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Background: Environmental reservoirs of antibiotic resistance pose a threat to human and animal health. Aquatic biofilms impacted by wastewater effluent (WW) are known environmental reservoirs for antibiotic resistance; however, the relative importance of biotic factors and abiotic factors from WW on the abundance of antibiotic resistance genes (ARGs) within aquatic biofilms remains unclear. Additionally, experimental evidence is limited within complex aquatic microbial communities as to whether genes bearing low sequence similarity to validated reference ARGs are functional as ARGs. Results: To disentangle the effects of abiotic and biotic factors on ARG abundances, natural biofilms were previously grown in flume systems with different proportions of stream water and either ultrafiltered or non-ultrafiltered WW. In this study, we conducted deep shotgun metagenomic sequencing of 75 biofilm, stream, and WW samples from these flume systems and compared the taxonomic and functional microbiome and resistome composition. Statistical analysis revealed an alignment of the resistome and microbiome composition and a significant association with experimental treatment. Several ARG classes exhibited an increase in normalized metagenomic abundances in biofilms grown with increasing percentages of non-ultrafiltered WW. In contrast, sulfonamide and extended-spectrum beta-lactamase ARGs showed greater abundances in biofilms grown in ultrafiltered WW compared to non-ultrafiltered WW. Overall, our results pointed toward the dominance of biotic factors over abiotic factors in determining ARG abundances in WW-impacted stream biofilms and suggested gene family-specific mechanisms for ARGs that exhibited divergent abundance patterns. To investigate one of these specific ARG families experimentally, we biochemically characterized a new beta-lactamase from the Planctomycetota (Phycisphaeraceae). This beta-lactamase displayed activity in the cleavage of cephalosporin analog despite sharing a low sequence identity with known ARGs. Conclusions: This discovery of a functional planctomycete beta-lactamase ARG is noteworthy, not only because it was the first beta-lactamase to be biochemically characterized from this phylum, but also because it was not detected by standard homology-based ARG tools. In summary, this study conducted a metagenomic analysis of the relative importance of biotic and abiotic factors in the context of WW discharge and their impact on both known and new ARGs in aquatic biofilms. DkJa1T-YWptaXXeH_hsDtB Video Abstract [ABSTRACT FROM AUTHOR]

When antimicrobial resistant bacteria (ARB) and genes (ARGs) reach novel habitats, they can become part of the habitat's microbiome in the long term if they are able to overcome the habitat's biotic resilience towards immigration. This process should become more difficult with increasing biodiversity, as exploitable niches in a given habitat are reduced for immigrants when more diverse competitors are present. Consequently, microbial diversity could provide a natural barrier towards antimicrobial resistance by reducing the persistence time of immigrating ARB and ARG. To test this hypothesis, a pan-European sampling campaign was performed for structured forest soil and dynamic riverbed environments of low anthropogenic impact. In soils, higher diversity, evenness and richness were significantly negatively correlated with relative abundance of >85% of ARGs. Furthermore, the number of detected ARGs per sample were inversely correlated with diversity. However, no such effects were present in the more dynamic riverbeds. Hence, microbiome diversity can serve as a barrier towards antimicrobial resistance dissemination in stationary, structured environments, where long-term, diversity-based resilience against immigration can evolve. A study based on a pan-European survey of soils and riverbeds of low-anthropogenic impact suggests that environmental microbiomes with a higher degree of bacterial diversity host less diverse and lower abundances of antibiotic resistance genes. [ABSTRACT FROM AUTHOR]

Microbial biodiversity is fundamental to maintain ecosystem functioning in seasonally variable ecosystems. However, it remains unclear how alterations in water availability caused by episodic drying compromise the ability of stream microbes to maintain multiple functions simultaneously (e.g., primary production and carbon cycling). Using data from 32 streams, we investigated how the phenology of annual drying influences stream sediment microbial biodiversity and their capacity to sustain multifunctionality. Our results showed that stream multifunctionality and most bacteria did not respond to changes in drying phenology. Only two bacterial groups, the drying‐resistant Sphingobacteriia and the drying‐sensitive Acidobacteria_Gp7, exhibited positive associations with multifunctionality; whereas, bacterial diversity showed a negative correlation with functions. Among these biodiversity aspects, Sphingobacteriia showed the strongest capacity to maintain multifunctionality at low and moderate performance levels. Our findings will help to better understand the mechanisms through which biodiversity sustains the functioning of seasonally variable streams and their responses to global change. [ABSTRACT FROM AUTHOR]

Aquatic fungi are highly diverse organisms that play a critical role in global biogeochemical cycles. Yet it remains unclear which assembly processes determine their co-occurrence and assembly patterns over gradients of drying intensity, which is a common stressor in fluvial networks. Although aquatic fungi possess drying-specific adaptations, little is known about how functional similarity influences co-occurrence probability and which functional traits are sorted by drying. Using field data from 15 streams, we investigated how co-occurrence patterns and assembly processes responded to drying intensity. To do so, we determined fungal co-occurrence patterns, functional traits that best explain species co-occurrence likelihood, and community assembly mechanisms explaining changes in functional diversity over the drying gradient. Our results identified 24 species pairs with positive co-occurrence probabilities and 16 species pairs with negative associations. The co-occurrence probability was correlated with species differences in conidia shape and fungal endophytic capacity. Functional diversity reduction over the drying gradient is generally associated with non-random abiotic filtering. However, the assembly processes changed over the drying gradient, with random assembly prevailing at low drying intensity and abiotic filtering gaining more importance as drying intensifies. Collectively, our results can help anticipate the impacts of global change on fungal communities and ecosystem functioning. [ABSTRACT FROM AUTHOR]

Summary: Microbes inhabiting intermittent streambeds are responsible for controlling and developing many biogeochemical processes essential for the ecosystem functions. Although streambed microbiota is adapted to intermittency the intensification of water scarcity and prolonged dry periods may jeopardise their capacity to cope with hydrological changes. This study aims to evaluate whether, and to what extent, the duration of dry periods affects streambed microbial density, diversity, composition (16S rRNA gene diversity) and functions (extracellular enzyme activities and respiration). Our results highlight the fact that hydrology modulates the community composition and, to some extent, the functions carried out under different environmental conditions. The relative abundance of certain taxa inhabiting the driest intermittent communities differs significantly from those found at sites with continuous flow. Microbial functional metrics revealed a progressive increase in recalcitrant carbon degradation activity at sites with an extended dry phase. In contrast, bacterial density and diversity were mainly influenced by the catchment land use, agriculture enhanced density but reduced diversity, and the presence of riparian vegetation supported greater streambed bacterial diversity. From this perspective, a combination of prolonged dryness with reduced riparian vegetation and increased agricultural land cover could compromise the ecosystem functioning by threaten microbially mediated processes linked to the carbon cycle. [ABSTRACT FROM AUTHOR]

Spatial and temporal widening of drought periods together with occurrence of flash storm events are consequences of global change affecting temporary stream ecosystems. Streambed heterotrophic microbes and the key biogeochemical processes they carry on could be endangered by the strengthening of drought episodes.Here, we performed a 165‐day experiment through 12 streambed sediment columns to study heterotrophic microbial functional and structural responses to long‐term drought. Two sediment depths (surface and hyporheic) and leaf litter were monitored under three treatments: control (maintained in wet, pool‐like conditions), dry (5 months of drought), and dry–storm (5 months of drought including two flash storms). All treatments were then followed by rewetting.Surface sediment followed by leaf litter was the most affected by the long‐term drought as shown by the reduction of polysaccharidic enzyme activities and litter decomposition rate, although lignin decomposition was not affected. This resulted in a greater use of recalcitrant compounds which might be due to reduced labile organic matter sources and the greater resistance of fungi than bacteria to drought. Moreover, in surface sediment, bacterial viability and algal biomass were reduced while the production of extracellular polymeric substances was intensified with dryness, suggesting its potential role as survival strategy.Hyporheic sediments appeared more resistant to long‐term drought, and this might be linked to its slightly higher water content (2.5%) than the surface (0.5%) during drying together with a greater content of fine material that allowed fungi and bacteria to survive and extracellular enzyme activities to be maintained.Microbial resilience to long‐term drought in sediment and leaf litter was promoted by the flash storms as shown by the fast recovery of enzyme activities, bacterial viability and leaf litter decomposition rate in the dry–storm treatment, once rewetted. Storms might liberate previously occluded carbon sources that fuel the fast microbial reactivation as well as providing sediment moisture for microbial survival.Our results highlight that long‐term drought may compromise stream biogeochemical processes linked to organic matter decomposition and that microbes are highly sensitive to minimal water content changes. Thus, long‐term drought consequences could be mitigated by occasional precipitations or by natural streambed moisture. [ABSTRACT FROM AUTHOR]