Sludge fermentation liquid addition attained advanced nitrogen removal in low C/N ratio municipal wastewater through short-cut nitrification-denitrification and partial anammox

• Sludge fermentation liquid addition resulted in a high NAR of 97.4%. • Extra NH₄⁺-N from SFL was removed by anammox in anoxic phase. • Nitrogen removal efficiency of 92.51% was achieved in municipal wastewater. • The novel system could efficiently treat low COD/N municipal wastewater. Biological nitrogen removal of wastewater with low COD/N ratio could be enhanced by the addition of wasted sludge fermentation liquid (SFL), but the performance is usually limited by the introducing ammonium. In this study, the process of using SFL was successfully improved by involving anammox process. Real municipal wastewater with a low C/N ratio of 2.8–3.4 was treated in a sequencing batch reactor (SBR). The SBR was operated under anaerobic-aerobic-anoxic (AOA) mode and excess SFL was added into the anoxic phase. Stable short-cut nitrification was achieved after 46d and then anammox sludge was inoculated. In the stable period, effluent total inorganic nitrogen (TIN) was less than 4.3 mg/L with removal efficiency of 92.3%. Further analysis suggests that anammox bacteria, mainly affiliated with Candidatus_Kuenenia, successfully reduced the external ammonia from the SFL and contributed approximately 28%–43% to TIN removal. Overall, this study suggests anammox could be combined with SFL addition, resulting in a stable enhanced nitrogen biological removal.     

Enhancing the efficiency of nitrogen removing bacterial population to a wide range of C:N ratio (1.5:1 to 14:1) for simultaneous C & N removal

• Simultaneous C & N removal in Methammox occurs at wide C:N ratio. • Biological Nitrogen Removal at wide C:N ratio of 1.5:1 to 14:1 is not reported. • Ammonia removal shifted from mixotrophy to heterotrophy at high C:N ratio. • Acetogenic population compensated for ammonia oxidizers at high C:N ratio. • Methanogens increase the plasticity of nitrogen removers at high C:N ratio. High C:N ratio in the wastewater limits biological nitrogen removal (BNR), especially in anammox based technologies. The present study attempts to improve the COD tolerance of the BNR process by associating methanogens with nitrogen removing bacterial (NRB) populations. The new microbial system coined as ‘Methammox’, was investigated for simultaneous removal of COD (C) and ammonia (N) at C:N ratio 1.5:1 to 14:1. The ammonia removal rate (11.5 mg N/g VSS/d) and the COD removal rates (70.6 mg O/g VSS/d) of Methammox was close to that of the NRB (11.1 mg N/g VSS/d) and the methanogenic populations (77.9 mg O/g VSS/d), respectively. The activities established that these two populations existed simultaneously and independently in ‘Methammox’. Further studies in biofilm reactor fetched a balanced COD and ammonia removal (55%–60%) at a low C:N ratio (≤2:1) and high C:N ratio (≥9:1). The population abundance of methanogens was reasonably constant, but the nitrogen removal shifted from mixotrophy to heterotrophy as the C:N ratio shifted from low (C:N≤2:1) to high (C:N≥9:1). The reduced autotrophic NRB (ammonia- and nitrite-oxidizing bacteria and Anammox) population at a high C:N ratio was compensated by the fermentative group that could carry out denitrification heterotrophically. The functional plasticity of the Methammox system to adjust to a broad C:N ratio opens new frontiers in biological nitrogen removal of high COD containing wastewaters.     

Reutilize tire in microbial fuel cell for enhancing the nitrogen removal of the anammox process coupled with iron-carbon micro-electrolysis

• MFC promoted the nitrogen removal of anammox with Fe-C micro-electrolysis. • Reutilize pyrolysis waste tire as micro-electrolysis and electrode materials. • Total nitrogen removal efficiency of modified MFC increased to 85.00%. • Candidatus kuenenia and SM1A02 were major genera responsible for nitrogen removal. In this study, microbial fuel cells (MFCs) were explored to promote the nitrogen removal performance of combined anaerobic ammonium oxidation (anammox) and Fe-C micro-electrolysis (CAE) systems. The average total nitrogen (TN) removal efficiency of the modified MFC system was 85.00%, while that of the anammox system was 62.16%. Additionally, the effective operation time of this system increased from six (CAE system alone) to over 50 days, significantly promoting TN removal. The enhanced performance could be attributed to the electron transferred from the anode to the cathode, which aided in reducing nitrate/nitrite in denitrification. The H⁺ released through the proton exchange membrane caused a decrease in the pH, facilitating Fe corrosion. The pyrolyzed waste tire used as the cathode could immobilize microorganisms, enhance electron transport, and produce a natural Fe-C micro-electrolysis system. According to the microbial community analysis, Candidatus kuenenia was the major genus involved in the anammox process. Furthermore, the SM1A02 genus exhibited the highest abundance and was enriched the fastest, and could be a novel potential strain that aids the anammox process.     

Electricity-driven ammonia oxidation and acetate production in microbial electrosynthesis systems

• MES was constructed for simultaneous ammonia removal and acetate production. • Energy consumption was different for total nitrogen and ammonia nitrogen removal. • Energy consumption for acetate production was about 0.04 kWh/g. • Nitrate accumulation explained the difference of energy consumption. • Transport of ammonia and acetate across the membrane deteriorated the performance. Microbial electrosynthesis (MES) is an emerging technology for producing chemicals, and coupling MES to anodic waste oxidation can simultaneously increase the competitiveness and allow additional functions to be explored. In this study, MES was used for the simultaneous removal of ammonia from synthetic urine and production of acetate from CO₂. Using graphite anode, 83.2%±5.3% ammonia removal and 28.4%±9.9% total nitrogen removal was achieved, with an energy consumption of 1.32 kWh/g N for total nitrogen removal, 0.45 kWh/g N for ammonia nitrogen removal, and 0.044 kWh/g for acetate production. Using boron-doped diamond (BDD) anode, 70.9%±12.1% ammonia removal and 51.5%±11.8% total nitrogen removal was obtained, with an energy consumption of 0.84 kWh/g N for total nitrogen removal, 0.61 kWh/g N for ammonia nitrogen removal, and 0.043 kWh/g for acetate production. A difference in nitrate accumulation explained the difference of total nitrogen removal efficiencies. Transport of ammonia and acetate across the membrane deteriorated the performance of MES. These results are important for the development of novel electricity-driven technologies for chemical production and pollution removal.     

Simultaneous Feammox and anammox process facilitated by activated carbon as an electron shuttle for autotrophic biological nitrogen removal

• The autotrophic nitrogen removal combining Feammox and Anammox was achieved. • Activated carbon can be used as an electron shuttle to enhance Feammox activity. • Fe(III) was reduced to Fe(II) and the secondary Fe(II) mineral (FeOOH) was obtained. • The iron-reducing bacteria and Anammox consortium was enriched simultaneously. Ferric iron reduction coupled with anaerobic ammonium oxidation (Feammox) is a novel ferric-dependent autotrophic process for biological nitrogen removal (BNR) that has attracted increasing attention due to its low organic carbon requirement. However, extracellular electron transfer limits the nitrogen transformation rate. In this study, activated carbon (AC) was used as an electron shuttle and added into an integrated autotrophic BNR system consisting of Feammox and anammox processes. The nitrogen removal performance, nitrogen transformation pathways and microbial communities were investigated during 194 days of operation. During the stable operational period (days 126–194), the total nitrogen (TN) removal efficiency reached 82.9%±6.8% with a nitrogen removal rate of 0.46±0.04 kg-TN/m³/d. The contributions of the Feammox, anammox and heterotrophic denitrification pathways to TN loss accounted for 7.5%, 89.5% and 3.0%, respectively. Batch experiments showed that AC was more effective in accelerating the Feammox rate than the anammox rate. X-ray photoelectron spectroscopy (XPS) analyses showed the presence of ferric iron (Fe(III)) and ferrous iron (Fe(II)) in secondary minerals. X-ray diffraction (XRD) patterns indicated that secondary iron species were formed on the surface of iron-AC carrier (Fe/AC), and Fe(III) was primarily reduced by ammonium in the Feammox process. The phyla Anaerolineaceae (0.542%) and Candidatus Magasanikbacteria (0.147%) might contribute to the Feammox process, and Candidatus Jettenia (2.10%) and Candidatus Brocadia (1.18%) were the dominative anammox phyla in the bioreactor. Overall, the addition of AC provided an effective way to enhance the autotrophic BNR process by integrating Feammox and anammox.     

Partial anammox achieved in full scale biofilm process for typical domestic wastewater treatment

• A full scale biofilm process was developed for typical domestic wastewater treatment. • The HRT was 8 h and secondary sedimentation tank was omitted. • Candidatus Brocadia were enriched in the HBR with an abundance of 2.89%. • Anammox enabled a stable ammonium removal of ~15% in the anoxic zone. The slow initiation of anammox for treating typical domestic wastewater and the relatively high footprint of wastewater treatment infrastructures are major concerns for practical wastewater treatment systems. Herein, a 300 m³/d hybrid biofilm reactor (HBR) process was developed and operated with a short hydraulic retention time (HRT) of 8 h. The analysis of the bacterial community demonstrated that anammox were enriched in the anoxic zone of the HBR process. The percentage abundance of Candidatus Brocadia in the total bacterial community of the anoxic zone increased from 0 at Day 1 to 0.33% at Day 130 and then to 2.89% at Day 213. Based upon the activity of anammox bacteria, the removal of ammonia nitrogen (NH₄⁺-N) in the anoxic zone was approximately 15%. This showed that the nitrogen transformation pathway was enhanced in the HBR system through partial anammox process in the anoxic zone. The final effluent contained 12 mg/L chemical oxygen demand (COD), 0.662 mg/L NH₄⁺-N, 7.2 mg/L total nitrogen (TN), and 6 mg/L SS, indicating the effectiveness of the HBR process for treating real domestic wastewater.     

Overlooked nitrogen-cycling microorganisms in biological wastewater treatment

• AOA and comammox bacteria can be more abundant and active than AOB/NOB at WWTPs. • Coupled DNRA/anammox and NO_x-DAMO/anammox/comammox processes are demonstrated. • Substrate level, SRT and stressors determine the niches of overlooked microbes. • Applications of overlooked microbes in enhancing nitrogen removal are promising. Nitrogen-cycling microorganisms play key roles at the intersection of microbiology and wastewater engineering. In addition to the well-studied ammonia oxidizing bacteria, nitrite oxidizing bacteria, heterotrophic denitrifiers, and anammox bacteria, there are some other N-cycling microorganisms that are less abundant but functionally important in wastewater nitrogen removal. These microbes include, but not limited to ammonia oxidizing archaea (AOA), complete ammonia oxidation (comammox) bacteria, dissimilatory nitrate reduction to ammonia (DNRA) bacteria, and nitrate/nitrite-dependent anaerobic methane oxidizing (NO_x-DAMO) microorganisms. In the past decade, the development of high-throughput molecular technologies has enabled the detection, quantification, and characterization of these minor populations. The aim of this review is therefore to synthesize the current knowledge on the distribution, ecological niche, and kinetic properties of these “overlooked” N-cycling microbes at wastewater treatment plants. Their potential applications in novel wastewater nitrogen removal processes are also discussed. A comprehensive understanding of these overlooked N-cycling microbes from microbiology, ecology, and engineering perspectives will facilitate the design and operation of more efficient and sustainable biological nitrogen removal processes.     

A heterotrophic nitrification-aerobic denitrification bacterium Halomonas venusta TJPU05 suitable for nitrogen removal from high-salinity wastewater

• H. venusta TJPU05 showed excellent HN-AD ability at high salinity. • Successful expression of AMO, HAO, NAR and NIR confirmed the HN-AD ability of TJPU05. • H. venusta TJPU05 could tolerate high salt and high nitrogen environment. • H. venusta TJPU05 is a promising candidate for the bio-treatment of AW. A novel salt-tolerant heterotrophic nitrification and aerobic denitrification (HN-AD) bacterium was isolated and identified as Halomonas venusta TJPU05 (H. venusta TJPU05). The nitrogen removal performance of H. venusta TJPU05 in simulated water (SW) with sole or mixed nitrogen sources and in actual wastewater (AW) with high concentration of salt and nitrogen was investigated. The results showed that 86.12% of NH₄⁺-N, 95.68% of NO₃^–-N, 100% of NO₂^–-N and 84.57% of total nitrogen (TN) could be removed from SW with sole nitrogen sources within 24 h at the utmost. H. venusta TJPU05 could maximally remove 84.06% of NH₄⁺-N, 92.33% of NO₃^–-N, 92.9% of NO₂^–-N and 77.73% of TN from SW with mixed nitrogen source when the salinity was above 8%. The application of H. venusta TJPU05 in treating AW with high salt and high ammonia nitrogen led to removal efficiencies of 50.96%, 47.28% and 43.19% for NH₄⁺-N, NO₃^–-N and TN respectively without any optimization. Furthermore, the activities of nitrogen removal–related enzymes of the strain were also investigated. The successful detection of high level activities of ammonia oxygenase (AMO), hydroxylamine oxidase (HAO), nitrate reductase (NAR) and nitrite reductase (NIR) enzymes under high salinity condition further proved the HN-AD and salt-tolerance capacity of H. venusta TJPU05. These results demonstrated that the H. venusta TJPU05 has great potential in treating high-salinity nitrogenous wastewater.     

Removal,distribution and plant uptake of perfluorooctane sulfonate (PFOS) in a simulated constructed wetland system

• PFOS was removed by soil adsorption and plant uptake in the VFCW. • Uptake of PFOS by E. crassipes was more than that of C. alternifolius. • PFOS in wastewater can inhibit the removal of nutrients. • Dosing with PFOS changed the soil microbial community in the VFCW. A vertical-flow constructed wetland (VFCW) was used to treat simulated domestic sewage containing perfluorooctane sulfonate (PFOS). The removal rate of PFOS in the domestic sewage was 93%–98%, through soil adsorption and plant uptake, suggesting that VFCWs can remove PFOS efficiently from wastewater. The removal of PFOS in the VFCW was dependent on soil adsorption and plant uptake; moreover, the percentage of soil adsorption was 61%–89%, and was higher than that of the plants uptake (5%–31%). The absorption capacity of Eichhornia crassipes (E. crassipes) (1186.71 mg/kg) was higher than that of Cyperus alternifolius (C. alternifolius) (162.77 mg/kg) under 10 mg/L PFOS, and the transfer factor of PFOS in E. crassipes and C. alternifolius was 0.04 and 0.58, respectively, indicating that PFOS is not easily translocated to leaves from roots of wetland plants; moreover, uptake of PFOS by E. crassipes was more than that of C. alternifolius because the biomass of E. crassipes was more than that of C. alternifolius and the roots of E. crassipes can take up PFOS directly from wastewater while C. alternifolius needs to do so via its roots in the soil. The concentration of 10 mg/L PFOS had an obvious inhibitory effect on the removal rate of total nitrogen, total phosphorus, chemical oxygen demand, and ammonia nitrogen in the VFCW, which decreased by 15%, 10%, 10% and 12%, respectively. Dosing with PFOS in the wastewater reduced the bacterial richness but increased the diversity in soil because PFOS stimulated the growth of PFOS-tolerant strains.     

Characterization of CANON reactor performance and microbial community shifts with elevated COD/N ratios under a continuous aeration mode

COD/N at low ratios (0–0.82) improved N removals of CANON. CANON performance decreased after COD/N up to 0.82. The relative abundance of AOB decreased continuously with increasing COD/N. AOB outcompeted at a high COD load led to CANON failure. The relative abundance of AnAOB decreased and increased with increasing COD/N. The effects of increasing COD/N on nitrogen removal performance and microbial structure were investigated in a SBR adopting a completely autotrophic nitrogen removal over nitrite process with a continuous aeration mode (DO at approximately 0.15–0.2 mg/L). As the COD/N increased from 0.1 to≤0.59, the nitrogen removal efficiency (NRE) increased from 88.7% to 95.5%; while at COD/N ratios of 0.59–0.82, the NRE remained at 90.7%–95.5%. As the COD/N increased from 0.82 to 1.07, the NRE decreased continuously until reaching 60.1%. Nitrosomonas sp. (AOB) and Candidatus Jettenia (anammox bacteria) were the main functional genera in the SBR. As the COD/N increased from 0.10 to 1.07, the relative abundance of Nitrosomonas decreased from 13.4% to 2.0%, while that of Candidatus Jettenia decreased from 35% to 9.9% with COD/N<0.82 then increased to 45.4% at a COD/N of 1.07. Aerobic heterotrophic bacteria outcompeted AOB at high COD loadings (650 mg/L) because of oxygen competition, which ultimately led to deteriorated nitrogen removal performance.     

Nitrate removal to its fate in wetland mesocosm filled with sponge iron: Impact of influent COD/N ratio

• CW-Fe allowed a high-performance of NO₃^‒-N removal at the COD/N ratio of 0. • Higher COD/N resulted in lower chem-denitrification and higher bio-denitrification. • The application of s-Fe⁰ contributed to TIN removal in wetland mesocosm. • s-Fe⁰ changed the main denitrifiers in wetland mesocosm. Sponge iron (s-Fe⁰) is a porous metal with the potential to be an electron donor for denitrification. This study aims to evaluate the feasibility of using s-Fe⁰ as the substrate of wetland mesocosms. Here, wetland mesocosms with the addition of s-Fe⁰ particles (CW-Fe) and a blank control group (CW-CK) were established. The NO₃^‒-N reduction property and water quality parameters (pH, DO, and ORP) were examined at three COD/N ratios (0, 5, and 10). Results showed that the NO₃^‒-N removal efficiencies were significantly increased by 6.6 to 58.9% in the presence of s-Fe⁰. NH₄⁺-N was mainly produced by chemical denitrification, and approximately 50% of the NO₃^‒-N was reduced to NH₄⁺-N, at the COD/ratio of 0. An increase of the influent COD/N ratio resulted in lower chemical denitrification and higher bio-denitrification. Although chemical denitrification mediated by s-Fe⁰ led to an accumulation of NH₄⁺-N at COD/N ratios of 0 and 5, the TIN removal efficiencies increased by 4.5%‒12.4%. Moreover, the effluent pH, DO, and ORP values showed a significant negative correlation with total Fe and Fe (II) (P<0.01). High-throughput sequencing analysis indicated that Trichococcus (77.2%) was the most abundant microorganism in the CW-Fe mesocosm, while Thauera, Zoogloea, and Herbaspirillum were the primary denitrifying bacteria. The denitrifiers, Simplicispira, Dechloromonas, and Denitratisoma, were the dominant bacteria for CW-CK. This study provides a valuable method and an improved understanding of NO₃^‒-N reduction characteristics of s-Fe⁰ in a wetland mesocosm.     

Taxonomic and functional variations in the microbial community during the upgrade process of a full-scale landfill leachate treatment plant – from conventional to partial nitrification-denitrification

• Upgrade process was investigated in a full-scale landfill leachate treatment plant. • The optimization of DO can technically achieve the shift from CND to PND process. • Nitrosomonas was mainly responsible for ammonium oxidation in PND system. • An obviously enrichment of Thauera was found in the PND process. • Enhanced metabolic potentials on organics was found during the process update. Because of the low access to biodegradable organic substances used for denitrification, the partial nitrification-denitrification process has been considered as a low-cost, sustainable alternative for landfill leachate treatment. In this study, the process upgrade from conventional to partial nitrification-denitrification was comprehensively investigated in a full-scale landfill leachate treatment plant (LLTP). The partial nitrification-denitrification system was successfully achieved through the optimizing dissolved oxygen and the external carbon source, with effluent nitrogen concentrations lower than 150 mg/L. Moreover, the upgrading process facilitated the enrichment of Nitrosomonas (abundance increased from 0.4% to 3.3%), which was also evidenced by increased abundance of amoA/B/C genes carried by Nitrosomonas. Although Nitrospira (accounting for 0.1%–0.6%) was found to stably exist in the reactor tank, considerable nitrite accumulation occurred in the reactor (reaching 98.8 mg/L), indicating high-efficiency of the partial nitrification process. Moreover, the abundance of Thauera, the dominant denitrifying bacteria responsible for nitrite reduction, gradually increased from 0.60% to 5.52% during the upgrade process. This process caused great changes in the microbial community, inducing continuous succession of heterotrophic bacteria accompanied by enhanced metabolic potentials toward organic substances. The results obtained in this study advanced our understanding of the operation of a partial nitrification-denitrification system and provided a technical case for the upgrade of currently existing full-scale LLTPs.     

Degradation of refractory organics in concentrated leachate by the Fenton process: Central composite design for process optimization

• 90% total COD, 95.3% inert COD and 97.2% UV₂₅₄ were removed. • High R² values (over 95%) for all responses were obtained with CCD. • Operational cost was calculated to be 0.238 €/g COD_removed for total COD removal. • Fenton oxidation was highly-efficient method for inert COD removal. • BOD₅/COD ratio of leachate concentrate raised from 0.04 to 0.4. The primary aim of this study is inert COD removal from leachate nanofiltration concentrate because of its high concentration of resistant organic pollutants. Within this framework, this study focuses on the treatability of leachate nanofiltration concentrate through Fenton oxidation and optimization of process parameters to reach the maximum pollutant removal by using response surface methodology (RSM). Initial pH, Fe²⁺ concentration, H₂O₂/Fe²⁺ molar ratio and oxidation time are selected as the independent variables, whereas total COD, color, inert COD and UV₂₅₄ removal are selected as the responses. According to the ANOVA results, the R² values of all responses are found to be over 95%. Under the optimum conditions determined by the model (pH: 3.99, Fe²⁺: 150 mmol/L, H₂O₂/Fe²⁺: 3.27 and oxidation time: 84.8 min), the maximum COD removal efficiency is determined as 91.4% by the model. The color, inert COD and UV₂₅₄ removal efficiencies are determined to be 99.9%, 97.2% and 99.5%, respectively, by the model, whereas the total COD, color, inert COD and UV₂₅₄ removal efficiencies are found respectively to be 90%, 96.5%, 95.3% and 97.2%, experimentally under the optimum operating conditions. The Fenton process improves the biodegradability of the leachate NF concentrate, increasing the BOD₅/COD ratio from the value of 0.04 to the value of 0.4. The operational cost of the process is calculated to be 0.238 €/g COD_removed. The results indicate that the Fenton oxidation process is an efficient and economical technology in improvement of the biological degradability of leachate nanofiltration concentrate and in removal of resistant organic pollutants.     

Removal of arsenic by pilot-scale vertical flow constructed wetland

• VFCWs are effective for the treatment of arsenic-containing wastewater. • Arsenic removal did not affect the removal of nutrients, except for TP in CW500. • Arsenic removal was highest when the temperature peaked and the reed was in bloom. • Substrate accumulation contributed more to arsenic removal than plant absorption. Four pilot-scale Vertical Flow Constructed Wetlands (VFCWs) filled with gravel and planted with Phragmites australis were operated for seven months in the field to study the efficiency of arsenic removal in contaminated wastewater. The average arsenic removal efficiency by the VFCWs was 52.0%±20.2%, 52.9%±21.3%, and 40.3%±19.4% at the theoretical concentrations of 50 μg/L (CW50), 100 μg/L (CW100), and 500 μg/L (CW500) arsenic in the wastewater, respectively. The results also showed no significant differences in the removal efficiency for conventional contaminants (nitrogen, phosphorus, or chemical oxygen demand) between wastewater treatments that did or did not contain arsenic (P>0.05), except for phosphorus in CW500. The highest average monthly removal rate of arsenic occurred in August (55.9%–74.5%) and the lowest in November (7.8%–15.5%). The arsenic removal efficiency of each VFCW was positively correlated with temperature (P<0.05). Arsenic accumulated in both substrates and plants, with greater accumulation associated with increased arsenic concentrations in the influent. The maximum accumulated arsenic concentrations in the substrates and plants at the end of the experiment were 4.47 mg/kg and 281.9 mg/kg, respectively, both present in CW500. The translocation factor (TF) of arsenic in the reeds was less than 1, with most of the arsenic accumulating in the roots. The arsenic mass balance indicated that substrate accumulation contributed most to arsenic removal (19.9%–30.4%), with lower levels in plants (3.8%–9.5%). In summary, VFCWs are effective for the treatment of arsenic-containing wastewater.     

Tracking Cryptosporidium in urban wastewater treatment plants in a cold region: Occurrence,species and infectivity

• Cryptosporidium in WWTPs in a cold region was investigated in different seasons. • The overall removal efficiency of Cryptosporidium in WWTPs was over 84%. • The infectivity rate declined below 53% in effluents mainly due to disinfection. • The infectivity of Cryptosporidium increased with a seasonal drop in temperature. • Low temperature promotes binding protein retention and virulence genes expression. This study investigated the occurrence, species, infectivity and removal efficiency of Cryptosporidium spp. across typical wastewater treatment train. Samples from different process units were collected seasonally and synchronously from four wastewater treatment plants (WWTPs) in Northeastern China. Live Cryptosporidium oocysts were identified in most samples from both influent (97.50%) and effluent (90.00%) wastewaters of the four WWTPs, at an average density of 26.34 and 4.15 oocysts/L, respectively. The overall removal efficiency was 84.25%, and oocysts were mainly removed (62.01%) by the modified secondary sedimentation process. Ten Cryptosporidium species were identified in the effluent samples. C. andersoni, C. bovis, and C. ryanae were the three most prevalent species. Oocyst viability assays indicated no reduction of excystation rate during the primary and secondary wastewater treatments (varied in the range of 63.08%–68.50%), but the excystation rate declined to 52.21% in the effluent after disinfection. Notably, the Cryptosporidium oocysts showed higher infection intensity in the cold season (winter and spring) than that in summer and autumn. The influences of environmental temperature on virulence factors of Cryptosporidium were further examined. It was observed that more extracellular secretory proteins were bound on the oocyst surface and several virulence genes were expressed relatively strongly at low temperatures, both of which could facilitate oocyst adhesion, invasion, and host immune evasion. This research is of considerable interest since it serves as an important step towards more accurate panoramic recognition of Cryptosporidium risk reduction in WWTPs, and especially highlights the potential health risk associated with Cryptosporidium in cold regions/seasons.     

Deciphering the effect of sodium dodecylbenzene sulfonate on up-flow anaerobic sludge blanket treatment of synthetic sulfate-containing wastewater

• UASB reactor can work efficiently with high COD/SO₄^2- ratios when SDBS exists. • Outcome of the competition between SRB and MPA was affected by SDBS. • Presence of SDBS makes methanogens with H₂/CO₂ as a substrate dominant. • Microbial diversity decreases in the presence of SDBS. In this study, the effects of organic sulfur on anaerobic biological processes were investigated by operating two up-flow anaerobic sludge blanket (UASB) reactors with sodium dodecylbenzene sulfonate (SDBS) as a representative of organic sulfur. The results indicated that the specific methanogenic activity (SMA) and chemical oxygen demand (COD) removal efficiency of R2 (with SDBS added) were higher than those of R1 (without SDBS) when the COD/SO₄²⁻ ratio was above 5.0. However, when the COD/SO₄²⁻ ratio was lower than 5.0, the sulfate reduction efficiency of R2 was higher than that of R1. These results and the observed SDBS transformation efficiency in anaerobic reactors indicate that low concentrations of SDBS accelerate methane production and the continuous accumulation of SDBS does not weaken the reduction of sulfate. Similarly, the calculated electron flux for a COD/SO₄²⁻ ratio of 1.0 indicates that the utilization intensity of electrons by sulfate-reducing bacteria (SRB) in R2 was 36.48% higher than that of SRB in R1 and exceeded that of methane-producing archaea (MPA) under identical working conditions. Moreover, the addition of SDBS in R2 made sulfidogenesis the dominant reaction at low COD/SO₄²⁻, and Methanobacterium and Methanobrevibacter with H₂/CO₂ as the substrate and Desulfomicrobium were the dominant MPA and SRB, respectively. However, methanogenesis was still the dominant reaction in R1, and Methanosaeta with acetic acid as the substrate and Desulfovibrio were the dominant MPA and SRB, respectively.     

Insights into simultaneous anammox and denitrification system with short-term pyridine exposure: Process capability,inhibition kinetics and metabolic pathways

• Short-term effect of the pyridine exposure on the SAD process was investigated. • The SAA at 150 mg/L pyridine reduced by 56.7% of the maximum value. • Inhibition kinetics models and inhibitory parameters were indicated. • Collaboration of AnAOB, HDB and PDB promoted the SAD. • Possible metabolic pathways of nitrogen and pyridine were proposed. In-depth knowledge on the role of pyridine as a bottleneck restricting the successful application of anammox-based process treating refractory coking wastewater remains unknown. In this study, the effect of short-term pyridine addition on a simultaneous anammox and denitrification (SAD) system fed with 25–150 mg/L pyridine was explored. The short-term operation showed that the highest total nitrogen (TN) removal efficiency was achieved at 25–50 mg/L of pyridine. As the pyridine addition increased, the contribution of the anammox pathway in nitrogen removal decreased from 99.3% to 79.1%, while the denitrification capability gradually improved. The specific anammox activity (SAA) at 150 mg/L pyridine decreased by 56.7% of the maximum SAA. The modified non-competitive inhibition model indicated that the 50% inhibitory concentration (IC₅₀) of pyridine on anammox was 84.18 mg/L and the substrate inhibition constant (K_i) of pyridine for self-degradation was 135.19 mg/L according to the Haldane model. Moreover, high-throughput sequencing confirmed the abundance of Candidatus Kuenenia as the amount of anammox species decreased, while the amounts of denitrifiers and pyridine degraders significantly increased as the pyridine stress increased. Finally, the possible pathways of nitrogen bioconversion and pyridine biodegradation in the SAD system were elucidated through metagenomic analysis and gas chromatography/mass spectrometry results. The findings of this study enlarge the understanding of the removal mechanisms of complex nitrogenous pyridine-containing wastewater treated by the SAD process.     

Removal of steroid hormones from mariculture system using seaweed Caulerpa lentillifera

•Steroid hormones could be removed efficiently from mariculture system using seaweed; • Caulerpa lentillifera was the most efficient seaweed for removal of steroid hormones; • More than 90% of E₂ or EE₂ were removed within 12 h using Caulerpa lentillifera; • The removal included the rapid biosorption and the slow bio-accumulation; •The hormones and nutrients in mariculture wastewater could be simultaneously removed. The removal of steroid hormones from the mariculture system using seaweeds (Caulerpa lentillifera, Ulva pertusa, Gracilaria lemaneiformis, and Codium fragile) was investigated. The results illustrated that both 17β-estradiol (E₂) and 17α-ethinylestradiol (EE₂) could be removed by the seaweeds at different levels, and the Caulerpa lentillifera was the most efficient one. More than 90% of E₂ or EE₂ at concentration of 10 μg/L was removed by Caulerpa lentillifera within 12 h. Processes including initial quick biosorption, the following slow accumulation, and biodegradation might explain the removal mechanisms of E₂/EE₂ by Caulerpa lentillifera. E₂/EE₂ removal was positively related to the nutrient level and the initial concentration of steroid hormone. A significant linear relationship for E₂ and EE₂ existed between the initial pollutant concentration and the average removal rate. The highest removal kinetic constant (k) value was obtained at 30°C as 0.34 /h for E₂ and at 20°C as 0.28 /h for EE₂, demonstrating the promising application potential of Caulerpa lentillifera in the water purification of the industrialized mariculture system with relatively high water temperature. Simultaneous and efficient removal of E₂ and EE₂ by Caulerpa lentillifera was still achieved after 3 cycles in the pilot-scale experiment. The steroid hormones and nutrients in mariculture wastewater could also be simultaneously removed using Caulerpa lentillifera. These findings demonstrated that Caulerpa lentillifera was the promising seaweed for the removal of steroid hormones in mariculture systems.     

Cultivation of Chlorella sp. HQ in inland saline-alkaline water under different light qualities

• Optimal growth of Chlorella in inland saline-alkaline water was achieved by blue LED. • Lipids of Chlorella sp. HQ were mainly composed of C16:0 and C18:2 under various LEDs. • The BiodieselAnalyzer^© software was used to evaluate the Chlorella biodiesel quality. • Chlorella sp. HQ was a high-quality feedstock for biodiesel production. Inland saline-alkaline water can be used for the low-cost cultivation of microalgae, but whether algal biomass under various light sources has the potential to produce biodiesel remains to be developed. Herein, the influence of different light-emitting diode (LEDs) light colors (blue, red, white, mixed blue-red, and mixed blue-white LED) on the growth performance, lipid accumulation, and fatty acid composition of Chlorella sp. HQ cultivated in inland saline-alkaline water was investigated. The highest algal density was obtained under blue LEDs at the end of cultivation, reaching 1.93±0.03 × 10⁷ cells/mL. White LEDs can improve biomass yield, total lipid yield, and triacylglycerol yield per algal cell. The main fatty acid components of Chlorella from inland saline-alkaline water were palmitic acid and linoleic acid. The BiodieselAnalyzer^© software was used to predict algal biodiesel quality by estimating different quality parameters. The cetane number, kinematic viscosity, and density of Chlorella biodiesel were 51.714–67.69, 3.583–3.845 mm²/s, and 0.834–0.863 g/cm³, respectively. This further proved that the Chlorella biomass obtained from inland saline-alkaline water has the potential to be used as a high-quality biodiesel feedstock.