A fungal CFEM-containing effector targets NPR1 regulator NIMIN2 to suppress plant immunity.

Colletotrichum fructicola causes a broad range of plant diseases worldwide and secretes many candidate proteinous effectors during infection, but it remains largely unknown regarding their effects in conquering plant immunity. Here, we characterized a novel effector CfEC12 that is required for the virulence of C. fructicola. CfEC12 contains a CFEM domain and is highly expressed during the early stage of host infection. Overexpression of CfEC12 suppressed BAX-triggered cell death, callose deposition and ROS burst in Nicotiana benthamiana. CfEC12 interacted with apple MdNIMIN2, a NIM1-interacting (NIMIN) protein that putatively modulates NPR1 activity in response to SA signal. Transient expression and transgenic analyses showed that MdNIMIN2 was required for apple resistance to C. fructicola infection and rescued the defence reduction in NbNIMIN2-silenced N. benthamiana, supporting a positive role in plant immunity. CfEC12 and MdNPR1 interacted with a common region of MdNIMIN2, indicating that CfEC12 suppresses the interaction between MdNIMIN2 and MdNPR1 by competitive target binding. In sum, we identified a fungal effector that targets the plant salicylic acid defence pathway to promote fungal infection.

Major quality regulation network of flavonoid synthesis governing the bioactivity of black wolfberry.

Black wolfberry (Lycium ruthenicum Murr.) contains various bioactive metabolites represented by flavonoids, which are quite different among production regions. However, the underlying regulation mechanism of flavonoid biosynthesis governing the bioactivity of black wolfberry remains unclear. Presently, we compared the bioactivity of black wolfberry from five production regions. Multi-omics were performed to construct the regulation network associated with the fruit bioactivity. The detailed regulation mechanisms were identified using genetic and molecular methods. Typically, Qinghai (QH) fruit exhibited higher antioxidant and anti-inflammatory activities. The higher medicinal activity of QH fruit was closely associated with the accumulation of eight flavonoids, especially Kaempferol-3-O-rutinoside (K3R) and Quercetin-3-O-rutinoside (rutin). Flavonoid biosynthesis was found to be more active in QH fruit, and the upregulation of LrFLS, LrCHS, LrF3H and LrCYP75B1 caused the accumulation of K3R and rutin, leading to high medicinal bioactivities of black wolfberry. Importantly, transcription factor LrMYB94 was found to regulate LrFLS, LrCHS and LrF3H, while LrWRKY32 directly triggered LrCYP75B1 expression. Moreover, LrMYB94 interacted with LrWRKY32 to promote LrWRKY32-regulated LrCYP75B1 expression and rutin synthesis in black wolfberry. Transgenic black wolfberry overexpressing LrMYB94/LrWRKY32 contained higher levels of K3R and rutin, and exhibited high medicinal bioactivities. Importantly, the LrMYB94/LrWRKY32-regulated flavonoid biosynthesis was light-responsive, showing the importance of light intensity for the medicinal quality of black wolfberry. Overall, our results elucidated the regulation mechanisms of K3R and rutin synthesis, providing the basis for the genetic breeding of high-quality black wolfberry.

Changes in planta K nutrient content altered the interaction pattern between Nicotiana benthamiana and Alternaria longipes.

Potassium (K) fertilisation has frequently been shown to enhance plant resistance against pathogens, though the mechanisms remain elusive. This study investigates the interaction dynamics between Nicotiana benthamiana and the pathogen Alternaria longipes under different planta K levels. On the host side, adding K activated the expressions of three NLR (nucleotide-binding domain and leucine-rich repeat-containing proteins) resistance genes, including NbRPM1, NbR1B23 and NbNBS12. Silencing these NLRs attenuated resistance in high-K (HK, 40.8 g/kg) plant, whereas their overexpression strengthened resistance in low-K (LK, 23.9 g/kg) plant. Typically, these NLRs mainly strengthened plant resistance via promoting the expression of pathogenesis-related genes (PRs), ROS burst and synthesis of antifungal metabolites in HK plant. On the pathogen side, the expression of effectors HKCSP1, HKCSP2 and LKCSP were shown to be related to planta K content. A. longipes mainly expressed effectors HKCSP1 and HKCSP2 in HK plant to interfere host resistance. HKCSP1 physically interacted with NbRPM1 to promote the degradation of NbRPM1, then attenuated related resistance in HK N. benthamiana. Meanwhile, HKCSP2 directly interacted with NbPR5 to suppress resistance in HK plant. In LK plant, A. longipes mainly deployed LKCSP that interacted with NbR1B23 to interfere reduce resistance in N. benthamiana. Overall, our research insights that both pathogen and host mobilise distinct strategies to outcompete each other during interactions in different K nutrient environments.

Soil water and nutrient availability interactively modify pollinator-mediated directional and correlational selection on floral display.

The individual and combined effects of abiotic factors on pollinator-mediated selection on floral traits are not well documented. To examine potential interactive effects of water and nutrient availability on pollinator-mediated selection on three floral display traits of Primula tibetica, we manipulated pollination and nutrient availability in a factorial experiment, conducted at two common garden sites with different soil water content (natural vs addition). We found that both water and nutrient availability affected floral trait expression in P. tibetica and that hand pollination increased seed production most when both nutrient content and water content were high, indicating joint pollen and resource limitation. We documented selection on all floral traits, and pollinators contributed to directional and correlational selection on plant height and number of flowers. Soil water and nutrient availability interactively influenced the strength of both pollinator-mediated directional and correlational selection, with significant selection observed when nutrient or water availability was high, but not when none or both were added. The results suggest that resource limitation constrains the response of P. tibetica to among-individual variation in pollen receipt, that addition of nutrients or water leads to pollinator-mediated selection and that effects of the two abiotic factors are nonadditive.

Labile carbon-induced soil organic matter turnover in a subtropical forest under different redox conditions.

Labile organic carbon (LOC) input strongly affects soil organic matter (SOM) dynamics, including gains and losses. However, it is unclear how redox fluctuations regulate these processes of SOM decomposition and formation induced by LOC input. The objective of this study was to explore the impacts of LOC input on SOM turnover under different redox conditions. Soil samples were collected in a subtropical forest. A single pulse of 13C-labeled glucose (i.e., LOC) was applied to the soil. Soil samples were incubated for 40 days under three redox treatments, including aerobic, anoxic, and 10-day aerobic followed by 10-day anoxic conditions. Results showed that LOC input affected soil priming and 13C-SOM accumulation differently under distinct redox conditions by altering the activities of various microorganisms. 13C-PLFAs (phospholipid fatty acids) were analyzed to determine the role of microbial groups in SOM turnover. Increased activities of fungi and gram-positive bacteria (i.e., the K-strategists) by LOC input could ingest metabolites or residues of the r-strategists (e.g., gram-negative bacteria) to result in positive priming. Fungi could use gram-negative bacteria to stimulate priming intensity via microbial turnover in aerobic conditions first. Reduced activities of K-strategists as a result of the aerobic to anoxic transition decreased priming intensity. The difference in LOC retention in SOM under different redox conditions was mainly attributable to 13C-particulate organic carbon (13C-POC) accumulation. Under aerobic conditions, fungi and gram-positive bacteria used derivatives from gram-negative bacteria to reduce newly formed POC. However, anoxic conditions were not conducive to the uptake of gram-negative bacteria by fungi and gram-positive bacteria, favoring SOM retention. This work indicated that redox-regulated microbial activities can control SOM decomposition and formation induced by LOC input. It is extremely valuable for understanding the contribution of soil affected by redox fluctuations to the carbon cycle.

Three-dimensional measurement method of color fringe projection based on an improved three-step phase-shifting method.

A three-dimensional (3D) measurement method of color fringe projection based on an improved three-step phase-shifting method is proposed. The color fringe pattern is encoded by two cosine fringe patterns with the same frequency but different shifting phase and a uniform gray flat image into three color channels R, G, and B. Although the measurement speed of the traditional three-step phase-shifting method can meet the requirements of measuring 3D objects, it makes the noise and inaccuracy of the captured images increase, and each image will cause measurement error. Therefore, we improve the three-step phase-shifting method and introduce the Hilbert transform into the three-step phase-shift method. The DC component of the fringe pattern is obtained by using the Hilbert transform principle, and the third fringe pattern in the three-step phase-shift method is replaced by the captured light intensity distribution of the DC component. The phase difference of the other two fringe patterns is fixed as π/2 by the Hilbert transform. The improved three-step phase-shifting method is used to obtain the phase information of the deformed color fringe image, and then the phase-unwrapping algorithm is used to obtain the phase distribution information of the whole field. The results show that the improved method can not only accurately calculate the phase information but also greatly improve the measurement speed and quality.

Fundus-simulating phantom for calibration of retinal vessel oximetry devices.

Retinal vessel oxygen supply is important for retinal tissue metabolism. Commonly used retinal vessel oximetry devices are based on dual-wavelength spectral measurement of oxyhemoglobin and deoxyhemoglobin. However, there is no traceable standard for reliable calibration of these devices. In this study, we developed a fundus-simulating phantom that closely mimicked the optical properties of human fundus tissues. Microchannels of precisely controlled topological structures were produced by soft lithography to simulate the retinal vasculature. Optical properties of the phantom were adjusted by adding scattering and absorption agents to simulate different concentrations of fundus pigments. The developed phantom was used to calibrate the linear correlation between oxygen saturation (SO2) level and optical density ratio in a dual-wavelength oximetry device. The obtained calibration factors were used to calculate the retinal vessel SO2 in both eyes of five volunteers aged between 24 and 27 years old. The test results showed that the mean arterial and venous SO2 levels after phantom calibration were coincident with those after empirical value calibration, indicating the potential clinical utility of the produced phantom as a calibration standard.

Transcriptome analysis of chrysanthemum (Dendranthema grandiflorum) in response to low temperature stress.

BACKGROUND: Chrysanthemum is one kind of ornamental plant well-known and widely used in the world. However, its quality and production were severely affected by low temperature conditions in winter and early spring periods. Therefore, we used the RNA-Seq platform to perform a de novo transcriptome assembly to analyze chrysanthemum (Dendranthema grandiflorum) transcription response to low temperature. RESULTS: Using Illumina sequencing technology, a total of 86,444,237 high-quality clean reads and 93,837 unigenes were generated from four libraries: T01, controls; T02, 4 °C cold acclimation (CA) for 24 h; T03, - 4 °C freezing treatments for 4 h with prior CA; and T04, - 4 °C freezing treatments for 4 h without prior CA. In total, 7583 differentially expressed genes (DEGs) of 36,462 annotated unigenes were identified. We performed GO and KEGG pathway enrichment analyses, and excavated a group of important cold-responsive genes related to low temperature sensing and signal transduction, membrane lipid stability, reactive oxygen species (ROS) scavenging and osmoregulation. These genes encode many key proteins in plant biological processes, such as protein kinases, transcription factors, fatty acid desaturase, lipid-transfer proteins, antifreeze proteins, antioxidase and soluble sugars synthetases. We also verified expression levels of 10 DEGs using quantitative real-time polymerase chain reaction (qRT-PCR). In addition, we performed the determination of physiological indicators of chrysanthemum treated at low temperature, and the results were basically consistent with molecular sequencing results. CONCLUSION: In summary, our study presents a genome-wide transcript profile of Dendranthema grandiflorum var. jinba and provides insights into the molecular mechanisms of D. grandiflorum in response to low temperature. These data contributes to our deeper relevant researches on cold tolerance and further exploring new candidate genes for chilling-tolerance and freezing-tolerance chrysanthemum molecular breeding.

Design of a portable phantom device to simulate tissue oxygenation and blood perfusion.

We propose a portable phantom system for calibration and validation of medical optical devices in a clinical setting. The phantom system comprises a perfusion module and an exchangeable tissue-simulating phantom that simulates tissue oxygenation and blood perfusion. The perfusion module consists of a peristaltic pump, two liquid storage units, and two pressure suppressors. The tissue-simulating phantom is fabricated by a three-dimensional (3D) printing process with microchannels embedded to simulate blood vessels. Optical scattering and absorption properties of biologic tissue are simulated by mixing graphite powder and titanium dioxide powder with clear photoreactive resin at specific ratios. Tissue oxygen saturation (StO2) and blood perfusion are simulated by circulating the mixture of blood and intralipid at different oxygenation levels and flow rates. A house-made multimodal imaging system that combines multispectral imaging and laser speckle imaging are used for non-invasive detection of phantom oxygenation and perfusion, and the measurements are compared with those of a commercial Moor device as well as numerical simulation. By acquiring multimodal imaging data from one phantom and applying the calibration factors in different settings, we demonstrate the technical feasibility to calibrate optical devices for consistent measurements. By simulating retina tissue vasculature and acquiring functional images at different tissue oxygenation and blood perfusion levels, we demonstrate the clinical potential to simulate tissue anomalies. Our experiments imply the clinical potential of a portable, low-cost, and traceable phantom standard to calibrate and validate medical optical devices for improved performance.

Fabrication of a multilayer tissue-mimicking phantom with tunable optical properties to simulate vascular oxygenation and perfusion for optical imaging technology.

Vast research has been carried out to fabricate tissue-mimicking phantoms, due to their convenient use and ease of storage, to assess and validate the performance of optical imaging devices. However, to the best of our knowledge, there has been little research on the use of multilayer tissue phantoms for optical imaging technology, although their structure is closer to that of real skin tissue. In this work, we design, fabricate, and characterize multilayer tissue-mimicking phantoms, with a morphological mouse ear blood vessel, that contain an epidermis, a dermis, and a hypodermis. Each tissue-mimicking phantom layer is characterized individually to match specific skin tissue layer characteristics. The thickness, optical properties (absorption coefficient and reduced scattering coefficient), oxygenation, and perfusion of skin are the most critical parameters for disease diagnosis and for some medical equipment. These phantoms can be used as calibration artifacts and help to evaluate optical imaging technologies.

Overexpression of a chrysanthemum transcription factor gene DgNAC1 improves the salinity tolerance in chrysanthemum.

KEY MESSAGE: DgNAC1, a transcription factor of chrysanthemum, was functionally verified to confer salt stress responses by regulating stress-responsive genes. NAC transcription factors play effective roles in resistance to different abiotic stresses, and overexpressions of NAC TFs in Arabidopsis have been proved to be conducive in improving salinity tolerance. However, functions of NAC genes in chrysanthemum continue to be poorly understood. Here, we performed physiology and molecular experiments to evaluate roles of DgNAC1 in chrysanthemum salt stress responses. In this study, DgNAC1-overexpressed chrysanthemum was obviously more resistant to salt over the WT (wild type). Specifically, the transgenic chrysanthemum showed a higher survival rate and lower EC (electrolyte conductivity) than WT under salt stress. The transgenic chrysanthemum also showed fewer accumulations of MDA (malondialdehyde) and reactive oxygen species (H2O2 and O2-), greater activities of SOD (superoxide dismutase), POD (peroxidase) and CAT (catalase), as well as more proline content than WT under salt stress. Furthermore, stress-responsive genes in transgenic chrysanthemum were greater up-regulated than in WT under salinity stress. Thus, all results revealed that DgNAC1 worked as a positive regulator in responses to salt stress and it may be an essential gene for molecular breeding of salt-tolerant plants.

Improved hydrogen production in the microbial electrolysis cell by inhibiting methanogenesis using ultraviolet irradiation.

Methanogenesis inhibition is essential for the improvement of hydrogen (H2) yield and energy recovery in the microbial electrolysis cell (MEC). In this study, ultraviolet (UV) irradiation was proposed as an efficient method for methanogenesis control in a single chamber MEC. With 30 cycles of operation with UV irradiation in the MEC, high H2 concentrations (>91%) were maintained, while without UV irradiation, CH4 concentrations increased significantly and reached up to 94%. In the MEC, H2 yields ranged from 2.87 ± 0.03 to 3.70 ± 0.11 mol H2/mol acetate with UV irradiation and from 3.78 ± 0.12 to 0.03 ± 0.004 mol H2/mol acetate without UV irradiation. Average energy efficiencies from the UV-irradiated MEC were 1.5 times of those without UV irradiation. Energy production from the MEC without UV irradiation was a negative energy yield process because of large amount of CH4 produced over time, which was mainly attributable to cathodic hydrogenotrophic methanogenesis. Our results clearly showed that UV irradiation could effectively inhibit methanogenesis and improve MEC performance to produce H2.

Autotrophic degradation of sulfamethoxazole using sulfate-reducing biocathode in microbial photo-electrolysis system.

Sulfamethoxazole is a representative of sulfonamide antibiotic pollutants. This study aims to investigate the degradation pathways of sulfamethoxazole and the response of microbial communities using the autotrophic biocathode in microbial photo-electrolysis systems (MPESs). Sulfamethoxazole with an initial concentration of 2 mg L-1 was degraded into small molecule propanol within 6 h with the biocathode. Elemental sulfur (S0) was detected in the cathode chamber, accounting for 57 % of the removed sulfate. The conversion from sulfate to S0 indicated that autotrophic microorganisms might adopt a novel pathway for sulfamethoxazole removal in the MPES. In the abiotic cathode, sulfamethoxazole degradation rate was 0.09 mg L-1 h-1 with the electrochemistry process. However, sulfamethoxazole was converted to products that still contain benzene rings, including p-aminothiophenol, 3-amino-5-methylisoxazole, and sulfonamide. The microbial community analysis indicated that the synergistic interaction of Desulfovibrio and Acetobacterium promoted the autotrophic degradation of sulfamethoxazole. The results suggested that autotrophic microorganisms may play an important role in the environmental transformation of sulfamethoxazole.

Natural-inspired hierarchic double-Janus solar evaporator for stable clean water production from high-salinity emulsions.

Interfacial solar evaporation shows great potential in clean water production, emulsions separation, and high-salinity brine treatment. However, it remains challenging for the evaporators to maintain a high evaporation rate in the high-salinity emulsions due to the co-pollution of salt and oil. Herein, we first proposed a hierarchic double-Janus solar evaporator (HDJE) with a hydrophobic salt-rejecting top layer and oil-rejecting bottom layer. Compared to the traditional one, HDJE could treat industrial high-salinity oil-in-water emulsions stably for over 70 h, with a stable average evaporation rate of 1.73 kg m-2 h-1 and a high purification efficiency of up to 99.8 % for oil and ions. It was also verified that HDJE could be used for high-efficiency purification of oily concentrated seawater outdoor. An average water production rate of 3.59 kg m-2 d-1 and a TOC removal ratio of over 98 % was obtained. In conclusion, this study provides a novel way to effectively dispose of high-salinity oily wastewater.

Structural Design and Energy and Environmental Applications of Hydrogen-Bonded Organic Frameworks: A Systematic Review.

Hydrogen-bonded organic frameworks (HOFs) are emerging porous materials that show high structural flexibility, mild synthetic conditions, good solution processability, easy healing and regeneration, and good recyclability. Although these properties give them many potential multifunctional applications, their frameworks are unstable due to the presence of only weak and reversible hydrogen bonds. In this work, the development history and synthesis methods of HOFs are reviewed, and categorize their structural design concepts and strategies to improve their stability. More importantly, due to the significant potential of the latest HOF-related research for addressing energy and environmental issues, this work discusses the latest advances in the methods of energy storage and conversion, energy substance generation and isolation, environmental detection and isolation, degradation and transformation, and biological applications. Furthermore, a discussion of the coupling orientation of HOF in the cross-cutting fields of energy and environment is presented for the first time. Finally, current challenges, opportunities, and strategies for the development of HOFs to advance their energy and environmental applications are discussed.

Enhanced metronidazole removal in seawater using a single-chamber bioelectrochemical system.

The aim of this study was to investigate the removal of metronidazole (MNZ) from seawater using a bioelectrochemical system (BES). Single-chamber BES (i.e., S-BES) and dual-chamber BES (i.e., D-BES) were constructed with carbon brush as the anode and cathode. With the inoculum of sea mud and 2 g/L of glucose as the substrate in seawater, S-BES and D-BES were acclimated to test the MNZ removal. Results showed that S-BES could remove almost 100 % of 200 mg/L MNZ within 120 h and remain stable within 10 cycles of operation (∼50 d) under the applied voltage of 0.8 V. The MNZ removal reached ∼100 % and 60.2 % in the cathodic and anodic chambers of D-BES fed by 100 mg/L MNZ under 0.8 V, respectively. The MNZ concentration of 200 mg/L significantly inhibited the sulfur metabolism, decreased the ratio of live to dead cells in the electrode biofilms, and thus reduced the SO42- removal in the S-BES. The MNZ degradation and S2- oxidation was mainly attributed to the cathodic and anodic biofilms of S-BES, respectively. Three degradation pathways of MNZ were proposed based on the identified intermediates and results of density functional theory calculations. The synergies among different genus species in the bacterial communities of biofilms, and between anodic and cathodic reactions could be responsible for the high performance of S-BES. Results from this study should be not only useful for the MNZ removal but also for effective MNZ inhibition of sulfate-reducing bacteria induced microbiologically influenced corrosion in seawater.

Highly efficient selective extraction of Li from spent LiNixCoyMnzO2 assisted with activated pyrite in a subcritical water system.

As the strategic importance of Li in the energy sector increases, selective Li extraction technology from spent lithium-ion batteries (LIBs) is attracting increasing attention. Current Li extraction processes typically suffer from lengthy procedures, high costs, and low efficiency. To improve the efficiency of Li extraction, a novel approach to achieve efficient Li recovery is proposed in this study, namely, reacting pyrite (FeS2) with LiNixCoyMnzO2 (NCM) powder in a subcritical water reduction (SWR) system. The reducing solvent environment created by the enhanced reaction of FeS2 with subcritical water converts the high-valent metals in NCM to a low-valent state, causing the collapse of the stable laminar structure and allowing Li+ to be released smoothly. After dual activation through mechanochemical and roasting processes, more than 99 % of Li is preferentially extracted under optimal conditions. Furthermore, Li+ in solution is converted into highly pure Li2CO3, while other metallic elements remain in the residue. Using inexpensive FeS2 for efficient Li extraction without adding additional chemical reagents is a promising approach for recovering spent LIBs.

Comprehensive profiles of sulfur cycling microbial communities along a mangrove sediment depth.

The sulfur (S) cycle is an important biogeochemical cycle with profound implications for both cellular- and ecosystem-level processes by diverse microorganisms. Mangrove sediments are a hotspot of biogeochemical cycling, especially for the S cycle with high concentrations of S compounds. Previous studies have mainly focused on some specific inorganic S cycling processes without paying specific attention to the overall S-cycling communities and processes as well as organic S metabolism. In this study, we comprehensively analyzed the distribution, ecological network and assembly mechanisms of S cycling microbial communities and their changes with sediment depths using metagenome sequencing data. The results showed that the abundance of gene families involved in sulfur oxidation, assimilatory sulfate reduction, and dimethylsulfoniopropionate (DMSP) cleavage and demethylation decreased with sediment depths, while those involved in S reduction and dimethyl sulfide (DMS) transformation showed an opposite trend. Specifically, glpE, responsible for converting S2O32- to SO32-, showed the highest abundance in the surface sediment and decreased with sediment depths; in contrast, high abundances of dmsA, responsible for converting dimethyl sulfoxide (DMSO) to DMS, were identified and increased with sediment depths. We identified Pseudomonas and Streptomyces as the main S-cycling microorganisms, while Thermococcus could play an import role in microbial network connections in the S-cycling microbial community. Our statistical analysis showed that both taxonomical and functional compositions were generally shaped by stochastic processes, while the functional composition of organic S metabolism showed a transition from stochastic to deterministic processes. This study provides a novel perspective of diversity distribution of S-cycling functions and taxa as well as their potential assembly mechanisms, which has important implications for maintaining mangrove ecosystem functions.

Overexpression of a novel chrysanthemum Cys2/His2-type zinc finger protein gene DgZFP3 confers drought tolerance in tobacco.

A drought stress-responsive Cys2/His2-type zinc finger protein gene DgZFP3 was previously isolated (Liu et al., Afr J Biotechnol 11:7781-7788, 2012b) from chrysanthemum. To assess roles of DgZFP3 in plant drought stress responses, we performed gain-of-function experiment. The DgZFP3-overexpression tobacco plants showed significant drought tolerance over the wild type (WT). The transgenic lines exhibited less accumulation of H2O2 under drought stress, more accumulation of proline and greater activities of peroxidase (POD) and superoxide dismutase than the WT under both control conditions and drought stress. In addition, there was greater up-regulation of the ROS-related enzyme genes (NtSOD and NtPOD) and stress-related genes (NtLEA5 and NtDREB) in transgenic lines under normal or drought conditons. Thus DgZFP3 probably plays a positive regulatory role in drought stress response and has the potential to be utilized in transgenic breeding to improve drought stress tolerance in plants.

A new oxic-settling-anaerobic (NOSA(®)) activated sludge process for minimizing excess sludge in secondary biological treatment plants: a pilot-scale evaluation of the absorption-biodegradation process.

This study compared the sludge reduction performance of a new oxic-settling-anaerobic (NOSA) process with that of a conventional adsorption-biodegradation process. A 50 m(3)/d pilot trial system with two different process configurations was operated for 6 months. The NOSA process functioned effectively in removing both chemical oxygen demand and nitrogen with the efficiencies of 86 and 92.5%, respectively, which reduced approximately 40% of the excess sludge. In this research, 0.77 kg volatile suspended solids/d sludge vanished in the anaerobic tank, which accounted for 58.9% of the total sludge loss in the NOSA process. Economic calculation suggests that the new process can dramatically upgrade the sludge reduction in wastewater treatment plants without a digestion device, and the investment for fundamental upgrading can be recovered in 5-6 years by cutting the costs of excess sludge dewatering and disposal treatment.