Isolation and characterization of a cold-tolerant heterotrophic nitrification-aerobic denitrification bacterium and evaluation of its nitrogen-removal efficiency.

With a view toward addressing the poor efficiency with which nitrogen is removed from wastewater below 10 °C, in this study, we isolated a novel cold-tolerant heterotrophic nitrification-aerobic denitrification (HN-AD) bacterium from a wetland and characterized its nitrogen removal performance and nitrogen metabolic pathway. On the basis of 16S rRNA gene sequencing, this strain was identified as a species of Janthinobacterium, designated J1-1. At 8 °C, strain J1-1 showed excellent removal efficiencies of 89.18% and 68.18% for single-source NH4+-N and NO3--N, respectively, and removal efficiencies of 96.23% and 79.64% for NH4+-N and NO3--N, respectively, when supplied with mixed-source nitrogen. Whole-genome sequence analysis and successful amplification of the amoA, napA, and nirK functional genes related to nitrogen metabolism provided further evidence in support of the HN-AD capacity of strain J1-1. The deduced HN-AD metabolic pathway of the strain was NH4+-N→NH2OH→NO2--N→NO3--N→NO2--N→NO→N2O. In addition, assessments of NH4+-N removal under different conditions revealed the following conditions to be optimal for efficient removal: a temperature of 20 °C, pH of 7, shaking speed of 150 rpm, sodium succinate as a carbon source, and a C/N mass ratio of 16. Given its efficient nitrogen removal capacity at 8 °C, the J1-1 strain characterized in this study has considerable application potential in the treatment of low-temperature wastewater.

Influence and mechanism of typical transition metal ions on the denitrification performance of heterotrophic nitrification-aerobic denitrification bacteria.

To investigate the inhibitory effects of various transition metal ions on nitrogen removal and their underlying mechanisms, the single and combined effects of Cu2+ Ni2+ and Zn2+ on Heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria Acinetobacter sp. TAC-1 were studied in a batch experiment system. The results revealed that increasing concentrations of Cu2+ and Ni2+ had a detrimental effect on the removal of ammonium nitrogen (NH4+-N) and total nitrogen (TN). Specifically, Cu2+ concentration of 10 mg/L, the TN degradation rate was 55.09%, compared to 77.60% in the control group. Cu2+ exhibited a pronounced inhibitory effect. In contrast, Zn2+ showed no apparent inhibitory effect on NH4+-N removal and even enhanced TN removal at lower concentrations. However, when the mixed ion concentration of Zn2++Ni2+ exceeded 5 mg/L, the removal rates of NH4+-N and TN were significantly reduced. Moreover, transition metal ions did not significantly impact the removal rates of chemical oxygen demand (COD). The inhibition model fitting results indicated that the inhibition sequence was Cu2+ > Zn2+ > Ni2+. Transcriptome analysis demonstrated that metal ions influence TAC-1 activity by modulating the expression of pivotal genes, including zinc ABC transporter substrate binding protein (znuA), ribosomal protein (rpsM), and chromosome replication initiation protein (dnaA) and DNA replication of TAC-1 under metal ion stress, leading to disruptions in transcription, translation, and cell membrane structure. Finally, a conceptual model was proposed by us to summarize the inhibition mechanism and possible response strategies of TAC-1 bacteria under metal ion stress, and to address the lack of understanding regarding the influence mechanism of TAC-1 on nitrogen removal in wastewater co-polluted by metal and ammonia nitrogen. The results provided practical guidance for the management of transition metal and ammonia nitrogen co-polluted water bodies, as well as the removal of high nitrogen.

Characteristics of a heavy metal resistant heterotrophic nitrification-aerobic denitrification bacterium isolated from municipal activated sludge.

The heterotrophic nitrification-aerobic denitrification (HNAD) is a new biological denitrification technology, the present study isolated a new HNAD strain named Cupriavidus metallidurans TX6 with heavy metal resistance. The gene expression, electron transport, enzyme activity and nitrogen removal property of strain TX6 were studied with different influencing factors. Strain TX6 has five nitrogen metabolism pathways (NH4+ → NH2OH → NO → NO2- → NH4+ → GOGAT/GDH; NH4+-N → NH2OH → NO → N2O → N2; NH4+ → NH2OH → NO → NO2- → NO3-; NO3- → NO2- → NH4+ → GOGAT/GDH; NO3-→ NO2- → NH4+ → GOGAT/GDH). Nitrogen balance analysis shows that 29 ± 4 mg/L of N was converted to intracellular nitrogen by assimilation and 50 ± 3 mg/L N loss may be attributed to aerobic denitrification. The results provide a theoretical basis for the HAND bacteria application in nitrogen removal from wastewaters containing heavy metals.

Research on the application of heterotrophic nitrification-aerobic denitrification bacteria in membrane bioreactor (MBR).

Inoculating heterotrophic nitrification-aerobic denitrification bacteria (HN-AD) to enhance membrane bioreactor (MBR) efficiency may result in the loss of functional bacteria. Therefore, this study compares the application results of enhancing MBR with a self-designed biological amplifier coupled with HN-AD against the performance of conventional MBR. After enhancement, the MBR achieved a removal efficiency of 96.7% for NH4+-N (100 mg/L) and 96.4% for COD (400 mg/L) in synthetic wastewater. There was a 33% increase in TN (100 mg/L) removal efficiency. The dominant bacteria in the MBR were Alcaligenes (48.4%) and Thauera (15.2%). Additionally, the abundance of denitrification genes (nirK, norB, nosZ) increased in the enhanced MBR, contributing to improved TN removal efficiency. The use of a biological amplifier effectively solved the problem of HN-AD loss in sewage treatment.

Effects of Cd(II) on nitrogen removal by a heterotrophic nitrification aerobic denitrification bacterium Pseudomonas sp. XF-4.

Simultaneous heterotrophic nitrification and aerobic denitrification (SND) is gaining tremendous attention due to its high efficiency and low cost in water treatment. However, SND on an industrial scale is still immature since effects of coexisting pollutants, for example, heavy metals, on nitrogen removal remains largely unresolved. In this study, a HNAD bacterium (Pseudomonas sp. XF-4) was isolated. It could almost completely remove ammonium and nitrate at pH 5-9 and temperature 20 ℃-35 ℃ within 10 h, and also showed excellently simultaneous nitrification and denitrification efficiency under the coexistence of any two of inorganic nitrogen sources with no intermediate accumulation. XF-4 could rapidly grow again after ammonium vanish when nitrite or nitrate existed. There was no significant effects on nitrification and denitrification when Cd(II) was lower than 10 mg/L, and 95 % of Cd(II) was removed by XF-4. However, electron carrier and electron transport system activity was inhibited, especially at high concentration of Cd(II). Overall, this study reported a novel strain capable of simultaneous nitrification and denitrification coupled with Cd(II) removal efficiently. The results provided new insights into treatment of groundwater or wastewater contaminated by heavy metals and nitrogen.

Electrospun nanofibers electrostatically adsorb heterotrophic nitrifying and aerobic denitrifying bacteria to degrade nitrogen in wastewater.

Nanofibers were prepared by electrospinning a mixture of polycaprolactone and silica, and modified to improve the hydrophilicity and stability of the material and to degrade nitrogenous wastewater by adsorbing heterotrophic nitrifying aerobic denitrifying (Ochrobactrum anthropic). The immobilized bacteria showed highly efficient simultaneous nitrification-denitrification ability, which could convert nearly 90 % of the initial nitrogen into gaseous nitrogen under aerobic conditions, and the average TN removal rate reached 5.59 mg/L/h. The average ammonia oxidation rate of bacteria immobilized by modified nanofibers was 7.36 mg/L/h, compared with 6.3 mg/L/h for free bacteria and only 4.23 mg/L/h for unmodified nanofiber-immobilized bacteria. Kinetic studies showed that modified nanofiber-immobilized bacteria complied with first-order degradation kinetics, and the effects of extreme pH, temperature, and salinity on immobilized bacteria were significantly reduced, while the degradation rate of free bacteria produced larger fluctuations. In addition, the immobilized bacterial nanofibers were reused five times, and the degradation rate remained stable at more than 80 %. At the same time, the degradation rate can still reach 50 % after 6 months of storage at 4 °C. It also demonstrated good nitrogen removal in practical wastewater treatment.

A novel derivative synchronous fluorescence method for the rapid, non-destructive and intuitive differentiation of denitrifying bacteria.

It is challenging to differentiate bacteria residing in the same habitat by direct observation. This difficulty impedes the harvest, application and manipulation of functional bacteria in environmental engineering. In this study, we developed a novel method for rapid differentiation of living denitrifying bacteria based on derivative synchronous fluorescence spectroscopy, as exemplified by three heterotrophic nitrification-aerobic denitrification bacteria having the maximum nitrogen removal efficiencies greater than 90%. The intact bacteria and their living surroundings can be analyzed as an integrated target, which eliminates the need for the complex pre-processing of samples. Under the optimal synchronous scanning parameter (Δλ = 40 nm), each bacterium possesses a unique fluorescence spectral structure and the derivative synchronous fluorescence technique can significantly improve the spectral resolution compared to other conventional fluorescence methods, which enables the rapid differentiation of different bacteria through derivative synchronous fluorescence spectra as fast as 2 min per spectrum. Additionally, the derivative synchronous fluorescence technique can extract the spectral signals contributed by bacterial extracellular substances produced in the biological nitrogen removal process. Moreover, the results obtained from our method can reflect the real-time denitrification properties of bacteria in the biological nitrogen removal process of wastewater. All these merits highlight derivative synchronous fluorescence spectroscopy as a promising analytic method in the environmental field.

Mechanisms of ammonia, calcium and heavy metal removal from nutrient-poor water by Acinetobacter calcoaceticus strain HM12.

An Acinetobacter calcoaceticus strain HM12 capable of heterotrophic nitrification-aerobic denitrification (HN-AD) under nutrient-poor conditions was isolated, with an ammonia nitrogen (NH4+-N) removal efficiency of 98.53%. It can also remove heavy metals by microbial induced calcium precipitation (MICP) with a Ca2+ removal efficiency of 75.91%. Optimal conditions for HN-AD and mineralization of the strain were determined by kinetic analysis (pH = 7, C/N = 2.0, Ca2+ = 70.0 mg L-1, NH4+-N = 5.0 mg L-1). Growth curves and nitrogen balance elucidated nitrogen degradation pathways capable of converting NH4+-N to gaseous nitrogen. The analysis of the bioprecipitation showed that Zn2+ and Cd2+ were removed by the MICP process through co-precipitation and adsorption (maximum removal efficiencies of 93.39% and 80.70%, respectively), mainly ZnCO3, CdCO3, ZnHPO4, Zn3(PO4)2 and Cd3(PO4)2. Strain HM12 produces humic and fulvic acids to counteract the toxicity of pollutants, as well as aromatic proteins to increase extracellular polymers (EPS) and promote the biomineralization process. This study provides a experimental evidence for the simultaneous removal of multiple pollutants from nutrient-poor waters.

The response of structure and nitrogen removal function of the biofilm on submerged macrophytes to high ammonium in constructed wetlands.

The ammonium exceedance discharge from sewage treatment plants has a great risk to the stable operation of subsequent constructed wetlands (CWs). The effects of high ammonium shocks on submerged macrophytes and epiphytic biofilms on the leaves of submerged macrophytes in CWs were rarely mentioned in previous studies. In this paper, the 16S rRNA sequencing method was used to investigate the variation of the microbial communities in biofilms on the leaves of Vallisneria natans plants while the growth characteristics of V. natans plants were measured at different initial ammonium concentrations. The results demonstrated that the total chlorophyll and soluble sugar synthesis of V. natans plants decreased by 51.45% and 57.16%, respectively, and malondialdehyde content increased threefold after 8 days if the initial NH4+-N concentration was more than 5 mg/L. Algal density, bacterial quantity, dissolved oxygen, and pH increased with high ammonium shocks. The average removal efficiencies of total nitrogen and NH4+-N reached 73.26% and 83.94%, respectively. The heat map and relative abundance analysis represented that the relative abundances of phyla Proteobacteria, Cyanobacteria, and Bacteroidetes increased. The numbers of autotrophic nitrifiers and heterotrophic nitrification aerobic denitrification (HNAD) bacteria expanded in biofilms. In particular, HNAD bacteria of Flavobacterium, Hydrogenophaga, Acidovorax, Acinetobacter, Pseudomonas, Aeromonas, and Azospira had higher abundances than autotrophic nitrifiers because there were organic matters secreted from declining leaves of V. natans plants. The analysis of the nitrogen metabolic pathway showed aerobic denitrification was the main nitrogen removal pathway. Thus, the nitrification and denitrification bacterial communities increased in epiphytic biofilms on submerged macrophytes in constructed wetlands while submerged macrophytes declined under ammonium shock loading.

Heterotrophic nitrification-aerobic denitrification performance of a novel strain, Pseudomonas sp. B-1, isolated from membrane aerated biofilm reactor.

A heterotrophic nitrification-aerobic denitrification (HN-AD) strain isolated from membrane aerated biofilm reactor (MABR) was identified as Pseudomonas sp. B-1, which could effectively utilize multiple nitrogen sources and preferentially consume NH4-N. The maximum degradation efficiencies of NO3-N, NO2-N and NH4-N were 98.04%, 94.84% and 95.74%, respectively. The optimal incubation time, shaking speed, carbon source, pH, temperature and C/N ratio were 60 h, 180 rpm, sodium succinate, 8, 30 °C and 25, respectively. The strain preferred salinity of 1.5% and resisted heavy metals in the order of Mn2+ > Co2+ > Zn2+ > Cu2+. It can be preliminarily speculated from the results of enzyme assay that the strain removed nitrogen via full nitrification-denitrification pathway. The addition of strain into the conventional MABR significantly intensified the HN-AD performance of the reactor. The relative abundance of the functional bacteria including Flavobacterium, Pseudomonas, Paracoccus, Azoarcus and Thauera was obviously increased after the bioaugmentation. Besides, the expression of the HN-AD related genes in the biofilm was also strengthened. Thus, strain B-1 had great application potential in nitrogen removal process.

Long-term stability of reactor microbiome through bioaugmentation with Alcaligenes aquatilis AS1 promotes nitrogen removal of piggery wastewater.

Bioaugmentation is considered as an attractive method for nitrogen removal in water treatment, but its effectiveness in actual high-strength piggery wastewater has not been adequately verified and the mechanism of bioaugmentation in actual wastewater treatment system is not very clear especially from the perspectives of microbial communities and functional genes. This study investigated the mechanisms of a heterotrophic nitrifying-aerobic denitrifying strain Alcaligenes aquatilis AS1 in the bioaugmentation of continuous biological nitrogen removal of actual piggery wastewater at laboratory scale. The addition of strain AS1 significantly improved the nitrogen removal efficiency (more than 95% of NH4+-N and 75% of TN were removed) and raised the activated sludge resistance to shock loading. AS1 addition also significantly shifted the microbiota structure and interactions among microbial networks were enhanced to obtain the stable bacterial communities. Moreover, strain AS1 achieved effective proliferation and long-term colonization in activated sludge with a relative abundance of genus Alcaligenes more than 70% during the whole operation process and played a dominant role in biological nitrogen removal, while different genera were respectively enriched and involved in pollutants removal at different stages in the control group. In addition, the abundances of most functional genes involved in carbon (C) degradation, carbon fixation and nitrogen (N), phosphorus (P), sulfur (S) cycling in activated sludge were significantly increased in reactor AS1, indicating that strain AS1 not only relied on its unique C and N metabolic activities, but also recruited microorganisms with diverse functions to jointly remove pollutants in wastewater, which could be a common bioaugmentation mechanism in open reactors. This study proves the promising application prospect of strain AS1 in the treatment of high-strength piggery wastewater and shows great importance for guiding bioaugmentation application of functional strains in practical wastewater treatment systems.

Low C/N promotes stable partial nitrification by enhancing the cooperation of functional microorganisms in treating high-strength ammonium landfill leachate.

Partial nitrification is an effective process for treating high-strength ammonium landfill leachate with low C/N ratio, for the cooperation with denitrification can save almost 40% carbon addition in biological nitrogen removal. However, high ammonia loading often causes the instability of partial nitrification process. Less carbon addition can promote the stability of partial nitrification and increase the nitrite accumulation ratio (NAR). Nevertheless, the microbial mechanisms within remain further elusive. In this study, two laboratory-scale sequencing batch reactors were constructed and operated for 125 days, which were fed with ammonia synthetic wastewater with C/N of 0.6 (CN system) and C/N of 0.0 as the control (N system). CN system performed more stably and had the highest NAR of 100%. Extracellular polymeric substances (EPS) generated from carbon source provided spatial and nutrient niches to tighten the cooperation of functional microorganisms, thus, enhanced the stability and efficiency of partial nitrification. Thauera was the dominant denitrifier in CN system. Nitrosomonas was one of the most important autotrophic ammonia oxidizing bacteria, while Paracoccus and Flavobacterium were the main heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria in CN system. The enrichment of HN-AD bacteria outcompeted nitrite oxidizing bacteria (NOB), therefore leaded to higher nitrite accumulation in CN system. The findings of this study may be conducive to increasing the understanding of the microbial collaboration mechanisms of partial nitrification, thereby provides theoretical support for the improvement of biological nitrogen removal technology.

Bioaugmentation performance for moving bed biofilm reactor (MBBR) treating mariculture wastewater by an isolated novel halophilic heterotrophic nitrification aerobic denitrification (HNAD) strain (Zobellella B307).

Moving bed biofilm reactor (MBBR) demonstrates weak nitrogen removal for mariculture wastewater treatment under high salinity environment. An isolated novel halophilic heterotrophic nitrification aerobic denitrification (HNAD) strain (Zobellella B307) was applied in MBBR process to enhance nitrogen removal. Results showed that strain Zobellella B307 could remove 90.9% ammonia nitrogen (NH4+-N) and 97.1% nitrate nitrogen (NO3--N) after 10 h cultivation, and strong resistance to salinity variation (high growth and nitrogen removal efficiency with salinity of 65‰) was observed. Besides, the chemical oxygen demand (COD), NH4+-N and NO3--N removal reached 95.6%, 94.4% and 85.7% with the strain added into MBBR process. In addition, microbial community structure analysis reflected that the strain Zobellella B307 successfully proliferated (the relative abundance increased to 2.33%). The HNAD bacteria abundance increased and dominated during the nitrogen removal process with the strain inoculation. A microbial functional analysis revealed that the main dominant functional categories (carbohydrate metabolism and amino acid metabolism) increased with the bioaugmentation of strain Zobellella B307, thus improving the nitrogen removal.

Effect mechanism of individual and combined salinity on the nitrogen removal yield of heterotrophic nitrification-aerobic denitrification bacteria.

One of the biggest challenges of applying heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria to treat high salt organic wastewater lies in the inhibitory effect exerted by salinity. To study the inhibition effect and underlying mechanism induced by different ion types and ion composition, the individual and combined effects of NaCl, KCl and Na2SO4 on HN-AD bacteria Acinetobacter sp. TAC-1 were systematically investigated by batch experiments. Results indicated that the ammonia nitrogen removal yield and TAC-1 activity decreased with increased salt concentration. NaCl, KCl and Na2SO4 exerted different degrees of inhibition on TAC-1, with half concentration inhibition constant values of 0.205, 0.238 and 0.110 M, respectively. A synergistic effect on TAC-1 was found with the combinations of NaCl + KCl, NaCl + Na2SO4 and NaCl + KCl + Na2SO4. The whole RNA resequencing suggested that transcripts of denitrification genes (nirB and nasA) were significantly downregulated with increased Na2SO4 concentration. Simultaneously, Na2SO4 stress disrupted cell respiration, DNA replication, transcription, translation, and induced oxidative stress. Finally, we proposed a conceptual model to summarize the inhibition mechanisms and possible response strategies of TAC-1 bacteria under Na2SO4 stress.

Role of heterotrophic nitrifiers and aerobic denitrifiers in simultaneous nitrification and denitrification process: a nonconventional nitrogen removal pathway in wastewater treatment.

Bacterial species capable of performing both nitrification and denitrification in a single vessel under similar conditions have gained significance in the wastewater treatment scenario considering their unique character of performing the above reactions under heterotrophic and aerobic conditions respectively. Such a novel strategy often referred to as simultaneous nitrification and denitrification (SND) has a tremendous potential in dealing with various wastewaters having low C : N content, considering that the process needs very little or no external carbon source and oxygen supply thus adding to its cost-effective and environmentally friendly nature. Though like other micro-organisms, heterotrophic nitrifiers and aerobic denitrifiers convert inorganic or organic nitrogen-containing substances into harmless dinitrogen gas in the wastewater, their ecophysiological role in the global nitrogen cycle is still not fully understood. Attempts to highlight the role played by the heterotrophic nitrifiers and aerobic denitrifiers in dealing with nitrogen pollution under various environmental operating conditions will help in developing a mechanistic understanding of the SND process to address the issues faced by the traditional methods of aerobic autotrophic nitrification-anaerobic heterotrophic denitrification.

Enhanced nitrogen removal from low-temperature wastewater by an iterative screening of cold-tolerant denitrifying bacteria.

The biological process to remove nitrogen in winter effluent is often seriously compromised due to the effect of low temperatures (< 13 °C) on the metabolic activity of microorganisms. In this study, a novel heterotrophic nitrifying-aerobic denitrifying bacterium with cold tolerance was isolated by iterative domestication and named Moraxella sp. LT-01. The LT-01 maintained almost 60% of its maximal growth activity at 10 °C. Under initial concentrations of 100 mg/L, the removal efficiencies of ammonium, nitrate, nitrite by LT-01 were 70.3%, 65.4%, 61.7% respectively for 72 h incubation at 10 °C. Nitrogen balance analysis showed that about 46% of TN was released as gases and 16% of TN was assimilated for cell growth. The biomarker genes involved in nitrification and denitrification pathways were identified by gene-specific PCR and revealed that the LT-01 has nitrite reductase (NirS) but not hydroxylamine reductase (HAO), which implies the involvement of other genes in the process. The study indicates that LT-01 has the potential for use in low-temperature regions for efficient sewage treatment.

Characteristics and mechanism of heterotrophic nitrification/aerobic denitrification in a novel Halomonas piezotolerans strain.

A strain was isolated from an activated sludge system and identified as Halomonas piezotolerans HN2 in this study, which is the first strain in H. piezotolerans with the capability of heterotrophic nitrification and aerobic denitrification. Strain HN2 showed the maximum nitrogen removal rate of 9.10 mg/L/h by utilizing ammonium at the salinity of 3.0%. Under saline environment, HN2 could remove nitrogen efficiently in neutral and slightly alkaline environments, with the carbon sources of sodium succinate and sodium citrate and the C/N ratio of 15-20, and the maximum removal efficiencies of ammonium, nitrite, and nitrate were 100%, 96.35%, and 99.7%, respectively. The genomic information revealed the presence of amoA, napA, and nosZ genes in strain HN2, and the target bands of nirS were obtained via a polymerase chain reaction. Therefore, we inferred that ammonium was mainly utilized for the growth of strain HN2 through assimilation, and another part of the initial ammonium was converted into nitrate through nitrification, and then into gaseous nitrogen through denitrification. This report indicated the potential application of strain HN2 and other nitrifying and denitrifying Halomonas strains in the removal of nitrogen pollution in marine-related environments and also implies the important role of Halomonas in the nitrogen cycle process of the ocean.

Heterotrophic nitrification-aerobic denitrification characteristics and antibiotic resistance of two bacterial consortia from Marinomonas and Halomonas with effective nitrogen removal in mariculture wastewater.

Heterotrophic nitrification-aerobic denitrification (HNAD) characteristics and antibiotic resistance of two bacterial consortia, Marinomonas communis & Halomonas titanicae (MCH) and Marinomonas aquimarina & Halomonas titanicae (MAH), and their single isolates (MC, MA, and H) were determinated in this study. When cultured in sole and mixed N-source media (NH4+-N and/or NO2--N of 10 mg/L), MCH and MAH exhibited greater efficiency and stability of inorganic-N removal than single isolates, and these strains preferred to remove NH4+-N by simultaneous HNAD in mixed N-source media. Meanwhile, 45%-70% of NH4+-N and/or NO2--N was mainly converted to organic nitrogen (15%-25%) and gaseous nitrogen (30%-40%) by these strains, and more inorganic-N was transformed to intracellular-N by MCH and MAH via assimilation instead of gaseous-N production by denitrification. Both isolates and their consortia had the maximal NH4+-N or NO2--N removal efficiency above 95% under the optimum conditions including temperature of 20-30 °C, C/N ratios of 15-20, and sucrose as carbon source. Interestingly, bacterial consortia performed greater nitrogen removal than single isolates under the low temperature of 10 °C or C/N ratios of 2-5. In real mariculture wastewater, MCH and MAH also showed higher NH4+-N removal efficiency (65%-68%) and more stable cell quantity (4.2-5.2 × 108 CFU/mL) than single strains, due to the interspecific coexistence detected by bacterial quantitation with indirect immunoassay. Additionally, these isolates and consortia had stronger resistances to polypeptides, tetracyclines, sulfonamides, furanes, and macrolides than other antibiotics. These findings will be conducive to the applications of HNAD bacteria of Marinomonas and Halomonas on reducing nitrogen pollution in mariculture or other saline environments.

Role of heterotrophic aerobic denitrifying bacteria in nitrate removal from wastewater.

With the increase in industrial and agricultural activities, a large amount of nitrogenous compounds are released into the environment, leading to nitrate pollution. The perilous effects of nitrate present in the environment pose a major threat to human and animal health. Bioremediation provides a cost-effective and environmental friendly method to deal with this problem. The process of aerobic denitrification can reduce nitrate compounds to harmless dinitrogen gas. This review provides a brief view of the exhaustive role played by aerobic denitrifiers for tackling nitrate pollution under different ecological niches and their dependency on various environmental parameters. It also provides an understanding of the enzymes involved in aerobic denitrification. The role of aerobic denitrification to solve the issues faced by the conventional method (aerobic nitrification-anaerobic denitrification) in treating nitrogen-polluted wastewaters is elaborated.

Simultaneous nitrification-denitrification by phosphate accumulating microorganisms.

Nitrogen and phosphorous are important inorganic water pollutants that pose a major threat to the environment and health of both humans and animals. The physical and chemical ways to remove these pollutants from water and soil are expensive and harsh, so biological removal becomes the method of choice to alleviate the problem without any side effects. The identification of microorganisms capable of simultaneous heterotrophic nitrification and aerobic denitrification has greatly simplified the sequestration of nitrogen from ammonium (NH4+) into dinitrogen (N2). Further, the discovery of phosphorous accumulating organisms offers greater economic benefits because these organisms can favourably and simultaneously remove both nitrogen and phosphorous from wastewaters hence reducing the nutrient burden. The stability of the system and removal efficiency of inorganic pollutants can be enhanced by the use of immobilized organisms. However, limited work has been done so far in this direction and there is a need to further the efforts towards refining process efficiency by testing low-cost substrates and diverse microbial populations for the total eradication of these contaminants from wastewaters.