Accumulation of nitrite after reclaimed water recharge due to the disinfection byproduct chlorite.

Due to its toxicity, the disinfection byproduct chlorite in drinking water is strictly regulated to be ≤ 1.0 mg/L, but in reclaimed, non-drinking water chlorite is unregulated and rarely considered. However, chlorite is cytotoxic and has a high oxidation potential. Therefore, as reclaimed water infiltrates soil and groundwater, it may alter the soil environment and microbial community, which may affect the degradation of organic matter and the transformation of the N element. In this study, the effects of reclaimed water containing chlorite on soil microorganisms were investigated by simulating subsurface infiltration. It was found that chlorite improved the conversion of nitrate nitrogen to nitrite nitrogen, but inhibited further conversion of nitrite nitrogen. The nitrite nitrogen in the effluent reached 4.61 mg/L when chlorite was present, while only 0.16 mg/L was found in the control system. The chlorite produced obvious oxidative stress reactions in cells, inhibited the EPSs production, in which the contents of polysaccharides and proteins reduced by nearly 41% and 62%, respectively. Besides, chlorite resulted in the enrichment of efflux resistance genes in the microbial community, mainly adeF and cmlB1. Self-protection against chlorite is achieved mainly using efflux pump related genes. Metagenomics data analysis showed that Delftia became the dominant genus when exposed to chlorite, with the greatest abundance at 17.9%. Chlorite also resulted in the upregulated expression of nar genes (by more than 149%) and downregulation of nir gene expression (by more than 62%). This study reveals the effects of the disinfection byproduct chlorite on a soil microecosystem, providing important information for the management and reuse of reclaimed water.

Attenuated Porcine Reproductive and Respiratory Syndrome Virus Regains Its Fatal Virulence by Serial Passaging in Pigs or Porcine Alveolar Macrophages To Increase Its Adaptation to Target Cells.

Porcine reproductive and respiratory syndrome (PRRS) is a globally important disease threatening the pork industry, and modified live-virus (MLV) vaccines are widely used for its prevention. However, PRRS MLV shows high potential for reversion to virulence, leading to a major concern about its safety. Yet the revertant mechanism is still poorly understood. Here, attenuated virus JXwn06-P80, derived from the highly pathogenic PRRS virus (PRRSV) strain JXwn06 by serial passaging in MARC-145 cells, was reversely passaged in pigs through intranasal inoculation to mimic natural infection for 13 rounds, and the pathogenicity of viruses at the 3rd, 5th, 9th, 10th, and 11th passages was evaluated in pigs. From the 9th passage, the viruses caused mortality, which was related to their increased adaptability and replication efficiency (100 times higher than those of JXwn06-P80) in porcine alveolar macrophage (PAM) target cells. Similarly, JXwn06-P80 could also regain fatal virulence through reverse passage in PAMs for 25 or more passages, indicating that the increased adaptability in PAMs directly contributes to its regained fatal virulence. Next, the full-genome sequences were analyzed to explore the genetic evolutionary processes during adaptation both in vivo and in vitro. Finally, by a reverse genetic operation, four reverse mutation sites, NSP12-W121R, ORF2b (open reading frame 2b)-H9D, ORF5-H15L, and ORF5-V189L, were finally identified to partially contribute to the ability of the virus to adapt to PAMs, which may be related to virulence reversion during reverse passage. These findings provided direct scientific evidence for the virulence reversion of PRRS MLV and provided valuable clues for exploring its molecular mechanism. IMPORTANCE Reversion to virulence of a live attenuated vaccine is a public concern; however, direct scientific evidence is limited, and the mechanism is still poorly understood. Here, we present direct evidence for the reversion to virulence of PRRS MLV after serial passaging in pigs or target cells and found a correlation between virulence reversion and increased replication fitness in primary PAMs. The genetic evolutionary process during adaptation will provide valuable clues for exploring the molecular mechanism of PRRS MLV virulence reversion and offer important implications for understanding the reversion mechanisms of other vaccines.

Effects of trace PFOA on microbial community and metabolisms: Microbial selectivity, regulations and risks.

Perfluorooctanoic acid (PFOA), a "forever chemical", is continuously discharged and mitigated in the environment despite its production and use being severely restricted globally. Due to the transformation, attachment, and adsorption of PFOA in aquatic environments, PFOA accumulates in the porous media of sediments, soils, and vadose regions. However, the impact of trace PFOA in the porous media on interstitial water and water safety is not clear. In this work, we simulated a porous media layer using a sand column and explored the effects of µg-level PFOA migration on microbial community alternation, microbial function regulation, and the generation and spread of microbial risks. After 60 days of PFOA stimulation, Proteobacteria became the dominant phylum with an abundance of 91.8%, since it carried 71% of the antibiotic resistance genes (ARGs). Meanwhile, the halogen-related Dechloromonas abundance increased from 0.4% to 10.6%. In addition, PFOA significantly stimulated protein (more than 1288%) and polysaccharides (more than 4417%) production by up-regulating amino acid metabolism (p< 0.001) and membrane transport (p < 0.001) to accelerate the microbial aggregation. More importantly, the rapidly forming biofilm immobilized and blocked PFOA. The more active antioxidant system repaired the damaged cell membrane by significantly up-regulating glycerophospholipid metabolism and peptidoglycan biosynthesis. It is worth noting that PFOA increased the abundance of antibiotic resistance genes (ARGs) and human bacterial pathogens (HBPs) in porous media by 30% and 106%. PFOA increased the proportion of vertical transmission ARGs (vARGs), and co-occurrence network analysis (r ≥ 0.8, p ≤ 0.01) verified that vARGs were mainly mediated by HBPs. A comprehensive understanding of PFOA interactions with its microecological environment is provided.

Therapeutic effects of myocardin-related transcription factor A (MRTF-A) knockout on experimental mice with nonalcoholic steatohepatitis induced by high-fat diet.

OBJECTIVE: To explore the effects of myocardin-related transcription factor A (MRTF-A) knockout on mice with nonalcoholic steatohepatitis (NASH) induced by high-fat diet (HFD). METHODS: Normal-fat diet (NFD) or HFD was fed to MRTF-A-knockout (MRTF-A-/-) and wild-type (WT) mice for 16 weeks. Liver histopathological status was observed using Hematoxylin and Eosin (HE) staining, Oil Red O staining, Sirius Red staining, and Immunohistochemical staining. The mRNA and protein levels in liver tissues were measured through quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blot. RESULTS: Compared with WT + HFD group, mice in MRTF-A-/- + HFD group were decreased in body weight, blood glucose, plasma insulin, liver TG and NAFLD activity score (NAS), with liver function recovery. Besides, compared with HFD-fed WT mice, HFD-fed MRTF-A-/- mice were improved in hepatic fibrosis, accompanied by decreased collagen content (%) and down-regulated expressions of α-SMA, COL1A2, TGFß1, and SMAD3. In mice fed with HFD, the expression of MCP-1, CCR2, F4/80 and CD68 declined in liver tissues of MRTF-A-/- mice as compared with WT mice. Besides, in hepatic macrophages isolated from HFD-fed mice, the observed increased expression of TNF-α, IL-1ß, MCP-1, as well as decreased expression of CCR2. Compared with WT + HFD group, MRTF-A-/- + HFD group mice were decreased regarding NF-κB p65 in liver tissues. CONCLUSION: MRTF-A knockout reduced macrophage infiltration, down-regulated NF-κB p65 expression, and ameliorated inflammation and fibrosis of liver tissues in mice, thereby becoming a potential therapeutic target for NASH treatment.

High-Performance, Free-Standing Symmetric Hybrid Membranes for Osmotic Separation.

The internal concentration polarization (ICP) of asymmetric osmotic membranes with support layers greatly reduced membrane water permeability, therefore compromising membrane performance. In this study, a series of free-standing symmetric hybrid forward osmosis (FO) membranes without experiencing ICP were fabricated by covalently linking metal-organic framework (MOF) nanofillers with a polymer matrix. Owing to the introduction of MOFs, which allow only water permeation but reject salts by steric hindrance, the prepared hybrid membranes could approach the empirical permeability-selectivity trade-off. The optimized hybrid membrane displayed an outstanding water/Na2SO4 selectivity of ∼1208.4 L mol-1, compared with that of conventional membranes of ∼375.6 L mol-1. Additionally, the fabricated hybrid membranes showed excellent mechanical robustness, maintaining structural integrity during the long-term FO separation of high-salinity solution. This work provides an effective methodology to fabricate high-performance, symmetric MOF-based membranes for osmotic separation processes such as seawater desalination and water purification.

A Decoupled Unified Observation Method of Stochastic Multidimensional Vibration for Wind Tunnel Models.

Active vibration control is the most effective method for stochastic multidimensional vibration in wind tunnel tests, in which vibration monitoring is the core foundation. Vibrations are induced by the disturbances of several complex air flow instabilities under extreme test conditions with high attack angles. Here, a decoupled unified observation method is proposed in order to fully monitor stochastic multidimensional vibration. First, stochastic multidimensional vibration is explained using the Cartesian coordinate system. Then, the multidimensional vibration decoupling of the pitch plane and the yaw plane is realized according to the proposed decoupling design principle of the long cantilever sting. A unified observation method is presented, based on inertial force theory, to observe multidimensional vibration due to acceleration in each decoupling plane. Verification experiments were conducted in lab and a transonic wind tunnel, using an established real-time monitoring system. The results of lab experiments indicate that, in the frequency region of 0-120 Hz, three vibration modes of a selected stochastic vibration can be decoupled and observed through the vibration components in pitch plane and yaw plane. In addition, wind tunnel tests were carried out according to the working conditions (α = -4~10° with γ = 45°) at Ma = 0.6 and Ma = 0.7, respectively. The results show that six vibration modes of two selected stochastic vibrations can be decoupled and observed through the vibration components in pitch plane and yaw plane. The experimental results prove that stochastic vibration can be fully monitored in multiple dimensions through the vibration components in pitch plane and yaw plane using the proposed decoupled unified observation method. Therefore, these results lay the foundation for active vibration control.

Exogenous refractory protein enhances biofilm formation by altering the quorum sensing system: A potential hazard of soluble microbial proteins from WWTP effluent.

Soluble microbial refractory proteins are major components of effluent from wastewater treatment plants that utilize a biological wastewater treatment process. The remaining proteins could negatively affect downstream treatment processes by altering the bacterial quorum sensing system. In this work, we elaborated the effects of exogenous refractory protein on biofilm formation. The results showed a linear relationship between biofilm formation and experimental protein concentrations at the range typically found in effluent, 0-8.0 mg/L. Micro-observation revealed that the exogenous refractory protein stimulated extracellular polysaccharide secretion to promote biofilm maturation. Extracellular polysaccharides increased by ~200% with the addition of only 2.0 mg/L protein. In addition, exogenous refractory proteins altered the quorum sensing system gene expression and polysaccharide gene expression. This work found that exogenous protein accelerated biofilm formation by influencing the quorum sensing system, thus providing new insight into the potential harm of soluble microbial refractory products.

Acceleration of saturated porous media clogging and silicon dissolution due to low concentrations of Al(III) in the recharge of reclaimed water.

The recharge of reclaimed water is an effective strategy for addressing the issues of water quality deterioration and groundwater level decline simultaneously. Residual Al coagulants are normally remained in the recovered water at low concentrations, and may induce clogging problems during the recharging process. However, this issue has been ignored in the past. In this study, we investigated the mechanisms of Al(III)-induced aquifer bio-clogging, the role of Al(III) in quartz sand media (SiO2) dissolution and re-precipitation in the series of aquifer columns. We determined that Al(III) resulted in serious clogging in ∼140 h at low concentrations that satisfied the national drinking water standard of China. The corresponding hydraulic conductivity decreased by more than ∼90% in the bacteria-containing aquifer, which was ∼30% greater than that for the bacteria-free trials. The enhanced Al(III)-related clogging was caused by modifying quartz sand to form Si-O-Al(OH)n and improving microbes attachment. Microbes retention kinetic coefficients (k) of the Al recharged simulated aquifer could increase by 3.0-8.3 times. The Al(III) also enhanced biomass production and clogging by binding to microbial extracellular polymeric substances. In turn, the greater amount of biomass accelerated the Si dissolution and re-precipitation, this may potentially damage the stability of aquifer structure. The results showed that reclaimed water treated with Al coagulation should be employed with caution for recharging.

Low concentrations of Al(III) accelerate the formation of biofilm: Multiple effects of hormesis and flocculation.

Residual Al(III) (at low concentration) is common in water treatment plants (WTPs) and is associated with bacteria. We hypothesize that Al(III) accelerate biofouling due to its hydrolysis and hormesis characteristics, as compared with other cations. To verify this, we elaborated the roles of Al(III) at low concentrations on the biofilm formation. Al(III) hormesis (<2.0mg/L) stimulated bacteria growth increased by ~3.7 times, and extracellular polymeric substances production also enhanced. Al(III) flocculation resulted in the suspended cells precipitation instantly, for Al(III) dosages of 0.6 and 2.0mg/L and the concentration of Al(III) decreased by 0.07 and 0.14mg/L, respectively. Al(III) poisoned the bridged bacterial cells and decreased their ATP by 22.36% and 55.91%, respectively. Al(III) formed polymer presented strong affinity with bacterial outer membrane, and this damaged the bacterial outer membrane. This caused proteins to leak at the combined point. Al-polymer bound to NH2 and/or NH on the leaked protein, contributed to biofilm formation. Biofilm maturity was aided by polysaccharides, which shielded Al(III) toxicity for the formed biofilm. Thus, the biofilm exhibited a distinguished double-layer microstructure, principally with proteins and inactivated cells at the bottom, polysaccharides and activated cells at the top. Thus, hormesis and flocculation caused by low concentration Al(III) mutually promoted each other, and together accelerated biofilm formation.

Bioresources inner-recycling between bioflocculation of Microcystis aeruginosa and its reutilization as a substrate for bioflocculant production.

Bioflocculation, being environmental-friendly and highly efficient, is considered to be a promising method to harvest microalgae. However, one limitation of this technology is high expense on substrates for bioflocculant bacteria cultivation. In this regard, we developed an innovative method for the inner-recycling of biomass that could harvest the typical microalgae, Microcystis aeruginosa, using a bioflocculant produced by Citrobacter sp. AzoR-1. In turn, the flocculated algal biomass could be reutilized as a substrate for Citrobacter sp. AzoR-1 cultivation and bioflocculant production. The experimental results showed that 3.4 ± 0.1 g of bioflocculant (hereafter called MBF-12) was produced by 10 g/L of wet biomass of M. aeruginosa (high-pressure steam sterilized) with an additional 10 g/L of glucose as an extra carbon source. The efficiency of MBF-12 for M. aeruginosa harvesting could reach ~95% under the optimized condition. Further analysis showed that MBF-12, dominated by ~270 kDa biopolymers, contributed the bioflocculation mechanisms of interparticle bridging and biosorption process. Bioflocculant synthesis by Citrobacter sp. AzoR-1 using microalga as a substrate, including the polyketide sugar unit, lipopolysaccharide, peptidoglycan and terpenoid backbone pathways. Our research provides the first evidence that harvested algae can be reutilized as a substrate to grow a bioflocculant using Citrobacter sp. AzoR-1.

Responses of the Microalga Chlorophyta sp. to Bacterial Quorum Sensing Molecules (N-Acylhomoserine Lactones): Aromatic Protein-Induced Self-Aggregation.

Bacteria and microalgae often coexist during the recycling of microalgal bioresources in wastewater treatment processes. Although the bacteria may compete with the microalgae for nutrients, they could also facilitate microalgal harvesting by forming algal-bacterial aggregates. However, very little is known about interspecies interactions between bacteria and microalgae. In this study, we investigated the responses of a model microalga, Chlorophyta sp., to the typical quorum sensing (QS) molecules N-acylhomoserine lactones (AHLs) extracted from activated sludge bacteria. Chlorophyta sp. self-aggregated in 200 µm bioflocs by secreting 460-1000 kDa aromatic proteins upon interacting with AHLs, and the settling efficiency of Chlorophyta sp. reached as high as 41%. However, Chlorophyta sp. cells were essentially in a free suspension in the absence of AHLs. Fluorescence intensity of the aromatic proteins had significant (P < 0.05) relationship with the Chlorophyta sp. settleability, and showed a positive correlation, indicating that aromatic proteins helped aggregate microalga. Transcriptome results further revealed up-regulation of synthesis pathways for aromatic proteins from tyrosine and phenylalanine that was assisted by anthranilate accumulation. To the best of our knowledge, this is the first study to confirm that eukaryotic microorganisms can sense and respond to prokaryotic QS molecules.

The effectiveness of coagulation for water reclamation from a wastewater treatment plant that has a long hydraulic and sludge retention times: A case study.

Coagulation is a feasible process to reclaim municipal wastewater, however, the role of coagulation in removing effluent organic matter (EfOM) from underutilized wastewater treatment plants (WWTPs) has not been fully explored. We identified the characteristics of the EfOM from a typical underutilized WWTP (i.e., the ratio of actual capacity to design capacity is 50%-70%), and investigated the performance of coagulation on suspended solids (SS) and dissolved organic matter (DOM) removal. The effluent could even satisfy the highest national standard of China (Class 1 A) for WWTP effluent, as evaluated by the traditional parameters such as SS and chemical oxygen demand (COD). However, the DOM in the EfOM we studied contained considerable biomass-associated products (BAPs), which were dominated by proteins with a molecular weight of approximately 150 kDa. In addition, protein also dominated the DOM after coagulation. Fulvic acid and humic-like acid organics were poorly removed by either AlCl3 or polyaluminum chloride (PAC) coagulation, even with a dosage as high as 24 mg Al L(-1). Biodegradability was very poor, as the ratio of biological oxygen demand (BOD5) to COD was less than 0.17. After coagulation the typical BAPs, protein and polysaccharide, remained as high as 1.6 mg L(-1) and 1.2 mg L(-1) respectively. In this study we found coagulation was ineffective for removal of recalcitrant BAPs.

Application of ultrasound and quartz sand for the removal of disinfection byproducts from drinking water.

To the best of our knowledge, little information is available on the combined use of ultrasound (US) and quartz sand (QS) in the removal of disinfection byproducts (DBPs) from drinking water. This study investigates the removal efficiency for 12 DBPs from drinking water by 20 kHz sonolytic treatment, QS adsorption, and their combination. Results indicate that DBPs with logKow≤1.12 could not be sonolysized; for logKow≥1.97, more than 20% removal efficiency was observed, but the removal efficiency was unrelated to logKow. DBPs containing a nitro group are more sensitive to US than those that comprise nitrile, hydrogen, and hydroxyl groups. Among the 12 investigated DBPs, 9 could be adsorbed by QS adsorption. The adsorption efficiency ranged from 12% for 1,1-dichloro-2-propanone to 80% for trichloroacetonitrile. A synergistic effect was found between the US and QS on DBPs removal, and all the 12 DBPs could be effectively removed by the combined use of US and QS. In the presence of US, part of the QS particles were corroded into small particles which play a role in increasing the number of cavitation bubbles and reducing cavitation bubble size and then improve the removal efficiency of DBPs. On the other hand, the presence of US enhances the DBP mass transfer rate to cavitation bubbles and quartz sand. In addition, sonolytic treatment led to a slight decrease of pH, and TOC values decreased under all the three treatment processes.

Pose measurement method and experiments for high-speed rolling targets in a wind tunnel.

High-precision wind tunnel simulation tests play an important role in aircraft design and manufacture. In this study, a high-speed pose vision measurement method is proposed for high-speed and rolling targets in a supersonic wind tunnel. To obtain images with high signal-to-noise ratio and avoid impacts on the aerodynamic shape of the rolling targets, a high-speed image acquisition method based on ultrathin retro-reflection markers is presented. Since markers are small-sized and some of them may be lost when the target is rolling, a novel markers layout with which markers are distributed evenly on the surface is proposed based on a spatial coding method to achieve highly accurate pose information. Additionally, a pose acquisition is carried out according to the mentioned markers layout after removing mismatching points by Case Deletion Diagnostics. Finally, experiments on measuring the pose parameters of high-speed targets in the laboratory and in a supersonic wind tunnel are conducted to verify the feasibility and effectiveness of the proposed method. Experimental results indicate that the position measurement precision is less than 0.16 mm, the pitching and yaw angle precision less than 0.132° and the roll angle precision 0.712°.