Genome-wide identification of the MED25 BINDING RING-H2 PROTEIN gene family in foxtail millet (Setaria italica L.) and the role of SiMBR2 in resistance to abiotic stress in Arabidopsis.

MAIN CONCLUSION: The SiMBR genes in foxtail millet were identified and studied. Heterologous expression of SiMBR2 in Arabidopsis can improve plant tolerance to drought stress by decreasing the level of reactive oxygen species. Foxtail millet (Setaria italica L.), a C4 crop recognized for its exceptional resistance to drought stress, presents an opportunity to improve the genetic resilience of other crops by examining its unique stress response genes and understanding the underlying molecular mechanisms of drought tolerance. In our previous study, we identified several genes linked to drought stress by transcriptome analysis, including SiMBR2 (Seita.7G226600), a member of the MED25 BINDING RING-H2 PROTEIN (MBR) gene family, which is related to protein ubiquitination. Here, we have identified ten SiMBR genes in foxtail millet and conducted analyses of their structural characteristics, chromosomal locations, cis-acting regulatory elements within their promoters, and predicted transcription patterns specific to various tissues or developmental stages using bioinformatic approaches. Further investigation of the stress response of SiMBR2 revealed that its transcription is induced by treatments with salicylic acid and gibberellic acid, as well as by salt and osmotic stresses, while exposure to high or low temperatures led to a decrease in its transcription levels. Heterologous expression of SiMBR2 in Arabidopsis thaliana enhanced the plant's tolerance to water deficit by reducing the accumulation of reactive oxygen species under drought stress. In summary, this study provides support for exploring the molecular mechanisms associated with drought resistance of SiMBR genes in foxtail millet and contributing to genetic improvement and molecular breeding in other crops.

Electrocoagulation enhanced gravity driven membrane bioreactor for advanced treatment of rural sewage.

The daily discharge of rural sewage in China occupies 30 % of the national wastewater discharge, and developing an energy-efficient, easy to operate, and decentralized rural sewage treatment technology becomes an important task. In this work, a novel rural sewage treatment technology, Electrocoagulation enhanced Gravity-Driven Membrane Bioreactor (EC-GDMBR) was exploited for the rural sewage treatment under long-term operation (160 days). Two EC-GDMBRs with various module structures of ceramic membrane (horizontal module and side module) not only displayed the desirable effluent quality, but also sustained the stable flux (8-13 LMH). The electrocoagulation, electrooxidation, biodegradation, and separation in EC-GDMBRs were able to synergistically remove the particle matter, organic (CODCr effluent <11.6 ± 1.2 mg/L) and nutrients (NH3-N effluent <0.1 mg/L, TN effluent <8.5 mg/L, TP effluent <0.05 mg/L). Besides, the high permeability of ceramic membrane and large porosity of biofilm on its surface improved the sustainability of stable flux during the long-term operation. Moreover, by analyzing bacterial abundance, Extracellular Polymeric Substances, Adenosine Tri-Phosphate and Confocal Laser Scanning Microscopy, a large number of microorganisms grew and accumulated on the carrier, as well as formed the biofilm (23.46-659.9 µm), while Nitrobacteria (1.6-4.1 %) and Nitrate (0.01-0.06 %) exited in the carrier biofilms, promoting the nitrogen removal. Compared with EC-GDMBR with side module of ceramic membrane, EC-GDMBR with horizontal module of ceramic membrane has advantages in flux behavior, organic/nutrient removal, microbial abundance/activity, abundance of nitrogen removal functional bacteria and water permeability of biofilm, because the ceramic membrane of horizontal module can promote the uniform growth of biofilm and improve the uniformity of flow penetration distribution. In general, the findings of this work verify the reliability of EC-GDMBR for the sustainable operation of wastewater treatment and improve its application value of rural sewage treatment.

Impacts of dibutyl phthalate on bacterial community composition and carbon and nitrogen metabolic pathways in a municipal wastewater treatment system.

Dibutyl phthalate (DBP) is a typical toxic and hazardous pollutant in pharmaceutical wastewater, affecting the metabolism of microbial flora, leading to decreased treatment efficiency, and deteriorated effluent quality in municipal wastewater treatment plants (WWTPs). This study conducted a long-term experiment with 6 operational stages in a pilot-scale A2O-MBR system, analyzing the effect of DBP on the bacterial community and their carbon and nitrogen metabolic pathways. 16S rRNA gene amplicon sequencing analysis and principal components analysis (PCA) showed that DBP at 8 mg/L significantly influenced the structure of bacterial community (P < 0.05), resulting in reduced bacterial community diversity. Metagenomic analysis was used to explore the embedded carbon and nitrogen metabolic pathways. At the presence of DBP, the metabolism of saccharides, lipids, and aromatic compounds were blocked owing to the vanishment of key enzyme (such as acetylaminohexosyltransferase (EC 2.4.1.92) and UDP-sugar pyro phosphorylase (EC 2.7.7.64)) encoding genes, resulting in weakened carbon metabolism, and thus reduced COD removal performance. The resultant deficiency of the genes such as those encoding hydroxyproline dehydrogenase (EC 1.5.5.3) gave rise to interrupted metabolic pathways of amino acid (arginine, proline, tyrosine, and tryptophan), resulting in declined function of nitrogen metabolism and thus reduced TN removal efficiency. The uncovery of the mechanisms by which DBP affects wastewater treatment system efficiency and microbial metabolism is of theoretical importance for the efficient operation of municipal and pharmaceutical wastewater treatment systems.

Investigation on molecular characteristics of organic compounds during a full-scale landfill leachate treatment process based on non-targeted analysis.

In this study, a new methodology for evaluating full-scale landfill leachate treatment processes by non-targeted analysis using comprehensive two-dimensional gas chromatography quadrupole time-of-flight mass spectrometry (GC × GC-QTOF-MS) was proposed. The method revealed the chemical complexity of organic compounds in landfill leachate samples at the molecular level and evaluated the removal efficiency of the anaerobic-anoxic-oxic (A2O) - membrane bioreactor (MBR) - nanofiltration (NF) treatment process in conjunction with multi-level classification of organic compounds. Results showed that the results of non-targeted analysis combined with multi-level classification of organic compounds had a significant correlation with the conventional water quality parameters and can be used to evaluate the treatment process. A total of 2508 organic compounds were detected in 6 samples. 17 emerging contaminants (ECs) with known potentially hazards were detected, including Diisobutyl Phthalate (DIBP), which is toxic to male reproduction and development, and 4-Tert-Butylphenol, which causes endocrine disruption in animals. The removal rate of organic compounds by this full-scale landfill leachate treatment processes reached 79.14%. The anaerobic tank played a crucial role with 64.98% contribution. For compounds, the removal rate of heterocyclics was as high as 94.67%, and the removal rate of aliphatics was poor, only 63.49%. This treatment process had almost perfect removal effect on the steroids in alicyclics and phenols in aromatics, but poor treatment effect on saturated alkanes in aliphatics and naphthenes in alicyclics. This study provides a methodology for accurate assessment of the molecular level of treatment processes, new insights for process optimization in waste treatment plants, and data support for the detection of emerging contaminants. The environmental hazards of landfill leachate can be further evaluated in the future in conjunction with ecotoxicity assessment studies.

Respirometric tests in a combined UASB-MBR system treating wastewater containing emerging contaminants at different OLRs and temperatures: Biokinetic analysis.

This research focuses on the application of respirometric techniques to provide new insights into the biokinetic behaviour of bacterial species developed in an Upflow Anaerobic Sludge Blanked -UASB reactor combined with a membrane bioreactor -MBR, treating urban wastewater with emerging contaminants frequently found in this kind of effluents. The lab-scale pilot plant was operated at different metabolic and operational conditions by limiting the organic loading rate- OLR of the influent. In a first stage, the MBR was performed with suspended biomass, while in a second stage bio-supports were introduced to operate coexisting suspended and fixed biomass. From the results of the microscopic monitoring of sludge, it was concluded that the decrease in OLR resulted in a greater disintegration of the floc structure, more dispersed growth, and a low presence of inter-floccular bonds. However, no effect of toxicity or inhibition of microorganisms caused by the presence of emerging contaminants -ECs was determined. Kinetic modelling was carried out to study the behaviour of the system. The results showed a slowing down of biomass degradative capacity at low OLR stages and operating at low temperatures of mixed liquor. In addition, a decrease in oxygen consumption was observed with decreasing biodegradable substrate, resulting in lower degradation of organic matter. Mean values of specific oxygen uptake rate and heterotrophic biomass yield at low OLR were SOUR end = 1.49 and 1.15 mg O2· g MLVSS-1 h-1 and YH,MLSSV end = 0.48 and 0.28 mg MLVSS· mg COD-1substrate at stage 1 (suspended biomass) and stage 2 (suspended and supported biomass), respectively. From the analysis of the endogenous decomposition constant (kd), a higher cell lysis was determined operating with suspended biomass (kd = 0.03 d-1) in comparison to the operation coexisting suspended and supported biomass (kd = 0.01 d-1). Heterotrophic biomass yield values (YH, MLVSS = 0.48 ± 0.06, 0.40 ± 0.01 and 0.29 ± 0.01 mg MLVSS· mg COD-1substrate at high, medium and low OLR) showed lower sludge production at low OLR due to the influence of substrate limitation on cell growth.

Quantitative and qualitative variations of biopolymers in a pilot-scale membrane bioreactor treating municipal wastewater throughout 3 years of operation.

In this study, the fouling potential of mixed liquor suspension samples collected from a pilot-scale membrane bioreactor (MBR) that treated municipal wastewater was monitored for more than 3 years. The fouling potential was assessed by batch filtration experiments using the same type of membrane as equipped in the MBR. The fouling potential increased when the temperature of the mixed liquor suspension in the MBR decreased. However, the polysaccharide and protein concentrations in the mixed liquor suspension, which have been focused on many previous studies, did not correlate with the fouling potential (R2 = 0.15 and 0.39, respectively). In contrast, the concentration of biopolymers, quantified by liquid chromatography-organic carbon detection (LC-OCD), exhibited a marked correlation with the fouling potential (R2 = 0.89). A high concentration of biopolymers with large molecular weight (>1 million Da) was likely responsible for the high fouling potential. Fourier transform infrared (FTIR) analysis of the dissolved organic matter in the mixed liquor suspension indicated that the chemical properties of the biopolymers considerably varied with the seasonal temperature variation, which has rarely been reported and gives insights into fouling in MBRs. The effect of temperature on the biopolymer concentration and molecular weight of biopolymers was also investigated in a separate bench-scale experiment in which temperature was controlled. It was clearly shown that a low temperature induced an increase in the biopolymer concentration and an associated increase in the fouling potential of the mixed liquor suspension.

Effect of quorum quenching on biofouling control and microbial community in membrane bioreactors by Brucella sp. ZJ1.

Quorum quenching (QQ) has been demonstrated to be a novel technique for controlling biofouling in membrane bioreactors (MBRs), as it can significantly inhibit biofilm formation by disrupting quorum sensing (QS). The exploration of new QQ bacterial strains and the evaluation of their performance in mitigating membrane fouling in MBR systems is significant. In this study, an efficient QQ strain, Brucella sp. ZJ1 was encapsulated in alginate beads and evaluated for its ability to mitigate biofouling. The findings revealed that MBR with QQ beads extended the operation time by 2-3 times without affecting pollutant degradation. QQ beads maintained approximately 50% QQ activity after more than 50 days operation, indicating a long-lasting and endurable QQ effect. The QQ effect reduced extracellular polymeric substance (EPS) production especially in terms of polysaccharide and protein by more than 40%. QQ beads in the MBR also reduced the cake resistance and the irreversible resistance of membrane biofouling. Metagenomic sequencing suggests that QQ beads suppressed the QS effect and increased the abundance of QQ enzyme genes, ultimately inducing efficient membrane biofouling control.

Dynamic modeling of a full-scale membrane bioreactor performance for landfill leachate treatment.

Leachate treatment is crucial in landfill management. As landfill ages, inert constituents and ammonia nitrogen concentration in leachate increases, which results in a decrease in biological treatability. In this study, a full-scale MBR treating leachate was dynamically modeled using ASM1. The investigated landfill has been serving for more than 25 years; thus, a decrease in biodegradable organic content and an increase in nitrogen content of the leachate is expected in the years ahead. The calibrated model predicted MLSS, effluent COD, and effluent TN concentrations with high accuracy. Following the calibration study, it was found that soluble inert COD and soluble inert organic nitrogen fractions were the primary reasons of high COD and TKN concentrations in the effluent, respectively. The validated model of the full-scale MBR system treating leachate can be a useful tool to understand the limitations of the system. Soluble inert constituents of the leachate that pass through the membrane necessiate additional treatment processes for discharge into surface water bodies.

Long-term impacts of polyethylene terephthalate (PET) microplastics in membrane bioreactor.

Due to the mass production and daily use of plastic products, the potential toxicity of microplastics to the water environment has attracted worldwide attention. In this work, the effect of typical microplastics (PET) on the performance of activated sludge from membrane bioreactors (MBR) was evaluated. The impacts on biological removal efficiency were unconspicuous with continuous dosing of 60 particles/L. However, further investigations revealed that PET particle accumulation caused adverse impacts on settleability and dewaterability. The SVI value increased from 53.3 ml/g MLSS to 69.9 ml/g MLSS and the CST in the PET reactor increased by 22%. Nevertheless, hydrophobicity was reduced by 49.2%. Mechanism studies exposed that the PET microplastics accumulation improved extracellular polymeric substances (EPS) from 116.96 mg/L to 138.70 mg/L and caused cell membrane damage. The abundance and diversity of microbial community reduced in activated sludge in PET reactor compared with control reactor. These phenomena revealed a possible hypothesis that the microplastic particles increased EPS and cytotoxicity of activated sludge. However, the rate of transmembrane pressure (TMP) build-up was significantly mitigated in PET-MBR compared to that in a control-MBR (1.27 folds), which attributes that physical scour of particles may still alleviate membrane contamination in MBR.

Sensitizing multidrug-resistant leukemia cells to common cytostatics by an aluminium-salen complex that has high-apoptotic effects in leukemia, lymphoma and mamma carcinoma cells.

We investigated the aluminium-salen complex MBR-8 as a potential anti-cancer agent. To see apoptotic effects induced by MBR-8, alone and in combination with common cytostatic drugs, DNA-fragmentations were studied using the flow cytometric analysis. Western blot analysis and measurement of the mitochondrial membrane potential with a JC-1 dye were employed to identify the pathway of apoptosis. An impressive overcoming of multidrug-resistance in leukemia (Nalm6) cells was observed. Additionally, solid tumor cells including Burkitt-like lymphoma (BJAB) and mamma carcinoma cells (MCF-7) are affected by MBR-8 in the same way. Western blot analysis revealed activation of caspase-3. MBR-8 showed very pronounced selectivity with regard to tumor cells and high synergistic effects in Nalm6 and daunorubicin-resistant Nalm6 cells when administered in combination with vincristine, daunorubicin and doxorubicin. The aluminium-salen complex MBR-8 showed very promising anti-cancer properties which warrant further development towards a cytostatic agent for future chemotherapy. Studies on aluminium compounds for cancer therapy are rare, and our report adds to this important body of knowledge.

SARS-CoV-2 in environmental perspective: Occurrence, persistence, surveillance, inactivation and challenges.

The unprecedented global spread of the severe acute respiratory syndrome (SARS) caused by SARS-CoV-2 is depicting the distressing pandemic consequence on human health, economy as well as ecosystem services. So far novel coronavirus (CoV) outbreaks were associated with SARS-CoV-2 (2019), middle east respiratory syndrome coronavirus (MERS-CoV, 2012), and SARS-CoV-1 (2003) events. CoV relates to the enveloped family of Betacoronavirus (ßCoV) with positive-sense single-stranded RNA (+ssRNA). Knowing well the persistence, transmission, and spread of SARS-CoV-2 through proximity, the faecal-oral route is now emerging as a major environmental concern to community transmission. The replication and persistence of CoV in the gastrointestinal (GI) tract and shedding through stools is indicating a potential transmission route to the environment settings. Despite of the evidence, based on fewer reports on SARS-CoV-2 occurrence and persistence in wastewater/sewage/water, the transmission of the infective virus to the community is yet to be established. In this realm, this communication attempted to review the possible influx route of the enteric enveloped viral transmission in the environmental settings with reference to its occurrence, persistence, detection, and inactivation based on the published literature so far. The possibilities of airborne transmission through enteric virus-laden aerosols, environmental factors that may influence the viral transmission, and disinfection methods (conventional and emerging) as well as the inactivation mechanism with reference to the enveloped virus were reviewed. The need for wastewater epidemiology (WBE) studies for surveillance as well as for early warning signal was elaborated. This communication will provide a basis to understand the SARS-CoV-2 as well as other viruses in the context of the environmental engineering perspective to design effective strategies to counter the enteric virus transmission and also serves as a working paper for researchers, policy makers and regulators.

Initial behaviors and removal of extracellular plasmid gene in membrane bioreactor.

Extracellular antibiotic resistance genes (eARG) are considered to play an important role in spread of antimicrobial resistance (AMR) in wastewater treatment and water environment. Membrane bioreactor (MBR) reportedly has better removal of ARGs in wastewater than conventional activated sludge process. However, removal of eARG is possibly limited because eARG is small to pass through microfiltration (MF) membranes. To evaluate potential removal of eARG in MBR, this study aimed to understand the initial behaviors of eARG received in MBR. The recombinant plasmid with artificial marker gene was spiked in lab-scale MBR to trace fate of eARG in MBR. Among 10 10 copies/L of the spiked gene, 2.6 × 109 copies/L was adsorbed on sludge particles at 6 h after spiking, while only 2.2 × 108-3.6 × 108 copies/L of the spiked gene was remained but constant in sludge liquid phase from 6 until 48 h. This result suggests that adsorption on sludge particles served as the main mechanism to govern the initial fate of eARG in MBR. Meanwhile, the spiked gene concentrations in membrane permeate was lower than sludge liquid phase and decreased overtime, suggesting retention of eARG in membrane filtration. Total LRV of the spiked extracellular gene were 3.4 ± 0.8 log at 48 h after spiking. LRV by adsorption corresponded to 1.7 ± 0.7 log constantly since 3 h after spiking, while LRV by membrane filtration increased from 0 to 1.7 ± 0.6 log. Linear correlation of LRV by membrane filtration with transmembrane pressure (TMP) suggested that foulant deposition on membrane governs removal of eARG by membrane filtration in MBR.

Influence of activated sludge dissolved oxygen concentration on a membrane bioreactor performance with intermittent aeration.

This study measured the effect of low activated sludge dissolved oxygen (DO) concentration on a membrane bioreactor (MBR) treating real urban wastewater with respect to organic matter and nitrogen removal efficiency and transmembrane pressure evolution. For this purpose, a full-scale experimental pre-denitrification MBR system was operated at a constant permeate flow rate of Q = 0.45 m3h-1 with intermittent aeration. The experimental installation worked at high hydraulic retention time, variable sludge retention time and with activated sludge temperatures of between 22.0 to 31.3 °C. Mean DO concentrations in the activated sludge were gradually decreased from 1.25 mgO2L-1 to less than 0.20 mgO2L-1. Variations in DO set points did not affect the main operational parameters of the MBR system and no clear relation was shown between DO concentration decrease and membrane fouling. At DO concentrations lower than 0.2 mgO2L-1, a deterioration in MBR effluent quality was observed, mainly with respect to chemical oxygen demand, biochemical oxygen demand at five days and NH4+, however, the opposite effect was observed for NO3-. These results indicate that employing low DO set points is a promising strategy for application in MBR systems.

Transcriptional response of Pseudomonas chenduensis strain MBR to cadmium toxicity.

Cadmium (Cd) contamination has resulted in serious environmental pollution and threatens human health and ecosystems. Our recent studies have demonstrated that Pseudomonas chenduensis strain MBR can decrease Cd bio-availability and reduce Cd accumulation in rice; however, the transcriptional mechanisms underlying the bacterial response during and particularly after Cd bioremediation are still unclear. In this study, we used RNA-Seq to investigate the transcriptional profiles of strain MBR during and after Cd bioremediation. During Cd bioremediation, MBR removed all Cd2+ ions in solution within 24 h, accompanied by 564 upregulated and 363 downregulated expressed genes compared with that of the control (without Cd supplementation). Specifically, under Cd stress, the upregulation of czc (czcA, czcB, and czcC) and mer (merA, merT, merC, and merP) genes enabled Cd efflux from the cytoplasm and conferred resistance of MBR to Cd toxicity. The upregulation of genes (algK, algX, and alg44) related to biofilm formation enabled Cd absorption and contributed to Cd bioremediation. After Cd bioremediation, MBR was transferred to non-Cd medium, and the genes related to histidine metabolism and flagellar assembly still showed similar expression patterns as those during bioremediation (defined as Cd legacy effects). However, the genes involved in Cd resistance and bioremediation were not influenced by Cd legacy effects. This study provides new and thorough insights into the molecular mechanisms underlying Cd bioremediation by a functional microbe. KEY POINTS: • The upregulation of czc and mer genes is responsible for MBR resistance to Cd. • The upregulation of genes related to biofilm formation contributes to Cd bioremediation. • Cd effects on genes involved in histidine metabolism and flagellar assembly are long-lasting.

Analysis of biosorption and biotransformation mechanism of Pseudomonas chengduensis strain MBR under Cd(II) stress from genomic perspective.

Microbial treatment of heavy metal-polluted sites is considered an environmentally friendly bioremediation technology with high potential. This study shows that Pseudomonas chengduensis strain MBR, a bacterium that can potentially be applied in the treatment of heavy metal pollution, is most affected by Cd(II) stress at the beginning of its growth. Up to 100% of total Cd(II) adsorption occurs in the first 48 h after treatment of stationary phase cells with Cd(II). A biofilm forms on the cell surface, Cd(II) adsorbs, and is reduced to Cd (0) in the form of nanoscale particles. The genome of strain MBR was sequenced, annotated and analyzed. We identified various genes potentially related to cadmium resistance, transport and metabolism. Analysis of the strain MBR genome is helpful to explore the mechanism of Cd(II) resistance, and can provide new ideas for cadmium pollution control.

Removal of sulfadiazine from simulated industrial wastewater by a membrane bioreactor and ozonation.

Ozonation can be used as a polishing treatment for degrading low-concentration pharmaceutical compounds recalcitrant to biological treatment, when large amounts of biodegradable organics have been previously removed by biological processes. Nevertheless, a systematic investigation has not yet been carried out for the coupled MBR + O3 process through an experimental design approach. Thereby, the purpose of this study is to evaluate the performance of different processes (membrane bioreactor-MBR, ozonation; and integrated MBR + O3) for removing the antibiotic sulfadiazine (SDZ) from a synthetic wastewater matrix of industrial interest. The MBR behavior was monitored over seven months for different parameters (pH, temperature, permeate flow, transmembrane pressure, biological oxygen demand-BOD5, chemical oxygen demand-COD, total organic carbon-TOC, solids, and SDZ concentration). Additionally, the amount of SDZ sorbed onto the sludge was characterized, an issue which is scarcely addressed in most research works. Ozonation experiments were conducted in batch mode in a 2-L glass reactor provided with openings for gas flow. For the MBR + O3 process, the effects of gas flow rate (0.1-1.5 L min-1) and inlet ozone concentration (4-12 mg L-1) on SDZ removal from the MBR permeate were systematically assessed using a Doehlert experimental design and response surface methodology. The results indicated that the MBR system showed good performance regarding organic matter removal efficiency, evaluated in terms of BOD5 (91.5%), COD (93.1%) and TOC (96.3%). In contrast, SDZ was partially removed (33%) by the MBR; in that case, the results indicated that the antibiotic was moderately removed with the sludge and partially biodegraded. In turn, the MBR + O3 system showed excellent performance for removing SDZ (100%), TOC (97%), BOD5 (94%) and COD (97%). The statistical analysis confirmed that the influence of ozone gas flow rate upon the SDZ removal rate was more important than that exhibited by inlet ozone concentration. Therefore, coupling MBR and ozone can be considered a promising alternative for point source treatment of antibiotic production wastewater.

Ibuprofen removal by a microfiltration membrane bioreactor during the startup phase.

The behavior of ibuprofen (IBU) during the startup phase of a microfiltration membrane bioreactor (MBR) was determined. A full-scale experimental installation treating real urban wastewater was used for the study. The MBR was composed of an anoxic and an aerobic bioreactors working in pre-denitrification configuration, followed of a membrane reactor. A full mass balance was carried out to estimate the contribution of biotransformation and sorption to biomass to the overall removal of the IBU. During the startup phase of the MBR system there were significant oscillations of the operational variables, mainly of the sludge retention time (SRT); nevertheless, the capacity of the system for IBU removal was very high, with yields of over 94%, despite reaching minimum SRT values of 4.15 d. The main IBU removal occurs in the aerobic reactor, both in the liquid phase and the one associated with the sludge, while in the anoxic bioreactor the removal was scarce, although a certain transfer of IBU from the liquid phase to the sludge took place under anoxic conditions. Despite the high IBU removal yields during startup, the SRT was the most influential variable in IBU removal, an effect observed in all bioreactors, particularly in the anoxic one.

A fully online sensor-equipped, disposable multiphase microbioreactor as a screening platform for biotechnological applications.

A new disposable, multiphase, microbioreactor (MBR; with a working volume of 550 µl) equipped with online sensors is presented for biotechnological screening research purposes owing to its high-throughput potential. Its design and fabrication, online sensor integration, and operation are described. During aerobic cultivation, sufficient oxygen supply is the most important factor that influences growth and product formation. The MBR is a microbubble column bioreactor (µBC), and the oxygen supply was realized by active pneumatic bubble aeration, ensuring sufficient volumetric liquid-phase mass transfer (k L a) and proper homogenization of the cultivation broth. The µBC was equipped with miniaturized sensors for the pH, dissolved oxygen, optical density and glucose concentration that allowed real-time online monitoring of these process variables during cultivation. The challenge addressed here was the integration of sensors in the limited available space. The MBR was shown to be a suitable screening platform for the cultivation of biological systems. Batch cultivations of Saccharomyces cerevisiae were performed to observe the variation in the process variables over time and to show the robustness and operability of all the online sensors in the MBR.

Functional Response of MBR Microbial Consortia to Substrate Stress as Revealed by Metaproteomics.

Bacterial consortia have a primary role in the biological degradations occurring in activated sludge for wastewater treatment, for their capacities to metabolize the polluting matter. Therefore, the knowledge of the main metabolic pathways for the degradation of pollutants becomes critical for a correct design and operation of wastewater treatment plants. The metabolic activity of the different bacterial groups in activated sludge is commonly investigated through respirometry. Furthermore, in the last years, the development of "omic" approaches has offered more opportunities to integrate or substitute the conventional microbiological assays and to deeply understand the taxonomy and dynamics of complex microbial consortia. In the present work, an experimental membrane bioreactor (MBR) was set up and operated for the treatment of municipal wastewater, and the effects of a sudden decrease of the organic supply on the activated sludge were investigated. Both respirometric and metaproteomic approaches revealed a resistance of autotrophic bacteria to the substrate stress, and particularly of nitrifying bacteria. Furthermore, metaproteomics allowed the identification of the taxonomy of the microbial consortium based on its protein expression, unveiling the prevalence of Sorangium and Nitrosomonas genera both before and after the organic load decrease. Moreover, it confirmed the results obtained through respirometry and revealed a general expression of proteins involved in metabolism and transport of nitrogen, or belonging to nitrifying species like Nitrosomonas europeae, Nitrosomonas sp. AL212, or Nitrospira defluvii.

Quorum sensing: an emerging link between temperature and membrane biofouling in membrane bioreactors.

Lab-scale membrane bioreactors (MBRs) were investigated at 12, 18, and 25 °C to identify the correlation between quorum sensing (QS) and biofouling at different temperatures. The lower the reactor temperature, the more severe the membrane biofouling measured in terms of the transmembrane pressure (TMP) during filtration. More extracellular polymeric substances (EPSs) that cause biofouling were produced at 18 °C than at 25 °C, particularly polysaccharides, closely associated with QS via the production of N-acyl homoserine lactone (AHL). However, at 12 °C, AHL production decreased, but the release of EPSs due to deflocculation increased the soluble EPS concentration. To confirm the temperature effect related to QS, bacteria producing AHL were isolated from MBR sludge and identified as Aeromonas sp., Leclercia sp., and Enterobacter sp. through a 16S rDNA sequencing analysis. Batch assays at 18 and 25 °C showed that there was a positive correlation between QS through AHL and biofilm formation in that temperature range.