Accounting for the microbial assembly of each process in wastewater treatment plants (WWTPs): study of four WWTPs receiving similar influent streams.

We evaluated a unique model in which four full-scale wastewater treatment plants (WWTPs) with the same treatment schematic and fed with similar influent wastewater were tracked over an 8-month period to determine whether the community assembly would differ in the activated sludge (AS) and sand filtration (SF) stages. For each WWTP, AS and SF achieved an average of 1-log10 (90%) and <0.02-log10 (5%) reduction of total cells, respectively. Despite the removal of cells, both AS and SF had a higher alpha and beta diversity compared to the influent microbial community. Using the Sloan neutral model, it was observed that AS and SF were individually dominated by different assembly processes. Specifically, microorganisms from influent to AS were predominantly determined by the selective niche process for all WWTPs, while the microbial community in the SF was relatively favored by a stochastic, random migration process, except two WWTPs. AS also contributed more to the final effluent microbial community compared with the SF. Given that each WWTP operates the AS independently and that there is a niche selection process driven mainly by the chemical oxygen demand concentration, operational taxonomic units unique to each of the WWTPs were also identified. The findings from this study indicate that each WWTP has its distinct microbial signature and could be used for source-tracking purposes.IMPORTANCEThis study provided a novel concept that microorganisms follow a niche assembly in the activated sludge (AS) tank and that the AS contributed more than the sand filtration process toward the final microbial signature that is unique to each treatment plant. This observation highlights the importance of understanding the microbial community selected by the AS stage, which could contribute toward source-tracking the effluent from different wastewater treatment plants.

The antibiotic crisis: On the search for novel antibiotics and resistance mechanisms.

In the relentless battle for human health, the proliferation of antibiotic-resistant bacteria has emerged as an impending catastrophe of unprecedented magnitude, potentially driving humanity towards the brink of an unparalleled healthcare crisis. The unyielding advance of antibiotic resistance looms as the foremost threat of the 21st century in clinical, agricultural and environmental arenas. Antibiotic resistance is projected to be the genesis of the next global pandemic, with grim estimations of tens of millions of lives lost annually by 2050. Amidst this impending calamity, our capacity to unearth novel antibiotics has languished, with the past four decades marred by a disheartening 'antibiotic discovery void'. With nearly 80% of our current antibiotics originating from natural or semi-synthetic sources, our responsibility is to cast our investigative nets into uncharted ecological niches teeming with microbial strife, the so-called 'microbial oases of interactions'. Within these oases of interactions, where microorganisms intensively compete for space and nutrients, a dynamic and ever-evolving microbial 'arms race' is constantly in place. Such a continuous cycle of adaptation and counter-adaptation is a fundamental aspect of microbial ecology and evolution, as well as the secrets to unique, undiscovered antibiotics, our last bastion against the relentless tide of resistance. In this context, it is imperative to invest in research to explore the competitive realms, like the plant rhizosphere, biological soil crusts, deep sea hydrothermal vents, marine snow and the most modern plastisphere, in which competitive interactions are at the base of the microorganisms' struggle for survival and dominance in their ecosystems: identify novel antibiotic by targeting microbial oases of interactions could represent a 'missing piece of the puzzle' in our fight against antibiotic resistance.

Nitrogen removal intensification of biofilm through bioaugmentation with Methylobacterium gregans DC-1 during wastewater treatment.

The increasing concern for environmental remediation has led to a search for effective methods to remove eutrophic nutrients. In this study, Methylobacterium gregans DC-1 was utilized to improve nitrogen removal in a sequencing batch biofilm reactor (SBBR) via aerobic denitrification. This bacterium has the extraordinary characteristics of strong auto-aggregation and a high ability to remove nitrogen efficiently, making it an ideal candidate for enhanced treatment of nitrogen-rich wastewater. This strain was used for the bioassessment of a test reactor (SBBRbio), which showed a shorter biofilm formation time compared to a control reactor (SBBRcon) without this strain inoculation. Moreover, the enhanced biofilm was enriched in TB-EPS and had a wider variety of protein secondary structures than SBBRcon. During the stabilization phase of SBBRbio, the EPS molecules showed the highest proportion of intermolecular hydrogen bonding. It is possible that bioaugmentation with this strain positively affects the structural stability of biofilm. At influent ammonia loadings of 100 and 150 mg. L-1, the average reduction of ammonia and nitrate-nitrogen was higher in the experimental system compared to the control system. Additionally, nitrite-N accumulation was lower and N2O production decreased compared to the control. Analysis of the microbial community structure demonstrated successful colonization in the bioreactor by a highly nitrogen-tolerant strain that efficiently removed inorganic nitrogen. These results illustrate the great potential of this type of denitrifying bacteria in the application of bioaugmentation systems.

Enhanced chromium and nitrogen removal by constructing a biofilm reaction system based on denitrifying bacteria preferential colonization theory.

Understanding the developmental characteristics of microbial communities in biofilms is crucial for designing targeted functional microbial enhancements for the remediation of complex contamination scenarios. The strong prioritization effect of microorganisms confers the ability to colonize strains that arrive first dominantly. In this study, the auto-aggregating denitrifying bacterial Pseudomonas stutzeri strain YC-34, which has both nitrogen and chromium removal characteristics, was used as a biological material to form a stable biofilm system based on the principle of dominant colonization and biofortification. The effect of the biofilm system on nitrogen and chromium removal was characterized by measuring the changes in the quality of influent and effluent water. The pattern of biofilm changes was analyzed by measuring biofilm content and thickness and characterizing extracellular polymer substances (EPS). Further analysis of the biofilm microbiota characteristics and potential functions revealed the mechanism of strain YC-34 biofortified biofilm. The results revealed that the biofilm system formed could achieve 90.56% nitrate-nitrogen removal with an average initial nitrate-nitrogen concentration of 51.9 mg/L and 40% chromium removal with an average initial hexavalent chromium Cr(VI) concentration of 7.12 mg/L. The biofilm properties of the system were comparatively analyzed during the biofilm formation period, the fluctuation period of Cr(VI)-stressed water quality, and the stabilization period of Cr(VI)-stressed water quality. The biofilm system may be able to increase the structure of hydrogen bonds, the type of protein secondary structure, and the abundance of amino acid-like components in the EPS, which may confer biofilm tolerance to Cr(VI) stress and allow the system to maintain a stable biofilm structure. Furthermore, microbial characterization indicated an increase in microbial diversity in the face of chromium stress, with an increase in the abundance of nitrogen removal-associated functional microbiota and an increasing trend in the abundance of nitrogen transfer pathways. These results demonstrate that the biofilm system is stable in nitrogen and chromium removal. This bioaugmentation method may provide a new way for the remediation of heavy metal-polluted water bodies and also provides theoretical and application parameters for the popularization and application of biofilm systems.

Pleurotus ostreatus polysaccharide-mediated modulation of skin damage caused by microcystin-LR in tadpoles.

In this study, we aimed to evaluate the effect of dietary supplementation with edible mushroom (Pleurotus ostreatus)-derived polysaccharides on microcystin leucine-arginine (MC-LR)-induced skin damage in Pelophylax nigromaculatus tadpoles. Tadpoles were exposed to 1 µg/L daily MC-LR, with or without 5.0 g/kg of dietary P. ostreatus polysaccharides, for 30 days. P. ostreatus polysaccharide supplementation significantly increased the dermal collagen fibrils, increased tight junction protein gene expression, decreased the amount of MC-LR accumulation in skin tissues, attenuated oxidative stress, downregulated apoptosis-associated gene transcription, decreased eosinophil numbers, and downregulated transcription of inflammation-related genes (e.g. TLR4, NF-κB, and TNF-α). The composition of the skin commensal microbiota of MC-LR-exposed tadpoles supplemented with P. ostreatus polysaccharides was similar to that of the no-treatment control group. Lipopolysaccharide (LPS) content was positively correlated with the abundance of Gram-negative bacteria, including Chryseobacterium and Thauera. Therefore, P. ostreatus polysaccharides may alleviate MC-LR-induced skin barrier damage in tadpoles in two ways: 1) attenuation of oxidative stress-mediated apoptosis mediated by increased glutathione (GSH) content and total superoxide dismutase activity; and 2) alteration of the skin commensal microbiota composition to attenuate the LPS/Toll-like receptor 4 inflammatory pathway response. Furthermore, P. ostreatus polysaccharides may increase skin GSH synthesis by promoting glycine production via the gut microbiota and may restore the MC-LR-damaged skin resistance to pathogenic bacteria by increasing antimicrobial peptide transcripts and lysozyme activity. This study highlights for the first time the potential application of P. ostreatus polysaccharides, an ecologically active substance, in mitigating the skin damage induced by MC-LR exposure, and may provide new insights for its further development in aquaculture.

Predicting Anaerobic Membrane Bioreactor Performance Using Flow-Cytometry-Derived High and Low Nucleic Acid Content Cells.

Having a tool to monitor the microbial abundances rapidly and to utilize the data to predict the reactor performance would facilitate the operation of an anaerobic membrane bioreactor (AnMBR). This study aims to achieve the aforementioned scenario by developing a linear regression model that incorporates a time-lagging mode. The model uses low nucleic acid (LNA) cell numbers and the ratio of high nucleic acid (HNA) to LNA cells as an input data set. First, the model was trained using data sets obtained from a 35 L pilot-scale AnMBR. The model was able to predict the chemical oxygen demand (COD) removal efficiency and methane production 3.5 days in advance. Subsequent validation of the model using flow cytometry (FCM)-derived data (at time t - 3.5 days) obtained from another biologically independent reactor did not exhibit any substantial difference between predicted and actual measurements of reactor performance at time t. Further cell sorting, 16S rRNA gene sequencing, and correlation analysis partly attributed this accurate prediction to HNA genera (e.g., Anaerovibrio and unclassified Bacteroidales) and LNA genera (e.g., Achromobacter, Ochrobactrum, and unclassified Anaerolineae). In summary, our findings suggest that HNA and LNA cell routine enumeration, along with the trained model, can derive a fast approach to predict the AnMBR performance.

Targeting cytohesin-1 suppresses acute myeloid leukemia progression and overcomes resistance to ABT-199.

Adhesion molecules play essential roles in the homeostatic regulation and malignant transformation of hematopoietic cells. The dysregulated expression of adhesion molecules in leukemic cells accelerates disease progression and the development of drug resistance. Thus, targeting adhesion molecules represents an attractive anti-leukemic therapeutic strategy. In this study, we investigated the prognostic role and functional significance of cytohesin-1 (CYTH1) in acute myeloid leukemia (AML). Analysis of AML patient data from the GEPIA and BloodSpot databases revealed that CYTH1 was significantly overexpressed in AML and independently correlated with prognosis. Functional assays using AML cell lines and an AML xenograft mouse model confirmed that CYTH1 depletion significantly inhibited the adhesion, migration, homing, and engraftment of leukemic cells, delaying disease progression and prolonging animal survival. The CYTH1 inhibitor SecinH3 exerted in vitro and in vivo anti-leukemic effects by disrupting leukemic adhesion and survival programs. In line with the CYTH1 knockdown results, targeting CYTH1 by SecinH3 suppressed integrin-associated adhesion signaling by reducing ITGB2 expression. SecinH3 treatment efficiently induced the apoptosis and inhibited the growth of a panel of AML cell lines (MOLM-13, MV4-11 and THP-1) with mixed-lineage leukemia gene rearrangement, partly by reducing the expression of the anti-apoptotic protein MCL1. Moreover, we showed that SecinH3 synergized with the BCL2-selective inhibitor ABT-199 (venetoclax) to inhibit the proliferation and promote the apoptosis of ABT-199-resistant leukemic cells. Taken together, our results not only shed light on the role of CYTH1 in cell-adhesion-mediated leukemogenesis but also propose a novel combination treatment strategy for AML.

Immobilization of BoPAL3 Phenylalanine Ammonia-Lyase on Electrospun Nanofibrous Membranes of Polyvinyl Alcohol/Nylon 6/Chitosan Crosslinked with Dextran Polyaldehyde.

Phenylalanine ammonia-lyase (PAL, EC 4.3.1.24) is common in plants and catalyzes the formation of trans-cinnamic acid and ammonia via phenylalanine deamination. Recombinant Bambusa oldhamii BoPAL3 protein expressed in Escherichia coli was immobilized on an electrospun nanofibrous membrane using dextran polyaldehyde as a crosslinker. The immobilized BoPAL3 protein exhibited comparable kinetic properties with the free BoPAL3 protein and could be recycled for six consecutive cycles compared with the free BoPAL3 protein. The residual activity of the immobilized BoPAL3 protein was 84% after 30 days of storage at 4 °C, whereas the free BoPAL3 protein retained 56% residual activity in the same storage conditions. Furthermore, the resistance of the immobilized BoPAL3 protein to chemical denaturants was greatly increased. Therefore, the BoPAL3 protein can be immobilized using the natural dextran polyaldehyde crosslinker in place of the conventional chemical crosslinker. Nanofibrous membranes made from polyvinyl alcohol (PVA), nylon 6, and chitosan (CS) are incredibly stable and useful for future industrial applications.

Legacy and Emerging Per- and Polyfluoroalkyl Substances Surveillance in Bufo gargarizans from Inlet Watersheds of Chaohu Lake, China: Tissue Distribution and Bioaccumulation Potential.

Amphibians are sensitive biomonitors of environmental pollutants but reports regarding per- and polyfluoroalkyl substances (PFAS), a class of synthetic organofluorine substances, are limited. In this study, samples of water and Chinese toads (Bufo gargarizans) were collected in Chaohu Lake, China. Tissue-specific bioaccumulation characteristics of 39 PFAS, including 19 perfluoroalkyl acids (PFAAs), 8 emerging PFAS, and 12 PFAA precursors, were investigated, and the levels of some biochemical indicators were determined. The highest PFAS concentrations were found in the liver [215.97 ng/g dry weight (dw)] of Chinese toads, followed by gonads (135.42 ng/g dw) and intestine (114.08 ng/g dw). A similar tissue distribution profile was found between legacy and emerging PFAS in the toads, and the occurrence of two emerging PFAS, 2,3,3,3-tetrafluoro-2-propanoate (HFPO-DA) and 6:2 hydrogen-substituted polyfluorooctane ether sulfonate (6:2 H-PFESA) in the amphibians were for the first time reported. Field-based bioaccumulation factors of HFPO-DA were higher than perfluorooctanoic acid, indicating the higher bioaccumulation potential of this emerging PFAS than the legacy C8 compound. Males had significantly higher gonad PFAS levels than females while estradiol levels in gonads increased with increasing concentrations of certain PFAS (e.g., 6:2 H-PFESA), implying that PFAS may trigger estrogenic effects in the toads, especially for male toads.

Evaluation of protein extraction methods to improve meta-proteomics analysis of treated wastewater biofilms.

Metaproteomics can be used to study functionally active biofilm-based bacterial populations in reclaimed water distribution systems, which in turn result in bacterial regrowth that impacts the water quality. However, existing protein extraction methods have differences in their protein recovery and have not been evaluated for their efficacies in reclaimed water biofilm samples. In this study, we first evaluated six different protein extraction methods with diverse chemical and physical properties on a mixture of bacterial cell culture. Based on a weighting scores-based evaluation, the extraction protocols in order of decreasing performance are listed as B-PER > RIPA > PreOmics > SDS > AllPrep > Urea. The highest four optimal methods on cell culture were further tested against treated wastewater non-chlorinated and chlorinated effluent biofilms. In terms of protein yield, our findings showed that RIPA performed the best; however, the highest number of proteins were extracted from SDS and PreOmics. Furthermore, SDS and PreOmics worked best to rupture gram-positive and gram-negative bacterial cell walls. Considering the five evaluation factors, PreOmics obtained highest weighted score, indicating its potential effectiveness in extracting proteins from biofilms. This study provides the first insight into evaluating protein extraction methods to facilitate metaproteomics for complex reclaimed water matrices.

Systematic investigation of simultaneous copper biosorption and nitrogen removal from wastewater by an aerobic denitrifying bacterium of auto-aggregation.

Finding effective methods for simultaneous removal of eutrophic nutrients and heavy metals has attracted increasing concerns for the environmental remediation. Herein, a novel auto-aggregating aerobic denitrifying strain (Aeromonas veronii YL-41) was isolated with capacities for copper tolerance and biosorption. The denitrification efficiency and nitrogen removal pathway of the strain were investigated by nitrogen balance analysis and amplification of key denitrification functional genes. Moreover, the changes in the auto-aggregation properties of the strain caused by extracellular polymeric substances (EPS) production were focused on. The biosorption capacity and mechanisms of copper tolerance during denitrification were further explored by measuring changes in copper tolerance and adsorption indices, as well as by variations in extracellular functional groups. The strain showed extremely strong total nitrogen removal ability, with 67.5%, 82.08% and 78.48% of total nitrogen removal when NH4+-N, NO2--N, and NO3--N were used as the only initial nitrogen source, respectively. The successful amplification of napA, nirK, norR, and nosZ genes further demonstrated that the strain accomplished nitrate removal through a complete aerobic denitrification pathway. The production of protein-rich EPS of up to 23.31 mg/g and an auto-aggregation index of up to 76.42% may confer a strong biofilm-forming potential to the strain. Under the stress of 20 mg/L copper ions, the removal of nitrate-nitrogen was still as high as 71.4%. In addition, the strain could achieve an efficient removal of 96.9% of copper ions at an initial concentration of 80 mg/L. Scanning electron microscopy and deconvolution analysis of characteristic peaks confirmed that the strains encapsulate heavy metals by secreting EPS and, meanwhile, form strong hydrogen bonding structures to enhance intermolecular forces to resist copper ion stress. This study provides an innovative and effective biological approach for the synergistic bioaugmentation removal of eutrophic substances and heavy metals from aquatic environments.

Promoting heterotrophic denitrification of Pseudomonas hunanensis strain PAD-1 using pyrite: A mechanistic study.

Denitrification is critical for removing nitrate from wastewater, but it typically requires large amounts of organic carbon, which can lead to high operating costs and secondary environmental pollution. To address this issue, this study proposes a novel method to reduce the demand for organic carbon in denitrification. In this study, a new denitrifier, Pseudomonas hunanensis strain PAD-1, was obtained with properties for high efficiency nitrogen removal and trace N2O emission. It was also used to explore the feasibility of pyrite-enhanced denitrification to reduce organic carbon demand. The results showed that pyrite significantly improved the heterotrophic denitrification of strain PAD-1, and optimal addition amount was 0.8-1.6 g/L. The strengthening effect of pyrite was positively correlated with carbon to nitrogen ratio, and it could effectively reduce demand for organic carbon sources and enhance carbon metabolism of strain PAD-1. Meanwhile, the pyrite significantly up-regulated electron transport system activity (ETSA) of strain PAD-1 by 80%, nitrate reductase activity by 16%, Complex III activity by 28%, and napA expression by 5.21 times. Overall, the addition of pyrite presents a new avenue for reducing carbon source demand and improving the nitrate harmless rate in the nitrogen removal process.

The succession of gut microbiota in the concave-eared torrent frog (Odorrana tormota) throughout developmental history.

The gut microbiota of amphibians plays a crucial role in maintaining health and adapting to various developmental stages. The composition of gut microbial community is influenced by the phylogeny, habitat, diet, and developmental stage of the host. The present study analyzed the microbiota in the intestine of O. tormota at 11 developmental stages (from the tadpole at Gosner stage 24 to the 3-year-old adult) using high-throughput 16S rRNA sequencing. Alpha diversity index analysis of the microbiota revealed that the index decreased from tadpole at Gosner stage 24 to adult frog stage, remained stable during the adult frog stages, but increased significantly at the early metamorphosis and hibernation preparation stages. The gut microbiota structure is similar in adult frogs but differs significantly in other developmental stages. Furthermore, the dominant phyla of gut microbiota in tadpoles were Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes, whereas those in adult frogs were Proteobacteria, Firmicutes, Bacteroidetes, and Verrucomicrobia. Host and environmental factors jointly affected the gut microbial diversity and community composition of O. tormota, but developmental stage, feeding habit, and habitat type had a more significant influence. The microbial community in the gut varies with the developmental stage of the host and constantly adapts to the survival requirements of the host. These findings advance our understanding of the evolutionary mechanism of amphibian gut microbiota in maintaining health homeostasis and adaptation.

Novel Use of a Ferric Salt to Enhance Mainstream Nitrogen Removal from Anaerobically Pretreated Wastewater.

This study aims to demonstrate a new technology roadmap to support the ongoing paradigm shift in wastewater management from pollutant removal to resource recovery. This is achieved by developing a novel use of an iron salt (i.e., FeCl3) in an integrated anaerobic wastewater treatment and mainstream anammox process. FeCl3 was chosen to be dosed in a proposed sidestream unit rather than in a primary settler or a mainstream reactor. This causes acidification of returned activated sludge and enables stable suppression of nitrite-oxidizing bacterial activity and excess sludge reduction. A laboratory-scale system, which comprised an anaerobic baffled reactor, a continuous-flow anoxic-aerobic (A/O) reactor, and a secondary settler, was designed to treat real domestic wastewater, with the performance of the system comprehensively monitored under a steady-state condition. The experimental assessments showed that the system had good effluent quality, with total nitrogen and phosphorus concentrations of 12.6 ± 1.3 mg N/L and 0.34 ± 0.05 mg P/L, respectively. It efficiently retained phosphorus in excess sludge (0.18 ± 0.03 g P/g dry sludge), suggesting its potential for further recovery. About half of influent organic carbon was recovered in the form of bioenergy (i.e., methane). This together with low energy consumption revealed that the system could produce a net energy of about 0.11 kWh/m3-wastewater, assessed by an energy balance analysis.

Understanding the role of sorption and biodegradation in the removal of organic micropollutants by membrane aerated biofilm reactor (MABR) with different biofilm thickness.

The role of sorption and biodegradation in a membrane aerated biofilm reactor (MABR) were investigated for the removal of 10 organic micropollutants (OMPs) including endocrine disruptors and pharmaceutical active compounds. The influence of the biofilm thickness on the mechanisms of removal was analyzed via kinetic test at three different stages. At all biofilm stages, biodegradation was demonstrated to dominate the removal of selected OMPs. Higher OMPs rates of removal via biodegradation (Kbiol) were achieved when biofilm increased its thickness from (stage T1) 0.26 mm, to (stage T2) 0.58 mm and (stage T3) 1.03 mm. At stage T1 of biofilm, heterotrophs contribute predominantly to OMPs degradation. Hydrophilic compounds removal (i.e., acetaminophen) continue to be driven by heterotrophic bacteria at the next stages of biofilm thickness. However, for medium hydrophobic neutral and charged OMPs, the combined action of heterotrophic and enriched nitrifying activity at stages T2 and T3 enhanced the overall removal. A degradation pathway based on heterotrophic activity for acetaminophen and combined action of nitrifiers-heterotrophs for estrone was proposed based on identified metabolites. Although biodegradation dominated the removal of most OMPs, sorption was also observed to be essential in the removal of biologically recalcitrant and lipophilic compounds like triclosan. Furthermore, sorption capacity of apolar compound was enhanced as the biofilm thickness grew and increased in EPS protein fraction. Microbial analysis confirmed the higher abundance of nitrifying and denitrifying activity at stage T3 of biofilm, which not only facilitated near complete ammonium removal but also enhanced degradation of OMPs.

SARS-CoV-2 wastewater-based epidemiology in an enclosed compound: A 2.5-year survey to identify factors contributing to local community dissemination.

Long-term (>2.5 years) surveillance of SARS-CoV-2 RNA concentrations in wastewater was conducted within an enclosed university compound. This study aims to demonstrate how coupling wastewater-based epidemiology (WBE) with meta-data can identify which factors contribute toward the dissemination of SARS-CoV-2 within a local community. Throughout the pandemic, the temporal dynamics of SARS-CoV-2 RNA concentrations were tracked by quantitative polymerase chain reaction and analyzed in the context of the number of positive swab cases, the extent of human movement, and intervention measures. Our findings suggest that during the early phase of the pandemic, when strict lockdown was imposed, the viral titer load in the wastewater remained below detection limits, with <4 positive swab cases reported over a 14-day period in the compound. After the lockdown was lifted and global travel gradually resumed, SARS-CoV-2 RNA was first detected in the wastewater on 12 August 2020 and increased in frequency thereafter, despite high vaccination rates and mandatory face-covering requirements in the community. Accompanied by a combination of the Omicron surge and significant global travel by community members, SARS-CoV-2 RNA was detected in most of the weekly wastewater samples collected in late December 2021 and January 2022. With the cease of mandatory face covering, SARS-CoV-2 was detected in at least two of the four weekly wastewater samples collected from May through August 2022. Retrospective Nanopore sequencing revealed the presence of the Omicron variant in the wastewater with a multitude of amino acid mutations, from which we could infer the likely geographical origins through bioinformatic analysis. This study demonstrated that long-term tracking of the temporal dynamics and sequencing of variants in wastewater would aid in identifying which factors contribute the most to SARS-CoV-2 dissemination within the local community, facilitating an appropriate public health response to control future outbreaks as we now live with endemic SARS-CoV-2.

Impact of electrochemically generated iron on the performance of an anaerobic wastewater treatment process.

Anaerobic treatment of domestic wastewater has the advantages of lower biomass yield, lower energy demand and higher energy recover over the conventional aerobic treatment process. However, the anaerobic process has the inherent issues of excessive phosphate and sulfide in effluent and superfluous H2S and CO2 in biogas. An electrochemical method allowing for in-situ generation of Fe2+ in the anode and hydroxide ion (OH-) and H2 in the cathode was proposed to overcome the challenges simultaneously. The effect of electrochemically generated iron (e­iron) on the performance of anaerobic wastewater treatment process was explored with four different dosages in this work. The results showed that compared to control, the experimental system displayed an increase of 13.4-28.4 % in COD removal efficiency, 12.0-21.3 % in CH4 production rate, 79.8-98.5 % in dissolved sulfide reduction, 26.0-96.0 % in phosphate removal efficiency, depending on the e­iron dosage between 40 and 200 mg Fe/L. Dosing of the e­iron significantly upgraded the quality of produced biogas, showing a much lower CO2 and H2S contents in biogas in experimental reactor than that in control reactor. The results thus demonstrated that e­iron can significantly improve the performance of anaerobic wastewater treatment process, bringing multiple benefits with the increase of its dosage regarding effluent and biogas quality.

Efficient in planta production of amidated antimicrobial peptides that are active against drug-resistant ESKAPE pathogens.

Antimicrobial peptides (AMPs) are promising next-generation antibiotics that can be used to combat drug-resistant pathogens. However, the high cost involved in AMP synthesis and their short plasma half-life render their clinical translation a challenge. To address these shortcomings, we report efficient production of bioactive amidated AMPs by transient expression of glycine-extended AMPs in Nicotiana benthamiana line expressing the mammalian enzyme peptidylglycine α-amidating mono-oxygenase (PAM). Cationic AMPs accumulate to substantial levels in PAM transgenic plants compare to nontransgenic N. benthamiana. Moreover, AMPs purified from plants exhibit robust killing activity against six highly virulent and antibiotic resistant ESKAPE pathogens, prevent their biofilm formation, analogous to their synthetic counterparts and synergize with antibiotics. We also perform a base case techno-economic analysis of our platform, demonstrating the potential economic advantages and scalability for industrial use. Taken together, our experimental data and techno-economic analysis demonstrate the potential use of plant chassis for large-scale production of clinical-grade AMPs.

Biomass generation and heterologous isoprenoid milking from engineered microalgae grown in anaerobic membrane bioreactor effluent.

Wastewater (WW) treatment in anaerobic membrane bioreactors (AnMBR) is considered more sustainable than in aerobic reactors. However, outputs from AnMBR are a mixed methane and carbon dioxide gas stream as well as ammonium- (N) and phosphate- (P) containing waters. Using AnMBR outputs as inputs for photoautotrophic algal cultivation can strip the CO2 while removing N and P from effluent which feed algal biomass generation. Recent advances in algal engineering have generated strains that produce high-value side products concomitant with biomass, although only shown in heavily domesticated, lab-adapted strains. Here, it was investigated whether engineered Chlamydomonas reinhardtii could be grown directly in AnMBR effluent with CO2 concentrations found in AnMBR off-gas. The strain was found to proliferate over bacteria in the non-sterile effluent, consume N and P to levels that meet general discharge or reuse limits, and tolerate cultivation in modelled (extreme) outdoor environmental conditions prevalent along the central Red Sea coast. In addition to ∼2.4 g CDW L-1 biomass production in 96 h, a high-value heterologous sesquiterpene co-product could be obtained from 'milking' up to 837 µg L-1 culture in 96 h. This is the first demonstration of a combined bio-process that employs a heavily engineered algal strain to enhance the product generation potentials from AnMBR effluent treatment. This study shows it is possible to convert waste into value through use of engineered algae while also improving wastewater treatment economics through co-product generation.