Oligotrophic deep-water lakes are unique and sensitive ecosystems with limited nutrient availability. Understanding bacterial communities within these lakes is crucial for assessing ecosystem health, biogeochemical cycling, and responses to environmental changes. In this study, we investigated the seasonal and vertical dynamics of both free-living (FL) and particle-attached (PA) bacteria in Lake Fuxian, a typical oligotrophic deep freshwater lake in southeast China. Our findings revealed distinct seasonal and vertical dynamics of FL and PA bacterial communities, driven by similar physiochemical environmental factors. PA bacteria exhibited higher α- and β-diversity and were enriched with Proteobacteria, Cyanobacteria, Firmicutes, Patescibacteria, Planctomycetota, and Verrucomicrobiota, while FL bacteria were enriched with Actinobacteria and Bacteroidota. FL bacteria showed enrichment in putative functions related to chemoheterotrophy and aerobic anoxygenic photosynthesis, whereas the PA fraction was enriched with intracellular parasites (mainly contributed by Rickettsiales, Chlamydiales, and Legionellales) and nitrogen metabolism functions. Deterministic processes predominantly shaped the assembly of both FL and PA bacterial communities, with stochastic processes playing a greater role in the FL fraction. Network analysis revealed extensive species interactions, with a higher proportion of positively correlated edges in the PA network, indicating mutualistic or cooperative interactions. Cyanobium, Comamonadaceae, and Roseomonas were identified as keystone taxa in the PA network, underscoring potential cooperation between autotrophic and heterotrophic bacteria in organic particle microhabitats. Overall, the disparities in bacterial diversity, community composition, putative function, and network characteristics between FL and PA fractions highlight their adaptation to distinct ecological niches within these unique lake ecosystems.IMPORTANCEUnderstanding the diversity of microbial communities, their assembly mechanisms, and their responses to environmental changes is fundamental to the study of aquatic microbial ecology. Oligotrophic deep-water lakes are fragile ecosystems with limited nutrient resources, rendering them highly susceptible to environmental fluctuations. Examining different bacterial types within these lakes offers valuable insights into the intricate mechanisms governing community dynamics and adaptation strategies across various scales. In our investigation of oligotrophic deep freshwater Lake Fuxian in China, we explored the seasonal and vertical dynamics of two bacterial types: free-living (FL) and particle-attached (PA). Our findings unveiled distinct patterns in the diversity, composition, and putative functions of these bacteria, all shaped by environmental factors. Understanding these subtleties provides insight into bacterial interactions, thereby influencing the overall ecosystem functioning. Ultimately, our research illuminates the adaptation and roles of FL and PA bacteria within these unique lake environments, contributing significantly to our broader comprehension of ecosystem stability and health.

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.

The shelled pteropod Creseis acicula is a marine pelagic shellfish widely distributed from temperate to tropical seas around the world. From June to July 2020, a C. acicula bloom first happened in the Daya Bay, southern China, and its density reached the highest value (5600 ind. m-3) ever recorded around the world. However, few studies have investigated the responses of bacterial communities to the C. acicula bloom. In the present study, we examined the community profiles of three communities of bacteria including the free-living and particle-attached bacteria in the blooming and reference waters, and bacteria attached to the whole body and shell of C. acicula using a high-throughput sequencing method. The results indicated that the C. acicula bloom had a greater impact on particle-attached bacteria than free-living bacteria. Among the bloom-sensitive particle-attached bacteria, the predominant bacterial phyla were Pseudomonadota, Bacteroidota and Verrucomicrobiota in the blooming areas, whereas they were Actinomycetota and Planctomycetota in the reference areas. Specifically, fecal bacteria Haloferula and Halioglobus spp. were significantly enriched in the blooming waters and accumulated on C. acicula shells. Conversely, the significantly lower relative abundance of Nocardioides sp. in the blooming area and accumulated on the whole body of C. acicula indicated their attachment to particles consumed by C. acicula. Overall, our results suggested that the C. acicula bloom influenced marine bacteria, particularly particle-attached bacteria, by increasing (e.g. providing shells and feces) or decreasing (e.g. filter-feeding the suspended particles) the abundance of available substances.

BACKGROUND: Marine microalgae (phytoplankton) mediate almost half of the worldwide photosynthetic carbon dioxide fixation and therefore play a pivotal role in global carbon cycling, most prominently during massive phytoplankton blooms. Phytoplankton biomass consists of considerable proportions of polysaccharides, substantial parts of which are rapidly remineralized by heterotrophic bacteria. We analyzed the diversity, activity, and functional potential of such polysaccharide-degrading bacteria in different size fractions during a diverse spring phytoplankton bloom at Helgoland Roads (southern North Sea) at high temporal resolution using microscopic, physicochemical, biodiversity, metagenome, and metaproteome analyses.
RESULTS: Prominent active 0.2-3 µm free-living clades comprised Aurantivirga, \"Formosa\", Cd. Prosiliicoccus, NS4, NS5, Amylibacter, Planktomarina, SAR11 Ia, SAR92, and SAR86, whereas BD1-7, Stappiaceae, Nitrincolaceae, Methylophagaceae, Sulfitobacter, NS9, Polaribacter, Lentimonas, CL500-3, Algibacter, and Glaciecola dominated 3-10 µm and > 10 µm particles. Particle-attached bacteria were more diverse and exhibited more dynamic adaptive shifts over time in terms of taxonomic composition and repertoires of encoded polysaccharide-targeting enzymes. In total, 305 species-level metagenome-assembled genomes were obtained, including 152 particle-attached bacteria, 100 of which were novel for the sampling site with 76 representing new species. Compared to free-living bacteria, they featured on average larger metagenome-assembled genomes with higher proportions of polysaccharide utilization loci. The latter were predicted to target a broader spectrum of polysaccharide substrates, ranging from readily soluble, simple structured storage polysaccharides (e.g., laminarin, α-glucans) to less soluble, complex structural, or secreted polysaccharides (e.g., xylans, cellulose, pectins). In particular, the potential to target poorly soluble or complex polysaccharides was more widespread among abundant and active particle-attached bacteria.
CONCLUSIONS: Particle-attached bacteria represented only 1% of all bloom-associated bacteria, yet our data suggest that many abundant active clades played a pivotal gatekeeping role in the solubilization and subsequent degradation of numerous important classes of algal glycans. The high diversity of polysaccharide niches among the most active particle-attached clades therefore is a determining factor for the proportion of algal polysaccharides that can be rapidly remineralized during generally short-lived phytoplankton bloom events. Video Abstract.

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; > 3 µm) and free-living (FL; 0.2-3.0 µm). 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.

In regions close to the continental shelf edge, the transfer of particulate organic carbon (POC) within the food web and with downward fluxes contributes to the long-term storage of carbon in deep basins. The degradation of POC by heterotrophic prokaryotes is one of the key indices of biological productivity and ecosystem functioning. Here we present data on the spatial and vertical distribution of the total bacterial abundance, fraction of high DNA-content and particle-associated bacteria, and POC concentration. Additionally we determined environmental parameters (temperature, salinity, turbidity, fluorescence and dissolved oxygen concentration). The data were collected in the Vilkitsky Strait and along two transects from the outer shelf of the Laptev Sea to the adjacent continental slope region of the Arctic Ocean in August-September 2018. Principal component analysis was used to reveal relationships between environmental factors and bacterial characteristics. Total bacterial abundance (BA) was positively correlated with seawater fluorescence, dissolved oxygen concentration and POC, but negatively linked to depth and salinity. The percentage of both particle-associated bacteria (%PAB) and bacteria with high DNA concentration (%HDNA) were positively linked to depth and salinity. The data obtained may be useful for a comprehensive assessment of the state of ecosystems in the vicinity of the Arctic continental slope under changing environmental conditions, as well as for studying microbial input in the carbon cycle.

Diatom-bacteria interactions and the associated bloom dynamics have not been fully understood in the coastal oceans. Here, we focus on the polyunsaturated aldehydes (PUAs) produced by diatoms in the post-bloom phase and look into their roles in microbial phosphorus (P) recycling outside of a P-limited estuary. The phytoplankton community in the bloom was dominated by PUAs-producing diatoms (Skeletonema costatum, Thalassiosira spp., and Pesudonitzschia delicates) with elevated concentrations of biogenic particulate PUAs. In addition, there were micromolar levels of particle-adsorbed PUAs hotspots with distinct compositions in and out of the bloom determined by a combining large-volume filtration and on-site derivation method. Field experiments were conducted to further assess the responses of particle-attached bacteria (PAB) to different PUAs amendments. We found no differences in the alkaline phosphatase (APase) activity and the abundance of PAB between inside and outside the bloom at a low PUAs dosage (<30 μM). However, for a high PUAs dosage (300 μM), APase activity and PAB growth were reduced significantly outside the bloom but no influences within the bloom. Our findings indicate that the hotspot-level oxylipins may play essential roles in bacterial P-remineralization in P-limited coastal areas. PAB can adapt to the high level of PUAs released by diatoms (or their resulting detritus) and potentially maintain a high rate of organic P recycling during the late stages of diatom blooms. Consequently, the interaction between oxylipin-rich diatoms and bacteria may affect phytoplankton blooms and carbon sequestration in the coastal oceans.

Organic aggregates (OA) are the important circulation hub of matter and energy in aquatic ecosystems. However, the comparison studies on OA in lakes with different nutrient levels are limited. In this study, spatio-temporal abundances of OA and OA-attached bacteria (OAB) in oligotrophic Lake Fuxian, mesotrophic Lake Tianmu, middle-eutrophic Lake Taihu, and hyper-eutrophic Lake Xingyun were investigated in different seasons during 2019-2021 using a scanning electron microscope, epi-fluorescence microscope, and flow cytometry. The results showed that:① the annual average abundances of OA in Lake Fuxian, Lake Tianmu, Lake Taihu, and Lake Xingyun were 1.4×104, 7.0×104, 27.7×104, and 16.0×104 ind·mL-1, whereas the annual average abundances of OAB in the four lakes were 0.3×106, 1.9×106, 4.9×106, and 6.2×106 cells·mL-1. The ratios of OAB:total bacteria (TB) in the four lakes were 30%, 31%, 50%, and 38%, respectively. ② OA abundance in summer was significantly higher than that in autumn and winter; however, the ratio of OAB:TB in summer was approximately 26%, which was significantly lower than that in the other three seasons. ③ Lake nutrient status was the most important environmental factor that affected the abundance variations of OA and OAB, accounting for 50% and 68% of the spatio-temporal variations in OA and OAB abundances. ④ Nutrient and organic matters were enriched in OA, especially in Lake Xingyun; the proportions of particle phosphorous, particle nitrogen, and organic matters in this lake were as high as 69%, 59%, and 79%, respectively. Under the circumstance of future climate change and the expansion of lake algal blooms, the effects of algal-originated OA in the degradation of organic matters and nutrient recycling would be increased.

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.

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.