main river in the low‐ and high‐water periods of each floodplain between 2011 and 2012. Different biological groups were analysed across three hierarchical spatial scales (fine, intermediate and subcontinental) within and between floodplain systems. We applied a series of partial redundancy analyses to estimate the relative contributions of environmental factors, spatial factors and hydrological period for each biological group. Results At the fine spatial scale, predominantly environmental factors and hydrological period structure metacommunities, although less so for microorganisms than for micro‐invertebrates and macro‐organisms. The relative importance of environmental factors increased at the intermediate spatial scale. At the subcontinental scale, the relative importance of spatial factors increased for all biological groups, but environmental factors remained the primary regulators of microorganisms even at the largest scale. Main conclusions This study design allowed us to make more robust inferences about the mechanisms responsible for regulating community structure of distinct biological groups at different spatial scales. Our results suggest that biological groups displaying distinct body size likely determine the spatial extent at which environmental, spatial and hydrological processes prevail as the primary regulators of community structure. These findings are important in guiding the conservation and management of floodplain biodiversity because these systems are naturally highly heterogeneous in space and time.

Food webs include complex ecological interactions that define the flow of matter and energy, and are fundamental in understanding the functioning of an ecosystem. Temporal variations in the densities of communities belonging to the planktonic food web (i.e., microbial: bacteria, flagellate, and ciliate; and grazing: zooplankton and phytoplankton) were investigated, aiming to clarify the interactions between these organisms and the dynamics of the planktonic food web in a floodplain lake. We hypothesized that hydrological pulse determines the path of matter and energy flow through the planktonic food web of this floodplain lake. Data were collected monthly from March 2007 to February 2008 at three different sites in Guaraná Lake (Mato Grosso do Sul State, Brazil). The path analysis provided evidence that the dynamics of the planktonic food web was strongly influenced by the hydrological pulse. The high-water period favored interactions among the organisms of the microbial loop, rather than their relationships with zooplankton and phytoplankton. Therefore, in this period, the strong interaction among the organisms of the grazing food chain suggests that the microbial loop functions as a sink of matter and energy. In turn, in the low-water period, higher primary productivity appeared to favor different interactions between the components of the grazing food chain and microorganisms, which would function as a link to the higher trophic levels.

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The seagrass Zostera marina is a widespread foundational species in temperate coastal ecosystems that supports diverse communities of epiphytes and grazers. Bacteria link the production of seagrass to higher trophic levels and are thought to influence seagrass biology and health. Yet, we lack a clear understanding of the factors that structure the seagrass microbiome, or whether there is a consistent microbial community associated with seagrass that underpins functional roles. We sampled surface microbiome (epibiota) from new and old growth seagrass leaves and the surrounding seawater in eight meadows among four regions along the Central Coast of British Columbia, Canada to assess microbiome variability across space and as leaves age. We found that the seagrass leaf microbiome differs strongly from seawater. Microbial communities in new and old growth leaves are different from each other and from artificial seagrass leaves we deployed in one meadow. The microbiome on new leaves is less diverse and there is a small suite of core OTUs (operational taxonomic units) consistently present across regions. The overall microbial community for new leaves is more dispersed but with little regional differentiation, while the epiphytes on old leaves are regionally distinct. Many core OTUs on old leaves are commonly associated with marine biofilms. Together these observations suggest a stronger role for host filtering in new compared to old leaves, and a stronger influence of the environment and environmental colonization in old leaves. We found 11 core microbial taxa consistently present on old and new leaves and at very low relative abundance on artificial leaves and in the water column. These 11 taxa appear to be strongly associated with Z. marina. These core taxa may perform key functions important for the host such as detoxifying seagrass waste products, enhancing plant growth, and controlling epiphyte cover.

Anthropogenic disturbances change the trophic structure of streams, ultimately affecting ecosystem functioning. We investigated the effects of human disturbances, mainly organic pollution, on ciliate functional feeding groups (FFG) in 10 tropical streams near agricultural and urban habitats, in the dry and rainy seasons. We hypothesised that the organic pollution would affect the ciliate composition and that the richness and abundance of ciliate FFG would be associated with different disturbances, such that an increase in the load of organic matter would result in an increase in the percentage of bacterivores ciliates, while streams with low organic matter concentration and wide canopy openness will determine a higher contribution of algivorous ciliates. Our results corroborate our hypothesis of an increased development of bacterivorous ciliates with increasing organic pollution, but only in the abundance of this FFG. Also, algivorous ciliates were found to be related to riparian vegetation clearing. Thus, ciliate FFG accurately reflected different anthropogenic disturbances, revealing a change in the trophic structure of the streams. In addition, we found that organic pollution can lead to both taxonomic and functional homogenization of the ciliate community, which implies serious consequences for ecosystem functioning.

With its network of lotic and lentic habitats that shift during changes in seasonal connection, the tropical and subtropical large-river systems represent possibly the most dynamic of all aquatic environments. Pelagic water samples were collected from Brazilian floodplain lakes (total n = 58) in four flood-pulsed systems (Amazon [n = 21], Araguaia [n = 14], Paraná [n = 15], and Pantanal [n = 8]) in 2011–2012 and sequenced via 454 for bacterial environmental DNA using 16S amplicons; additional abiotic field and laboratory measurements were collected for the assayed lakes. We report here a global comparison of the bacterioplankton makeup of freshwater systems, focusing on a comparison of Brazilian lakes with similar freshwater systems across the globe. The results indicate a surprising similarity at higher taxonomic levels of the bacterioplankton in Brazilian freshwater with global sites. However, substantial novel diversity at the family level was also observed for the Brazilian freshwater systems. Brazilian freshwater bacterioplankton richness was relatively average globally. Ordination results indicate that Brazilian bacterioplankton composition is unique from other areas of the globe. Using Brazil-only ordinations, floodplain system differentiation most strongly correlated with dissolved oxygen, pH, and phosphate. Our data on Brazilian freshwater systems in combination with analysis of a collection of freshwater environmental samples from across the globe offers the first regional picture of bacterioplankton diversity in these important freshwater systems.Electronic supplementary materialThe online version of this article (doi:10.1007/s00248-016-0834-5) contains supplementary material, which is available to authorized users.

After much discussion about the cosmopolitan nature of microbes, the great issue nowadays is to identify at which spatial extent microorganisms may display biogeographic patterns and if temporal variation is important in altering those patterns. Here, planktonic ciliates were sampled from shallow lakes of four Neotropical floodplains, distributed over a spatial extent of ca. 3000 km, during high and low water periods, along with several abiotic and biotic variables potentially affecting the ciliate community. We found that common ciliate species were more associated with environmental gradients and rare species were more related to spatial variables; however, this pattern seemed to change depending on the temporal and spatial scales considered. Environmental gradients were more important in the high waters for both common and rare species. In low waters, common species continued to be mainly driven by environmental conditions, but rare species were more associated with the spatial component, suggesting dispersal limitation likely due to differences in dispersal ability and ecological tolerance of species. We also found that common and rare species were related to different environmental variables, suggesting different ecological niches. At the largest spatial extents, rare species showed clear biogeographic patterns.