Response of the mainstream anammox process to the biodegradable carbon sources in the granule-based systems: The difference in self-stratification of the microbial community.

The inevitable introduction of biodegradable carbon sources (such as monosaccharides and volatile fatty acids) originating from pretreatment units might affect the performance of the mainstream anaerobic ammonium oxidation (anammox) process. Two model carbon sources (glucose and acetate) were selected to investigate their effects on granule-based anammox systems under mainstream conditions (70 mg total nitrogen (TN) L-1, 15 °C). At a nitrogen loading rate of 2.87 ± 0.80 kg N m-3 d-1, a satisfactory effluent quality (TN < 10 mg L-1) was achieved in the presence of glucose or acetate at a chemical oxygen demand (COD/N) ratio of 0.5. The contribution of anammox to nitrogen removal decreased with increasing COD/N ratio to 1.0 because the expression of anammox functional genes was inhibited, whereas the expression of denitrifying functional genes was promoted. However, the nitrogen removal efficiency of the two considered reactors was maintained above 80 %. Self-stratification of the microbial community along the reactor height facilitated a functional balance through the retention of anammox bacteria in granules but resulted in washout of denitrifying bacteria in flocs under a high-flow pattern. These findings highlighted the advantages of granule-based systems in the mainstream anammox process due to their inherent biomass self-segregation property and the need for the development of targeted biomass retention strategies.

Linear anionic surfactant (SDBS) destabilized anammox process through sludge disaggregation and metabolic inhibition.

The increase of emerging contaminants, such as surfactants, is one of the major challenges to biological wastewater treatment. However, the potential impact of linear alkylbenzene sulphonates (LAS), a major class of anionic surfactants, on anammox process is unclear. The long-term effects of sodium dodecyl benzene sulfonate (SDBS, as a model LAS) on reactor performance, microbial community and sludge properties were investigated in this study. The presence of 5 mg L-1 SDBS promoted the release of extracellular microbial products from anammox granules and the wash-out of anammox population via effluent. Despite sludge disaggregation, the reactor performance was robust to the exposure of 5 mg L-1 SDBS due to functional redundancy. With the further increase of SDBS to 10 mg L-1, the metabolic activity of anammox biomass and the transcription and post-translation of hydrazine dehydrogenase were significantly decreased. The potential mechanism might be associated with the damage on cell membrane that induced the leakage of intracellular matrix. These results highlight the need to consider the potential risk of LAS to operation of anammox process in biological wastewater treatment plant.

Anammox Granules Acclimatized to Mainstream Conditions Can Achieve a Volumetric Nitrogen Removal Rate Comparable to Sidestream Systems.

The implementation of mainstream anammox has gained increasing attention. In this study, the feasibility of using sidestream anammox granules to start up mainstream reactors was investigated by comparing two switching strategies. A maximum nitrogen removal potential of 3.6 ± 0.2 kg N m-3 d-1 was obtained for the reactor after direct switching to mainstream conditions (70 mg TN L-1, 15 °C). Nevertheless, the reactor preacclimatized to 25 °C (Ma) exhibited a higher nitrogen removal potential of 7.0 ± 0.3 kg N m-3 d-1 at 15 °C, which is the highest volumetric nitrogen removal rate of mainstream anammox reactors to date. Candidatus Kuenenia stuttgartiensis was identified as the dominant anammox bacterium, and its relative abundance in two reactors remained stable throughout the whole operation (200 days). Moreover, with the aid of acclimatization, the activation energy was reduced and the specific growth rate became higher. These results indicated that the physiological evolution of the dominant anammox bacterium instead of interspecies selection was the main reason for the high potential during the switch to mainstream conditions. Therefore, using sidestream anammox granules as seed sludge to start up mainstream reactors was demonstrated to be feasible, and a switching strategy of acclimatization at 25 °C was recommended.

Resistance of anammox granular sludge to copper nanoparticles and oxytetracycline and restoration of performance.

Nanoparticles and antibiotics, the two most frequently detected emerging pollutants from different wastewater sources, are eventually discharged into wastewater treatment plants. In this study, the widely used materials CuNPs and oxytetracycline (OTC) were selected as target pollutants to investigate their joint effects on anaerobic ammonium oxidation (anammox). The results indicated that the environmental concentration slightly inhibited the performance of the reactors, while the performance rapidly deteriorated within a week under high-level combined shocks (5.0 mg L-1 CuNPs and 2.0 mg L-1 OTC). After the second shock (2.5 mg L-1 CuNPs and 2.0 mg L-1 OTC), the resistance of anammox bacteria was enhanced, with an elevated relative abundance of Candidatus Kuenenia and absolute abundance of hzsA, nirS, and hdh. Moreover, the extracellular polymeric substance (EPS) content and specific anammox activity (SAA) showed corresponding changes. Improved sludge resistance was observed with increasing CuNP and OTC doses, which accelerated the recovery of performance.

Long-term effects of copper nanoparticles on granule-based denitrification systems: Performance, microbial communities, functional genes and sludge properties.

The widespread use of copper nanoparticles (CuNPs) has attracted increasing concern because of their potential effects on biological wastewater treatment. However, their effect on granule-based denitrification systems is unclear. Hence, the effects of CuNPs on denitrifying granules were investigated during long-term operation. The results showed that 51.9% of nitrogen removal capacity was lost after exposure to 5 mg L-1 CuNPs, with the amount of Cu(II) gradually increasing with elevating CuNP levels. Moreover, the relative abundance of denitrifying bacteria (Castellaniella) and denitrifying functional genes (nirK, napA, narG and nosZ) obviously decreased. Meanwhile, the specific denitrification activity, the content of extracellular polymeric substances and dehydrogenase activity decreased by 44.0%, 15.2% and 99.9%, respectively, compared to their values in the initial sludge. Considering the downtrend in the abundance of copper resistance genes, it was deduced that the toxicity of CuNPs was mainly or at least partially due to the release of Cu(II).

Effects of ZnO nanoparticles on high-rate denitrifying granular sludge and the role of phosphate in toxicity attenuation.

The increasing release of engineered nanoparticles (NPs) from consumer products has raised great concerns about their impacts on biological wastewater treatment. In this study, the widely-used ZnO NP was selected as a model NP to investigate its impact on high-rate denitrifying granular sludge in terms of sludge properties and community structure. A hormesis effect was observed during short-term exposure, in which the specific denitrification activity (SDA) was stimulated by 10% at 1 mg L-1 ZnO NPs, but inhibited by 23% at 5.0 mg L-1 ZnO NPs. When continuously exposed to 2.5 mg L-1 ZnO NPs, the nitrogen removal capacity of the denitrification reactor was nearly deprived within 15 days, and the relative abundance of the dominant denitrifying bacterium (Castellaniella) was decreased from 51.0 to 8.0%. Meanwhile, the dehydrogenase activity (DHA) and the content of extracellular polymeric substance (EPS) significantly decreased to 22.3 and 61.1%, respectively. Nevertheless, the presence of phosphate substantially weakened the adverse effects of ZnO NPs on the SDA, EPS, DHA and the relative abundance of functional genes even exposed to 6.25 mg L-1 ZnO NPs, which was associated with the fact that the level of Zn(II) released from ZnO NPs was significantly reduced in the presence of phosphate. Therefore, the toxicity of ZnO NPs may be mainly attributed to the release of toxic Zn(II) and could be attenuated in the presence of phosphate. Overall, this study provided further reference and meaningful insights into the impact of engineered NPs on biological wastewater treatment.

Evaluating the effects of Zn(II) on high-rate biogranule-based denitrification: Performance, microbial community and sludge characteristics.

High-rate denitrification is a popular and efficient process for treatment of nitrate-rich wastewater. Knowing the effect of heavy metals on denitrification is essential for industrial development. In the present study, the long-term impacts of Zn(II) on denitrifying granular sludge were investigated. The suppression threshold of Zn(II) on denitrifying bacteria was 10 mg L-1 for long-term exposure. The nitrogen removal rate was decreased by long-term addition of 10 mg L-1 Zn(II). Castellaniella and Klebsiella were the two dominant genera under Zn(II) stress. The relative abundance of Klebsiella sharply decreased to 4.64% after the addition of 10 mg L-1 Zn(II), whereas Castellaniella was susceptible to 2 mg L-1 Zn(II), revealing that Castellaniella mainly was devoted to denitrification under no or low Zn(II) stress conditions, whereas Klebsiella was effective under high Zn(II) stress.

Deciphering the evolution characteristics of extracellular microbial products from autotrophic and mixotrophic anammox consortia in response to nitrogen loading variations.

Extracellular microbial products (EMP) in biological wastewater treatment systems vary with operational conditions and in turn indicate the metabolic status of functional bacteria. In this study, the response of EMP from autotrophic and mixotrophic anammox consortia (AAC and MAC) to the variation of total nitrogen loading rates (TNLR) were investigated as well as their correlations with the community evolution. The variation of TNLR showed a significantly negative correlation with the production of bound microbial products (BMP) but a significantly positive correlation with the production of soluble microbial products (SMP). The presence of organic matters with COD/TN ratio of 0.15 limited the abundance of anammox bacteria in MAC at the full-load phase and suppressed their proliferation at the restart phase. Due to the improved abundance of carbohydrate metabolism genes, MAC with lower abundance of anammox bacteria produced lower soluble polysaccharides than AMC at the full-load phase. Furthermore, four components (C1-4) were identified on the excitation-emission matrix fluorescence spectra of SMP using parallel factor analysis. C1 exhibited a relative higher proportion at the full-load phase, whereas C4 was generated only at the light-load phase or empty-load phase. At the restart phase, C2 and C3 appeared simultaneously and accounted for a high proportion. The information of four components also suggested the metabolic status of AC as revealed by the specific anammox activity, which therefore provided a novel complementary but direct approach for monitoring the operation status of anammox bioreactors.

Achieving completely anaerobic ammonium removal over nitrite (CAARON) in one single UASB reactor: Synchronous and asynchronous feeding regimes of organic carbon make a difference.

At least 11% of total nitrogen (TN) remains in the anammox effluent, making it difficult to meet increasingly stringent discharge standards. To overcome this bottleneck, an innovative process to achieve completely anaerobic ammonium removal over nitrite (CAARON) in one single up-flow anaerobic sludge blanket reactor was proposed in this study. The synchronous feeding of acetate at a C/N (nitrite) ratio of 0.6 significantly reduced the nitrogen removal capacity of anammox reactor by limiting the abundance and metabolism of anammox bacteria. In contrast, the asynchronous feeding of acetate optimized the partition of the reactor column into two specific compartments: the lower half favoring anammox and the upper half dominated by DEAMOX (DEnitrifying AMmonium Oxidation). A high TN removal efficiency of 96.2±0.4% and a low effluent TN concentration of 9.3±0.9mgL-1 were obtained under a high TN loading rate of 9.0kgNm-3d-1. The dominant functional microbes in the CAARON process were identified as Candidatus Kuenenia and Thauera, which were responsible for the anammox and denitratation reactions, respectively. Overall, the results in this study provide valuable insight into the coupling of anammox with denitratation, which is a cost-efficient approach for treating ammonium-rich wastewaters.

Anammox granule as new inoculum for start-up of anaerobic sulfide oxidation (ASO) process and its reverse start-up.

The feasibility of implementing anaerobic ammonium oxidation (anammox) granules to start up high-loading anaerobic sulfide oxidation (ASO) in an upflow anaerobic sludge bed (UASB) reactor was investigated. An innovation method of the reverse start-up of anammox was also validated. Firstly, the reactor was operated to treat sulfide-rich wastewaters into which nitrite was introduced as an electron acceptor. An high-rate performance with sulfide and nitrate removal rates of 105.5 ±â€¯0.11 kg S m-3 d-1 and 28.45 ±â€¯3.40 kg N m-3 d-1, respectively, was accomplished. Sulfurovum were enriched with the increase of the substrate load and then conquered Candidatus Kuenenia to be the predominant bacteria. Excitation-emission matrix (EEM) spectroscopy showed that the intensities of fluorescence decreased and protein-like substrates were the main components associated with the process of start-up. FT-IR analysis found that the main functional groups indicator were O-H groups. Secondly, the reverse start-up of anammox (achieving 90% TN removal) was achieved immediately when the substrate changed. 16S rRNA analysis indicated the successfully enrichment of anammox bacteria (Candidatus Kuenenia). These results suggest that anammox granules can act as inoculum of high-loading ASO process and the reverse start-up provides a new perspective for the fast initiation of anammox process.

Lactobacillus reuteri F-9-35 Prevents DSS-Induced Colitis by Inhibiting Proinflammatory Gene Expression and Restoring the Gut Microbiota in Mice.

Probiotics are considered to be a potential treatment for ulcerative colitis (UC). The aim of this study was to compare the preventive effect of a space flight-induced mutant L. reuteri F-9-35 and its wild type on UC in vivo. Female mice were randomly assigned to five groups: one normal and four colitic. Mice from colitis groups were daily gavaged with 0.2 mL 12% (w/v) skim milk containing the mutant or wild type (1 × 1011 CFU/mL), skim milk alone or distilled water for the whole experiment period, starting 7 days before colitis induction. UC was induced by administrating mice with 3.5% (w/v) dextran sulfate sodium (DSS) in drinking water for 7 days, after which DSS was removed and maintained for 3 days as a recovery phase. The results showed that the mice fed with L. reuteri F-9-35 had less inflammatory phenotype according to macroscopic and histological analysis, reduced myeloperoxidase activity, and lower expression of proinflammatory genes (Tumor necrosis factor-α, cyclooxygenase-2 and interleukin-6) in colonic tissue compared with control. Furthermore, L. reuteri F-9-35 protected the mice from gut microbiota dysbiosis from DDS induced colitis. Neither wild type nor the milk alone had such beneficial effects. From above we conclude that L. reuteri F-9-35 has great potential in the prevention of UC as a dietary supplement. PRACTICAL APPLICATION: Ulcerative colitis (UC) is the most common inflammatory bowel diseases and there is still a lack of safe and effective treatments. Consumption of L. reuteri F-9-35 may effective in preventing human UC.

Discrepant effects of metal and metal oxide nanoparticles on anammox sludge properties: A comparison between Cu and CuO nanoparticles.

Metal and metal oxide nanoparticles (NPs) show differences in antimicrobial activity due to different chemical and physical properties. Using copper as a representative example, this study compared the NP effects on anaerobic ammonium oxidation (anammox) bacteria in wastewater treatment. Long-term exposure to 5 mgCu L-1 CuNPs reduced the physiological activity and abundance of anammox bacteria, thereby causing deterioration of reactor performance. However, anammox granules exhibited stronger resistance and resilience to perturbation by 1-160 mgCu L-1 CuONPs, and no adverse effects on performance were observed. Moreover, the level of Cu(II) released from NPs in the influent exhibited good correlations with variations of the community structure and sludge properties. Therefore, the effects of Cu-based NPs on anammox sludge properties are dependent on their forms and levels, and their discrepant effects are partially attributed to their ability to release ionic copper.

Increased salinity improves the thermotolerance of mesophilic anammox consortia.

While the application of anammox-based process for mesophilic sidestream treatment is at present the state of the art and mainstream treatment at ambient temperature is also in development, the feasibility of thermophilic anammox process is still unclear. This study investigated the effects of salinity on the thermotolerance of mesophilic anammox sludge. In batch activity tests, 45 °C seems to be the critical temperature for the tolerance of mesophilic anammox consortia without acclimatization or amendments. The optimal anammox activity at 40, 42.5, and 45 °C can be achieved with the amendment of salt at 5-8, 8-10, and ~12 g NaCl L-1, respectively. However, this improvement effect was limited at 50 °C or when the shock duration was longer than 24 h even at 45 °C. In continuous-flow bioreactors, mesophilic anammox consortia could gradually adapt to 40-50 °C under a transition of 2.5 °C, and the performance was enhanced by an increase in salinity, which may be associated with the increase in extracellular polymeric substances. A nitrogen removal rate of 0.53 kgN m-3 d-1 was finally obtained at 50 °C. Overall, these interesting results facilitate further opportunities for thermophilic anammox process.

Evaluating the effects of metal oxide nanoparticles (TiO2, Al2O3, SiO2 and CeO2) on anammox process: Performance, microflora and sludge properties.

The increasing use of engineered metal oxide nanoparticles (MONPs) in consumer products raises great concerns about their environmental impacts, but their potential impacts on anaerobic ammonium oxidation (anammox) process in wastewater treatment remain unclear. In this study, the presence of MONPs (1, 50, 200 mg L-1) exhibited no visible effects on the nitrogen removal performance of anammox reactors, but high levels (200 mg L-1) of SiO2NPs, Al2O3NPs and CeO2NPs had a distinct effect on shaping the anammox community. Long-term exposure of MONPs caused different responses in the relative abundance of Ca. Kuenenia, the level of functional gene HzsA and the activities of three key enzymes involved in anammox metabolism, but no significant inhibition effects on specific anammox activity were detected. Overall, the effects of MONPs on anammox community structure and sludge properties depended on their types and levels and followed the order SiO2 > CeO2 > Al2O3 > TiO2.

Short-term effects of nanoscale Zero-Valent Iron (nZVI) and hydraulic shock during high-rate anammox wastewater treatment.

The stability and resilience of an anaerobic ammonium oxidation (anammox) system under transient nanoscale Zero-Valent Iron (nZVI) (50, 75 and 100 mg L-1), hydraulic shock (2-fold increase in flow rate) and their combination were studied in an up-flow anaerobic sludge blanket reactor. The response to the shock loads can be divided into three phases i.e. shock, inertial and recovery periods. The effects of the shock loads were directly proportional to the shock intensity. The effluent quality was gradually deteriorated after exposure to high nZVI level (100 mg L-1) for 2 h. The higher effluent sensitivity index and response caused by unit intensity of shock was observed under hydraulic and combined shocks. Notably, the specific anammox activity and the content of heme c were considerably reduced during the shock phase and the maximum loss rates were about 30.5% and 24.8%, respectively. Nevertheless, the extracellular polymeric substance amount in the shock phase was enhanced in varying degrees and variation tendency was disparate at all the tested shock loads. These results suggested that robustness of the anammox system was dependent on the magnitude shocks applied and the reactor resistance can be improved by reducing hydraulic retention time with the increase of nZVI concentration under these circumstances.

Anammox granules show strong resistance to engineered silver nanoparticles during long-term exposure.

The increasing use of engineered Ag nanoparticles (NPs) in consumer products raises great concerns about their environmental impacts, but their potential impact on anaerobic ammonium oxidation (anammox) bacteria in wastewater treatment remains unclear. In this study, the response of anammox consortia inhabiting in granules to long-term exposure of AgNPs was investigated. The gradually increased stress of AgNPs (1, 10, 50 mg L-1) showed no adverse effects on the nitrogen removal performance and on the relative abundance of Ca. Kuenenia inhabiting in granules. Notably, in response to the escalating burden of AgNPs, anammox consortia tended to secrete more protein but less polysaccharide, and meanwhile increase the relative abundances of Ag+ efflux protein SilA and regulatory protein SilR. Overall, these results suggested that the strong resistance of anammox granules to AgNPs might be mainly attributed to the defense role of extracellular polymeric substances and gene-based regulation of ion transport.

Transient disturbance of engineered ZnO nanoparticles enhances the resistance and resilience of anammox process in wastewater treatment.

The increasing use of engineered nanoparticles (NPs) in consumer and industrial products raises concerns about their environmental impacts, but their potential influence on anaerobic ammonium oxidation (anammox) process in wastewater treatment remains unknown. In this study, the response of granule-based anammox reactor to different loads of ZnONPs was investigated. The introduction of 1-5mgL-1 ZnONPs did not affect reactor performance, but 90% of the nitrogen removal capacity was deprived by a shock of 10mgL-1 ZnONPs within 3days. Anammox activity was significantly inhibited, but no significant stimulation of intracellular reactive oxygen species (ROS) production or extracellular lactate dehydrogenase (LDH) activity was observed. The inhibition was thus mainly due to the accumulation of toxic Zn(II) ions in anammox biomass. However, the resistance and resilience of this anammox reactor to ZnONPs were enhanced by intermittent perturbations in the mode of "shock-recovery". The up-regulated abundance of Zn(II)-exporter ZntA might contribute to the enhanced resistance. In addition, these repeated transient disturbances improved the functional specificity of the anammox community despite the reduction of its diversity. Overall, these results may provide useful references for evaluating and controlling the risk of NPs to anammox process.

Enhanced effects of maghemite nanoparticles on the flocculent sludge wasted from a high-rate anammox reactor: Performance, microbial community and sludge characteristics.

Magnetic nanoparticles (NPs) have been widely applied in environmental remediation, biomass immobilization and wastewater treatment, but their potential impact on anaerobic ammonium oxidation (anammox) biomass remains unknown. In this study, the short-term and long-term impacts of maghemite NPs (MHNPs) on the flocculent sludge wasted from a high-rate anammox reactor were investigated. Batch assays showed that the presence of MHNPs up to 200 mg L-1 did not affect anammox activity, reactive oxygen species production, or cell membrane integrity. Moreover, long-term addition of 1-200 mg L-1 MHNPs had no adverse effects on reactor performance. Notably, the specific anammox activity, the abundance of hydrazine synthase structural genes and the content of extracellular polymeric substance were increased with elevated MHNP concentrations. Meanwhile, the community structure was shifted to higher abundance of Candidatus Kuenenia indicated by high-throughput sequencing. Therefore, MHNPs could be applied to enhance anammox flocculent sludge due to their favorable biocompatibility.

Mass transfer characteristics, rheological behavior and fractal dimension of anammox granules: The roles of upflow velocity and temperature.

In this study, the mass transfer, rheological behavior and fractal dimension of anaerobic ammonium oxidation (anammox) granules in upflow anaerobic sludge blanket reactors at various temperatures (8.5-34.5°C) and upflow velocities (0.06, 0.18mh-1) were investigated. The results demonstrated that a lower temperature increased the external mass transfer coefficient and apparent viscosity and impaired the performance of anammox granules. The external mass transfer coefficient was decreased, but efficient nitrogen removal of up to 96% was achieved under high upflow velocity, which also decreased the apparent viscosity. Furthermore, a fractal dimension of up to 2.93 achieved at low temperature was higher than the previously reported values for mesophilic anammox granules. A higher upflow velocity was associated with the lower fractal dimension. Because of the disturbance in granule flaking, the effectiveness factor was less suitable than the external mass transfer coefficient for characterization of mass transfer resistance.

Unraveling the impact of nanoscale zero-valent iron on the nitrogen removal performance and microbial community of anammox sludge.

Given the increasing use of nanoscale zero-valent iron (NZVI) particles for environmental remediation and wastewater treatment, their potential impact on anaerobic ammonium oxidation (anammox) bacteria was investigated in this study using anammox sludge. Batch assays showed that NZVI concentrations up to 200mgL-1 did not affect anammox activity, reactive oxygen species production, and cell membrane integrity. The nitrogen removal efficiency of the continuous-flow reactor fluctuated in the presence of 20 or 50mgL-1 NZVI, but it could return to normal over time, even at 200mgL-1 NZVI. 16S rDNA-based high-throughput sequencing indicated that although the presence of 10, 20, 50, and 200mgL-1 NZVI to some extent affected microbial composition, the anammox bacteria (Candidatus Kuenenia) never lost its dominance. The abundance of gene families that are related to the assimilation and utilization of iron was down-regulated in response to the stress of high-level NZVI.