Recognizing Functional Groups of MES/APG Mixed Surfactants for Enhanced Solubilization toward Benzo[a]pyrene.

Benzo[a]pyrene is difficult to remove from soil due to its high octanol/water partition coefficient. The use of mixed surfactants can increase solubility but with the risk of secondary soil contamination, and the compounding mechanism is still unclear. This study introduced a new approach using environmentally friendly fatty acid methyl ester sulfonate (MES) and alkyl polyglucoside (APG) to solubilize benzo[a]pyrene. The best result was obtained when the ratio of MES/APG was 7:1 under 6 g/L total concentration, with an apparent solubility (Sw) of 8.58 mg/L and a molar solubilization ratio (MSR) of 1.31 for benzo[a]pyrene, which is comparable to that of Tween 80 (MSR, 0.95). The mechanism indicates that the hydroxyl groups (-OH) in APG form "O-H···OSO2-" hydrogen bonding with the sulfonic acid group (-SO3-) of MES, which reduces the electrostatic repulsion between MES molecules, thus facilitating the formation of large and stable micelles. Moreover, the strong solubilizing effect on benzo[a]pyrene should be ascribed to the low polarity of ester groups (-COOCH3) in MES. Functional groups capable of forming hydrogen bonds and having low polarity are responsible for the enhanced solubilization of benzo[a]pyrene. This understanding helps choose suitable surfactants for the remediation of PAH-contaminated soils.

Removal of benzo[a]pyrene from the soil by adsorption coupled with degradation on saponin-modified bentonite immobilized crude enzymes.

Bentonite is a non-metallic mineral with montmorillonite as the main component. It is an environmentally friendly mineral material with large reserves, wide distribution, and low price. Bentonite can be easily modified organically using the surfactant saponin to obtain saponin-modified bentonite (Sap-BT). This study investigates the immobilization of crude enzymes obtained from Trametes versicolor by physical adsorption with Sap-BT. Thus, saponin-modified bentonite immobilized crude enzymes (CE-Sap-BT) were developed to remove benzo[a]pyrene. Immobilization improves the stability of free enzymes. CE-Sap-BT can maintain more than 80% of activity at 45 °C and after storage for 15 d. Additionally, CE-Sap-BT exhibited a high removal rate of benzo[a]pyrene in soil, with 65.69% after 7 d in highly contaminated allotment soil and 52.90% after 6 d in actual soil contaminated with a low concentration of benzo[a]pyrene at a very low laccase dosage (0.1 U/3 g soil). The high catalytic and removal performance of CE-Sap-BT in contaminated sites showed more excellent practical application value.

Bioremediation of PAHs-contaminated site in a full-scale biopiling system with immobilized enzymes: Removal efficiency and microbial communities.

Bioremediation of PAHs-contaminated soil by immobilized enzymes is a promising technology. Nevertheless, the practical implementation of highly efficient enzymatic remediation remains confined to laboratory settings, with limited experience in full-scale applications. In this study, the extracellular enzymes from white rot fungi are fully applied to treat sites contaminated with PAHs by combining a new hydrogel microenvironment and a biopiling system. The full-scale project was conducted on silty loam soil contaminated with PAHs. In line with China's guidelines for construction land, 7 out of the 12 PAHs identified are considered to be a threat to the soil quality of construction sites, with benzo[a]pyrene levels reaching 1.50 mg kg-1, surpassing the acceptable limit of 0.55 mg kg-1 for the first type of land. After 7 days of remediation, the benzo[a]pyrene level decreased from 1.50 mg kg-1 to 0.51 mg kg-1, reaching the remediation standard of Class I screening values, with a removal rate of 66%. Microbiomes were utilized to assess the microbial biodiversity and structure analyses for PAHs biodegradation. The remediation enhanced the abundance of dominant bacterium (Marinobacter, Pseudomonas, and Truepera) and fugin (Thielavia, Neocosmospora, and Scedosporium). The research offers further insights into the exploration of soil remediation on the full-scale of the immobilized enzyme and biopiling technology.

Bioremediation of 2,4,6-trichlorophenol by extracellular enzymes of white rot fungi immobilized with sodium alginate/hydroxyapatite/chitosan microspheres.

Hydroxyapatite, a calcium phosphate biomass material known for its excellent biocompatibility, holds promising applications in water, soil, and air treatment. Sodium alginate/hydroxyapatite/chitosan (SA-HA-CS) microspheres were synthesized by cross-linking sodium alginate with calcium chloride. These microspheres were carriers for immobilizing extracellular crude enzymes from white rot fungi through adsorption, facilitating the degradation of 2,4,6-trichlorophenol (2,4,6-TCP) in water and soil. At 50 °C, the immobilized enzyme retained 87.2% of its maximum activity, while the free enzyme activity dropped to 68.86%. Furthermore, the immobilized enzyme maintained 68.09% of its maximum activity at pH 7, surpassing the 51.16% observed for the free enzyme. Under optimal conditions (pH 5, 24 h), the immobilized enzymes demonstrated a remarkable 94.7% removal rate for 160 mg/L 2,4,6-TCP, outperforming the 62.1% achieved by free crude enzymes. The degradation of 2,4,6-TCP by immobilized and free enzymes adhered to quasi-first-order degradation kinetics. Based on LC-MS, the plausible biodegradation mechanism and reaction pathway of 2,4,6-TCP were proposed, with the primary degradation product identified as 1,2,4-trihydroxybenzene. The immobilized enzyme effectively removed 72.9% of 2,4,6-TCP from the soil within 24 h. The degradation efficiency of the immobilized enzyme varied among different soil types, exhibiting a negative correlation with soil organic matter content. These findings offer valuable insights for advancing the application of immobilized extracellular crude enzymes in 2,4,6-TCP remediation.

Mulches assist degraded soil recovery via stimulating biogeochemical cycling: metagenomic analysis.

Soil degradation of urban greening has caused soil fertility loss and soil organic carbon depletion. Organic mulches are made from natural origin materials, and represent a cost-effective and environment-friendly remediation method for urban greening. To reveal the effects of organic mulch on soil physicochemical characteristics and fertility, we selected a site that was covered with organic mulch for 6 years and a nearby lawn-covered site. The results showed that soil organic matter, total nitrogen, and available phosphorus levels were improved, especially at a depth of 0-20 cm. The activities of cellulase, invertase, and dehydrogenase in soil covered with organic mulch were 17.46%, 78.98%, and 283.19% higher than those under lawn, respectively. The marker genes of fermentation, aerobic respiration, methanogenesis, and methane oxidation were also enriched in the soil under organic mulch. Nitrogen cycling was generally repressed by the organic mulch, but the assimilatory nitrate and nitrite reduction processes were enhanced. The activity of alkaline phosphatase was 12.63% higher in the mulch-covered soil, and functional genes involved in phosphorus cycling were also enriched. This study presents a comprehensive investigation of the influence of organic mulch on soil microbes and provides a deeper insight into the recovery strategy for soil degradation following urban greening. KEY POINTS: • Long-term cover with organic mulches assists soil recovery from degradation • Soil physical and chemical properties were changed by organic mulches • Organic mulches enhanced genes involved in microbially mediated C and P cycling • Soil organic matter was derived from decomposition of organic mulch and carbon fixation • N cycling was repressed by mulches, except for assimilatory NO2- and NO3- reductions.

Enhance PD/A biofilm formation via a novel biochar/tourmaline modified-biocarriers to treat low-strength contaminated surface water: Initial adhesion and high-substrate microenvironment.

In this work, a novel polyurethane carrier modified with biochar and tourmaline/zeolite powder at ratio of 1:1 and 1:2 was developed to promote the formation of biofilms and the synergy of overall bacterial activity for Partial Denitrification/Anammox to treat low-nitrogen contaminated surface water. Based on the batch experiment, the modified biocarrier, BTP2 (biochar: tourmaline = 2: 1), exhibited the highest total nitrogen removal efficiency (83.63%) under influent total nitrogen of 15 mg/L and COD/NO3- of 3. The dense biofilm was formed in inner side of biocarrier owing to the increased surface roughness and various functional groups suggested by scanning electron microscopy and Fourier-transform infrared analysis. The EPS content increased from 200.15 to 220.26 mg/g VSS in BTP2 system. Besides, the rapid NH4+ capture and organics release of the modified carrier fueled the growth of anammox and denitrification bacteria, with the activity of 2.13 ± 0.52 mg N/gVSS/h and 6.70 ± 0.52 mg N/gVSS/h (BTP2). High-throughput sequencing unraveled the increased abundances of Candidatus_Competibacter (0.82%), Thauera (0.60%) and Candidatus_Brocadia (0.55%) which was responsible for the synergy of incomplete reduction of NO3- to NO2- and NH4+ oxidation. Overall, this study provided a valid and simple-control guide for biofilm formation towards rapid enrichment and great collaboration of Anammox and denitrification bacteria.

Enhanced biodegradation of benzo[a]pyrene with Trametes versicolor stimulated by citric acid.

Polycyclic aromatic hydrocarbons (PAHs) are a class of persistent organic pollutants with carcinogenic, mutagenic and teratogenic effects. The white-rot fungi in the fungal group have significant degradation ability for high molecular weight organic pollutants. However, exogenous fungi are easily antagonized by indigenous microorganisms. Low molecular weight organic acids, a small molecular organic matter secreted by plants, can provide carbon sources for soil microorganisms. Combining organic acids with white rot fungi may improve the nutritional environment of fungi. In this study, immobilized Trametes versicolor was used to degrade benzo[a]pyrene in soil, and its effect on removing benzo[a]pyrene in soil mediated by different low molecular weight organic acids was investigated. The results showed that when the degradation was 35 days, the removal effect of the experimental group with citric acid was the best, reaching 43.7%. The degradation effect of Trametes versicolor on benzo[a]pyrene was further investigated in the liquid medium when citric acid was added, and the effects of citric acid on the biomass, extracellular protein concentration and laccase activity of Trametes versicolor were investigated by controlling different concentrations of citric acid. In general, citric acid can act as a carbon source for Trametes versicolor and promote its extracellular protein secretion and laccase activity, thereby accelerating the mineralization of benzo[a]pyrene by Trametes versicolor. Therefore, citric acid can be used as a biostimulant in the remediation of PAHs contaminated soil with Trametes versicolor.

Biodegradation of Trichloroethylene by Trametes versicolor and its Physiological Response to Contaminant Stress.

Trichloroethylene (TCE) poses a potentially toxic threat to humans and the environment and widely exists in contaminated sites. White rot fungi effectively degrade refractory pollutants, while a few research studies use white rot fungi to degrade TCE. In this study, we investigated TCE biodegradation by white rot fungi and the potential influencing factors in the environment and attempted to research the effect of TCE on the physiological characteristics of white rot fungi. White rot fungi (Trametes versicolor, Pseudotrametes gibbosa, Pycnoporus sanguines and Pleurotus ostreatus) were added to the liquid medium for shock culture. The results revealed that T. versicolor exhibited the most pronounced efficacy in removing TCE, with a degradation rate of 81.10% within a 7 d period. TCE induces and is degraded by cytochrome P450 enzymes. High pH and Cr(VI) adversely affected the effectiveness of the biodegradation of TCE, but the salinity range of 0-1% had less effect on biodegradation. Overall, the effectiveness of degradation of TCE by T. versicolor has been demonstrated, and it provides a reference for the application prospects of white rot fungi in TCE-contaminated soils.

Mediative Mechanism of Freezing/Thawing on Greenhouse Gas Emissions in an Inland Saline-Alkaline Wetland: a Metagenomic Analysis.

Inland saline-alkaline wetlands distributed in the mid-high latitude have repeatedly experienced freezing and thawing. However, the response of greenhouse gas (GHG) emission and microbially-mediated carbon and nitrogen cycle to freezing and thawing remains unclear. We monitored the GHG flux in an inland saline-alkaline wetland and found that, compared with the growth period, the average CO2 flux decreased from 171.99 to 76.61-80.71 mg/(m2‧h), the average CH4 flux decreased from 10.72 to 1.96-3.94 mg/(m2‧h), and the average N2O flux decreased from 56.17 to - 27.14 to - 20.70 µg/(m2‧h). Freezing and thawing significantly decreased the relative abundance of functional genes involved in carbon and nitrogen cycles. The aceticlastic methanogenic pathway was the main methanogenic pathway, whereas the Candidatus Methylomirabilis oxyfera was the most abundant methane oxidizer in the wetland. Ammonia-oxidizing archaea and denitrifier belonging to proteobacteria was the major microbial N2O source, while bacteria within clade II nosZ was the major microbial N2O sink. Freezing and thawing reduced the relative abundance of these genes, leading to a decrease in GHG flux.

Efficient nitrogen removal from mature landfill leachate by single-stage partial-nitritation anammox using expanded granular sludge bed.

The effective retention of anaerobic ammonia oxidizing (anammox) bacteria and its high sensitivity to toxic substances and oxygen posed a major challenge to the application of partial nitrification combined with anammox (PN/A) in mature landfill leachate treatment, although it is a promising and efficient nitrogen removal process. In this study, a single-stage PN/A process based on expanded granular sludge bed was proposed to treat the mature landfill leachate. During the last phase, when the NH+ 4-N concentration of mature landfill leachate in influent was 1150.0 mg/L, the nitrogen removal efficiency (NRE) was 83.64% with 1.07 kg N/(m3·d) nitrogen removal rate (NRR). The activity of anammox bacteria (AnAOB) and ammonia oxidizing bacteria (AOB) was 9.21 ± 0.22 mg N/(gVSS·h) and 14.34 ± 0.65 mg N/(gVSS·h), respectively. The bacteria produced a high amount of tightly bound extracellular polymeric substance (TB-EPS) i.e., 4071.79 mg/(g·VSS). This helped to create granular sludge and provided favorable spatial conditions for the distribution of functional bacteria that were adapted to different environments. Due to the efficient retention of functional bacteria by the granular sludge, the relative abundance of Ca.Brocadia and Ca.Kuneneia was 1.71% and 0.31%, respectively. Redundancy analysis (RDA) and microbial correlation network diagram showed that the relative abundance of Ca. Kuenenia, Nitrosomonas and Truepera had a stronger positive correlation with the increase of the proportion of mature landfill leachate added to the influent. Overall, the PN/A process based on granular sludge provides an effective method for autotrophic biological nitrogen removal from mature landfill leachate.

Bioremediation of Diesel-Contaminated Soil by Fungal Solid-State Fermentation.

To address the poor removal of diesel in soil by indigenous microorganisms, we proposed a fungal solid-state fermentation (SSF) method for bioremediation. We screened Pycnoporus sanguineus 5.815, Trametes versicolor 5.996, and Trametes gibbosa 5.952 for their diesel-degrading abilities, with Trametes versicolor 5.996 showing the most promise. The fungal inoculum was obtained through SSF using wood chips and bran. Trametes versicolor 5.996 was applied to two treatments: natural attenuation (NA, diesel-contaminated soil) and bioremediation (BR, 10% SSF added to diesel-contaminated soil). Over 20 days, NA removed 12.9% of the diesel, while BR achieved a significantly higher 38.3% degradation rate. BR also increased CO2 and CH4 emissions but reduced N2O emissions. High-throughput sequencing indicated SSF significantly enriched known diesel-degrading microorganisms like Ascomycota (83.82%), Proteobacteria (46.10%), Actinobacteria (27.88%), Firmicutes (10.35%), and Bacteroidota (4.66%). This study provides theoretical support for the application of fungal remediation technology for diesel and improves understanding of microbiologically mediated diesel degradation and soil greenhouse gas emissions.

Landfill leachate treatment by graphite engineered anaerobic membrane bioreactor: Performance enhancement and membrane fouling mitigation.

Low efficiency of anaerobic digestion and membrane fouling, treating landfill leachate, are big barriers in the application of anaerobic membrane bioreactor (AnMBR). Anaerobic digestion enhancement and membrane fouling mitigation of AnMBR with graphite addition, treating landfill leachate, were investigated in this study. The effect of graphite on organics removal, biogas production, methane content in biogas, membrane fouling, microbial responses and foulant compositions were analyzed. With the graphite addition, chemical oxygen demand (COD) removal of 78% was achieved for influent COD concentration of 3000 mg/l, which was significantly higher than the stage without graphite addition (65%) for influent COD concentration of 2000 mg/l. Similarly, methane content in biogas with graphite addition was 56%, while without graphite addition it was 46%. These digestion improvements were due to the promotion of organics degradation, facilitated by direct interspecies electron transfer (DIET) mechanism via graphite addition in AnMBR. The graphite addition prolonged membrane cleaning cycle from 13 days to 30 days. Protein content in loosely bound extracellular polymeric substance (LB-EPS) was the main fouling agent, which decreased with the graphite addition. The main mechanism behind membrane fouling mitigation was the protein content reduction in LB-EPS, which was biodegraded by Trichococcus being increased in relative abundance with the graphite addition. Furthermore, abundance of Denitratisoma decreased in anaerobic sludge and its accumulation reduced on membrane surface, subsequently membrane fouling was mitigated. Overall, graphite addition in AnMBR is a potential eco-innovative approach that efficiently removes pollutants from landfill leachate, enhances biogas quality and mitigates membrane fouling.

Effect of exogenous hydrazine on metabolic process of anammox bacteria.

The effect of N2H4 (hydrazine) on AnAOB (anaerobic ammonia oxidizing bacteria) metabolic pattern is unknown. Therefore, the main purpose of this paper was to explore the effects of exogenous N2H4 on the SAA (specific anammox activity), characteristics and metabolic pathway of AnAOB. The results showed that low N2H4 concentration (1-5 mg/L) continuous dosing can promote SAA. The promoting effect was found to be more obvious within the dosage of 3-5 mg/L N2H4. It was also indicated that high N2H4 concentration dosing (5-10 mg/L) can trigger the self-protection mechanism of AnAOB granular sludge by secreting a large amount of B-PN (binding polymeric protein). Intermittent addition of N2H4 at low concentration is conducive to the long-term stable operation of anammox process. Exogenous N2H4 can be directly oxidized by AnAOB to promote the consumption of NO2--N and NH4+-N. In addition, excess electrons can also drive the process of NO3--N reduction and NO2--N disproportionation. Theoretically, these reaction processes need two and ten extra electrons respectively, which is not easy to occur compared with the anammox process.

Combining high pressure homogenization with free nitrous acid pretreatment to improve anaerobic digestion of sewage sludge.

Single pretreatment of sewage sludge, either physical, chemical or biological, has its own drawbacks in term of poor sanitization, energy intensity and high operational and capital cost. To tackle these drawbacks, combined high pressure homogenization (HPH) and free nitrous acid (FNA) pretreatment for sludge solubilization and further biodegradation in anaerobic digestion was investigated. Synergistic effect of combined HPH (40 MPa) and FNA (2.49 mg/L) pretreatment (HPH-FNA) for improving anaerobic digestion was evaluated, and its effect on archaeal and bacterial community structure was analyzed. Compared with single HPH and FNA pretreatments, HPH-FNA pretreatment efficiently solubilized wasted activated sludge (WAS), subsequently improved anaerobic digestion. Cumulative biogas production from sewage sludge pretreated with HPH-FNA was 154%, 108% and 284% more than that with single pretreatment of FNA, HPH and raw sludge, respectively. In addition, volumetric biogas production of combined pretreatment system (815 ml) was more than the sum from single pretreatment (710 ml). Methane content in biogas for raw sludge, FNA, HPH and HPH-FNA pretreated sludge was 45%, 51%, 55% and 65%, respectively. Illumina MiSeq sequencing analysis revealed that HPH-FNA pretreatment promoted bacterial growth of phyla Bacteroidetes, Firmicutes and Synergistetes and archaeal genera Methanospirillum and Methanosaeta. Overall, combined HPH-FNA pretreatment of sewage sludge, prior to anaerobic digestion, is an environmentally-friendly and potentially economic technology.

A comprehensive review on the use of conductive materials to improve anaerobic digestion: Focusing on landfill leachate treatment.

Globally, around 70% of waste is disposed of in open dumps or landfill sites, with the leachate generated from these sites containing high concentrations of organic and inorganic compounds, which will adversely affect aquatic environments if discharged without proper treatment. Anaerobic digestion of landfill leachate is an environmentally-friendly method that efficiently converts organic compounds into methane-rich biogas. However, the widespread application of anaerobic digestion has been hindered by poor system stability, low methanogenic activity and a high level of volatile fatty acids (VFAs) accumulation, increasing the operational costs of treatment. Conductive materials can be added to the digester to improve the performance of anaerobic digestion in landfill leachate treatment systems and studies reporting the use of conductive materials for this purpose are hereby thoroughly reviewed. The mechanism of microbial growth and enrichment by conductive materials is discussed, as well as the subsequent effect on waste metabolism, methane production, syntrophic relationships and interspecies electron transfer. The porous structure, specific surface area and conductivity of conductive materials play vital roles in the facilitation of syntrophic relationships between fermentative bacteria and methanogenic archaea. In addition, the mediation of direct interspecies electron transfer (DIET) by conductive materials increases the methane content of biogas from 16% to 60% as compared to indirect interspecies electron transfer (IIET) in conventional anaerobic digestion systems. This review identifies research gaps in the field of material-amended anaerobic systems, suggesting future research directions including investigations into combined chemical-biological treatments for landfill leachate, microbial management using conductive materials for efficient pollutant removal and the capacity for material reuse. Moreover, findings of this review provide a reference for the efficient and large-scale treatment of landfill leachate by anaerobic digestion with conductive materials.

Current and emerging trends in bioaugmentation of organic contaminated soils: A review.

Organic contaminated soils constitute an important environmental problem, whereas field applicability of existing physical-chemical methods has encountered numerous obstacles, such as high chemical cost, large energy consumption, secondary pollution, and soil degradation. Bioaugmentation is an environmentally friendly and potentially economic technology that efficiently removes toxic pollutants from organic contaminated soils by microorganisms or their enzymes and bioremediation additives. This review attempted to explore the recent advances in bioaugmentation of organic contaminated soils and provided a comprehensive summary of various bioaugmentation methods, including bacterial, fungus, enzymes and bioremediation additives. The practical application of bioaugmentation is frequently limited by soil environmental conditions, microbial relationships, enzyme durability and remediation cycles. To tackle these problems, the future of bioaugmentation can be processed from sustainability of broad-spectrum bioremediation carriers, microbial/enzyme agents targeting combined contaminants, desorption of environmentally friendly additives and small molecular biological stimulants. Findings of this research are expected to provide new references for bioaugmentation methods that are practically feasible and economically potential.

Phytoremediation of Three Herbaceous Plants to Remove Metals from Urban Runoff.

Phytoremediation experiments were carried out to assess the ability of different herbaceous buffer strips to absorb metals. Results revealed the Medicago sativa buffer strip had the best removal capacity for copper, lead, and cadmium in the runoff and seepage flow. Copper and lead content in M. sativa were significantly higher than in Lolium perenne and Poa pratensis (p < 0.05). Cadmium content in P. pratensis was significantly higher than in L. perenne and M. sativa(p < 0.05). Total accumulation of copper and lead in M. sativa buffer strips was 13.45 mg kg-1 and 1.01 mg kg-1, respectively. Total cadmium accumulation was approximately 0.50 mg kg-1 in both M. sativa and P. pratensis. Overall, results indicated that using M. sativa in the buffer strips was optimal for the remediation of copper, lead, and cadmium in sewage water.

Dynamics of Archaeal and Bacterial Communities in Response to Variations of Hydraulic Retention Time in an Integrated Anaerobic Fluidized-Bed Membrane Bioreactor Treating Benzothiazole Wastewater.

An integrated anaerobic fluidized-bed membrane bioreactor (IAFMBR) was investigated to treat synthetic high-strength benzothiazole wastewater (50 mg/L) at a hydraulic retention time (HRT) of 24, 18, and 12 h. The chemical oxygen demand (COD) removal efficiency (from 93.6% to 90.9%), the methane percentage (from 70.9% to 69.27%), and the methane yield (from 0.309 m3 CH4/kg·CODremoved to 0.316 m3 CH4/kg·CODremoved) were not affected by decreasing HRTs. However, it had an adverse effect on membrane fouling (decreasing service period from 5.3 d to 3.2 d) and benzothiazole removal efficiency (reducing it from 97.5% to 82.3%). Three sludge samples that were collected on day 185, day 240, and day 297 were analyzed using an Illumina® MiSeq platform. It is striking that the dominant genus of archaea was always Methanosaeta despite of HRTs. The proportions of Methanosaeta were 80.6% (HRT 24), 91.9% (HRT 18), and 91.2% (HRT 12). The dominant bacterial genera were Clostridium in proportions of 23.9% (HRT 24), 16.4% (HRT 18), and 15.3% (HRT 12), respectively.

Egalitarianism in Biofilms.

Microbial biofilms are multicellular communities of sessile microorganisms encased by the hydrated polymeric matrix. They have significant influences on both aquatic/terrestrial ecosystem and anthropogenic activities. Taking advantage of the governing features of selective stress (Tan and Ng in Water Res 42:1122-1132, 2008; Wei in Water Res 45:863-871, 2011; Dereli in Water Res 59C:11-22, 2014), the evenness of microbial communities in a membrane-centered mesocosm was successfully manipulated. By measuring the biofilm growing rates under different evenness levels of communities, an evenly distributed community favors the formation of biofilms was observed. This finding is not only a new evidence linking biofilm diversity to its functionality but also a clear suggestion on controlling a biofilm-based process via a simple and smart way.