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Global climate change has significantly increased the duration of droughts in intermittent rivers, impacting benthic microbial-mediated biogeochemical processes. However, the response mechanisms of biofilms on different substrate types to alternating dry and wet conditions and their related ecosystem functions remain poorly understood. This study uses high-throughput sequencing and enzyme assays to investigate the impact of gradient drought stress on microbial diversity and functional changes of biofilm communities inhabiting on gravel, cobblestone, and sediment. Results showed that the duration of drought significantly affects microbial diversity, with algal and bacterial α-diversity declining under extended drought across gravel, cobblestone, and sediment substrates. At the same time, fungal diversity was less affected, likely due to their distinct ecological niches and reproductive strategies. β-diversity analysis revealed significant changes in community heterogeneity, with algae and bacteria showing increased Bray-Curtis dissimilarities, indicating distinct adaptation strategies that may affect ecosystem functioning. Fungal communities, however, were less impacted by drought-induced heterogeneity changes. Network analysis showed that drought altered microbial network connectivity, with algal networks displaying decreased path distances, while bacterial networks remained stable, suggesting greater resilience to drought stress. Functional enzyme assays revealed significantly reduced denitrification rates across all substrates post-drought, with distinct denitrifying enzyme activity responses depending on substrate type. Partial least squares path modeling revealed that algal biodiversity were closely linked to the maintaining of enzyme activities, particularly denitrification rates of biofilms on cobblestone and gravel. These findings indicated the critical role of substrate types in shaping microbial responses to drought stress, with distinct microbial groups and diversity indices playing key roles in maintaining ecosystem functions. This study highlights the importance of understanding the interactions between microbial community dynamics and ecosystem functions under varying environmental stressors in river ecosystems., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The intensification of human activities has led to flow reduction and cut-off in most global rivers, seriously affecting riverine organisms and the biogeochemical processes. As key indicators of river ecosystems' structure and function, benthic biofilms play a critical role in driving primary production and material cycling in rivers. This research aimed to investigate the characteristics of microbial communities' complexity and stability during river flow reduction. Benthic biofilms were grown in artificial channels and subjected to eight gradients of flow reduction (represented by flow velocity from 0.4 to 110 cm/s). Biofilms' biodiversity, ecological networks and community assembly of bacteria, fungi and algae were investigated by high-throughput sequencing. Results showed significant differences in community composition and structure under different flow conditions. The eight flow gradients' microbial communities were divided into three groups: low, medium and high flows. The flow reduction led to significant decreases in bacterial and fungal communities' Chao1 index. Low flow conditions enriched the bacterial phyla Oxyphotobacteria, Alphaproteobacteria and Mollicutes, but significantly decreased the fungal phylum Chytridiomycota. Lowering flow reduced the fungal network's number of nodes and increased the algal network's number of edges. Cross-domain interactions network analysis showed a gradual increase in node and edge numbers with decreasing flow, while decreasing average path length. The neutral model predicted stochastic processes primarily drove biofilm community assembly, and that model's explanations decreased as the flow gradient decreased. The null model analysis revealed diffusion limitation as the most common stochastic ecological process for bacterial and algal communities, with reduced flow reducing heterogeneous selection and increasing diffusion-limited processes. This study provides an in-depth analysis of flow reduction's effects on biofilm communities' ecological networks and community assembly., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Microplastics (MPs) can disintegrate into smaller sized microplastics and even nanoplastics (NPs). The toxicity of nanoplastics and microplastics on freshwater organisms have been well explored recently, however, very little is known about the potential impacts of NPs on freshwater biofilms, which are essential for primary production and nutrient cycling in aquatic ecosystems. In this study, we studied the acute effects (3 h of exposure) of polystyrene beads (PS, with diameter range from 100 nm to 9 μm) on five biological endpoints targeting community and ecosystem-level processes in biofilms: chlorophyll a, photosynthetic yield, and three extracellular enzyme activities. The results showed that the large size PS beads (500 nm, 1 μm, and 9 μm) exhibited negligible effects on the determined biological endpoints in biofilms within the range of concentrations (5-100 mg/L) in this study. However, high concentration of PS beads (100 nm, 100 mg/L) significantly decreased the content of chlorophyll a, and the functional enzyme activities of β-glucosidase and leucine aminopeptidase, suggesting negative effects on the carbon and nitrogen cycling of freshwater biofilms. Moreover, the influences of PS NPs (100 nm) on biofilms strongly depended on the surface modification of PS particles, with the positively charged PS NPs (amide-modified) exhibiting the highest toxicity to biofilms. The excess generation of reactive oxygen species (ROS) in this study indicated oxidative stress induced by PS NPs, which might lead to the observed nano-toxic effects on biofilms. In response, the antioxidant activity of biofilm was enhanced as indicated by the increased total antioxidant capacity (T-AOC). Overall, our findings highlight nanoplastics have potential to disrupt the basic ecological functions of biofilms in aquatic environments., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The widespread application of commercial TiO 2 NPs inevitably leads to their release into environmental waters through various ways. TiO 2 NPs released into water might be absorbed by and react with periphytic biofilms, which are a kind of aquatic environmental media of important ecological significance, and influence the physiological activity and ecological function of periphytic biofilms. This study investigated the effects of exposure to 1 mg/L and 5 mg/L of TiO 2 NPs on periphytic biofilms cultured indoors. After a 10-day exposure to TiO 2 NPs, the growth (measured by chlorophyll-a content) of microalgal community was inhibited greatly (more than 60%); however, the primary production (indicated by quantum yield) of periphytic biofilms maintained changeless. As for bacteria, TiO 2 NP-exposure increased the bacterial diversity and altered the composition structure. Significant changes were observed in the bacterial communities at the class level, mainly including Alphaproteobacteria, Gammaproteobacteria, Cytophagia, Flavobacteriia, Sphingobacteriia, Synechococcophycideae and Oscillatoriophycideae. The enhancement of metabolic activities (the production of extracellular polymeric substances, especially proteins content increased by 48.51%) of periphytic biofilms was a resistance mechanism to toxicity of NPs. As for extracellular enzyme activities of periphytic biofilms, alkaline phosphatase activity was inhibited (22.43%) after exposed to 5 mg/L of TiO 2 NPs, which posed a threat to phosphorus metabolism of periphytic biofilms. Overall, this study demonstrated that 1 mg/L and 5 mg/L of TiO 2 NPs negatively influenced physiological activities and ecological functions of periphytic biofilms, highlighting that the ecological risks of TiO 2 NPs should be paid attention to., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

In this paper, the effects of cerium oxide nanoparticles (CeO 2 NPs) on the group bacterial behaviors were elaborated. After 36-h cultivation, the biofilm biomass was enhanced by the sub-lethal concentrations of 0.5 and 2 mg/L CeO 2 NP exposure. Meanwhile, the promoted production of total amino acids in microbes further resulted in the increased surface hydrophobicity and percentage aggregation. To resist the CeO 2 NPs stress, the biofilm exhibited a double-layer microstructure, with the protein (PRO) and living cells occupying the bottom, the polysaccharide (PS), and dead cells dominating the top. The bacterial diversity was highly suppressed and Citrobacter and Pseudomonas from the phylum of γ-Proteobacteria strongly dominated the biofilm, indicating the selective and enriched effects of CeO 2 NPs on resistant bacteria. The stimulated inherent resistance of biofilm was reflected by the reduced adenosine triphosphate (ATP) content after 4 h exposure. The increased levels of reactive oxygen species (ROS) in the treatments of 8 h CeO 2 NP exposure led to the upregulated quorum sensing signals of acylated homoserine lactone (AHL) and autoinducer 2 (AI-2), beneficial to mitigating the environmental disturbance of CeO 2 NPs. These results provide evidences for the accelerating effects of CeO 2 NPs on biofilm formation through oxidative stress, which expand the understanding of the ecological effects of CeO 2 NPs.

In this study, we investigated the effects of copper oxide (CuO) NPs on freshwater sediment biofilms in terms of the functional properties and microbial community structure. Biofilms were incubated in microcosms and CuO NPs (1 mg/L uncoated and humic-acid-coated) were exposed with Cu 2+ (Cu(NO 3 ) 2 ) as the positive control. As determined from DO (dissolved oxygen) microelectrodes measurements, a high-DO region emerged inside the biofilms after 5-day exposure to CuO NPs compared with those before NP additions, which suggested CuO NPs inhibit the oxygen respiration activity. These results were consistent with the decreased heterotrophic respiration. CuO NPs significantly altered the bacterial community composition and decreased the abundances of Anaerolineaceae, Acidobacteria, Aminicenantes, and Anaerolinea. Functional analysis from PICRUSt (Phylogenetic Investigation of Communities by Reconstruction of Unobserved States)-predicted metagenomes indicated that bacterial genera depleted by CuO NP treatments were related to carbohydrate and glycan biosynthesis and metabolism, and biosynthesis of other secondary metabolites. These functional profiles combined with the decreased activities of extracellular enzymes, β-glucosidase (GLU) and l-leucine aminopeptidase (LAP), suggested that the introduction of CuO NPs exhibit negative effects on the biogeochemical processes and the cycling of carbon and nitrogen in biofilm systems. Whereas these toxic effects of CuO NPs could be mitigated when the aquatic environment is enriched with natural organic matters such as humic acid., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Microplastics are frequently detected in freshwater environments, serving as a new factitious substrate for colonization of biofilm-forming microorganisms. Distinct microbial assemblages between microplastics and surrounding waters have been well documented; however, there is insufficient knowledge regarding biofilm colonization of plastic and non-plastic substrates, despite the fact that microbial communities generally aggregate on natural solid surfaces. In this study, the effects of substrate type on microbial communities were evaluated by incubation of biofilms on microplastic substrates (polyethylene and polypropylene) and natural substrates (cobblestone and wood) for 21 days under controlled conditions. Results from high-throughput sequencing of 16S rRNA revealed that the alpha diversity (richness, evenness, and diversity) was lower in the microplastic-associated communities than in those on the natural substrates, indicating substrate-type-coupled species sorting. Distinct community structure and biofilm composition were observed between these two substrate types. Significantly higher abundances of Pirellulaceae, Phycisphaerales, Cyclobacteriaceae, and Roseococcus were observed on the microplastic substrates compared with the natural substrates. Simultaneously, the functional profiles (KEGG) predicted by Tax4Fun showed that the pathways of amino acid metabolism and metabolism of cofactors and vitamins were increased in biofilms on the microplastic substrates. The findings illustrate that microplastic acts as a distinct microbial habitat (compared with natural substrates) that could not only change the community structure but also affect microbial functions, potentially impacting the ecological functions of microbial communities in aquatic ecosystems., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In this study, the biofilm formation of Pseudomonas aeruginosa in the presence of cerium oxide nanoparticles (CeO 2 NPs) was investigated. With the addition of 0.1 mg/L and 1 mg/L CeO 2 NPs, the biofilm development was substantially enhanced. During the attachment process, the enhanced surface hydrophobicity and excess production of mannosan and rhamnolipids in CeO 2 NP treatments were detected, which were conductive to the colonization of bacterial cells. During the maturation period, the biofilm biomass was accelerated by the improved aggregation percentage as well as the secretion of extracellular DNA and pyocyanin. The reactive oxygen species (ROS) generated by CeO 2 NPs were found to activate the N-butyryl homoserine lactone (C 4 -HSL) and quinolone signals secreted by Pseudomonas aeruginosa. Moreover, the quorum sensing (QS) systems of rhl and pqs were initiated, reflected by the stimulated expression levels of biofilm formation-related genes rhlI-rhlR, rhlAB, and pqsR-pqsA. The addition of a quorum quencher, furanone C-30, significantly declined the activities of QS-controlled catalase and superoxide dismutase. A dose of antioxidant, ascorbic acid, effectively relieved the accelerating effects of NPs on biofilm formation. These results indicated that CeO 2 NPs could accelerate biofilm formation through the interference of QS system by generating ROS, which provides possible targets for controlling biofilm growth in the NP exposure environments.

Inhibitory effects of ceria nanoparticles (CeO 2 NPs) on biofilm were investigated individually and in combination with phosphate (P), ethylene diamine tetraacetic acid (EDTA), humic acid (HA) and citrate (CA) to further explore the toxicity alleviating solutions. Exposure to 20 mg/L CeO 2 NPs significantly decreased the performance of biofilm in nutrients removal. Distribution experiments suggested >98% of the CeO 2 NPs retained in microbial aggregates, leading to 51.26 μg/L Ce ions dissolution. The dissolved Ce IV and its further being reduced to Ce III stimulated the formation of O 2 and OH, which increased lipid peroxidation level to 130.93% in biofilms. However, P/EDTA/CA captured or precipitated Ce ions, whereas EDTA/HA/CA shielded NPs-bacteria direct contacts, both disturbing the NPs adsorption, intercepting the redox transition between Ce - and OH, which increased lipid peroxidation level to 130.93% in biofilms. However, P/EDTA/CA captured or precipitated Ce ions, whereas EDTA/HA/CA shielded NPs-bacteria direct contacts, both disturbing the NPs adsorption, intercepting the redox transition between Ce IV and Ce III , reducing the generation of O 2 - and OH, thus mitigating the toxicity of CeO 2 NPs. These results illustrate the main drivers of CeO 2 NPs biotoxicity and provide safer-by-design strategies., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

With the accelerated application of CeO 2 nanoparticles (NPs), wastewater treatment plants will increasingly receive CeO 2 NPs, thus inevitably causing CeO 2 NPs to encounter microaggregates. Here, we comprehensively elucidate the responses in the structural, physicochemical and microbial properties of wastewater biofilms to chronic exposure (75 days) to different CeO 2 NPs concentrations, with a particular emphasis on the protective mechanisms of stratified extracellular polymeric substances (EPSs). Chronic exposure to 0.1 mg/L CeO 2 NPs boosted the content and broadened the distribution of α-d-glucopyranose polysaccharides (PS), while the sharply increased production and breadth of β-d-glucopyranose PS, forming a formidable shield, was a response to 10 mg/L CeO 2 NPs. After the bacteria were exposed to CeO 2 NPs, loosely bound EPSs (LB-EPSs) aggregated into macromolecules (increasing in apparent molecular weight (AMW)) but at a lower abundance, whereas the average AMW in tightly bound EPSs (TB-EPSs) decreased. The acetyl content and (α-helix+3-turn helix)/β-sheet value of TB-EPSs increased to resist CeO 2 NPs. Furthermore, long-term exposure to CeO 2 NPs decreased cell viability, reduced microbial diversity and shifted the microbial composition. N-acylated-l-homoserine lactone concentrations increased with increased density of Pseudomonas, which was associated with PS-regulated control, thus promoting PS production in EPSs in response to CeO 2 NPs. These results expand the understanding of how microaggregates resist environmental stress caused by NPs., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The influences of cerium dioxide nanoparticles (CeO 2 NPs) on nitrogen removal in biofilm were investigated. Prolonged exposure (75d) to 0.1mg/L CeO 2 NPs caused no inhibitory effects on nitrogen removal, while continuous addition of 10mg/L CeO 2 NPs decreased the treatment efficiency to 53%. With the progressive concentration of CeO 2 NPs addition, the removal efficiency could nearly stabilize at 67% even with the continues spike of 10mg/L. The micro-profiles of dissolved oxygen, pH, and oxidation reduction potential suggested the developed protection mechanisms of microbes to progressive CeO 2 NPs exposure led to the less influence of microenvironment, denitrification bacteria and enzyme activity than those with continuous ones. Furthermore, high throughput sequencing illustrated the drastic shifted communities with gradual CeO 2 NPs spiking was responsible for the adaption and protective mechanisms. The present study demonstrated the acclimated microbial community was able to survive CeO 2 NPs addition more readily than those non-acclimated., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The short term (8h) influences of cerium oxide nanoparticles (CeO 2 NPs) on the process of phosphorus removal in biofilm were investigated. At concentration of 0.1mg/L, CeO 2 NPs posed no impacts on total phosphorus (TP) removal. While at 20mg/L, TP removal efficiency reduced from 85.16% to 59.62%. Results of P distribution analysis and 31 P nuclear magnetic resonance spectroscopy implied that the anaerobic degradation of polyphosphate (polyP) and the release of orthophosphate in extracellular polymeric substances (EPS) were inhibited. After aerobic exposure, the average chain length of polyP in microbial cells and EPS was shorter than control, and monoester and diester phosphates in cells were observed to release into EPS. Moreover, the EPS production and its contribution to P removal increased, while the capacity of EPS in P storage declined. X-ray diffraction analysis and saturation index calculation revealed that the formation of inorganic P precipitation in biofilm was inhibited., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The growing production and application of CuO nanoparticles increase the chance that these particles will be released into wastewater treatment plants (WWTPs) and interact with microorganisms. However, the toxicity response mechanism of biofilm to NP exposure may be different from that of activated sludge due to the denser and stronger microbial aggregate structure of biofilm. Thus, in this study, the response to CuO NPs of wastewater biofilm collected from a rotating biological contactor was investigated. Short-term exposure (24h) to CuO NPs led to a great loss in cell viability, and SEM-EDS images revealed that the nano-CuO aggregates were not transformed to Cu-S species in the biofilm samples. In response, more extracellular polymeric substance (EPS) (especially loosely bound-EPS) was produced in wastewater biofilm exposed to CuO NPs, with a higher content of protein compared to polysaccharides. The shifts of fluorescence intensity and peak locations in 3D-EEM fluorescence spectra indicated chemical changes of the EPS components. FT-IR analysis revealed that exposure to nano-CuO had more distinct effects on the functional groups of proteins and polysaccharides in LB-EPS. Illumina sequencing of 16S rRNA gene amplicons revealed that CuO NPs enhanced bacterial diversity. The bacterial community structure significantly shifted, with a significantly increased abundance of Comamonas, a slight increase in Zoogloea, and a notable decrease in Flavobacterium. The shifts of these dominant genera may be associated with altered EPS production, which might result in microbial community function fluctuations. In conclusion, exposure to high concentrations of CuO NPs has the potential to shape wastewater biofilm bacterial community structure., (Copyright © 2017 Elsevier B.V. All rights reserved.)

In this study, we examined the long-term effects of copper oxide nanoparticles (CuO NPs) on the production and properties of EPS and the resulting variations in surface physicochemical characteristics of biofilms in a sequencing batch biofilm reactor. After exposure to 50 mg/L CuO NPs for 45 days, the protein (PRO) and polysaccharide (PS) contents in loosely bound EPS (LB-EPS) decreased as the production of LB-EPS decreased from 34.4 to 30 mg TOC/g EPS. However, the production of tightly bound EPS (TB-EPS) increased by 16.47 % as the PRO and PS contents increased. The content of humic-like substances (HS) increased significantly, becoming the predominant constituent in EPS with the presence of 50 mg/L CuO NPs. Furthermore, the results of three-dimensional excitation-emission fluorescence spectra confirmed the various changes in terms of the LB-EPS and TB-EPS contents after exposure to CuO NPs. Fourier transform infrared spectroscopy showed that the -OH and -NH 2 groups of proteins in EPS were involved in the reaction with CuO NPs. Moreover, the chronic exposure to CuO NPs induced a negative impact on the flocculating efficiency of EPS and on the hydrophobicity and aggregation ability of microbial cells. The PRO/PS ratios of different EPS fractions were consistent with their hydrophobicities (R 2 >0.98) and bioflocculating efficiencies (R 2 >0.95); however, there was no correlation with aggregation ability. Additionally, the presence of bovine serum albumin (BSA) prevented the physical contact between CuO NPs and EPS as a result of NP aggregation and electrostatic repulsion.

The impacts of CuO nanoparticle (NP) exposure on total nitrogen (TN) removal in a sequencing batch biofilm reactor (SBBR) as well as the effects of natural organic matter (NOM) in wastewater were studied. Short-term exposure (8 h) to 1 and 50 mg/L CuO NPs induced negligible influence on the nitrogen removal efficiency, and biofilms could recover from the slight damage caused by the prolonged exposure (45 days) to 1 mg/L CuO NPs. On the other hand, long-term exposure to 50 mg/L CuO NPs notably decreased the nitrogen removal efficiencies to 47.74 and 59.04 % in the absence and presence of bovine serum albumin (BSA), much lower than those in the control (75.43 %), mainly as the suppressed denitrification process. Analysis of key enzyme activities showed that the activities of nitrite reductase and nitrate reductase were obviously reduced with 50 mg/L CuO NP exposure. Further studies revealed that the inhibited nitrite/nitrate reductase was related to the variations of microenvironment pH and decrease of nirS and nirK by microelectrode and fluorescent quantitative polymerase chain reaction (PCR) analysis. In addition, the presence of BSA mitigated the toxicity of CuO NPs due to the enhanced particle size and Cu 2+ release, electrostatic repulsion, and surface coating of CuO NPs, which indicated that lower inhibition effects of CuO NPs in NOM-rich wastewater is of importance when evaluating the environmental risk of NPs to wastewater treatment plants.

The presence of ZnO nanoparticles (ZnO NPs) in natural waters has raised concerns about their environmental impacts, but the potential influences of ZnO NPs on fluvial biofilm have not been reported. In this study, the utility of antioxidant enzyme activities (AEA) as biomarkers of fluvial biofilm to ZnO NPs toxicity and a method that combines AEA into an index of "Integrated Biomarker Responses (IBR)" were studied. Compared with the absence of ZnO NPs, scanning electron microscopy (SEM) images revealed that a large amount of ZnO NPs were adsorbed onto biofilm and these NPs exerted adverse effects on the viability of bacteria in biofilm. The production of reactive oxygen species (ROS) with high concentrations (30 and 100mg/L) of ZnO NPs exposure reached to 184% and 244% of the control, while no cell leakage and membrane damage were observed. After exposure to ZnO NPs for 0.25 and 3 days, the activities of catalase (CAT), superoxide dismutase (SOD) and glutathione reductase (GR), glutathione peroxidase (GSH-Px) were significantly increased, respectively. At the end of exposure period (21 days), the AEA with the presence of 1mg/L ZnO NPs exposure were comparable to the control, while most of those in high concentrations of ZnO NPs were decreased. The results of IBR showed that the biofilm can adapt to 1mg/L ZnO NPs exposure, while be seriously damaged by 30 and 100mg/L ZnO NPs after 3 and 0.25 days. IBR can be used as an appropriate evaluation system of the toxicity effects of ZnO NPs on fluvial biofim., (Copyright © 2016. Published by Elsevier Inc.)

Zinc oxide nanoparticles (ZnO NPs) used in consumer products are largely released into the environment through the wastewater stream. The health hazard of ZnO NPs and the contribution of dissolved Zn(2+) in toxicity of ZnO NPs has attracted extensive worldwide attention. In this study, the toxic effects of ZnO nanoparticles (ZnO NPs) and the effects of dissolved Zn(2+) on fluvial biofilms were investigated. At the end of the exposure time (21 days), scanning electron microscopy (SEM) images and bioaccumulation experiments revealed that large quantities of ZnO NPs were adsorbed on the biofilm. The algal biomasses were significantly decreased by six- and eleven-fold compared with the control (1.43 μg/L) by exposure to concentrations of 100mg/L ZnO NPs and 7.85 mg/L Zn(2+), respectively. Moreover, under the same exposure conditions, the quantum yields presented contents of 53.33 and 33.33% relative to the control, and a shift in the community composition that manifested as a strong reduction in diatoms was observed from 7 days and reached 15.63 and 6.25% of the control after 21 days of exposure, respectively. The reductions in bacteria viability and reactive oxygen species (ROS) production were noticeably enhanced following exposure to 100mg/L ZnO NPs and 7.85 mg/L Zn(2+), respectively. Additionally, the acute and rapid toxicity of Zn(2+) and the increasing toxicity of the ZnO NPs with increased bioaccumulation were noted in the exposure experiment., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The effects of CeO2 nanoparticles (CeO2 NPs) exposure on biological nitrogen removal in a sequencing batch biofilm reactor (SBBR) were investigated. At low concentration (1 mg/L), no significant effect was observed on total nitrogen (TN) removal. However, at high concentrations (10 and 50 mg/L), the TN removal efficiency reduced from 74.09% to 64.26% and 55.17%, respectively. Scanning electron microscope imaging showed large amounts of CeO2 NPs adsorbed on the biofilm, which increased the production of reactive oxygen species. The exposure at only 50 mg/L CeO2 NPs measurably affected the lactate dehydrogenase release. Confocal laser scanning microscopy showed that high concentrations of CeO2 NPs reduced bacterial viability. Moreover, after a short-term exposure, extracellular polymeric substances (EPS) were observed to increase, forming a compact matrix to protect the bacteria. The activities of nitrate reductase and ammonia monooxygenase were inhibited, but there was no significant impact on the activity of nitrite oxidoreductase., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The surfaces of natural sediments are ubiquitously coated by biofilms that increase the content of organic matter in sediments. However, it is less understood whether the biofilms act as a sorbent or a barrier of mass transfer from water column to sediment phase. This study focused on the role of biofilms coverage on sediments in the sorption of bisphenol A (BPA), 17α-ethinyl estradiol (EE2), and 4-nonylphenols (4-NP) as model compounds for endocrine-disrupting chemicals (EDCs). The OC-normalized distribution coefficients (k OC) for BPA, EE2 and 4-NP ranged from 10(1.87) to 10(3.09) l/kg, the k OC of EE2 was slightly higher (10(2.23) l/kg) for sediment after H2O2 oxidation than before (10(1.93) l/kg). A two-stage model with a fast section and slow section was employed to describe the sorption process (r (2) > 0.95). The model results showed that the fast sorption section played a main role in the sorption process, while the slow section determined the extent of the reaction (the second-phase partition coefficient (k p2) ranged from 11.7 to 118.9 l/kg). The ratios of the mass transfer rate constant of the two stages for the natural sediment ranged from 6.0 to 7.2, which were somewhat lower than those for soil samples. These results indicated that the biofilm coverage on sediment may act as a barrier in mass transfer from water to sediment and scarcely increased the sorption capacity of sediments.

The presence of ZnO NPs in waste streams can negatively affect the efficiency of biological nutrient removal from wastewater. However, details of the toxic effects of ZnO NPs on microbial activities of wastewater biofilms have not yet been reported. In this study, the temporal and spatial inhibitory effects of ZnO NPs on the O2 respiration activities of aerobic wastewater biofilms were investigated using an O2 microelectrode. The resulting time-course microelectrode measurements demonstrated that ZnO NPs inhibited O2 respiration within 2h. The spatial distributions of net specific O2 respiration were determined in biofilms with and without treatment of 5 or 50mg/L ZnO NPs. The results showed that 50mg/L of nano-ZnO inhibited the microbial activities only in the outer layer (∼200μm) of the biofilms, and bacteria present in the deeper parts of the biofilms became even more active. Scanning electron microscopy (SEM) analysis showed that the ZnO NPs were adsorbed onto the biofilm, but these NPs had no adverse effects on the cell membrane integrity of the biofilms. It was found that the inhibition of O2 respiration induced by higher concentrations of ZnO NPs (50mg/L) was mainly due to the release of zinc ions by dissolution of the ZnO NPs., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Flume experiments were conducted to investigate the distribution and composition of extracellular polymeric substances (EPS) in biofilms and the detachment mechanism of biofilms grown under different flow velocity conditions. The results of biofilm growth kinetics showed that the growth trends were coincident with the logistic growth model. The growth kinetics parameters reached their maximum under intermediate velocity (IV) condition. Biofilms exhibited different profiles of EPS composition and distribution, depending on the flow conditions in which the biofilms were grown. The amounts of total polysaccharide and total protein in the thin biofilm (high velocity condition 2 - HV2) were both generally greater than those in the thick biofilm (IV). As compared to the heterogeneous distribution of EPS in the thick biofilms (IV), the EPS in the thin biofilms (HV2) exhibited more homogeneous distribution, and the bound EPS in the thin biofilms (HV2) were much greater than those in the thick biofilms (IV). From the detachment tests, an inverse relationship was observed between the proportion of detached biomass and the value of flow velocity during growth. Biofilms grown under higher velocities showed stronger cohesion than those grown under lower velocities. Therefore, water velocity during biofilm growth conditioned the distribution and composition of EPS, as well as its detachment characteristics under higher shear stress.

With the wide application of selenium (Se) in industrial production, different Se-based compounds (selenate and selenite) are produced and released into aquatic environments. The potential impacts of such Se compounds on the biofilms (a complex microbial aggregate in aquatic systems) need to be substantially explored. Herein, we investigated the responses of bacterial community diversity, composition and structure, and function of biofilms after 21 days of exposure to low concentrations (100 µg/L) and high concentrations (1 mg/L) of sodium selenate and sodium selenite, respectively. Distinct effects of selenium in different valences on the community structure and microbial functions of biofilms were observed. Compared with the controls, the addition of selenate and selenite solutions altered the richness of biofilms but not the diversity, which is dependent on the concentration and valences, with sodium selenite (1 mg/L) exhibiting a strong inhibition effect on community richness. Significant changes of community composition and structure were observed, with a significant increase in Proteobacteria (31.08–58.00%) and a significant decrease in Bacteroidetes (32.15–11.45%) after exposure to sodium selenite with high concentration. Also, different responses of gamma-Proteobacteria and alpha-Proteobacteria were observed between the sodium selenite and sodium selenate treatments. Moreover, results showed that sodium selenite could strengthen the function of the metabolism of biofilms, and the higher the concentration is, the more apparent the enhancement effect is. All these results suggested that the effects of different valence states of selenium were obvious, and sodium selenite with high concentration strongly changed the diversity, structure and function of biofilms. [ABSTRACT FROM AUTHOR]

Periphytic biofilms have been widely used in wastewater purification and water ecological restoration, and artificial substrates have been progressively used for periphyton immobilisation to substitute natural substrates. However, there is insufficient knowledge regarding the interaction network structure and microbial functions in biofilm communities on artificial substrates, which are essential attribute affecting their applications in biofilm immobilisation. This study compared the community structure, co-occurrence network, and metabolic functions of bacterial and microeukaryotic periphytic biofilms during a 35-day indoor cultivation on artificial substrates, such as artificial carbon fibre (ACF) and polyvinyl chloride (PVC), and natural substrates, such as pebble and wood. Results demonstrated that different types of artificial substrates could affect the community composition and functional diversity of bacterial and microeukaryotic biofilms. The bacterial and microeukaryotic community on ACF and PVC showed significantly higher Simpson index compared to those on wood. Bacterial networks on artificial substrates were more complex than those on natural substrates, while the keystone species on natural substrates were more abundant, indicating that the bacterial communities on artificial substrates had stronger stability and resistance to external interference. Furthermore, the functional metabolic profiles predicted showed the abilities of bacterial communities to metabolise nitrogen and carbon sources colonised on artificial substrates were stronger than those on natural substrates. These findings demonstrated that artificial substrates could be special niches for microbial colonisation, possibly altering microbial compositions, interactions, and functions. Therefore, this study provides a powerful theoretical basis for choosing suitable artificial substrates for microbial aggregation and immobilisation technology. [ABSTRACT FROM AUTHOR]

The widespread use of microplastics (MPs) inevitably lead to their release into aquatic environment, where they are likely to encounter the periphytic biofilms, leading to changes in microbial community structure and function. However studies on the toxicity of MPs with different particle sizes and concentrations to biofilms are still lacking. Here, the effects of polyethylene (PE) MPs with three sizes (10, 40 and 120 µm) and two concentrations (2 and 20 mg/L) on biofilm communities were investigated in microcosms over a 28-day incubation period. High-throughput sequencing showed that the alpha and beta diversity of biofilm community were significantly altered by MPs, depending on their size and concentration. Typically, larger particle sizes and higher concentrations resulted in more significant changes in biofilm community structure, indicating a stronger shading effect of larger particles. Compared to the controls, 120 µm MPs treatments significantly reduced the Chao1 and Shannon index of biofilm community regardless of the concentration, while 40 µm MPs reduced the Shannon index only at high concentrations. Moreover, MPs resulted in the changes in the abundances of bacterial communities, with the abundance of Cyanobacteria increasing significantly while that of Proteobacteria and Bacteroidetes decreasing in all treatments. Interestingly, functional analysis by FAPROTAX suggested that a decrease in the abundance of chemoheterotrophy genes and an increase in photoautotrophic- and nitrification-associated genes were observed after exposure to MPs, leading to potential changes of the carbon and nitrogen metabolism of biofilms. These results highlighted that MPs exposure had negative impacts on microbial community structure and function, which in turn affects microbial-mediated processes. [Display omitted] • Effects of MPs with different particle sizes on biofilms were studied. • The responses were dependent on the particle sizes and concentrations of MPs. • MPs of larger sizes resulted in more significant changes in community structure. [ABSTRACT FROM AUTHOR]

Sulfamethoxazole (SMX) is an extensively applied antibiotic frequently detected in municipal wastewater, which cannot be efficiently removed by conventional biological wastewater processes. In this work, an intimately coupled photocatalysis and biodegradation (ICPB) system consisting of Fe3+-doped graphitic carbon nitride photocatalyst and biofilm carriers was fabricated to remove SMX. The results of wastewater treatment experiments showed that 81.2 ± 2.1% of SMX was removed in the ICPB system during the 12 h, while only 23.7 ± 4.0% was removed in the biofilm system within the same time. In the ICPB system, photocatalysis played a key role in removing SMX by producing hydroxyl radicals and superoxide radicals. Besides, the synergism between photocatalysis and biodegradation enhanced the mineralization of SMX. To understand the degradation process of SMX, nine degradation products and possible degradation pathways of SMX were analyzed. The results of high throughput sequencing showed that the diversity, abundance, and structure of the biofilm microbial community remained stable in the ICPB system at the end of the experiments, which suggested that microorganisms had accommodated to the environment of the ICPB system. This study could provide insights into the application of the ICPB system in treating antibiotic-contaminated wastewater. [Display omitted] • An ICPB system consisting of Fe3+-doped g-C 3 N 4 and bio-carriers was constructed. • 57.5% more SMX was removed in the ICPB system than in the biofilm system. • Synergism of photocatalysis and biodegradation in mineralizing SMX was found. • The microbial community structure was kept relatively stable in the ICPB system. [ABSTRACT FROM AUTHOR]

Because of the high production rates, low recycling rates, and poor waste management of plastics, an increasing amount of plastic is entering the aquatic environment, where it can provide new ecological niches for microbial communities and form a so-called plastisphere. Recent studies have focused on the one-way impact of plastic substrata or biofilm communities. However, our understanding of the two-way interactions between plastics and biofilms is still limited. This review first summarizes the formation process and the co-occurrence network analysis of the aquatic plastisphere to comprehensively illustrate the succession pattern of biofilm communities and the potential consistency between keystone taxa and specific environmental behavior of the plastisphere. Furthermore, this review sheds light on mutual interactions between plastics and biofilms. Plastic properties, environmental conditions, and colonization time affect biofilm development. Meanwhile, the biofilm communities, in turn, influence the environmental behaviors of plastics, including transport, contaminant accumulation, and especially the fragmentation and degradation of plastics. Based on a systematic literature review and cross-referencing from these disciplines, the current research focus, and future challenges in exploring aquatic plastisphere development and biofilm-plastic interactions are proposed. [Display omitted] • Interaction between plastics and biofilms in aquatic environments were reviewed. • Microbial structure and co-occurrence network in plastisphere were discussed. • Impacts of plastic properties and environmental conditions on biofilm were reviewed. • Impacts of biofilm on the environmental behavior of plastics were analyzed. [ABSTRACT FROM AUTHOR]

Extracellular polymeric substances (EPS) are a major component of biofilms that act as a gel-like matrix, binding the cells together to form their three-dimensional structure. The effects of ceria nanoparticles (CeO 2 NPs) on the production and physicochemical characteristics of EPS in biofilms in a sequencing batch biofilm reactor were investigated. Total EPS production, including loosely bound EPS (LB-EPS) and tightly bound EPS (TB-EPS), increased by 35.41% compared to in control tests without CeO 2 NPs. Protein production increased by 47.02% (LB-EPS) and 58.83% (TB-EPS) after 50 mg/L CeO 2 NP exposure. Three-dimensional excitation–emission fluorescence spectra revealed that tyrosine (LB-EPS) and aromatic (TB-EPS) protein-like substances formed after CeO 2 NP exposure. Fourier transform infrared spectroscopy results indicated the susceptibility of –OH and –NH 2 in EPS hydroxyl and amine groups to CeO 2 NPs. Exposure to 50 mg/L CeO 2 NPs reduced the flocculating capacity of LB-EPS (51.78%) and TB-EPS (17.14%), consistent with the decreased zeta potential. [ABSTRACT FROM AUTHOR]

Microbial biofilms can rapidly colonize plastic debris in aquatic environments and subsequently, accumulate chemical pollutants from the surrounding water. Here, we studied the microbial colonization of different plastics, including polyethylene terephthalate (PET), polypropylene (PP), polyvinyl chloride (PVC), and polyethylene (PE) exposed in three freshwater systems (the Qinhuai River, the Niushoushan River, and Donghu Lake) for 44 days. We also assessed the biofilm mass and associated metals attached to plastics. The plastics debris characteristics, such as contact angle and surface roughness, greatly affected the increased biofilm biomass. All types of metal accumulation onto the plastic substrate abundances significantly higher than the concentrations of heavy metal in the water column, such as Ba (267.75 μg/g vs. 42.12 μg/L, Donhu Lake), Zn (254 μg/g vs. 0.023 μg/L the Qinhuai River), and Cr (93.75 μg/g vs. 0.039 μg/L, the Niushoushan River). Compared with other metals, the heavy metal Ba, Cr and Zn accumulated easily on the plastic debris (PET, PP, PVC, and PE) at all incubation sites. Aquatic environmental factors (total nitrogen, total phosphorus, and suspended solids concentrations) largely shaped metal accumulation onto plastic debris compared with plastic debris properties. [Display omitted] • The heavy metal accumulation onto biofilm covered plastic debris were studied. • Distinct biomasses of biofilms were found on various plastic debris. • Ba, Cr and Zn showed highest accumulation on the plastic debris. • Aquatic environmental factors largely shaped metal accumulation on plastic debris. Aquatic environmental factors (i.e., TN, TP and SS) largely shaped metal accumulation on plastic debris. [ABSTRACT FROM AUTHOR]

The transport behaviors of copper oxide (CuO) NPs in wastewater matrix and their possible impacts on microbial activities of stable wastewater biofilms cultivated in a lab scale rotating biological contactor (RBC) were investigated. Significant aggregation of CuO NPs was observed in the wastewater samples, depending on their mass concentrations. Extracellular polymeric substance (EPS)-adsorbed copper accounted for a large proportion of the total copper accumulated in biofilms. The microelectrode profiles showed that a single pulse exposure to 50 mg/L CuO resulted in a deeper penetration depth of oxygen in biofilms compared to the CuO NP free biofilms. The maximum oxygen consumption rate shifted to the deeper parts of biofilms, indicating that the respiration activities of bacteria in the top region of the biofilms was significantly inhibited by CuO NPs. Biofilms secreted more EPS in response to the nano-CuO stress, with higher production of proteins compared to polysaccharides. [ABSTRACT FROM AUTHOR]

Periphytic biofilms have served as a potentially important environmental media of contaminants removal and an important indicator of the health of aquatic systems. When the nanoparticles (NPs) released, they are likely to encounter periphytic biofilms, and then their ecosystem functioning might be changed. Here, we explored the long-term (28 days) impacts of CuO NPs (μM magnitude) on the microbial biomass, metabolic activities, community structure and associated ecosystem-level processes in multi-species biofilms inoculated from lake water. Slight aggregation of CuO NPs was observed during the exposure experiment. All of the algal, fungal, and bacterial community structure in biofilms were already altered by seven days of exposure to 0.781 and 7.81 μM CuO NPs, but a series of functional endpoints did not show clear differences, suggesting the functional redundancy within biofilm communities. After 28 days, however, a wide range of microbial endpoints that are specific to autotrophs (photosynthetic yield) or heterotrophs (basal respiration) or enzymes that represent the target communities were significantly affected. These decreases matched with the total copper content in biofilms, and indicated the negative effects of CuO NPs on the primary production and nutrient cycling of biofilms. Overall, the response of biofilms to CuO NPs produced a slow and delayed effect on microbial-mediated functions over community structure. These results highlight the highly sensitive responses of biofilms in fresh water to CuO NP, suggesting that CuO NPs (at environmentally relevant doses) would, although changes are slow, affect the biofilms' functioning and services, such as their sustainability for purification of polluted surface water. Image 1 • 28-day exposure of CuO nanoparticles (∼μM) negatively affected the multi-species biofilm. • Community structure shifted rapidly while microbial functions had a delayed response. • Algal, fungal, and bacterial community structure were already altered at day 7. • Functional endpoints specific to autotrophs or heterotrophs or the target communities decreased at day 28. [ABSTRACT FROM AUTHOR]

Abstract: Vibrio parahaemolyticus AphA and OpaR are the two master regulators of quorum sensing (QS) that are abundantly produced and operate at low cell density (LCD) and high cell density (HCD), respectively, with an outcome of reciprocally gradient production of these two proteins with transition between LCD and HCD. The cpsQ-mfpABC gene cluster is transcribed as two operons cpsQ-mfpABC and mfpABC in V. parahaemolyticus. MfpABC is a putative membrane fusion transporter that contributes to biofilm development. CpsQ is a c-di-GMP-binding regulator that activates the expression of capsular polysaccharide genes and mfpABC and, thus, induces biofilm development. As shown in this study, OpaR and AphA bind to the promoter region of mfpABC to enhance and repress its transcription, respectively. In contrast, the positive and negative regulation of cpsQ-mfpABC by AphA and OpaR, respectively, achieves probably through acting of AphA or OpaR on additional unknown regulator(s) of cpsQ-mfpABC. The transcriptional levels of cpsQ-mfpABC and mfpABC enhance gradually with transition from LCD to HCD due to the above reciprocal regulatory action of OpaR and AphA. Data presented here present a novel paradigm of combined action of the two master QS regulators in controlling expression of the QS regulon members. [Copyright &y& Elsevier]

There is an increasing concern regarding the potential effects of nanoplastics (NPs) on freshwater ecosystems. Considering the functional values of biofilms in freshwater, knowledge on whether and to what extent NPs can influence the ecosystem processes of biofilms were still limited. Herein, the freshwater biofilms cultured in lab were exposed to 100 nm polystyrene NPs (PS-NPs) of different dosages (1 and 10 mg/L) for 14 days. Confocal laser scanning microscope observation indicated that biofilms were dominated by filamentous, and spiral algae species and the intensity of extracellular polymeric substances increased under PS-NPs exposure. GeoChip 5.0 analysis revealed that PS-NPs exposure triggered a significant increase in functional genes α diversity (p < 0.05) and altered biofilms' functional structure. Furthermore, the abundance of genes involved in the total carbon and nitrogen cycling were increased under PS-NPs exposure. The abundance of nitrogen fixation genes experienced the most pronounced increase (24.4%) under 1 mg/L PS-NPs treatment, consistent with the increase of ammonium in overlying water. Whereas antibiotic resistance genes and those related to photosynthetic pigments production were suppressed. These results provided direct evidence for PS-NPs' effects on the biofilm functions in terms of biogeochemical cycling, improving our understanding of the potentials of NPs on freshwater ecosystems. [Display omitted] • Effects of PS NPs on freshwater biofilms were studied by Geochip 5 microarrays. • PS NPs exposure significantly altered the microbial functional diversity. • Genes of total carbon and nitrogen cycling were increased under NPs exposure. • 1 mg/L NPs treatment increased the nifH gene consistent with the increase of NH 4 +-N. • PS NPs negatively influenced the ecosystem processes of biofilm communities. [ABSTRACT FROM AUTHOR]

Microplastics (MPs) have frequently been detected in freshwater environments, and there is growing concern about their ecological effects, especially the influence of the "plastisphere" on the freshwater ecosystems. The colonization of microbes on MPs would significantly alter their transport behavior, i.e., buoyancy, in fresh water. In this research, we studied the effects of biofilm colonization on the sinking and floating of three MPs, i.e., polyethylene terephthalate (PET), polypropylene (PP), and polyvinyl chloride (PVC), after 44 days of incubation in three freshwater systems (the Niushoushan River, the Qinhuai River, and East Lake) in China. The results showed that the biofilms attached to the three MPs contained different biomass and chlorophyll-a levels were related to water environmental conditions and physicochemical properties of MPs, based on redundancy analysis. Generally, PET and PVC sinking, with density higher than water, tended to increase after biofilm formation. Thereafter, the settling velocity of biofouled PET and PVC squares became faster than that of the virgin ones. In summary, our study suggested that biofouling does affect the sinking of MPs in fresh water and consequently influences the transport behavior and the distribution characteristics of MPs in freshwater environments, and this issue deserves more scientific attention. [Display omitted] • Effects of biofilm colonization on the transport behaviors of MPs were studied. • Distinct biomasses of biofilms were found on various MPs in the three fresh waters. • Biofouling increased PET and PVC density. • Biofouling favored the sinking of MPs with original density higher than water. [ABSTRACT FROM AUTHOR]

• Microbial functions of biofilms on microplastic and inert substrates were studied. • Distinct carbon metabolic patterns of biofilms colonizing microplastic substrates were found. • Water physicochemical properties affected the carbon metabolism of biofilms. • Microplastics may influence microbial-mediated carbon cycling in aquatic ecosystems. Concerns are growing about the increasing amounts of microplastics (MPs) and their ecological impacts, especially the influences of "plastisphere" in the freshwater ecosystems. Although the microbial structure and composition of biofilms are investigated, knowledge of their microbial functions remains limited. Herein, we investigated the functional diversity of carbon metabolism in biofilms colonizing one inert (glass) and two MPs as polyvinyl chloride (PVC) and polyethylene terephthalate (PET) substrates incubated for 44 days in situ in the Niushoushan River, the Qinhuai River, and Donghu Lake. 2D confocal laser scanning microscopy images visualized distinct micro-structures and biofilm compositions on three substrates. BIOLOG ECO microplates indicated variation on carbon utilization capacities of biofilms of inert and MPs in three freshwater ecosystems. Biofilms on PET showed lower capacities and carbon metabolism rates than those on glass and PVC, indicating the presence of substrate-specific functional diversity. The Shannon-Wiener diversity, Simpson diversity and Shannon evenness indices for the Niushoushan River and Donghu Lake were ordered as glass > PVC > PET. Besides to MPs-specific factors, environmental factors including nutrient (i.e., TN and TP) and turbidity largely shaped biofilm carbon metabolism. Overall findings demonstrated that as specific niches, MPs influenced microbial-mediated carbon cycling in the freshwater ecosystems and MPs-promoted microbial communities posed ecological significance. [ABSTRACT FROM AUTHOR]

• Microbial functioning of biofilms on plastics and natural substrates were studied. • Biofilms on plastics showed higher biomasses than that of natural substrates. • Distinct carbon metabolic activities of biofilms inhabited on plastics were observed. • Environmental factors dominated the metabolism differentiation over substrate types. • Plastic debris are new microbial niche affecting carbon cycling in aquatic ecosystems. As an artificial type of microbial carrier, plastic debris has been widely detected in freshwater habitats, and the potential impacts of the plastisphere (biofilms colonized on plastics) in aquatic ecosystems have drawn increasing attention. Distinct community compositions and structures of biofilms in plastic and natural substrates have been recorded in freshwater environments. However, the microbial metabolic functioning of the plastisphere was underestimated, especially in freshwater environments. In this study, the effects of substrate types on the carbon metabolic functions of biofilms were studied by in situ cultivation of biofilms on plastics (polyvinyl chloride, PVC and polyethylene, PE) and natural substrate (cobblestone) for 44 days in two rivers (the Niushoushan River and the Qinhuai River) and two lakes (Donghu Lake and Xuanwu Lake). Biofilms on plastics showed higher biomasses than those on natural substrates in all ecosystems. Variations in the micro-structure and compactness of biofilms developed under different substrates were observed from scanning electron microscope and confocal laser scanning microscope image analyses. The carbon metabolic activities of the biofilms evaluated by BIOLOG EcoPlate were different between plastics (PVC and PE) and natural substrate (cobblestone) in the four freshwater ecosystems. In the Niushoushan River, PE-associated biofilms had different capacity in using carbon sources from cobblestone-associated biofilms as illustrated by the Shannon-Wiener diversity index and Shannon evenness index. Additionally, the metabolic functional diversity profiles of biofilms on PVC were significantly different from those on cobblestone in the other three aquatic ecosystems. Moreover, results from variation partitioning analysis suggested that the impact of environmental factors (contribution: 21%) on microbial carbon metabolic functions was much greater than that of substrate types (contribution: 6%). These findings illustrated distinct microbial functions of biofilms inhabited on plastics, and environmental factors play a decisive role in the differentiation and specificity of carbon metabolism of the plastisphere. This study offers new insights that plastics serving as artificial microbial niches have the ability to affect the microbial-mediated carbon cycling process in aquatic ecosystems. [ABSTRACT FROM AUTHOR]