Free-living Bacterial Communities Research Articles

Beyond the Bloom: Unraveling the Diversity, Overlap, and Stability of Free-Living and Particle-Attached Bacterial Communities in a Cyanobacteria-Dominated Hypereutrophic Lake

In aquatic ecosystems with low nutrient levels, organic aggregates (OAs) act as nutrient hotspots, hosting a diverse range of microbial species compared to those in the water column. Lake eutrophication, marked by intensified and prolonged cyanobacterial blooms, significantly impacts material and energy cycling processes, potentially altering the ecological traits of both free-living (FL) and particle-attached (PA) bacteria. However, the extent to which observed patterns of FL and PA bacterial diversity, community assembly, and stability extend to hypereutrophic lakes remains understudied. To address this gap, we investigated bacterial diversity, composition, assembly processes, and stability within hypereutrophic Lake Xingyun. Our results revealed that FL bacterial communities exhibited higher α-diversity than PA counterparts, coupled with discernible taxonomic compositions. Both bacterial communities showed distinct seasonality, influenced by cyanobacterial bloom intensity. Environmental factors accounted for 71.1% and 54.2% of the variation among FL and PA bacteria, respectively. The assembly of the PA bacterial community was predominantly stochastic, while FL assembly was more deterministic. The FL network demonstrated greater stability, complexity, and negative interactions, indicative of competitive relationships, while the PA network showed a prevalence of positive correlations, suggesting mutualistic interactions. Importantly, these findings differ from observations in oligotrophic, mesotrophic, and eutrophic lakes. Overall, this research provides valuable insights into the interplay among bacterial fractions, enhancing our understanding of nutrient status and cyanobacterial blooms in shaping bacterial communities.

Organic matter composition fluctuations disrupt free-living bacterial communities more than particle-associated bacterial communities in coastal waters

Marine organic matter fuels the growth of microbial communities, shaping the composition of bacteria that specialize in its breakdown. However, responses of free-living (FL) and particle-associated (PA) bacterial communities to the changing pools of dissolved organic matter (DOM) and particulate organic matter (POM) remained unclear. This study investigates the composition of size-fractionated bacterial communities, DOM and POM in coastal waters over a 22-day period that includes a diatom bloom. Co-occurrence analysis showed that the FL bacterial communities were significantly less stable than PA communities. During the diatom bloom, we observed a significant increase in DOM molecules, particularly those derived from amino acids and peptides. In contrast, the relative intensities of major POM molecule classes remained stable despite the algal bloom's influence. Our study revealed a strong negative correlation between bacterial alpha-diversity and the amount of molecules in the organic matter pool. Similarly, bacterial community beta-diversity was found to be related to the composition of organic matter pool. However, the composition of organic matter was more strongly related to the composition of FL bacterial communities compared to PA communities. This suggests that FL bacteria exhibit greater variations in temporal dynamics and higher sensitivity to the specific structure of organic matter molecules.

Spatio-temporal changes of small protist and free-living bacterial communities in a temperate dimictic lake: insights from metabarcoding and machine learning.

Microbial communities, which include prokaryotes and protists, play an important role in aquatic ecosystems and influence ecological processes. To understand these communities, metabarcoding provides a powerful tool to assess their taxonomic composition and track spatio-temporal dynamics in both marine and freshwater environments. While marine ecosystems have been extensively studied, there is a notable research gap in understanding eukaryotic microbial communities in temperate lakes. Our study addresses this gap by investigating the free-living bacteria and small protist communities in Lake Roś (Poland), a dimictic temperate lake. Metabarcoding analysis revealed that both the bacterial and protist communities exhibit distinct seasonal patterns that are not necessarily shaped by dominant taxa. Furthermore, machine learning and statistical methods identified crucial amplicon sequence variants (ASVs) specific to each season. In addition, we identified a distinct community in the anoxic hypolimnion. We have also shown that the key factors shaping the composition of analysed community are temperature, oxygen, and silicon concentration. Understanding these community structures and the underlying factors is important in the context of climate change potentially impacting mixing patterns and leading to prolonged stratification.

Deciphering the pathogenic risks of microplastics as emerging particulate organic matter in aquatic ecosystem

Microplastics are accumulating rapidly in aquatic ecosystems, providing habitats for pathogens and vectors for antibiotic resistance genes (ARGs), potentially increasing pathogenic risks. However, few studies have considered microplastics as particulate organic matter (POM) to elucidate their pathogenic risks and underlying mechanisms. Here, we performed microcosm experiments with microplastics and natural POM (leaves, algae, soil), thoroughly investigating their distinct effects on the community compositions, functional profiles, opportunistic pathogens, and ARGs in Particle-Associated (PA) and Free-Living (FL) bacterial communities. We found that both microplastics and leaves have comparable impacts on microbial community structures and functions, enriching opportunistic pathogens and ARGs, which may pose potential environmental risks. These effects are likely driven by their influences on water properties, including dissolved organic carbon, nitrate, DO, and pH. However, microplastics uniquely promoted pathogens as keystone species and further amplified their capacity as hosts for ARGs, potentially posing a higher pathogenic risk than natural POM. Our research also emphasized the importance of considering both PA and FL bacteria when assessing microplastic impacts, as they exhibited different responses. Overall, our study elucidates the role and underlying mechanism of microplastics as an emerging POM in intensifying pathogenic risks of aquatic ecosystems in comparison with conventional natural POM.

Succession of particle-attached and free-living bacterial communities in response to microalgal dynamics induced by the biological cyanocide paucibactin A

Microalgae, including cyanobacteria and eukaryotic algae, are hotspots of primary production and play a critical role in global carbon cycling. However, these species often form blooms that poses a threat to aquatic ecosystems. Although the use of bacteria-derived cyanocides is regarded as an environmentally friendly method for controlling cyanobacterial blooms, only a few studies have examined their potential impact on ecosystems. This study is the first to explore the response of particle-attached (PA) and free-living (FL) bacteria to the dynamics of microalgal communities induced by the biological cyanocide paucibactin A. The microalgal community dynamics were divided into two distinct phases [phase I (days 0–2) and phase II (days 3–7)]. In phase I, paucibactin A caused a sudden decrease in the cyanobacterial concentration. Phase II was characterized by increased growth of eukaryotic microalgae (Scenedesmus, Pediastrum, Selenastrum, and Coelastrum). The stability of the bacterial community and the contribution of stochastic processes to community assembly were more pronounced in phase II than in phase I. The microalgal dynamics triggered by paucibactin A coincided with the succession of the PA and FL bacterial communities. The lysis of cyanobacteria in phase I favored the growth of microbial organic matter degraders in both the PA (e.g., Aeromonas and Rheinheimera) and FL (e.g., Vogesella) bacterial communities. In phase II, Lacibacter, Phycisphaeraceae, and Hydrogenophaga in the PA bacterial community and Lacibacter, Peredibacter, and Prosthecobacter in the FL bacterial community showed increased relative abundances. Overall, the FL bacterial community exhibited greater sensitivity to the two sequential processes compared with the PA bacterial community. These results highlight the need for studies evaluating the impact of biological cyanocides on aquatic ecosystems when used to control natural cyanobacterial blooms.

Cross-domain diversity effects: linking diatom species richness, intraspecific richness, and biomass production to host-associated bacterial diversity.

Interactions between bacteria and microalgae are important for the functioning of aquatic ecosystems, yet interactions based on the biodiversity of these two taxonomic domains have been scarcely studied. Specifically, it is unclear whether a positive biodiversity-productivity relationship in phytoplankton is largely facilitated by niche partitioning among the phytoplankton organisms themselves or whether associated bacterial communities play an additional role in modifying these diversity effects. Moreover, the effects of intraspecific diversity in phytoplankton communities on bacterial community diversity have not been tested. To address these points, we factorially manipulated both species and intraspecific richness of three diatoms to test the effects of diatom species/strain diversity on biomass production and bacterial diversity in algae-bacteria communities. The results show that diatom intraspecific diversity has significant positive effects on culture biomass and the diversity of the associated free-living bacterial community (0.2-3μm size fraction), which are comparable in magnitude to species diversity effects. However, there were little to no effects of diatom diversity on host-associated bacterial diversity (>3μm size fraction), or of bacterial diversity on biomass production. These results suggest a decoupling of bacterial diversity from the diatom diversity-productivity relationship and provide early insights regarding the relations between diversity across domains in aquatic ecosystems.

Response of particle-attached and free-living bacterial communities to Microcystis blooms.

The massive proliferation of Microcystis threatens freshwater ecosystems and degrades water quality globally. Understanding the mechanisms that contribute to Microcystis growth is crucial for managing Microcystis blooms. The lifestyles of bacteria can be classified generally into two groups: particle-attached (PA; > 3µm) and free-living (FL; 0.2-3.0µm). However, little is known about the response of PA and FL bacteria to Microcystis blooms. Using 16S rRNA gene high-throughput sequencing, we investigated the stability, assembly process, and co-occurrence patterns of PA and FL bacterial communities during distinct bloom stages. PA bacteria were phylogenetically different from their FL counterparts. Microcystis blooms substantially influenced bacterial communities. The time decay relationship model revealed that Microcystis blooms might increase the stability of both PA and FL bacterial communities. A contrasting community assembly mechanism was observed between the PA and FL bacterial communities. Throughout Microcystis blooms, homogeneous selection was the major assembly process that impacted the PA bacterial community, whereas drift explained much of the turnover of the FL bacterial community. Both PA and FL bacterial communities could be separated into modules related to different phases of Microcystis blooms. Microcystis blooms altered the assembly process of PA and FL bacterial communities. PA bacterial community appeared to be more responsive to Microcystis blooms than FL bacteria. Decomposition of Microcystis blooms may enhance cooperation among bacteria. Our findings highlight the importance of studying bacterial lifestyles to understand their functions in regulating Microcystis blooms. KEY POINTS: • Microcystis blooms alter the assembly process of PA and FL bacterial communities • Microcystis blooms increase the stability of both PA and FL bacterial communities • PA bacteria seem to be more responsive to Microcystis blooms than FL bacteria.

The Assembly Process of Free-Living and Particle-Attached Bacterial Communities in Shrimp-Rearing Waters: The Overwhelming Influence of Nutrient Factors Relative to Microalgal Inoculation.

The ecological functions of bacterial communities vary between particle-attached (PA) lifestyles and free-living (FL) lifestyles, and separately exploring their community assembly helps to elucidate the microecological mechanisms of shrimp rearing. Microalgal inoculation and nutrient enrichment during shrimp rearing are two important driving factors that affect rearing-water bacterial communities, but their relative contributions to the bacterial community assembly have not been evaluated. Here, we inoculated two microalgae, Nannochloropsis oculata and Thalassiosira weissflogii, into shrimp-rearing waters to investigate the distinct effects of various environmental factors on PA and FL bacterial communities. Our study showed that the composition and representative bacteria of different microalgal treatments were significantly different between the PA and FL bacterial communities. Regression analyses and Mantel tests revealed that nutrients were vital factors that constrained the diversity, structure, and co-occurrence patterns of both the PA and FL bacterial communities. Partial least squares path modeling (PLS-PM) analysis indicated that microalgae could directly or indirectly affect the PA bacterial community through nutrient interactions. Moreover, a significant interaction was detected between PA and FL bacterial communities. Our study reveals the unequal effects of microalgae and nutrients on bacterial community assembly and helps explore microbial community assembly in shrimp-rearing ecosystems.

Persistent response of the bottom free-living bacteria to typhoon events in a subtropical reservoir

Typhoon-induced perturbations can result in long-lasting effects on aquatic communities in subtropical lakes or reservoirs. However, the responses of bacterial communities and their related nutrient cycling to episodic typhoon events throughout the water column in deep waters remain largely unknown. Here, we conducted a four-year field study to reveal the depth-specific responses of both free-living (FL) and particle-attached (PA) bacteria to typhoon events in a subtropical deep reservoir from 2015 to 2018. By comparing the depth-specific responses of FL and PA bacteria, we found that typhoon-induced inputs of organic matter and microorganisms significantly increased FL bacterial diversity and changed FL bacterial community composition in bottom waters perhaps through the density current or undercurrent. Typhoon events had a more persistent effect on FL than PA bacterial communities, especially in bottom waters of the reservoir. Free-living bacteria were more associated with nutrient cycling in bottom waters than particle-attached bacteria. These findings provide deep understanding of how FL and PA bacteria respond to typhoon events at community level in subtropical deep reservoir and thus help us to improve reservoir management in a rapidly changing world.

Comparative microbiota characterization unveiled a contrasting pattern of floc-associated versus free-living bacterial communities in biofloc aquaculture

Bacterial communities thriving in various components determine the host health and water quality in biofloc technology (BFT)-based aquaculture system. Despite increasing utilization of BFT in aquaculture industries, studies distinguishing bioflocs (attached community) and rearing water (free-living community) microbiota compositions remain rudimentary. In this study, we compared floc-associated bacterial (FAB; size fraction ≥3 μm) and planktonic free-living bacterial (PFLB; 0.22–3.00 μm) communities from a commercial shrimp aquaculture system to investigate how their diversity, community compositions, inter-species interactions, and functional potentials varied. Based on 16S rRNA gene amplicon sequencing, the metataxonomic results indicated that though these communities share identical habitat, the FAB community possesses a higher bacterial richness and noticeably distinct community composition than its PFLB counterpart. The Alphaproteobacteria (mostly Rhodobacteraceae) preponderated in the FAB community, while Bacteroidetes (primarily Flavobacteriaceae) dominated the PFLB. By analyzing co-occurrence pattern, we portrayed a complex ecological network, strong inter-species interactions, and diverse keystone taxa in the FAB community. Intriguingly, Halieaceae and Rhodobacteraceae members were recognized as prominent keystone taxa in FAB and PFLB communities, respectively, highlighting their regulatory roles in sustaining stability of these communities. Furthermore, negative edges proportion varied considerably in ecological networks, suggesting that both communities experience distinct niche differentiation. PICRUSt2-based prediction demonstrated that functional pathways pertaining to membrane transporters, cell motility, and polymer degradation were enriched in the FAB community, suggesting their involvement in surface-adapted lifestyles. Altogether, we proved that attached and planktonic bacterial communities serve as two distinct microbial consortia, perform different tasks, and hence could have diverse ecological roles in biofloc aquaculture systems. Our findings advance scientific understanding of microbiota variations between planktonic and attached communities in aquatic environments, and strengthen the proposed species sorting hypothesis, which states that prevailing environmental factors determine bacterial community assembly.

Spatial patterns in host-associated and free-living bacterial communities across six temperate estuaries.

A major goal of microbial ecology is to establish the importance of spatial and environmental factors in driving community variation. Their relative importance likely varies across spatial scales, but focus has primarily been on free-living communities within well-connected aquatic environments rather than less connected island-like habitats such as estuaries, and key host-associated communities within these systems. Here we sampled both free-living (seawater and sediment) and host-associated (estuarine fish hindgut microbiome, Pelates sexlineatus) communities across six temperate Australian estuaries spanning ∼500km. We find that spatial and environmental factors have different influences on these communities, with seawater demonstrating strong distance-decay relationships (R = -0.69) and significant associations with a range of environmental variables. Distance-decay relationships were weak for sediment communities but became stronger over smaller spatial scales (within estuaries, R = -0.5), potentially reflecting environmental filtering across biogeochemical gradients or stochastic processes within estuary sediments. Finally, P. sexlineatus hindgut microbiome communities displayed weak distance-decay relationships (R = -0.36), and limited variation explained by environmental variables, indicating the significance of host-related factors in driving community variation. Our findings provide important ecological insights into the spatial distributions and driving forces of both free-living and host-associated bacterial patterns across temperate estuarine systems.

Free-living bacteria stimulate sugarcane growth traits and edaphic factors along soil depth gradients under contrasting fertilization

Free-living bacterial community and abundance have been investigated extensively under different soil management practices. However, little is known about their nitrogen (N) fixation abilities, and how their contributions to N budgets impact plant growth, yield, and carbon (C) and N cycling enzymes in a long-term consecutive sugarcane monoculture farming system, under contrasting amendments, along different soil horizons. Here, nifH gene amplicon was used to investigate diazotrophs bacterial community and abundance by leveraging high-throughput sequencing (HTS). Moreover, edaphic factors in three soil depths (0–20, 20–40, and 40–60 cm) under control (CK), organic matter (OM), biochar (BC), and filter mud (FM) amended soils were investigated. Our analysis revealed that β-glucosidase activity, acid phosphatase activity, ammonium (NH4+-N), nitrate (NO3–N), total carbon (TC), total nitrogen (TN), and available potassium (AK) were considerably high in 0–20 cm in all the treatments. We also detected a significantly high proportion of Proteobacteria and Geobacter in the entire sample, including Anabaena and Enterobacter in 0–20 cm soil depth under the BC and FM amended soils, which we believed were worthy of promoting edaphic factors and sugarcane traits. This phenomenon was further reinforced by network analysis, where diazotrophs bacteria belonging to Proteobacteria exhibited strong and positive associations soil electrical conductivity (EC), soil organic matter content (SOM) available phosphorus (AP), TN, followed by NH4+-N and NO3–N, a pattern that was further validated by Mantel test and Pearson’s correlation coefficients analyses. Furthermore, some potential N-fixing bacteria, including Burkholderia, Azotobacter, Anabaena, and Enterobacter exhibited a strong and positive association with sugarcane agronomic traits, namely, sugarcane stalk, ratoon weight, and chlorophyll content. Taken together, our findings are likely to broaden our understanding of free-living bacteria N-fixation abilities, and how their contributions to key soil nutrients such as N budgets impact plant growth and yield, including C and N cycling enzymes in a long-term consecutive sugarcane monoculture farming system, under contrasting amendments, along different soil horizons.

Abundance and composition of particles and their attached microbiomes along an Atlantic Meridional Transect

Particulate organic matter plays a significant role in the marine carbon cycle. Its sinking exports organic carbon from the surface to deep oceans. Using fractionated filtration, we analysed particles of 3 – 10 µm and >10 µm and their microbiomes in thirty-five stations along a latitudinal transect of the Atlantic Ocean and provide new insights into the composition, community dynamics, and catabolic potential of particle-attached bacteria. Samples were taken during an Atlantic Meridional Transect (AMT22), which traversed six distinctive ocean provinces. Using 16S rRNA amplicon sequencing and fluorescence in situ hybridisation, we could show a strong variation between particle-attached and free-living bacterial communities at each station and across the biogeographical provinces – a dynamic likely driven by chlorophyll a concentrations, temperature, and the oxygen content of the respective biogeographical provinces. Whereas the <3 µm fraction was primarily composed of SAR11, SAR86, Prochlorococcus and Bacteroidetes of the NS9 and NS5 clades, particle-attached communities were dominated by other Bacteroidetes (Polaribacter spp.), diverse Gammaproteobacteria including members of the genera Alteromonas and Vibrio, Alphaproteobacteria, Planctomycetes, OM27 and Verrucomicrobia. In three provinces, we quantified particle abundance and analysed their glycan composition using four lectins targeting fucose, galactose, N-acetylgalactosamine and mannose. Particles were mainly composed of fucose glycans with only a minor abundance of the other glycans, and particle abundance was directly correlated with the chlorophyll a concentrations. Functional analysis of 54 metagenome-assembled genomes retrieved from bacterial communities attached to small particles showed that particle-attached Bacteroidetes, Planctomycetes and Verrucomicrobia displayed key roles in the degradation of sulfated fucose-containing polysaccharides. We also identified gene clusters potentially encoding the utilisation of mannan and laminarin, suggesting an adaptation to the glycan composition of the particles, potentially resulting in niche diversification. Together, our results provide insights into particle-attached bacteria and their ecological strategies in the Atlantic.

Patterns of free-living and particle-attached bacteria along environmental gradients in Lake Taihu.

To elucidate the effects of environmental heterogeneity on diversity, composition, and degree of overlap between free-living (FL) and particle-attached (PA) bacteria, we sampled large, shallow, eutrophic Lake Taihu, China across gradients spanning riverine inflow, cyanobacterial blooms, and the open limnetic area. Using high-throughput sequencing of the 16S rRNA gene, we show that (i) bacterial communities near riverine inflow had high α-diversity and a high degree of overlap between FL and PA lifestyles, (ii) communities in cyanobacterial blooms have reduced α-diversity within the PA lifestyle, and (iii) communities from the limnetic area had the lowest bacterial α-diversity within the FL lifestyle and a medium degree of overlap between the FL and PA lifestyles. Redundancy analysis showed that the variation of the FL bacterial community was shaped by suspended solids and total phosphorous, while the variation of the PA bacterial community was shaped by suspended solids, dissolved oxygen, and the percentage of organic matter in suspended solids. This study highlights the importance of environmental heterogeneity, riverine input, cyanobacterial blooms, and nutrient status on the spatial distribution patterns of FL and PA bacterial communities in freshwater lakes.

Spatiotemporal variations, assembly processes, and co-occurrence patterns of particle-attached and free-living bacteria in a large drinking water reservoir in China.

Particle-attached (PA) and free-living (FL) bacterial communities are sensitive to pollutant concentrations and play an essential role in biogeochemical processes and water quality maintenance in aquatic ecosystems. However, the spatiotemporal variations, assembly processes, co-occurrence patterns, and environmental interactions of PA and FL bacteria in drinking water reservoirs remain as yet unexplored. To bridge this gap, we collected samples from 10 sites across four seasons in Lake Tianmu, a large drinking water reservoir in China. Analysis of 16S rRNA gene libraries demonstrated spatiotemporal variations in bacterial diversity and identified differences in bacterial community composition (BCC) between PA and FL lifestyles. Capacity for nitrogen respiration, nitrogen fixation, and nitrate denitrification was enriched in the PA lifestyle, while photosynthesis, methylotrophy, and methanol oxidation were enriched in the FL lifestyle. Deterministic processes, including interspecies interactions and environmental filtration, dominated the assembly of both PA and FL bacterial communities. The influence of environmental filtration on the FL community was stronger than that on the PA community, indicating that bacteria in the FL lifestyle were more sensitive to environmental variation. Co-occurrence patterns and keystone taxa differed between PA and FL lifestyles. The ecological functions of keystone taxa in the PA lifestyle were associated with the supply and recycling of nutrients, while those in FL were associated with the degradation of complex pollutants. PA communities were more stable than FL communities in the face of changing environmental conditions. Nutrients (e.g., TDN and NO3 -) and abiotic and biotic factors (e.g., WT and Chl-a) exerted positive and negative effects, respectively, on the co-occurrence networks of both lifestyles. These results improve our understanding of assembly processes, co-occurrence patterns, and environmental interactions within PA and FL communities in a drinking water reservoir.

The Ecological Differentiation of Particle-Attached and Free-Living Bacterial Communities in a Seasonal Flooding Lake-the Poyang Lake.

Particle-attached (PA) and free-living (FL) bacterial communities play essential roles in the biogeochemical cycling of essential nutrients in aquatic environments. However, little is known about the factors that drive the differentiation of bacterial lifestyles, especially in flooding lake systems. Here we assessed the compositional and functional similarities between the FL and PA bacterial fractions in a typical flooding lake-the Poyang Lake (PYL) of China. The results revealed that PA communities had significantly different compositions and functions from FL communities in every hydrological period, and the diversity of both PA and FL communities was affected mainly by the water regime rather than bacterial lifestyles. PA communities were more diverse and enriched with Proteobacteria and Bacteroidetes, while FL communities had more Actinobacteria. There was a higher abundance of photosynthetic and nitrogen-cycling bacterial groups in PA communities, but a higher abundance of members involved in hydrocarbon degradation, aromatic hydrocarbon degradation, and methylotrophy in FL communities. Water properties (e.g., temperature, pH, total phosphorus) significantly regulated the lifestyle variations of PA and FL bacteria in PYL. Collectively, our results have demonstrated a clear ecological differentiation of PA and FL bacterial communities in flooding lakes, suggesting that the connectivity between FL and PA bacterial fractions is water property-related rather than water regime-related.

Similar assembly mechanisms but distinct co-occurrence patterns of free-living vs. particle-attached bacterial communities across different habitats and seasons in shallow, eutrophic Lake Taihu

Eutrophication due to nitrogen and phosphorus input is an increasing problem in lake ecosystems. Free-living (FL) and particle-attached (PA) bacterial communities play a primary role in mediating biogeochemical processes in these lakes and in responding to eutrophication. However, knowledge of factors governing function, assembly mechanisms, and co-occurrence patterns of these communities remain poorly understood and are key challenges in microbial ecology. To address this knowledge gap, we collected 96 samples from Lake Taihu across four seasons and investigated the bacterial community using 16S rRNA gene sequencing. Our results demonstrate that the α-diversity, β-diversity, community composition, and functional composition of FL and PA bacterial communities exhibited differing spatiotemporal dynamics. FL and PA bacterial communities displayed similar distance-decay relationships across seasons. Deterministic processes (i.e., environmental filtering and species interaction) were the primary factors shaping community assembly in both FL and PA bacteria. Similar environmental factors shaped bacterial community structure while different environmental factors drove bacterial functional composition. Habitat filtering influenced enrichment of bacteria within specific functional groups. Among them, the FL bacterial community appeared to play a critical role in methane-utilization, whereas the PA bacteria contributed more to biogeochemical cycling of carbon. FL and PA bacterial communities exhibited distinct co-occurrence pattern across different seasons. In the FL network, Methylotenera and Methylophilaceae were identified as keystone taxa, while Burkholderiaceae and the hgcI clade were keystone taxa in the PA network. The PA bacterial community appeared to possess greater stability in the face of environmental change than did FL counterparts. These results broaden our knowledge of the driving factors, co-occurrence patterns, and assembly processes in FL and PA bacterial communities in eutrophic ecosystems and provide improved insight into the underlying mechanisms responsible for these results.

Stochastic processes shape the aggregation of free‐living and particle‐attached bacterial communities in the Yangtze River Estuary, China

An estuary plays an important role in material and energy exchange between the land and sea, where complex physical, chemical, and biological processes occur. Here, we investigated the assembly processes of free-living (FL) and particle-associated (PA) bacterial communities in two seawater layers at five stations in the Yangtze River Estuary (YRE) by using 16S rRNA sequencing methods. The results indicated that Proteobacteria was the most abundant phylum in the YRE. The α-diversity of PA community was significantly higher than FL community, and analysis of similarity showed significantly different (Global R = 0.2809, p < 0.005). RDA revealed that phosphate (PO4 3- ) was significantly correlated with PA bacterial community abundance (p < 0.05). An ecological null model showed that both PA and FL bacterial communities were mainly influenced by stochastic processes (PA: 100%, FL: 70%), which PA attached to nutrient particles and are less affected by environmental filtration. Dispersal limitation (50%) was the main assembly process of the PA community, while homogeneous selection (30%) and drift (30%) were important processes in the FL community assembly. The available substrate for colonization limits the transformation from FL to PA bacteria. This study would improve our understanding of FL and PA bacterial community structure and factors affecting assembly process in estuarine environments.

Response of a Coastal Microbial Community to Olivine Addition in the Muping Marine Ranch, Yantai

Spreading olivine powder in seawater to enhance alkalinity through weathering reactions has been proposed as a potential solution to control atmospheric CO2 concentration. Attention has usually been paid to the chemical properties of seawater after the addition of olivine within lab and modeling studies. However, both microbial acclimation and evolution in such manipulated natural environments are often overlooked, yet they are of great importance for understanding the biological consequences of whether olivine addition is a feasible approach to mitigating climate change. In this study, an olivine addition experiment was conducted to investigate variation in bacterial diversity and community composition in the surface and bottom seawater of a representative marine ranch area in the Muping, Yantai. The results show that the composition of the particle-attached microbial community was particularly affected by the application of olivine. The relative abundance of biofilm-forming microbes in particle-attached fraction increased after the addition of olivine, while no significant variation in the free-living bacterial community was observed. Our study suggests that olivine addition would reshape the bacterial community structure, especially in particle-attached microenvironments. Therefore, the risk evaluation of alkalinity enhancement should be further studied before its large-scale application as a potential ocean geoengineering plan.

A microbial perspective on the local influence of Arctic rivers and estuaries on Hudson Bay (Canada)

A clear divide typically exists between freshwater and marine microbial communities, with transitional communities found in estuarine zones. The estuarine communities can derive from inflowing rivers and the sea via tidal mixing and incursions or be comprised of unique brackish species, depending on flow regimes and retention time within an estuary. Only a few studies have been carried out in the Arctic, where moderate salinities associated with the influence of seasonal ice melt could potentially favor marine microbes adapted to lower salinities in fresh-to-saltwater transition zones irrespective of river flows and tidal mixing. To test this idea, we examined early summer microbial communities in 2 western Hudson Bay (Canada) river-to-sea systems: the Churchill and Nelson river systems. Both rivers originate from the same headwaters, suggesting similar catchment conditions, but differ in geomorphology and hydroelectric diversions that induce very different flow and stratification regimes. Using amplicons of the V4 region of the 16S rRNA gene, we identified distinct riverine bacterial communities that were significantly different from the 2 associated estuaries and offshore communities. In the much smaller Churchill Estuary, the microbial community showed a marked influence of freshwater microbial species, along with marine influences. In contrast, in the larger high-flow Nelson River Estuary, riverine bacterioplankton were less evident in the estuary, where the marine signal was much stronger. The marine samples in both systems differed somewhat consistently with the phenology of the phytoplankton bloom in the Bay and tended to harbor distinct attached and free-living bacterial communities. Our results highlight the relevance of river flow and estuarine circulation on selection of bacterial species in estuaries, with ecological implications for food web functionality and biogeochemical cycles in the Anthropocene, where flow regimes would be affected by larger climatic variability.