Aerobic Denitrifying Research Articles

Start-up and synergistic nitrogen removal of partial nitrification and anoxic/aerobic denitrification in membrane aerated biofilm reactor

To reduce energy consumption and improve operational stability of traditional biological nitrogen removal (BNR) system, partial nitrification and anoxic/aerobic denitrification were synergistically implemented in membrane aerated biofilm reactor (MABR) by regulating DO and pH. The results indicated that the optimal DO, pH and C/N ratio were 1–2 mg/L, 9.0 and 4–7, respectively. The corresponding average organic removal rate (ORR), total nitrogen removal rate (TNRR) and nitrite accumulation rate (NAR) reached 324 gCOD・m−3・d−1, 48 gN・m−3・d−1 and 77.70%, respectively. Extracellular polymeric substance (EPS) content in biofilm was more abundant than that in inoculated sludge. Multiple aerobic denitrifiers were detected in the biofilm with the relative abundance of 11.19%–22.71%. AQUASIM simulation implied that the distribution and proportion of substrates and bacteria were significantly affected by DO and pH regulation. Overall, this study provided some important insights in the start-up and operation of synergistic nitrogen removal process in BNR system.

Adaptive regulation of activated sludge's core functional flora based on granular internal spatial microenvironment

A great deal of efforts has been put into studying the influence of the external macroenvironment for activated sludge to survive on microbial community succession, while granular internal spatial microenvironment should be given equal attention, because it is more directly involved in the information exchange and material transfer among microorganisms. This study systematically investigated the effects of granular microenvironment on spatial colonization and composition of sludge's core functional flora, and the corresponding difference of biological treatment performance. High content of extracellular-proteins (67.53 mg/gVSS) or extracellular-polysaccharide (65.02 mg/gVSS) stimulated the microbial flocculation and aggregation of 0.5–1.5 mm granules (GS) or 1.5–3.0 mm granules (GM), respectively, which was resulted from excellent cell hydrophobicity (59.26%) or viscosity (3.47 mPa s), therefore, constituted relatively dense porous frame. More hollow space existed in 3.0–5.0 mm granules (GL), which formed loose skeleton with 0.213 mL/g of total pore volume and 17.21 nm of average pore size. Combining scanning electron microscope images and fluorescent in-situ hybridization based microbiological analysis, aerobic nitrifiers were observed to wrap or surround anaerobic bacteria, or facultative/anaerobic bacteria were self-encapsulated, which created granule's unique microenvironment with alternating aerobic and anaerobic zones. GS has the most rich organic matter degrading bacteria and anaerobic heterotrophic denitrifiers, while GM and GL presented the greatest relative abundance of facultative and aerobic denitrifiers, respectively. The activity of dehydrogenase and nitrogen invertase of GM showed be 1.32–3.09 times higher than those of GS and GL, contributing to its higher carbon and nitrogen removal. These findings highlight the importance of granular microenvironment to adaptive regulation of activated sludge's core functional flora and corresponding pollutant removal performance.

Isolation and Nitrogen Removal Efficiency of the Heterotrophic Nitrifying-Aerobic Denitrifying Strain K17 From a Rare Earth Element Leaching Site.

K17, an indigenous and heterotrophic nitrifying-aerobic denitrifying bacterium, was isolated from the soil of a weathered crust elution-deposited rare earth ore leaching site in Longnan County, China. Strain K17 was identified as Pseudomonas mosselii. In this study, the morphological characteristics of strain K17 were observed and the optimal ammonia nitrogen removal conditions for the strain were studied using a single-factor experiment. Key enzyme activities were determined, and we also explored the ammonia nitrogen removal process of strain K17 on simulated leaching liquor of the rare earth element leaching site. Based on the determination of ammonia nitrogen removal and enzyme activity, it was found that strain K17 has both heterotrophic nitrifying and aerobic denitrifying activities. In addition, single-factor experiments revealed that the most appropriate carbon source for strain K17 was sodium citrate with a C/N ratio of 10 and an initial NH4+-N concentration of 100 mg/l. Furthermore, the optimal initial pH and rotation speed were 7 and 165 r/min, respectively. Under optimal conditions, the ammonia nitrogen removal efficiency of strain K17 was greater than 95%. As an indigenous bacterium, strain K17 has great potential for treating residual ammonium leaching solutions from rare earth element leaching sites.

Mix-cultured aerobic denitrifying bacteria augmented carbon and nitrogen removal for micro-polluted water: Metabolic activity, coexistence and interactions, and immobilized bacteria for reservoir raw water treatment

Less attention has been paid on the oligotrophic water body nitrogen treatment with mix-cultured aerobic denitrifying bacteria (Mix-CADB). In this study, three Mix-CADB communities were screened from the sediments of reservoirs. The nitrate and dissolved organic carbon (DOC) removal efficiencies of Mix-CADB communities were higher than 92 % and 91 %, respectively. Biolog results suggested that Mix-CADB communities displayed excellent carbon source metabolic activity. The nirS gene sequencing indicated that Pseudomonas sp. and Pseudomonas stutzeri accounted for more proportions in the core species of three Mix-CADB communities. The network model revealed that Pseudomonas sp. and Pseudomonas stutzeri mainly drove the total nitrogen and DOC removal of Mix-CADB communities. More importantly, the immobilized Mix-CADB communities could reduce >91 % nitrate in the adjusted reservoir raw water. Overall, this study showed that the three Mix-CADB communities could be regarded as potential candidates for the nitrogen treatment in oligotrophic water body ecosystems.

Investigation of an intermittently-aerated moving bed biofilm reactor in rural wastewater treatment under low dissolved oxygen and C/N condition

An intermittently-aerated moving bed biofilm reactor (MBBR) was proposed for nitrogen and carbon removal from low C/N synthetic rural wastewater. In purposes of low energy consumption and costs, the intermittent aeration modes were changed and the dissolved oxygen was reduced gradually during the operation. The results showed that effluent concentrations of ammonia nitrogen and chemical oxygen demand were lower than 15 and 50 mg/L, respectively, even under microaerobic condition (0.1–1.0 mg/L). Meanwhile, the simultaneous nitrification–denitrification was achieved by intermittent aeration. The activity of functional bacteria was still high and the proportion of autotrophic biomass increased significantly under intermittent micro-aeration mode, which improved the nitrification performance. Aerobic denitrifier Hydrogenophaga, anoxic denitrifier Thiothrix, and heterotrophic nitrifier such as Rhodobacter were enriched in the intermittently micro-aerated MBBR, which will provide an applicable solution for rural wastewater treatment under low C/N and costs.

Cooperation triggers nitrogen removal and algal inhibition by actinomycetes during landscape water treatment: Performance and metabolic activity

The actinomycetes strain Streptomyces sp. XD-11-9-3 and Streptomyces sp. 5 were isolated and presented poor denitrification performance. Co-culture of actinomycetes triggers nitrogen removal capacity under aerobic conditions (reduced 96% of total nitrogen). Nitrogen balance analysis presented that 71% of initial nitrogen converted as gaseous nitrogen. Moreover, co-culture increased the concentrations of adenosine triphosphate (>2.1 folds) and electron-transmission system activity (>1.5 folds) significantly. The co-culture presented excellent carbon source metabolism activity (especially amines and carboxylic acids) compared with monoculture. The removal efficiency of total nitrogen in the micro-polluted landscape water water reached 61% in the co-culture system, and the algal survival could be inhibited significantly. However, the dominant niche of the co-culture system restrained the diversity of the indigenous nirS-type denitrifying bacterial community. This study provided a novel pathway to the research of co-culture inefficiency aerobic denitrifier and further application in the restoration of polluted water.

Effective bioactive systems for nitrate removal from building materials

Bioactive system for reducing nitrate content in brick and limestone was developed using mineral poultice and aerobic denitrifier Pseudomonas stutzeri cells. The efficiency of the systems with P. stutzeri D1 was considerably higher compared with the reference strain for the both examined materials. After 9 days of application of the bioactive systems, the biocleaning efficiency was approximately 70% for both strains on the brick surface, while the efficiency in the depth was 72% and 95% using the reference strain and natural isolate, respectively. Artificial neural network efficiency prediction showed good prediction according to material layer depth, time, and bacteria type.

Transcriptomics uncover the response of an aerobic denitrifying bacteria to zinc oxide nanoparticles exposure

ABSTRACT Zinc oxide nanoparticles (ZnO NPs) show adverse impacts on aerobic denitrifying bacteria, little is known about the response of these bacteria to ZnO NPs exposure at cellular level. This study assessed the multiple responses of Pseudomonas aeruginosa PCN-2 under ZnO NPs exposure. We demonstrated that ZnO NPs exposure could inhibit the intracellular metabolism and stimulate the antioxidant defence capability of PCN-2. At lower exposure concentration (5 mg/L), exogenous ROS generated and resulted in the inhibition of pyruvate metabolism and citrate cycle, which caused deficient energy for aerobic denitrification. At higher concentrations (50 mg/L), endogenous ROS additionally generated and triggered to stronger down-regulation of oxidative phosphorylation, which caused suppressed electron transfers for aerobic denitrification. Meanwhile, ZnO NPs exposure promoted EPS production and biofilm formation, and antioxidases was especially particularly stimulated at higher concentration. Our findings are significant for understanding of microbial bacterial susceptibility, tolerance and resistance under the exposure of ZnO NPs.

Application of aerobic denitrifier for simultaneous removal of nitrogen, zinc, and bisphenol A from wastewater

High concentrations of heavy metals and other pollutants affect microbial activity in the wastewater treatment system and impede biological denitrification process. In this study, a novel Zn(II)-resistant aerobic denitrifier (Pseudomonas stutzeri KY-37) was isolated with potential in Bisphenol A (BPA) biodegradation and removal. The capability of this denitrifier in removing nitrogen, zinc, and BPA was tested. Using 56 mg/L nitrate as the sole nitrogen source, its removal efficiency achieved 98.5% in 12 h. This novel denitrifier had a strong auto-aggregation (maximum 65.8%), a high hydrophobicity rate (maximum 88.2%), and a massive amount (maximum 41.1 mg/g cell dry weight) of extracellular polymeric substances (EPS) production. Moreover, Zn(II) removal efficiency reached more than 95% with the initial high concentrations of 200 mg/L. The maximum BPA removal efficiency reached 88.8% with initial 10 mg/L. The removal mechanism of BPA was further explored in terms of microbial degradation, EPS adsorption, and intermediate degradation products.

Bioelectrochemical reactor improved by assembling anode with rice husk for treating nitrate-contaminated groundwater

To maximize the hydrogenotrophic denitrification and minimize the negative impact of anode by-products, a rice husk-enhanced anode bioelectrochemical reactor (RABER) was constructed by assembling anode with rice husk, and the denitrification performances and mechanisms involved were investigated. It was demonstrated that rice husk can be used as a slow-release carbon source and microbial carrier in RABER and weakened the anodic by-products. RABER obtained the highest contribution of hydrogenotrophic denitrification as 33.7% and achieved the fastest nitrate removal rate of 0.27 mg-N/(L·d) at 200 mA/m2. The relative lower accumulation of dissolved oxygen (1.99 ± 0.11 mg/L) and hydroxyl radicals were obtained at 200 mA/m2. Moreover, stable extracellular polymeric substances and higher activities of denitrification-related enzymes, i.e., around 2.78-fold for nitrate reductase and 1.96-fold for nitrite reductase those of 0 mA/m2, promoted denitrification in RABER at 200 mA/m2. At an optimal current density of 200 mA/m2, the coexistence of cellulose-degrading bacteria (Cellvibrio), aerobic denitrifier (Brevundimonas), anaerobic denitrifier (Flavobacterium), and hydrogenotrophic denitrifier (Alicycliphilus) was achieved, and the inorganic carbon generated by heterotrophic denitrifiers and electrochemical mineralization was sufficient to meet the demands of hydrogenotrophic denitrifiers, conductive to the heterotrophic‑hydrogenotrophic synergistic denitrification. This study provided a theoretical basis for the enhancement of hydrogenotrophic denitrification in BER as well as minimizing the side effects of the anode and providing a new strategy for the disposal of agricultural waste.

Nitrogen Removal Characteristics of a Cold-Tolerant Aerobic Denitrification Bacterium, Pseudomonas sp. 41

Nitrogen pollution of surface water is the main cause of water eutrophication, and is considered a worldwide challenge in surface water treatment. Currently, the total nitrogen (TN) content in the effluent of wastewater treatment plants (WWTPs) is still high at low winter temperatures, mainly as a result of the incomplete removal of nitrate (NO3−-N). In this research, a novel aerobic denitrifier identified as Pseudomonas sp. 41 was isolated from municipal activated sludge; this strain could rapidly degrade a high concentration of NO3−-N at low temperature. Strain 41 completely converted 100 mg/L NO3−-N in 48 h at 15 °C, and the maximum removal rate reached 4.0 mg/L/h. The functional genes napA, nirS, norB and nosZ were successfully amplified, which provided a theoretical support for the aerobic denitrification capacity of strain 41. In particular, the results of denitrification experiments showed that strain 41 could perform aerobic denitrification under the catalysis of NAP. Nitrogen balance analysis revealed that strain 41 degraded NO3−-N mainly through assimilation (52.35%) and aerobic denitrification (44.02%), and combined with the gene amplification results, the nitrate metabolism pathway of strain 41 was proposed. Single-factor experiments confirmed that strain 41 possessed the best nitrogen removal performance under the conditions of sodium citrate as carbon source, C/N ratio 10, pH 8, temperature 15–30 °C and rotation speed 120 rpm. Meanwhile, the bioaugmentation test manifested that the immobilized strain 41 remarkably improved the denitrification efficiency and shortened the reaction time in the treatment of synthetic wastewater.

Advanced analytical, chemometric, and genomic tools to identify polymer degradation products and potential microbial consumers in wastewater environments

The worldwide growth in the production of plastics and lack of appropriate waste management has led to unsustainable environmental pollution of global concern. Hence, determining the key drivers characterizing the degradation of polymers in water is a rising topic. Here we propose a workflow based on advanced high throughput data analysis and processing tools aiming at the characterization of (a) the degradation products of polymers exposed to an aquatic environment, and (b) the taxonomical composition of the potential microbial consumers' consortia. To test this approach, polycaprolactonediol (PCLD) polymer probes were exposed to different wastewater environments (influent, aerobic, and anaerobic denitrifying reactors) in four different wastewater treatment plants (WWTPs). Probes were extracted and analyzed by non-target liquid chromatography-high resolution mass spectrometry (LC-HRMS) and the acquired data were processed using advanced chemometric tools (Regions of Interest-Multivariate Curve Resolution-Alternate Least Squares, ROIMCR-ALS). Up to 26 components explaining ca. 69% of the variance were resolved, and 20 of these were further identified. Degradation was dominated by two processes: (a) hydrolysis of the ester groups present in PCLD and (b) formation of cyclic polycaprolactone oligomers. These transformation pathways occurred regardless of what were the WWTPs and the treatment stages, varying only the respective concentrations of the formed by-products. The composition of the microbial communities attached to the polymer and free-living in the surrounding water was characterized by 16S rDNA amplicon sequencing analysis. The two communities showed taxonomic differences and were unique in each WWTP treatment stage. Polymer-attached communities were tentatively proposed as potential consumers.

Complete nitrogen removal from over-aeration treated black-odorous water via adding aerobic denitrifiers and iron-carbon micro-electrolysis carriers

Over-aeration is usually unavoidable for removing ammonia nitrogen from black-odorous water bodies, which consumes a large amount of organic matter and creates a long-term oxygen-rich environment, limiting the denitrification process in the water. This study aims to investigate the synergistic effect of aerobic denitrifying bacteria and iron-carbon micro-electrolysis (IC-ME) carriers on complete nitrogen removal from the over-aerated black-odorous water. For nitrification, aeration can completely oxidize ammonia nitrogen to nitrate. For denitrification, adding aerobic denitrifying bacteria Pseudomonas ATCC 9027 allowed the nitrate reduction to occur under conditions of excess oxygen and enriched indigenous denitrifying bacteria (1.45% increase in cumulative abundance). These changes brought by Pseudomonas ATCC 9027 reduced the total nitrogen concentration from 12.07 to 7.12 mg/L. Besides, research has shown that IC-ME carriers as electron donors can provide additional electrons and improve the electron transport system activity by 0.07 μg O2/(g protein∙min) in the denitrification process. Enzyme activity test and metagenome analysis revealed that enzymes and genes associated with denitrification were both upregulated with the addition of Pseudomonas ATCC 9027 and IC-ME carriers. Specifically, the improvement for activities of nitrate reductase (NAR), nitrite reductase (NIR), nitric oxide reductase (NOR), nitrous oxide reductase (NOS), and cumulative abundance of denitrification genes was 0.93 μmol NO2−-N/(g·d), 3.74 μmol NO2−-N/(g·d), 0.18 μmol N2O-N/(g·d) and 0.09 μmol N2O-N/(g·d) and 1.48%, respectively. These findings provide an understanding of the cooperative mechanism of aerobic denitrifying bacteria and IC-ME carriers in enhancing nitrogen removal from over-aerated black-odorous water bodies.

Microbial community and nitrogen transformation pathway in bioretention system for stormwater treatment in response to formulated soil medium

Bioretention system has been commonly recognized as an emerging management method to control urban rainwater runoff. However, nitrogen conversion mechanisms in bioretention system in response to formulated soil medium remains unknown. In this work, five bioretention columns with different formulated soil medium were established, and results indicated that all bioretention columns achieved high NH4+-N removal rate, but exhibited significantly different NO3--N removal performance. High-throughput sequencing results suggested that nitrification might be achieved by heterotrophic nitrifying bacteria, while Pseudomonas and Rubellimicrobium may be the main denitrifying bacteria, which served as anaerobic and aerobic denitrifier, respectively. Heterotrophic nitrification-anaerobic/aerobic denitrification may the microbial pathways to realize effective nitrogen removal performance in these bioretention systems. Bioretention system with moderate silt content (NC, 10%) displayed higher abundant potential denitrifier, leading to better nitrogen removal performance. Besides, soil adsorption played an important role in nitrogen removal, particularly when activated carbon was added. This work could provide guidance of formulated soil medium optimization for the application of bioretention systems.

Steel pickling rinse wastewater treatment by two-stage MABR system: Reactor performance, extracellular polymeric substances (EPS) and microbial community

A bench-scale two-stage membrane-aerated biofilm reactor (MABR) system was applied to treat steel pickling rinse wastewater with high salinity and refractory organic. The effects of salinity and aeration pressure on the treatment efficiency, extracellular polymeric substances (EPS) characteristics and microbial community structure were studied. The optimal removal efficiencies of COD, NH+ 4-N and TN reached to 62.84%, 99.57% and 51.65%, respectively. Shortcut nitrification was achieved at low aeration, and the salinity less than 4% did not remarkable affect system performance. Colorimetric determination, three-dimensional exaction-emission matrix (3D-EEM) and Fourier transform infrared spectrum (FTIR) were employed to characterize the content and composition of proteins (PN) and polysaccharides (PS) in EPS of the biofilm. The results indicated that PN, not PS, response to changes of environmental conditions played a key role. Moreover, EPS might alleviate intracellular and extracellular osmotic pressure imbalance induced by high salinity, which imparted the biofilm in MABR with prominent salt-tolerant. High-throughput sequencing displayed that nitrifiers (Nitrosomonas, Nitrospira), denitrifiers (Dechloromonas, Hyphomicrobium, Denitromonas, Denitratisoma, Candidatus_Competibacter) and aerobic denitrifiers (Pseudomonas, Thauera) were predominant salt-tolerant bacteria.

Effective co-treatment of synthetic acid mine drainage and domestic sewage using multi-unit passive treatment system supplemented with silage fermentation broth as carbon source

A multi-unit passive treatment system was constructed for co-treatment of synthetic acid mine drainage (AMD) and domestic sewage supplemented with silage fermentation broth as carbon source. AMD and domestic sewage mixing pretreatment (unit 1) improved influent quality with pH increase, metals removal and nutrients supplement. The generated metal-rich sludge in unit 1 retained the metals (69.95% of Fe, 97.36% of Cu, 96.53% of Cd, 72.52% of Zn, and 8.59% of Mn) of influent prior to entering subsequent bioreactors. Silage fermentation broth performed well to promote bacterial sulfate reduction in sulfate reducing bioreactor system (unit 2). Residual metals (Mn) and organic/nutrient pollutants were further polished in surface-flow aerobic wetland (unit 3), where relatively high pH (7.4–8.6), aerobic condition, potential Mn-oxidizing bacteria, limestone layer and low concentrations of Fe(II) (0.04–3.5 mg/L) favored the efficient removal of Mn. After 210-day continuous flow-through experiment, this passive treatment system demonstrated the efficient performance, increasing pH from 2.5 to 8.0 with removal of metals (99%), sulfate and organic/nutrient pollutants. Diverse sulfate reducing bacteria including complete organic oxidizers (e.g. Desulfobacter) and incomplete organic oxidizers (e.g. Desulfovibrio) promoted sulfate reduction and organic/nutrient pollutants removal. Ammonia oxidizing bacteria (e.g. Nitrosomonas) and nitrite oxidizing bacteria (e.g. unidentified_Nitrospiraceae) were the potential nitrifiers for ammonia removal. Collaboration of anaerobic denitrifiers (e.g. Denitratisoma) and potential heterotrophic nitrifying and aerobic denitrifiers (HN-AD) achieved effective nitrate removal. This multi-unit treatment system with domestic sewage and silage fermentation broth as stimulation substrates provided an attractive option for AMD treatment.

Impact of ciprofloxacin and copper combined pollution on activated sludge: Abundant-rare taxa and antibiotic resistance genes

This study aimed to explore the impacts of ciprofloxacin (CIP, 0.05–40 mg/L) and copper (3 mg/L) combined pollution on nitrification, microbial community and antibiotic resistance genes (ARGs) in activated sludge system during stress- and post-effect periods. Higher CIP concentration inhibited nitrification and an average of 50% total nitrogen removal occurred under 40 mg/L of CIP pressure. The stress- and post-effects on bacterial diversity and structure were obviously distinct. Abundant genera were more sensitive to combined pollution than rare genera based on full-scale classification and conditionally rare or abundant taxa were keystone taxa in their interactions. Ammonia oxidation genes were inhibited under high CIP level, but some aerobic denitrifying bacteria (Thauera, Comamonas and Azoarcus) and key genes increased. 96 ARG subtypes were detected with complex positive relationships and their potential hosts (abundant-rare-functional genera) changed in two periods. This study highlights the different stress- and post-effects of combined pollution on activated sludge.

Micronutrients and COD/N ratio as factors influencing granular size and SND in aerobic granular sequencing batch reactors operated at low organic loading

Simultaneous nitrification-denitrification (SND) depends on dissolved oxygen, granule size and COD/N ratio. Slow-growing nitrifying bacteria at low COD/N ratio lead to the formation of small-size granule. Anoxic denitrification is encouraged at higher granule size. In the present study, the effect of different COD/N ratios and addition of micronutrients (Fe, Zn, Mn, Cu, Co, Mo, B) on granule size and SND was studied in a laboratory-scale aerobic SBR at low organic loading (358.4–1075.2 mg COD.(L·day)−1). Synthetic cheese whey wastewater and ammonia was used. Small-size granules (0.28–0.7 mm) were developed at COD/TKN ratio of 8.6–13.7 with micronutrients and long famine phase (83–92% of the cycle). COD/TKN ratio of 13.7 achieved the best removal of inorganic nitrogen Ni (62%). At COD/TKN ratio of 8.6, without micronutrients, the low COD uptake rate caused shortening of the famine phase to 66%. Large-size granules (3.0 mm) with limitation of substrates diffusion showed low Ni removal and dominant slow-growing rhizobia (Bradyrhizobium) as detected by 16S rRNA gene amplicon sequencing. In small granules, without diffusion limitations and higher ammonia exposition, the energetically unfavorable heterotrophic nitrification occurred. Heterotrophic nitrifiers Lysobacter and Dyadobacter were found. Hydrogenophaga was a potential aerobic denitrifier. SND process was probably performed by AOB, heterotrophic nitrifiers, and anoxic and aerobic denitrifiers.

Physiological mechanism of the response to Cr(VI) in the aerobic denitrifying ectomycorrhizal fungus Pisolithus sp.1

Pisolithus sp. 1 (P sp. 1) is an ectomycorrhizal fungus (EMF) with a strong Cr(VI) tolerance and reduction ability. The noninvasive microttest technique (NMT), real-time quantitative PCR (qRT–PCR), and the three-dimensional excitation-emission matrix (3D-EEM) were used to deeply explore the physiological mechanism of the P sp. 1 response to Cr(VI) and investigate the relationship between Cr(VI) reduction and denitrification in P sp. Cr(VI) induced the strongest elevations in nitrate reductase (NR) activity and NO production in the mycelia after treatment with Cr(VI) for 48 h under aerobic conditions. The NR inhibitor tungstate significantly inhibited Cr(VI) reduction, proton efflux and the expression of the NR gene (niaD) and NiR gene (niiA). In addition, NO was generated via NR-regulated denitrification. Combined treatments with Cr(VI) and the NO scavenger carboxy-PTIO (cPTIO) significantly increased O2-, H2O2 and MDA contents and reduced SDH, CAT, GSH, GR and GSNOR activity. Therefore, the NR-driven aerobic denitrifying process requires protons, and the generated NO reduces the oxidative stress effect of Cr(VI) on mycelia by reducing ROS accumulation and lipid peroxidation, enhancing mycelial and CAT activity, and promoting GSH recycling and regeneration. Psp.1 can also secrete humic acid-like and protein-like substances to combine with Cr(III) in a culture system.

Transcriptomics and proteomics revealed the psychrotolerant and antibiotic-resistant mechanisms of strain Pseudomonas psychrophila RNC-1 capable of assimilatory nitrate reduction and aerobic denitrification

Aerobic denitrification has been proved to be profoundly affected by temperature and antibiotics, but little is known about how aerobic denitrifiers respond to temperature and antibiotic stress. In this study, the nitrate reduction performance and the intracellular metabolism by a psychrotolerant aerobic denitrifying bacteria, named Pseudomonas psychrophila RNC-1, were systematically investigated at different temperatures (10 °C, 20 °C, 30 °C) and different sulfamethoxazole (SMX) concentrations (0 mg/L, 0.1 mg/L, 0.5 mg/L, 1.0 mg/L, and 5.0 mg/L). The results showed that strain RNC-1 performed satisfactory nitrate removal at 10 °C and 20 °C, but its growth was significantly inhibited at 30 °C. Nitrate removal by strain RNC-1 was slightly promoted in the presence of 0.5 mg/L SMX, whereas it was significantly suppressed with 5.0 mg/L SMX. Nitrogen balance analysis indicated that assimilatory nitrate reduction and dissimilatory aerobic denitrification jointly dominated in the nitrate removal process of strain RNC-1, in which the inhibition effected on assimilation process was much higher than that on the aerobic denitrification process under SMX exposure. Further transcriptomics and proteomics analysis revealed that the psychrotolerant mechanism of strain RNC-1 could be attributed to the up-regulation of RNA translation, energy metabolism, ABC transporters and the over-expression of cold shock proteins, while the down-regulation of oxidative phosphorylation pathway was the primary reason for the deteriorative cell growth at 30 °C. The promotion of nitrate reduction with 0.5 mg/L SMX was related to the up-regulation of amino acid metabolism pathways, while the down-regulation of folate cycle, glycolysis/gluconeogenesis and bacterial chemotaxis pathways were responsible for the inhibition effect at 5.0 mg/L SMX. This work provides a mechanistic understanding of the metabolic adaption of strain RNC-1 under different stress, which is of significance for its application in nitrogen contaminated wastewater treatment processes.