Novel insights into aerobic duration control-based partial nitritation in source-separated blackwater treatment: Growth type, inoculation source, and comammox threat.

Aerobic duration control (ADC), whereby aeration is terminated before nitrite is extremely oxidized during the nitrification process, is an effective strategy to achieve partial nitritation (PN) for blackwater. This study evaluated the effects of microbial growth type, influent ammonia-oxidizing organisms (AOO), and comammox bacteria from seeding sludge to ADC-based PN. The long-term operation of lab-scale reactors and model simulations were implemented to select the best growth type. The biofilm formed on the inner wall of the activated sludge reactor decreased the nitrite accumulation ratio (NAR) from 99.2% to 77.2%. Meanwhile, the NAR of the pure-biofilm reactor decreased from 95.9% to 47.8%. The deteriorated PN of the biofilm-related reactors was due to the extended solid retention time and increased substrate saturation constants of AOOs compared with those of nitrite-oxidizing organisms (NOO). Periodic biofilm carrier regeneration and biofilm thickness control can recover PN performance but are difficult to implement. In contrast, the optimized activated sludge reactor exhibited high (NAR >94%) and stable (>3 months) PN performance when treating real blackwater. Nitrifiers were found in blackwater, and chemically enhanced high-rate activated sludge pretreatment removed more NOOs than AOOs (41.8% vs. 24.3%) and increased the influent AOO/NOO ratio. Interestingly, the influent AOOs supported fast PN start-up in the moving-bed biofilm reactor without the initial inoculation of activated sludge. Moreover, model simulations verified that high and stable PN could also be realized in an activated sludge reactor by the continuous inoculation of influent AOOs, which is a novel PN start-up strategy. Metagenomic analyses showed that the comammox bacteria from the seeding sludge eventually disappeared owing to their intrinsic specific growth rates and free ammonia inhibition. The findings of this study will provide insightful guidelines for PN application in decentralized and semi-centralized wastewater treatment systems.

Full-scale semi-centralized wastewater treatment facilities for resource recovery: operation, problems and resolutions.

The first full-scale semi-centralized wastewater treatment and resource recovery system based on source separation was implemented from 2014. To assess the operation performance, operating costs and resolve the problems faced in this system, the latest operation data from April 2017 to September 2018 was investigated. The results show that greywater and blackwater modules exhibited good removal performance for organics and nutrients, although misconnection between pipelines existed and influent loading rates fluctuated. The effluent could meet reuse standards. The biogas production rates of raw sludge could reach 7.27-10.9 m3 gas·per cubic raw sludge. The specific cost of treated water was higher than in a conventional treatment system. Power consumption made a major contribution to the total cost with a proportion of 55.3-94.2%. After optimizing and considering the comprehensive efficiencies, the costs would be affordable. The dewatered sludge of the anaerobic digestion module has been applied to agricultural and landscaping soil. It is suggested that organics in blackwater could be recovered as volatile fatty acids with high-efficiency anaerobic fermentation and used as an external carbon source for short-cut biological nitrogen removal. In conclusion, the semi-centralized system will be a feasible and sustainable alternative for conventional treatment systems in future.

Carbon capture for blackwater: chemical enhanced high-rate activated sludge process.

Blackwater has more benefits for carbon recovery than conventional domestic wastewater. Carbon capture and up-concentration are crucial prerequisites for carbon recovery from blackwater, the same as domestic wastewater. Both chemical enhanced primary treatment (CEPT) and high-rate activated sludge (HRAS) processes have enormous potential to capture organics. However, single CEPT is subject to the disruption of influent sulfide, and single HRAS has insufficient flocculation capacity. As a result, their carbon capture efficiencies are low. By combining CEPT and HRAS with chemical enhanced high rate activated sludge (CEHRAS) process, the limitations of single CEPT and single HRAS offset each other. The carbon mineralization efficiency was significantly influenced by SRT rather than iron salt dosage. An iron dosage significantly decreased chemical oxygen demand (COD) lost in effluent. Both SRT and iron dosage had a significant influence on the carbon capture efficiency. However, HRT had no great impact on the organic mass balance. CEHRAS allowed up to 78.2% of carbon capture efficiency under the best conditions. The results of techno-economic analysis show that decreasing the iron salt dosage to 10 mg Fe/L could promise profiting for blackwater treatment. In conclusion, CEHRAS is a more appropriate technology to capture carbon in blackwater.

A novel graphene oxide-carbon nanotubes anchored α-FeOOH hybrid activated persulfate system for enhanced degradation of Orange II.

Persulfate activation has been applied as one of the efficient advanced oxidation processes (AOPs) to remediate polluted environments. In this study, a novel α-FeOOH anchored by graphene oxide (GO)-carbon nanotubes (CNTs) aerogel (α-FeOOH@GCA) nanocomposite activated persulfate system (α-FeOOH@GCA + K2S2O8) was applied for decolorization of Orange II (OII). The decolorization of OII was remarkably enhanced to a level of ~99% in this system compared with that of pristine α-FeOOH (~44%) or GO-CNTs (~18%). The enhanced catalytic activity of α-FeOOH@GCA was due to the formation of a heterojunction by α-FeOOH and GO-CNTs as confirmed by the presence of Fe-O-C chemical bonds. The degradation intermediates of OII were comprehensively identified. The proposed degradation pathway of OII begins with the destruction of the conjugated structures of OII by the dominant reactive oxygen species, surface-bound SO4•-. The decolorization efficiency of OII by the α-FeOOH@GCA activated persulfate system decreased from the first to third cycle of recycling. Ultraviolet (UV) irradiation or introduction of a small amount of Fe2+ could restore the activation of this system. The results show that the α-FeOOH@GCA persulfate activation system promises to be a highly efficient environmental remediation method for organic pollutants.

Identification of human neutralizing antibodies against MERS-CoV and their role in virus adaptive evolution.

The newly emerging Middle East Respiratory Syndrome coronavirus (MERS-CoV) causes a Severe Acute Respiratory Syndrome-like disease with ∼43% mortality. Given the recent detection of virus in dromedary camels, zoonotic transfer of MERS-CoV to humans is suspected. In addition, little is known about the role of human neutralizing Ab (nAb) pressure as a driving force in MERS-CoV adaptive evolution. Here, we used a well-characterized nonimmune human Ab-phage library and a panning strategy with proteoliposomes and cells to identify seven human nAbs against the receptor-binding domain (RBD) of the MERS-CoV Spike protein. These nAbs bind to three different epitopes in the RBD and human dipeptidyl peptidase 4 (hDPP4) interface with subnanomolar/nanomolar binding affinities and block the binding of MERS-CoV Spike protein with its hDPP4 receptor. Escape mutant assays identified five amino acid residues that are critical for neutralization escape. Despite the close proximity of the three epitopes on the RBD interface, escape from one epitope did not have a major impact on neutralization with Abs directed to a different epitope. Importantly, the majority of escape mutations had negative impacts on hDPP4 receptor binding and viral fitness. To our knowledge, these results provide the first report on human nAbs against MERS-CoV that may contribute to MERS-CoV clearance and evolution. Moreover, in the absence of a licensed vaccine or antiviral for MERS, this panel of nAbs offers the possibility of developing human mAb-based immunotherapy, especially for health-care workers.

The simultaneous removal of methylene blue (MB) and Ca2+ by recyclable adsorbents based the scales derived from coal gasification system.

The transformation of solid wastes from industrial production into effective adsorbents could significantly contribute to wastewater treatment. In this study, after acidizing and burning soft scale (SS) from coal gasification system, two magnetic adsorbents (mag-ASS and mag-BASS) were prepared via the combination of magnetite with ultrasonic, respectively. The treatment effects of mag-ASS and mag-BASS were then investigated for simulated wastewater containing macromolecular organic matter [i.e., methylene blue (MB)] and Ca2+. The results indicated that the pseudo second order kinetic, Elovich, Freundlich, Langmuir and Temkin model could well describe the adsorption behavior of MB and Ca2+ onto mag-ASS and mag-BASS. The maximum adsorption capacities of mag-ASS for MB and mag-BASS for Ca2+ were 600.53 mg/g and 102.54 mg/g, respectively. Surprisingly, the adsorption abilities of mag-ASS for MB and mag-BASS for Ca2+ show significantly higher than the others. The adsorption mechanisms of MB mainly included electrostatic interaction, π-π conjugate interaction and cation exchange, while those of Ca2+ were mainly electrostatic interaction and cation exchange. The diffusion of MB and Ca2+ onto the magnetic adsorbents might be controlled by the combined effects of intraparticle and liquid film diffusion. There was no significant reduction in adsorption capacity after 8 cycles of adsorption and desorption, indicating that SS-based magnetic adsorbents had good recyclability and stability. Moreover, the removal efficiency of mag-BASS for total hardness and total organic carbon in real coal gasification gray water (CGGW) was 82.60 and 64.10%, respectively. The treatment of CGGW and the resource of wastes would significantly promote the reasonable disposal of coal gasification scales.

Impact of invertebrates on water quality safety and their sheltering effect on bacteria in water supply systems.

Invertebrates in drinking water not only affect human health, but also provide migration and shelter for pathogenic microorganisms. Their residues and metabolites also produce DBPs (disinfection by-products), which have adverse effects on the health of residents. In this study, the contributions of the rotifers and nematodes to the BDOC (biodegradable dissolved organic carbon), BRP (bacterial regrowth potential) and DBPs in drinking water were explored, and the sheltering effects of chlorine-resistant invertebrates on indigenous bacteria and pathogenic bacteria were studied, and the health and safety risk of invertebrates in drinking water was also assessed. The contributions of rotifer BAPs (biomass-associated products), UAPs (utilization-associated products) of rotifer, and nematode BAPs to the BRP were 46, 1240, and 24 CFU/mL. Nematodes were found to have a sheltering effect on indigenous bacteria and pathogenic bacteria, allowing them to resist chlorine disinfection and UV (ultraviolet) disinfection. When subjected to a UV dose of 40 mJ/cm2, the inactivation rates of indigenous bacteria and three pathogenic bacteria decreased by 85% and 39-50% when bacteria were sheltered by the living nematodes; while decreased by 66% and 15-41% when they were sheltered by nematode residue. The safety risk posed by invertebrates in the drinking water was mainly due to their ability to promote bacterial regeneration and carry bacteria. This study aims to provide a theoretical basis and technical support for the risk control of invertebrates' pollution, and provides references for ensuring the safety of drinking water and formulating standards for the levels of invertebrates in drinking water.

The pretreatment effects of various target pollutant in real coal gasification gray water by coupling pulse electrocoagulation with chemical precipitation methods.

To prevent the scale formation in the equipments and pipelines after pre-treated coal gasification gray water (CGGW) entering the reuse system and reduce the influence of various pollutants in the effluent on subsequent biochemical treatment, this study presented a coupled use of pulse electrocoagulation (PEC) and chemical precipitation (CP) coupling method for the pretreatment of coal gasification gray water (CGGW). In addition, the operation parameters of PEC and the reaction conditions of PEC-CP were optimized based on iron plate as electrode and total hardness, turbidity and sludge yield as assessment indicators. Due to the formation of multi-hydroxyl iron by several minutes of pulse current, and the addition of pH regulator and coagulant aid, the efficient removal of various ions, hardness and turbidity was significantly reduced via various mechanism such as redox, precipitation, adsorption and coagulation reaction. The result indicated that under the optimal operation conditions, the total hardness, turbidity, and Fen+ of PEC-CP effluents were 275.0 mg/L, 3.0 NTU and 5.6 mg/L, respectively and sludge amount was 0.88 kg/m3. The removal rates of Si, B, Mn, Ba, COD, NPOC and NH4+-N by PEC-CP reached 80.0%, 75.4%, 97.0%, 99.8%, 35.0%, 33.6% and 23.8%, respectively. The present results suggested that the CGGW pretreatment effluents could be not only reused directly, but also greatly alleviate the scaling problem of water pipeline and coal gasification production facilities.

Differential stepwise evolution of SARS coronavirus functional proteins in different host species.

BACKGROUND: SARS coronavirus (SARS-CoV) was identified as the etiological agent of SARS, and extensive investigations indicated that it originated from an animal source (probably bats) and was recently introduced into the human population via wildlife animals from wet markets in southern China. Previous studies revealed that the spike (S) protein of SARS had experienced adaptive evolution, but whether other functional proteins of SARS have undergone adaptive evolution is not known. RESULTS: We employed several methods to investigate selective pressure among different SARS-CoV groups representing different epidemic periods and hosts. Our results suggest that most functional proteins of SARS-CoV have experienced a stepwise adaptive evolutionary pathway. Similar to previous studies, the spike protein underwent strong positive selection in the early and middle phases, and became stabilized in the late phase. In addition, the replicase experienced positive selection only in human patients, whereas assembly proteins experienced positive selection mainly in the middle and late phases. No positive selection was found in any proteins of bat SARS-like-CoV. Furthermore, specific amino acid sites that may be the targets of positive selection in each group are identified. CONCLUSION: This extensive evolutionary analysis revealed the stepwise evolution of different functional proteins of SARS-CoVs at different epidemic stages and different hosts. These results support the hypothesis that SARS-CoV originated from bats and that the spill over into civets and humans were more recent events.

miR-487a promotes progression of gastric cancer by targeting TIA1.

Gastric cancer (GC) is one of the most common malignancies as well as the third leading cause for cancer-related death. Molecular basis of GC are essential and critical for its therapeutic treatment, but still remain poorly understood. T-cell intracellular antigen-1 (TIA1) extensively involves in cancer progression, whereas its role and regulation mechanism in GC have not been revealed. In the present study, we found that TIA-1 protein level was down-regulated in GC tissues and TIA1 inhibited proliferation and promoted apoptosis of GC cells. Then, we used bioinformatics to predict miR-487a as the upstream regulator of TIA1 and we also observed an inverse correlation between miR-487a level and TIA-1 protein level in GC tissues. Next, we demonstrated that miR-487a directly targeted TIA1 via binding to its 3'-untranslated region. Furthermore, we investigated the role of miR-487a-TIA1 pathway in the growth of GC cells both in vitro and in vivo. The repression of TIA-1 by miR-487a promoted cell proliferation and suppressed cell apoptosis in vitro, and the knockdown of miR-487a had the opposite effects. Finally, we demonstrated that miR-487a promoted the development of gastric tumor growth in xenograft mice by targeting TIA-1. These effects could be partially reversed by restoring the expression of TIA-1. Overall, our results reveal that TIA1 is a tumor suppressor gene and is directly regulated by miR-487a in GC, which may offer new therapeutic targets for GC treatment.

Evolutionary relationships between bat coronaviruses and their hosts.

Recent studies have suggested that bats are the natural reservoir of a range of coronaviruses (CoVs), and that rhinolophid bats harbor viruses closely related to the severe acute respiratory syndrome (SARS) CoV, which caused an outbreak of respiratory illness in humans during 2002-2003. We examined the evolutionary relationships between bat CoVs and their hosts by using sequence data of the virus RNA-dependent RNA polymerase gene and the bat cytochrome b gene. Phylogenetic analyses showed multiple incongruent associations between the phylogenies of rhinolophid bats and their CoVs, which suggested that host shifts have occurred in the recent evolutionary history of this group. These shifts may be due to either virus biologic traits or host behavioral traits. This finding has implications for the emergence of SARS and for the potential future emergence of SARS-CoVs or related viruses.

A mouse model for MERS coronavirus-induced acute respiratory distress syndrome.

Middle East respiratory syndrome coronavirus (MERS-CoV) is a novel virus that emerged in 2012, causing acute respiratory distress syndrome (ARDS), severe pneumonia-like symptoms and multi-organ failure, with a case fatality rate of ∼36%. Limited clinical studies indicate that humans infected with MERS-CoV exhibit pathology consistent with the late stages of ARDS, which is reminiscent of the disease observed in patients infected with severe acute respiratory syndrome coronavirus. Models of MERS-CoV-induced severe respiratory disease have been difficult to achieve, and small-animal models traditionally used to investigate viral pathogenesis (mouse, hamster, guinea-pig and ferret) are naturally resistant to MERS-CoV. Therefore, we used CRISPR-Cas9 gene editing to modify the mouse genome to encode two amino acids (positions 288 and 330) that match the human sequence in the dipeptidyl peptidase 4 receptor, making mice susceptible to MERS-CoV infection and replication. Serial MERS-CoV passage in these engineered mice was then used to generate a mouse-adapted virus that replicated efficiently within the lungs and evoked symptoms indicative of severe ARDS, including decreased survival, extreme weight loss, decreased pulmonary function, pulmonary haemorrhage and pathological signs indicative of end-stage lung disease. Importantly, therapeutic countermeasures comprising MERS-CoV neutralizing antibody treatment or a MERS-CoV spike protein vaccine protected the engineered mice against MERS-CoV-induced ARDS.

3B11-N, a monoclonal antibody against MERS-CoV, reduces lung pathology in rhesus monkeys following intratracheal inoculation of MERS-CoV Jordan-n3/2012.

Middle East Respiratory Syndrome Coronavirus (MERS-CoV) was identified in 2012 as the causative agent of a severe, lethal respiratory disease occurring across several countries in the Middle East. To date there have been over 1600 laboratory confirmed cases of MERS-CoV in 26 countries with a case fatality rate of 36%. Given the endemic region, it is possible that MERS-CoV could spread during the annual Hajj pilgrimage, necessitating countermeasure development. In this report, we describe the clinical and radiographic changes of rhesus monkeys following infection with 5×10(6) PFU MERS-CoV Jordan-n3/2012. Two groups of NHPs were treated with either a human anti-MERS monoclonal antibody 3B11-N or E410-N, an anti-HIV antibody. MERS-CoV Jordan-n3/2012 infection resulted in quantifiable changes by computed tomography, but limited other clinical signs of disease. 3B11-N treated subjects developed significantly reduced lung pathology when compared to infected, untreated subjects, indicating that this antibody may be a suitable MERS-CoV treatment.

Baculovirus-produced influenza virus-like particles in mammalian cells protect mice from lethal influenza challenge.

Influenza virus-like particles (VLPs) are effective vaccines against influenza infection, which can be produced either in insect cells by recombinant baculovirus (BV) infection or in mammalian cells by DNA plasmid transfection. However, VLPs produced from baculovirus/insect cells are difficult to purify due to baculovirus contamination; VLPs produced by plasmid transfection are limited by scale-up capability. In this study, a BacMam BV, in which three CMV-promoters drive the hemagglutinin, neuraminidase, and matrix of influenza virus was constructed. This baculovirus can deliver these genes into mammalian cells/hosts and subsequently influenza VLPs can be produced and secreted from transduced cells. Transduction conditions were optimized and influenza VLPs were purified from transduced 293T cells. Mice were vaccinated with BV transduction-produced VLPs, plasmid transfection-produced VLPs, and BacMam BV. Two vaccinations of each vaccine induced high hemagglutination-inhibition (HAI) titers and prevented influenza virus infection. In contrast, following a single vaccination, all mice vaccinated with each vaccine had significantly lower lung viral titers compared to unvaccinated mice. Remarkably, mice vaccinated with a single dose of BV transduction-produced VLPs survived challenge, whereas mice vaccinated with one dose of BacMam BV- or plasmid transfection-produced VLPs had 60-80% survival. This finding is particularly significant for producing easily purified VLPs. The BacMam system is an alternative strategy for VLP production, which is easy to scale up and purify. Besides, BacMam BV can be used as a gene delivery vector to produce VLPs in vivo, to stimulate immune responses.

Hemagglutinin displayed baculovirus protects against highly pathogenic influenza.

Baculovirus (BV) replicating in insect cells can express a foreign gene product as part of its genome. The influenza hemagglutinin (HA) can be expressed from BV and displayed on the surface of baculovirus (HA-DBV). In this study we first generated six recombinant baculoviruses that expressed chimeric HAs with segments of the BV glycoprotein (gp64). The signal peptide (SP) and cytoplasmic tail (CT) domains of gp64 can enhance the display of HA from A/PR8/34 on BV surface, while the transmembrane (TM) domain of gp64 impairs HA display. Different doses of either live or ß-propiolactone (BPL)-inactivated HA-DBV were administered to BALB/c mice. Live HA-DBV elicited higher hemagglutination-inhibition (HAI) titers than BPL-inactivated HA-DBV, and provided sterilizing protection. A second generation recombinant BV simultaneously displaying four HAs derived from four subclades of H5N1 influenza viruses was constructed. This tetravalent H5N1 HA-DBV vaccine elicited HAI titers against all four homologous H5N1 viruses, significantly decreasing viral lung titers of challenged mice and providing 100% protection against lethal doses of homologous H5N1 viruses. Moreover, mice vaccinated with HA-DBV had high levels of IFNγ-secreting and HA-specific CD8+ T cells. Taken together, this study demonstrates that HA-DBV can stimulate strong humoral, as well as cellular immune responses, and is an effective vaccine candidate for influenza.

Pivotal role of dogs in rabies transmission, China.

The number of dog-mediated rabies cases in China has increased exponentially; the number of human deaths has been high, primarily in poor, rural communities. We review the incidence of rabies in China based on data from 1950 and 2004, obtained mainly from epidemiologic bulletins published by the Chinese Ministry of Health.