There are a large number of simple landfills in hilly areas, and the results of previous studies have shown that pollutants in landfills can spread via interflow and cause surface source pollution. The hybrid activated sludge-membrane aerated bioreactor (H-MABR) developed in a previous study can be used for the treatment of interflow with a low chemical oxygen demand (COD)/total nitrogen (TN) ratio, and it has been shown to be effective in laboratory simulations. To investigate the effectiveness of the H-MABR in treating interflow around landfills in real-world applications, an in-situ pilot-scale evaluation of the effectiveness of H-MABR operation was conducted at a landfill. The results indicated that the removal efficiencies of COD, TN, and ammonia nitrogen in interflow by H-MABR were 87.1 ± 6.0%, 80.9 ± 7.9%, and 97.9 ± 1.4%, respectively. The removal rate of TN reached 148.6-205.6 g-N/m3·d. The concentration of each pollutant in the effluent was in accordance with China\'s \"Standard for pollution control on the landfill site of municipal solid waste (GB16889-2008),\" wherein the COD, TN, and ammonia nitrogen of effluent should be less than 100 mg/L, 40 mg/L, and 25 mg/L, respectively. The results of community composition analysis and PICRUSt analysis based on 16S rRNA gene sequencing showed that there were different dominant functional bacteria between the inner and outer rings, but functional genes involved in the nitrification-denitrification, assimilated nitrate reduction, and dissimilated nitrate reduction pathway were all detected. Furthermore, except for the nitrite oxidation gene narG, the abundance of which did not significantly differ between the inner and outer rings, the abundance of the other functional genes was higher in the outer ring than in the inner ring. An economic evaluation revealed that the operation cost of interflow treatment by the H-MABR was as low as ¥2.78/m3; thus, the H-MABR is a shock-load-resistant and cost-effective technology for interflow treatment.

The sludge fermentation-coupled denitrification process, utilized for sludge reduction and nitrogen removal from wastewater, is frequently hindered by its hydrolysis step\'s efficacy. This study addresses this limitation by extending the sludge retention time (SRT) to 120 days. As a result, the nitrate removal efficiency (NRE) of the nitrification-sludge fermentation coupled denitrification (NSFD) pilot system increased from 67.1 ± 0.2 % to 96.7 ± 0.1 %, and the sludge reduction efficiency (SRE) rose from 40.2 ± 0.5 % to 62.2 ± 0.9 %. Longer SRT enhanced predation and energy dissipation, reducing intact cells from 99.2 % to 78.0 % and decreasing particle size from 135.2 ± 4.6 μm and 19.4 ± 2.1 μm to 64.5 ± 3.5 μm and 15.5 ± 1.6 μm, respectively. It also created different niches by altering the biofilm\'s adsorption capacity, with interactions between these niches driving improved performance. In conclusion, extending SRT optimized the microbial structure and enhanced the performance of the NSFD system.

Emulsified vegetable oil (EVO), as a novel green slow-releasing substrate, has performed great potential in subsurface bioremediation due to its slow release and longevity. Nevertheless, the long time it takes to initiate this process still exposed some limitations. Herein, multiple enzyme-based EVOs (EN-EVOs) were developed to enhance the quick-acting effect in nitrate-contaminated bioremediation. This study demonstrated that EN-EVOs loaded with cellulose (c-EVO) and protein enzymes (p-EVO) performed best, not only did not change the advantages of traditional EVO, but also optimized the stability and particle size to the level of 0.8-0.9 and 247.95-252.25 nm, respectively. Nitrate (NO3-N) degradation further confirmed the superiority of c-EVO in rapidly initiating degradation and achieving stable denitrification. Compared with traditional EVO, the maximum start-up efficiency and the rapid achieving stable denitrification efficiency were improved by 37.6% and 1.71 times, respectively. In such situation, the corresponding NO3-N removal efficiency, kinetics rate constant (k1), and half-life period (t1/2) reached as high as 85.39%, as quick as 1.079 d-1, and as short as 0.64 d after 30-day cultivation. Meanwhile, the rapid conversion efficiency of NO2-N was observed (k2 = 0.083 d-1). High-throughput 16S rRNA gene sequencing indicated that the quick-acting process of NO3-N reduction coupled to c-EVO was mediated by microbial reducers (e.g., Ralstonia, Gulbenkiania, and Sphingobacterium) with regulations of narG, nirS and norB genes. Microorganisms with these genes could achieve quick-acting not only by enhancing microbial activity and the synthesis and metabolism of volatile fatty acids, but also by reducing the production and accumulation of loosely bound-extracellular polymeric substances (LB-EPS). These findings advance our understanding on fast-acting of NO3-N degradation supported by c-EVO and also offer a promising direction for groundwater remediation.

Further reducing total nitrogen (TN) and total phosphorus (TP) in the secondary effluent needs to be realized effectively and in an eco-friendly manner. Herein, four pyrite/sawdust composite-based biofilters were established to treat simulated secondary effluent for 304 days. The results demonstrated that effluent TN and TP concentrations from biofilters under the optimal hydraulic retention time (HRT) of 3.5 h were stable at <2.0 and 0.1 mg/L, respectively, and no significant differences were observed between inoculated sludge sources. The pyrite/sawdust composite-based biofilters had low N2O, CH4, and CO2 emissions, and the effluent\'s DOM was mainly composed of five fluorescence components. Moreover, mixotrophic denitrifiers (Thiothrix) and sulfate-reducing bacteria (Desulfosporosinus) contributing to microbial nitrogen and sulfur cycles were enriched in the biofilm. Co-occurrence network analysis deciphered that Chlorobaculum and Desulfobacterales were key genera, which formed an obvious sulfur cycle process that strengthened the denitrification capacity. The higher abundances of genes encoding extracellular electron transport (EET) chains/mediators revealed that pyrite not only functioned as an electron conduit to stimulate direct interspecies electron transfer by flagella but also facilitated EET-associated enzymes for denitrification. This study comprehensively evaluates the water-gas-biofilm phases of pyrite/sawdust composite-based biofilters during a long-term study, providing an in-depth understanding of boosted electron transfer in pyrite-based mixotrophic denitrification systems.

Removing selenium (Se) from mine effluent is a common challenge. A long-term, in situ experiment was conducted to bioremediate large volumes (up to 7500 mc d-1) of Se(VI)-contaminated water (mean 87 μg L-1) by injecting the water into a saturated waste rock fill (SRF) at a coal mining operation in Elk Valley, British Columbia, Canada. To stimulate/maintain biofilm growth in the SRF, labile organic carbon (methanol) and nutrients were added to the water prior to its injection. A conservative tracer (Br-) was also added to track the migration of injected water across the SRF, identify wells with minimal dilution and used to quantify the extent of bioreduction. The evolution of the Se species through the SRF was monitored in time and space for 201 d. Selenium concentrations of <3.8 μg L-1 were attained in monitoring wells located 38 m from the injection wells after 114 to 141 d of operation. Concentrations of Se species in water samples from complementary long-term (351-498 d) column experiments using influent Se(VI) concentrations of 1.0 mg L-1 were consistent with the results of the in situ experiment. Solid samples collected at the completion of the column experiments confirmed the presence of indigenous Se-reducing bacteria and that the sequestered Se was present as insoluble Se(0), likely in Se-S ring compounds. Based on the success of this ongoing bioremediation experiment, this technology is being applied at other mine sites.

This study investigates the nitrogen removal efficacy and microbial community dynamics in seawater aquaculture effluent treatment using three different substrate combinations of microscale laboratory aerated filters (MFs) - MF1 (LECA), MF2 (LECA/Fe-C), and MF3 (LECA/Pyrite). The findings indicated that the COD removal exceeded 95% across all MFs, with higher removal efficiencies in MF2 and MF3. In terms of nitrogen removal performance, MF2 exhibited the highest average nitrogen removal of 93.17%, achieving a 12.35% and 3.56% increase compared to MF1 (80.82%) and MF3 (89.61%), respectively. High-throughput sequencing analysis revealed that the Fe-C substrate significantly enhanced the diversity of the microbial community. Notably, in MF2, the salinophilic denitrifying bacterium Halomonas was significantly enriched, accounting for 42.6% of the total microbial community, which was beneficial for nitrogen removal. Moreover, an in-depth analysis of nitrogen metabolic pathways and microbial enzymes indicated that MF2 and MF3 possessed a high abundance of nitrification and denitrification enzymes, related to the high removal rates of NH4+-N and NO3--N. Therefore, the combination of LECA with iron-based materials significantly enhances the nitrogen removal efficiency from mariculture wastewater.

Soil nitrogen (N) transformation processes, encompassing denitrification, anaerobic ammonium oxidation (anammox), and anaerobic ammonium oxidation coupled with iron reduction (Feammox), constitute the primary mechanisms of soil dinitrogen (N2) loss. Despite the significance of these processes, there is a notable gap in research regarding the assessment of managed fertilization and irrigation impacts on anaerobic N transformations in paddy soil, crucial for achieving sustainable soil fertility management. This study addressed the gap by investigating the contributions of soil denitrification, anammox, and Feammox to N2 loss in paddy soil across varying soil depths, employing different fertilization and irrigation practices by utilizing N stable isotope technique for comprehensive insights. The results showed that anaerobic N transformation processes decreased with increasing soil depth under alternate wetting and drying (AWD) irrigation, but increased with the increasing soil depth under conventional continuous flooding (CF) irrigation. The denitrification and anammox rates varied from 0.41 to 2.12 mg N kg-1 d-1 and 0.062-0.394 mg N kg-1 d-1, respectively, which accounted for 84.3-88.1% and 11.8-15.7% of the total soil N2 loss. Significant correlations were found among denitrification rate and anammox rate (r = 0.986, p < 0.01), Fe (Ⅲ) reduction rate and denitrification rate (r = 0.527, p < 0.05), and Fe(Ⅲ) reduction rate and anammox rate (r = 0.622, p < 0.05). Moreover, nitrogen loss was more pronounced in the surface layer of the paddy soil compared to the deep layer. The study revealed that denitrification predominantly contributed to N loss in the surface soil, while Feammox emerged as a significant N loss pathway at depths ranging from 20 to 40 cm, accounting for up to 26.1% of the N loss. It was concluded that fertilization, irrigation, and soil depth significantly influenced anaerobic N transformation processes. In addition, the CF irrigation practice is best option to reduce N loss under managed fertilization. Furthermore, the role of microbial communities and their response to varying soil depths, fertilization practices, and irrigation methods could enhance our understanding on nitrogen loss pathways should be explored in future study.

Mixed carbon sources have been developed for denitrification to eliminate the \"carbon dependency\" problem of single carbon. The metabolic correlation between different carbon sources is significant as guidance for the development of novel mixed carbon sources. In this study, to explore the metabolic similarity of denitrifying carbon sources, we selected alcohols (methanol, ethanol, and glycerol) and saccharide carbon sources (glucose, sucrose, and starch). Batch denitrification experiments revealed that methanol-acclimated sludge improved the denitrification rate of both methanol (14.42 mg-N/gMLVSS*h) and ethanol (9.65 mg-N/gMLVSS*h), whereas ethanol-acclimated sludge improved the denitrification rate of both methanol (7.80 mg-N/gMLVSS*h) and ethanol (22.23 mg-N/gMLVSS*h). In addition, the glucose-acclimated sludge and sucrose-acclimated sludge possibly improved the denitrification rate of glucose and sucrose, and the glycerol-acclimated sludge improved the denitrification rate of volatile fatty acids (VFAs), alcohols, and saccharide carbon sources. Functional gene analysis revealed that methanol, ethanol, and glycerol exhibited active alcohol oxidation and glyoxylate metabolism, and glycerol, glucose, and sucrose exhibited active glycolysis metabolism. This indicated that the similarity in the denitrification metabolism of these carbon sources was based on functional gene similarity, and glycerol-acclimated sludge exhibited the most diverse metabolism, which ensured its good denitrification effect with other carbon sources.

The present study was conducted with the aim of isolation and identification of the biofilm-forming denitrifying Pseudomonas bacterial strains from eutrophic waters of Dal lake, India, followed by the study of inter-relation of biofilm formation and denitrification potential of Pseudomonas strains. The bacterial strains were characterized by morphological observations and identified using 16S rDNA sequencing followed by the quantification of biofilm formation of these st by crystal violet (CV) assay using 96-well microtiter plate and extracellular polymeric substance (EPS) extraction. Lastly, the nitrate-reducing potential of all Pseudomonas species was studied. Our evaluation revealed that four different Pseudomonas species were observed to have the biofilm-forming potential and nitrate-reducing properties and the species which showed maximum biofilm-forming potential and maximum EPS production exhibited higher nitrate-removing capacity. Moreover, P. otitis was observed to have the highest denitrification capacity (89%) > P. cedrina (83%) > P. azotoform (79%) and the lowest for P. peli (70%). These results clearly signify a positive correlation of biofilm-forming capacity and nitrate-removing ability of Pseudomonas species. This study has for the first time successfully revealed the bioremediation potential of P. otitis, P. cedrina, P. azotoform, and P. peli species, thus contributing to the growing list of known nitrate-reducing Pseudomonas species. Based upon the results, these strains can be extrapolated to nitrate-polluted water systems for combating water pollution.

This study outlines the development of an effective pilot-scale simultaneous denitrification and nitrification (SDN) system using intermittent aeration for the removal of carbon and nitrogen from real domestic sewage. Given the limited research in this area, the main objective was to evaluate the overall performance of the SBRIA system on a pilot scale and show its benefits in domestic wastewater treatment. The structured bed reactor with intermittent aeration (SBRIA) notably achieved 57% efficiency in removing total nitrogen without requiring external carbon sources. It also demonstrated impressive removal rates of 56% for total chemical oxygen demand (CODT) and 82% for biochemical oxygen demand (BOD5), indicating its effectiveness in degrading organic matter. In addition, the SBRIA showed high pH control and managed the consumption of alkalinity without the need for an alkalizer, maintaining consistent mean values of 7.7 ± 0.8 for pH and 166.8 ± 79.8 mg·L-1 for alkalinity. The system also proved resilient against toxic shocks caused by significant variations in influent characteristics. This study offers valuable insights and compelling results into a cost-effective and efficient treatment approach using an innovative technology not previously applied at the pilot scale. Its potential to remediate polluted water is substantial.