Effects of wastewater treatment plant effluent on microbial risks of pathogens and their antibiotic resistance in the receiving river.

The increase in effluent discharge from wastewater treatment plants (WWTPs) into urban rivers has raised concerns about the potential effects on pathogen risks. This study utilized metagenomic sequencing combined with flow cytometry to analyze pathogen concentrations and antibiotic resistance in a typical effluent-receiving river. Quantitative microbial risk assessment (QMRA) was employed to assess the microbial risks of pathogens. The results indicated obvious spatial-temporal differences (i.e., summer vs. winter and effluent vs. river) in microbial composition. Microcystis emerged as a crucial species contributing to these variations. Pathogen concentrations were found to be higher in the river than in the effluent, with the winter exhibiting higher concentrations compared to the summer. The effluent discharge slightly increased the pathogen concentrations in the river in summer but dramatically reduced them in winter. The combined effects of cyanobacterial bloom and high temperature were considered key factors suppressing pathogen concentrations in summer. Moreover, the prevalence of antibiotic resistance of pathogens in the river was inferior to that in the effluent, with higher levels in winter than in summer. Three high-concentration pathogens (Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa) were selected for QMRA. The results showed that the risks of pathogens exceeded the recommended threshold value. Escherichia coli posed the highest risks. And the fishing scenario posed significantly higher risks than the walking scenario. Importantly, the effluent discharge helped reduce the microbial risks in the receiving river in winter. The study contributes to the management and decision-making regarding microbial risks in the effluent-receiving river.

Deciphering solute transport, microbiota assembly patterns and metabolic functions in the hyporheic zone of an effluent-dominated river.

We lack a clear understanding of how anthropogenic pressures, exemplified by effluent discharge from wastewater treatment plants, destabilize microbial communities in the hyporheic zone (HZ) of receiving rivers. In this study, the spatiotemporal characteristics of hydrological parameters, and the physicochemical properties of surface and subsurface water in a representative effluent-dominated river were monitored. Sequencing of 16S rRNA amplicons and metagenomes revealed the microbial community structure in the HZ of both effluent discharge area and downstream region. The keystone taxa (taxa vital in determining the composition of each microbial cluster) and the keystone functions they controlled were subsequently identified. Effluent discharge amplified the depth of the oxic/suboxic zone and the hyporheic exchange fluxes in the effluent discharge area, which was 50-120% and 40-300% higher than in the downstream region, respectively. Microbial community structure pattern analysis demonstrated an enhancement in the rate of dispersal, an increase in microbial diversity, and an improved community network complexity in the effluent discharge area. By contrast, the number of keystone taxa in the effluent discharge area was 50-70% lower than that of the downstream region, resulting in reduced community network stability and functionality. The keystone taxa controlling metabolic functions in the networks categorized to effluent discharge area were comprised of more genera related to nitrogen and sulfur cycling, e.g., Dechloromonas, Desulfobacter, Flavobacterium, Nitrosomonas, etc., highlighting a research need in monitoring species associated with nutrient element cycling in the HZ of receiving waterbodies. The results showed that the keystone taxa could contribute positively to network stability, which was negatively correlated to hyporheic exchange fluxes and redox gradients. This study provides valuable insights that will improve our understanding of how river ecosystems respond to changes in anthropogenic pressures.

How redox gradient potentially influences nitrate reduction coupled with sulfur cycling: A new insight into nitrogen cycling in the hyporheic zone of effluent-dominated rivers.

The coupled N and S cycling in variable redox gradients in the hyporheic zone (HZ) of the rivers receiving effluents from wastewater treatment plants is unclear. Using two representative effluent-dominated rivers as model systems, a metagenome approach was employed to explore the spatiotemporal redox zonation of the HZ and the N/S cycling processes within the system. The results manifested that nitrate reduction represented the fundamental nitrogen pathway in the HZ. Interestingly, DNRA coupled with sulfur reduction, and denitrification coupled with sulfur oxidation were respectively abundant in the oxic and anoxic zone. Lower nitrate concentration (0-2.72 mg-N/L) and more abundant genes involved in denitrification (napB, NarGHI) and sulfur oxidation (sseA, glpE) were detected in the anoxic zone. Contrarily, the nitrate concentration (0.07-4.87 mg-N/L) and the abundance of genes involved in sulfur reduction (ttrB, sudA) and DNRA (nirBD) were observed more abundant in the oxic zone. Therefore, the results verified the oxygen-limited condition did not suppress but rather facilitated the denitrification process in the presence of active S cycling. The high relative abundances of nosZ gene encoding sequence (3-5 % relative to all nitrogen-cycling processes) in both the effluent-discharging area and downstream area highly confirmed that HZ was capable of alleviating the N2O emission in the region. The functional keystone taxa were revealed through co-occurrence network analysis. The structural equation model shows that the genes of N/S cycling were positively impacted by functional keystone taxa, especially the N cycling genes. Functional keystone taxa were proven driven by the redox gradient, demonstrating their positive roles in mediating N/S cycling processes. The promoting effect on nitrate reduction coupled with sulfur cycling was clarified when redox conditions oscillated, providing a new perspective on mitigating nitrogen pollution and greenhouse gas emissions in effluent-receiving rivers.

Prevalence of antibiotic resistance genes and virulence factors in the sediment of WWTP effluent-dominated rivers.

In the context of increasing aridity due to climate changes, effluent from wastewater treatment plants (WWTPs) became dominant in some rivers. However, the prevalence of antibiotic resistance genes (ARGs) and virulence factors (VFs) in effluent-dominated rivers was rarely investigated. In this study, the profiles of ARGs and VFs in the sediment of two effluent-dominated rivers were revealed through the metagenomic sequencing technique. In each river, samples from the effluent discharge point (P site) and approximately 500 m downstream (D site) were collected. Results showed that the abundances of ARGs and VFs were both higher in D sites than those in P sites, indicating higher risks in the downstream areas. The compositions of ARGs were similar in the P sites of two rivers while being distinct in the D sites. The same was true for changes in the VFs compositions. Microbial community structure variations were the main driver for the changes in ARGs and VFs. Network analysis revealed that the interaction of ARGs and VF genes (VFGs) in sediment was intense. Two VFGs and eleven ARGs were identified to play important roles in the network. Metagenome-assembled genomes (MAGs) were generated to evaluate the coexistence of ARGs and VFGs at the single genome level. It was found that 38.4 % of the MAGs contained both ARGs and VFGs, and two MAGs were from pathogenic genera. These results suggested that high microbiological risks existed in effluent-dominated rivers, and necessary measures should be taken to prevent the potential threat to public health.

Deep insight into oriented propionate production from food waste: Microbiological interpretation and design practice.

Using mixed microbial cultures (MMCs) for oriented volatile fatty acids (VFAs) refining in an open environment is a typical challenge due to the microbial diversiform and the process complexity. Especially for carbohydrate-rich waste (such as food waste), butyrate-type fermentation is usually dominant in a single-stage MMCs anaerobic process, while the production of odd-carbon VFAs (such as propionate) is difficult although it plays a significant role in chemicals industries. In this study, firstly, we gave a new perspective on the rationality of the oriented propionate production using MMCs with lactate as feedstock by conducting in-depth microbial informatics and reaction analysis. Secondly, we verified the feasibility of the "food waste-lactate-propionate" route to reverse the original butyrate-type fermentation situation and explore mechanisms for maintaining stability. In the first stage, a defined lactate fermentation microbiome was used to produce lactate-containing broth (80% of total chemical oxygen demand) at pH=4. In the second stage, an undomesticated undefined anaerobic microbiome was used to drive propionate production (45.26% ± 2.23% of total VFAs) under optimized conditions (C/N = 100:1-200:1 and pH=5.0). The low pH environment in the first stage enhanced the lactic acid bacteria to resist the invasion of non-functional flanking bacteria, making the community stable. In the second stage, the system maintained the propionate-type fermentation due to the absence of the ecological niche of the invasive lactic acid bacteria; The selection of propionate-producing specialists was a necessary but not sufficient condition for propionate-type fermentation. At last, this study proposed an enhanced engineering strategy framework for understanding elaborate MMCs fermentation.

Effect of pH on the mitigation of extracellular/intracellular antibiotic resistance genes and antibiotic resistance pathogenic bacteria during anaerobic fermentation of swine manure.

Effects of various initial pH values (i.e., 3, 5, 7, 11) during anaerobic fermentation of swine manure on intracellular and extracellular antibiotic resistance genes (iARGs and eARGs) and ARG-carrying potential microbial hosts were investigated. The abundance of almost all iARGs and eARGs decreased by 0.1-1.7 logs at pH 3 and pH 5. The abundance of only three iARGs and eARGs decreased by 0.1-0.9 logs at pH 7 and pH 11. Under acidic initial fermentation conditions (pH 3 and pH 5), the ARG removal effect was more pronounced. Acidic conditions (pH 3 and pH 5) significantly reduced the diversity and abundance of the microbial community, thereby eliminating many potential ARG hosts and antibiotic-resistant pathogenic bacteria (ARPB). Therefore, the study results contribute to the investigation of the effects of swine manure anaerobic fermentation on the removal and risk of contamination of ARGs and ARPB.

Insights into antibiotic stewardship of lake-rivers-basin complex systems for resistance risk control.

Antibiotic stewardship is hindered by a lack of consideration for complicated environmental fate of antibiotics and their role in resistance development, while the current methodology of eco-toxicological risk assessment has not been fully protective against their potential to select for antibiotic resistance. To address this problem, we established a novel methodologic framework to perform comprehensive environmental risk assessment of antibiotics in terms of resistance development, which was based on selection effect, phenotype resistance level, heteroresistance frequency, as well as prevalence and stability of antibiotic resistance genes. We tracked the contribution of antibiotic load reduction to the mitigation of environmental risk of resistance development by fate and transport modeling. The method was instantiated in a lake-river network-basin complex system, taking the Taihu Basin as a case study. Overall, antibiotic load posed no eco-toxicological risk but an average medium-level environmental risk for resistance development in Taihu Lake. The effect of antibiotic load on resistance risk was both seasonal-dependent and category-dependent, while quinolones posed the greatest environmental risk for resistance development. Mass-flow analysis indicated that temporal-spatial variation in hydrological regime and antibiotic fate together exerted a significant effect on antibiotic load in the system. By apportioning antibiotic load to riverine influx, we identified the hotspots for load reduction and predicted the beneficial response of resistance risk under load-reduction scenarios. Our study proposed a risk-oriented strategy of basin-scaled antibiotic load reduction for environmental risk control of resistance development.

Comprehensive Techno-environmental Evaluation of a Pilot-Scale PHA Production from Food Waste in China.

Polyhydroxyalkanoates (PHAs), a biodegradable plastic that might replace petroleum-based plastics, can be recovered from organic waste using mixed microbial cultures (MMCs). Research in this field has been ongoing for about 25 years and is now in a critical commercialization period. However, few pilot-scale studies are available to analyze its technical feasibility and environmental impact. We ran an MMC PHA production pilot plant for 6 months using local food waste as the feedstock. The traditional three-stage process achieved PHA content of 47.91 ± 1.91% dry cell weight and volumetric productivity of 9.94 ± 0.01 g/L·d, while a novel rapid proliferation stage was built in, the PHA content and productivity could reach 41.39 ± 2.39% cell dry weight and 20.02 ± 0.01 g/L·d, respectively. Life cycle assessment using field data showed that greenhouse warming potential was much more than five times that of the known literature, and the fossil depletion potential was 10.30 (scenario #1)/7.59 (scenario #2) times higher than petroleum-based polyethylene (PE) plastic. However, establishing a resource-energy-water union instead of an isolated plant could achieve environmental benefits compared to PE plastic. This techno-environmental analysis provides emerging MMC PHA producers worldwide with a valuable reference for further development opportunities and market planning.

Deciphering the extracellular and intracellular antibiotic resistance genes in multiple environments reveals the persistence of extracellular ones.

The extracellular and intracellular antibiotic resistance genes (eARGs and iARGs) together constitute the entire resistome in environments. However, the systematic analysis of eARGs and iARGs was still inadequate. Three kinds of environments, i.e., livestock manure, sewage sludge, and lake sediment, were analyzed to reveal the comprehensive characteristics of eARGs and iARGs. Based on the metagenomic data, the diversities, relative abundances, and compositions of eARGs and iARGs were similar. The extracellular and intracellular integrons and insertion sequences (ISs) also did not show any significant differences. However, the degree and significance of the correlation between total relative abundances of integrons/ISs and ARGs were lower outside than inside the cells. Gene cassettes carried by class 1 integron were amplified in manure and sludge samples, and sequencing results showed that the identified ARGs extracellularly and intracellularly were distinct. By analyzing the genetic contexts, most ARGs were found located on chromosomes. Nevertheless, the proportion of ARGs carried by plasmids increased extracellularly. qPCR was employed to quantify the absolute abundances of sul1, sul2, tetO, and tetW, and their extracellular proportions were found highest in sludge samples. These findings together raised the requirements of considering eARGs and iARGs separately in terms of risk evaluation and removal management.

Anaerobic digestion reduces extracellular antibiotic resistance genes in waste activated sludge: The effects of temperature and degradation mechanisms.

The fate of extracellular antibiotic resistance genes (eARGs) in waste activated sludge during anaerobic digestion (AD) remained unclear. The current study investigated the changes in seven eARGs (sulI, sulII, tet(A), tet(O), tet(X), blaTEM, and blaSHV) and intI1 during sludge AD at 35 °C and 55 °C. First, the extracellular DNA (eDNA) extraction method from sludge was optimized by adding sodium dodecyl sulfonate, and the eDNA recovery nearly doubled. Second, analysis via qPCR showed that eARGs ranged from 1.5% to 85.1% of the total ARGs, stressing the importance of eARGs in sludge. Besides, the abundances of all detected eARGs decreased following AD, where removal rates ranged from 22.8% to 93.9% at 35 °C and 52.7% to 96.6% at 55 °C. Further analysis showed that the removal rates of eARGs were negatively correlated with their initial abundances (P < 0.05). Last, the degradation characteristics of eARGs under AD conditions were determined. The first-order degradation rate constants for different eARGs did not vary significantly, indicating that gene sequences did not cause a removal distinction, and fitted Michaelis-Menten equation confirmed the higher eARGs degradation ability at the higher temperature. Overall, this study firstly uncovered the decrease of eARGs in sludge during AD treatment, and advanced the understanding of the positive effect of AD on eARGs dissemination control.

Functional structures of soil microbial community relate to contrasting N2O emission patterns from a highly acidified forest.

Nitrous oxide (N2O) is an important greenhouse gas contributing to global climate change. Emissions of N2O from acidic forests are increasing rapidly; however, little is known about the mechanisms driving these emissions. We analyzed soil samples from a high N2O emission area (HEA, 224-601 µg N m-2 h-1) and an adjacent low emission area (LEA, 20-30 µg N m-2 h-1) of a highly acidified forest. HEA showed similar carbon and nitrogen (N) pools and microbial biomass to LEA, but significantly higher moisture and extractable nutrients than LEA did. GeoChip 4 detected 298 gene families (unadjusted P < 0.05; 94, adjusted P < 0.05) showing significantly different structures between HEA and LEA. Both areas had highly diverse N cycling functional genes. However, HEA had higher relative abundances of nor, P450nor, and archaeal nitrifier nirK, which provided evidence for the importance of denitrifiers in N2O emission. HEA also showed significantly higher relative abundances of lignin- and cellulose-degrading genes, oxygen-limitation-response genes and denitrifier ppk, but lower abundances of N- and phosphorus (P) -limitation-response genes especially denitrifier pstS, corresponding to the higher moisture and extractable nutrients conducive to denitrification. The moisture, extractable nutrients and pH explained over 50% variation in microbial communities, and extractable P appeared as the key factor driving community variation and consequently regulated N2O production. CAPSULE ABSTRACT: N2O emission in highly acidified forest soils was related to the diverse N functional genes, especially denitrification genes, and was affected by soil properties.

Reducing the effect of non-volatile fatty acids (non-VFAs) on polyhydroxyalkanoates (PHA) production from fermented thermal-hydrolyzed sludge.

Thermal-hydrolyzed sludge (THS) can be fermented to produce volatile fatty acids (VFAs) rich liquids. These fermentative liquids are considered as a potential feedstock for polyhydroxyalkanoates (PHA) production. However, the presence of high levels of non-VFAs organics supporting the growth response instead of PHA accumulation hindered an efficient culture selection in a feast and famine regime. Lowering the non-VFAs content can compromise activities of microorganisms to take up these external carbon sources and improve the selective pressure; thus two enhanced strategies were tested to optimize the selection process: 1) increasing the proportion of VFAs in the original substrate or 2) removing most of the non-VFAs at the end of the feast phase. Results showed that the strategies resulted in PHA yields on VFAs respectively of 0.62 and 0.54 Cmol/Cmol, significantly higher than that in original SBR (0.16-0.35 Cmol/Cmol), confirming that reducing the adverse effect of non-VFAs can impose effective internal limitation and induce high PHA storage responses. In PHA accumulation tests, cultures selected with synthetic substrates accumulated a maximum PHA content of 61.4 wt%, which is the highest as ever reported among PHA production from THS. In summary, the study provided valuable references for improving PHA production from complex substrates.

New insight into fates of sulfonamide and tetracycline resistance genes and resistant bacteria during anaerobic digestion of manure at thermophilic and mesophilic temperatures.

This study investigated the variations in antibiotic (sulfonamide and tetracycline) resistance genes (ARGs) and resistant bacteria (ARB) during manure anaerobic digestion (AD) at 35 ℃ and 55 ℃, and discussed the mechanisms of variations in ARGs. The AD lasted for 60 days, five ARGs and intI1 each decreased in abundance after AD at the thermophilic temperature, while only half decreased at the mesophilic temperature. On days 10, 30, and 60, sulfonamide and tetracycline ARB were screened on selective media. During thermophilic AD, ARB numbers reduced by 4-log CFUs per gram dry manure, but only by approximately 1-log CFU at the mesophilic temperature. However, ARB composition analysis showed that at either temperature, no significant reduction in identified ARB species was observed. Furthermore, 72 ARB clones were randomly selected to detect the ARGs they harbored, and the results showed that each ARG was harbored by various hosts, and no definitive link existed between ARGs and bacterial species. In addition, by comparison with the identified host by culture method, the host prediction results based on the correlation analysis between ARGs and the bacterial community was proven to be unreliable. Overall, these findings indicated that relationships between ARB and ARGs were intricate.

Impacts of aeration management and polylactic acid addition on dissolved organic matter characteristics in intensified aquaponic systems.

Aquaponics as a potential alternative for conventional aquaculture industry has increasingly attracted worldwide attention in recent years. However, the sustainable application of aquaponics is facing a growing challenge. In particular, there is a pressing need to better understand and control the accumulation of dissolved organic matter (DOM) in aquaponics with the aim of optimizing nitrogen utilization efficiency. This study was aiming for assessing the characteristics of DOM in the culture water and the relationship with the nitrogen transformations in different intensified aquaponic systems with hydroponic aeration supplement and polylactic acid (PLA) addition. Two enhancing attempts altered the quantity of DOM in aquaponic systems significantly with a varying DOM content of 21.98-45.65 mg/L. The DOM could be represented by four identified fluorescence components including three humic -like materials (83-86%) and one tryptophan-like substance (14-17%). The fluorescence intensities of humic acid-like components were decreased significantly after the application of intensifying strategies, which indicating that two enhancing attempts possibly affected humic acid-like fluorescence. Variation of optical indices also suggested the reductions of water DOM which could be impacted by the enhancing nitrogen treatment processes. These findings will benefit the potential applications and sustainable operation of these strategies in aquaponics.

Mitigation of N2O Emission from Aquaponics by Optimizing the Nitrogen Transformation Process: Aeration Management and Exogenous Carbon (PLA) Addition.

N2O production in aquaponics is an inevitable concern when aquaponics is developed as a future production system. In the present study, two attempts were applied to mitigate N2O emission from aquaponics, i.e., aeration in hydroponic bed (HA) and addition of polylactic acid (PLA) into fillers (PA). Results showed that N2O emission from HA and PA was decreased by 47.1-58.1% and 43.2-74.9% respectively compared with that in control. Denitrification was proved to be the main emission pathway in all treatments, representing 62.4%, 86.4%, and 75.8% of the total N2O emission in HA, PA, and control, respectively. However, production of plants in HA was severely impaired, which was only 3.04 ± 0.39 kg/m2, while in PA and control, plants yields were 4.87 ± 0.56 kg/m2 and 4.33 ± 0.58 kg/m2. Combining the environmental and economic benefits, adding PLA in aquaponics may have a better future when developing and applying aquaponics systems.

Improving nitrogen utilization efficiency of aquaponics by introducing algal-bacterial consortia.

Aquaponics is a promising technology combining aquaculture with hydroponics. In this study, algal-bacterial consortia were introduced into aquaponics, i.e., algal-bacterial based aquaponics (AA), to improve the nitrogen utilization efficiency (NUE) of aquaponics. The results showed that the NUE of AA was 13.79% higher than that of media-based aquaponics (MA). In addition, higher NO3- removal by microalgae assimilation led to better water quality in AA, which made up for the deficiencies of poor aquaponic management of nitrate. As a result of lower NO3- concentrations and dramatically higher dissolved oxygen (DO) concentrations caused by microalgae photosynthesis in the photobioreactor, the N2O emission of AA was 89.89% lower than that of MA, although nosZ gene abundance in MA's hydroponic bed was approximately 30 times over that in AA. Considering the factors mentioned above, AA would improve the sustainability of aquaponics and have a good application foreground.

Improvement of aquaponic performance through micro- and macro-nutrient addition.

Aquaponics is one of the "zero waste" industry in the twenty-first century, and is considered to be one of the major trends for the future development of agriculture. However, the low nitrogen utilization efficiency (NUE) restricted its widely application. To date, many attempts have been conducted to improve its NUE. In the present study, effect of micro- and macro-nutrient addition on performance of tilapia-pak choi aquaponics was investigated. Results showed that the addition of micro- and macro-nutrients improved the growth of plant directly and facilitated fish physiology indirectly, which subsequently increased NUE of aquaponics from 40.42 to 50.64%. In addition, remarkable lower total phosphorus concentration was obtained in aquaponics with micro- and macro-nutrient addition, which was attributed to the formation of struvite. Most of the added micro-nutrients were enriched in plant root, while macro-nutrients mainly existed in water. Moreover, no enrichment of micro- and macro-nutrients in aquaponic products (i.e., fish and plant leaves) was observed, indicating that it had no influence on food safety. The findings here reported manifest that appropriate addition of micro- and macro-nutrients to aquaponics is necessary, and would improve its economic feasibility.

Effects of pH on nitrogen transformations in media-based aquaponics.

To investigate the effects of pH on performance and nitrogen transformations in aquaponics, media-based aquaponics operated at pH 6.0, 7.5 and 9.0 were systematically examined and compared in this study. Results showed that nitrogen utilization efficiency (NUE) reached its maximum of 50.9% at pH 6.0, followed by 47.3% at pH 7.5 and 44.7% at pH 9.0. Concentrations of nitrogen compounds (i.e., TAN, NO2(-)-N and NO3(-)-N) in three pH systems were all under tolerable levels. pH had significant effect on N2O emission and N2O conversion ratio decreased from 2.0% to 0.6% when pH increased from 6.0 to 9.0, mainly because acid environment would inhibit denitrifiers and lead to higher N2O emission. 75.2-78.5% of N2O emission from aquaponics was attributed to denitrification. In general, aquaponics was suggested to maintain pH at 6.0 for high NUE, and further investigations on N2O mitigation strategy are needed.

Attempts to improve nitrogen utilization efficiency of aquaponics through nitrifies addition and filler gradation.

Aquaponics has attracted worldwide attention in recent years and is considered as an alternative technology for conventional aquaculture. In this study, common carp (Cyprinus carpio) and pakchoi (Brassica chinensis) were cultured in lab-scale aquaponics, and attempts were conducted to enhance its nitrogen utilization efficiency (NUE) through two optimization methods, i.e., nitrifies addition (NA) and filler gradation (FG). Results showed that NA and FG could improve the NUE of aquaponics by 8.8 and 16.0%, respectively, compared with control. The total ammonia (TAN) and nitrite (NO2(-)) concentrations in NA and FG systems were maintained at relatively low level (TAN < 0.5 mg/L, NO2(-) < 0.1 mg/L), which demonstrated that both the NA and FG could provide non-toxic water environment for fish culture. Nitrous oxide conversion ratio of the control, NA, and FG were 0.8, 1.2, and 1.7%, respectively, indicating that media-based aquaponics also contributed to global warming. Although the two proposed attempts in this study caused more N2O emission, they made new breakthrough in improving the NUE of aquaponics.

Effect of phosphorus load on nutrients removal and N2O emission during low-oxygen simultaneous nitrification and denitrification process.

Three laboratory scale anaerobic-aerobic (low-oxygen) SBRs (R1, R2 and R3) were conducted at different influent phosphorus concentration to evaluate the impacts of phosphorus load on nutrients removal and nitrous oxide (N2O) emission during low-oxygen simultaneous nitrification and denitrification (SND) process. The results showed that TP and TN removals were enhanced simultaneously with the increase in phosphorus load. It was mainly caused by the enrichment of polyphosphate accumulating organisms (PAOs) under high phosphorus load and low COD/P ratio (<50), which could use nitrate/nitrite as electron acceptors to take up the phosphorus. N2O emission was reduced with increasing phosphorus load. N2O-N emission amount per cycle of R3 was 24.1% lower than that of R1. It was due to the decrease of N2O yield by heterotrophic denitrification. When the phosphorus load increased from R1 to R3, heterotrophic denitrification (D) ranged from 42.6% to 36.6% of the N2O yield.