Hydroxylamine supplementation accelerated the rates of cell growth, aerobic denitrification and nitrous oxide emission of Pseudomonas taiwanensis EN-F2.

Hydroxylamine can disrupt the protein translation process of most reported nitrogen-converting bacteria, and thus hinder the reproduction of bacteria and nitrogen conversion capacity. However, the effect of hydroxylamine on the denitrification ability of strain EN-F2 is unclear. In this study, the cell growth, aerobic denitrification ability, and nitrous oxide (N2O) emission by Pseudomonas taiwanensis were carefully investigated by addition of hydroxylamine at different concentrations. The results demonstrated that the rates of nitrate and nitrite reduction were enhanced by 2.51 and 2.78 mg/L/h after the addition of 8.0 and 12.0 mg/L hydroxylamine, respectively. The N2O production from nitrate and nitrite reaction systems were strongly promoted by 4.39 and 8.62 mg/L, respectively, through the simultaneous acceleration of cell growth and both of nitrite and nitrate reduction. Additionally, the enzymatic activities of nitrate reductase and nitrite reductase climbed from 0.13 and 0.01 to 0.22 and 0.04 U/mg protein when hydroxylamine concentration increased from 0 to 6.0 and 12.0 mg/L. This may be the main mechanism for controlling the observed higher denitrification rate and N2O release. Overall, hydroxylamine supplementation supported the EN-F2 strain cell growth, denitrification and N2O emission rates.

Exopolysaccharide production by salt-tolerant bacteria: Recent advances, current challenges, and future prospects.

Natural biopolymers derived from exopolysaccharides (EPSs) are considered eco-friendly and sustainable alternatives to available traditional synthetic counterparts. Salt-tolerant bacteria inhabiting harsh ecological niches have evolved a number of unique adaptation strategies allowing them to maintain cellular integrity and assuring their long-term survival; among these, producing EPSs can be adopted as an effective strategy to thrive under high-salt conditions. A great diversity of EPSs from salt-tolerant bacteria have attracted widespread attention recently. Because of factors such as their unique structural, physicochemical, and functional characteristics, EPSs are commercially valuable for the global market and their application potential in various sectors is promising. However, large-scale production and industrial development of these biopolymers are hindered by their low yields and high costs. Consequently, the research progress and future prospects of salt-tolerant bacterial EPSs must be systematically reviewed to further promote their application and commercialization. In this review, the structure and properties of EPSs produced by a variety of salt-tolerant bacterial strains isolated from different sources are summarized. Further, feasible strategies for solving production bottlenecks are discussed, which provides a scientific basis and direct reference for more scientific and rational EPS development.

Effects of continuous low-speed biogas agitation on anaerobic digestion of high-solids pig manure: Performance and microbial community.

This study aimed to investigate effects of continuous low-speed biogas agitation on the anaerobic digestion (AD) performance and microbial community of high-solids pig manure (total solids content of 10%). Our results reveal that at a biogas agitation intensity of 1.10 L/g feed VS/d, CH4 production increased by 16.67% compared to the non-agitated condition, the removal efficiency of H2S reached 63.18%, and the abundance of Methanosarcina was the highest. The presence of Hungateiclostridiaceae was associated with H2S concentrations. An increasing biogas agitation intensity led to an elevated pH and a decreased oxidation-reduction potential (ORP). Acetate concentrations, pH, and ORP values indicated changes in H2S concentrations. Sedimentibacter demonstrates the potential to indicate biogas agitation intensity and pH. We demonstrate that continuous low-speed biogas agitation effectively increases CH4 production and reduces H2S concentrations in AD of high-solids pig manure, offering a potential technical pathway for developing AD processes for high-solids pig manure, it also demonstrates that AD process can reduce the risk of pathogen and parasite transmission.

Review of the mechanisms involved in dissimilatory nitrate reduction to ammonium and the efficacies of these mechanisms in the environment.

Dissimilatory nitrate reduction to ammonium (DNRA) is currently of great interest because it is an important method for recovering nitrogen from wastewater and offers many advantages, over other methods. A full understanding of DNRA requires the mechanisms, pathways, and functional microorganisms involved to be identified. The roles these pathways play and the effectiveness of DNRA in the environment are not well understood. The objectives of this review are to describe our current understanding of the molecular mechanisms and pathways involved in DNRA from the substrate transfer perspective and to summarize the effects of DNRA in the environment. First, the mechanisms and pathways involved in DNRA are described in detail. Second, our understanding of DNRA by actinomycetes is reviewed and gaps in our understanding are identified. Finally, the effects of DNRA in the environment are assessed. This review will help in the development of future research into DNRA to promote the use of DNRA to treat wastewater and recover nitrogen.

Efficient transformation of hydroxylamine from wastewater after supplementation with sodium carbonate or calcium bicarbonate.

Hydroxylamine is a highly reactive inorganic nitrogen compound that not only has a toxic effect on microorganisms, but also makes wastewater treatment more difficult, which in turn damages the environment and even endangers human health. This study reported a new method for converting of hydroxylamine by adding sodium carbonate or calcium bicarbonate to the hydroxylamine-polluted wastewater. The conversion efficiency of hydroxylamine was more than 99% in the presence of sodium carbonate or calcium bicarbonate under the reaction conditions of 25 °C, C/N ratio 15, and dissolved oxygen 7.4 mg/L. And its maximal conversion rate can reach 3.49 mg/L/h. This method overcomes various shortcomings of the reported hydroxylamine removal technologies that require a large material dosage and high cost. The technology in this report has many advantages: low cost, 'green' environmental protection, easy market promotion, and high economic benefits.

Bacillus thuringiensis EM-A1: A novel bacterium for high concentration of ammonium elimination with low nitrite accumulation.

The biological elimination of high concentration of ammonium from wastewater has attracted increasing attention in recent years. However, few studies on the efficient elimination of high concentration of ammonium by a single bacterium have been reported. Here, the efficient elimination of NH4+-N (>99%) and total nitrogen (TN) (>77%) were attained by Bacillus thuringiensis EM-A1 under 150 rpm at pH 7.2 with sodium succinate and a carbon/nitrogen ratio of 15 at 30 °C with an inoculum size (as measured by absorbance at 600 nm) of 0.2. Strain EM-A1 effectively eliminated 100 mg/L of inorganic nitrogen with maximal NH4+-N, NO3--N, and NO2--N elimination rates of 4.88, 2.57, and 3.06 mg/L/h, respectively. The elimination efficiencies of NH4+-N were 99.87% and 97.13% at initial concentrations of 500 and 1000 mg/L, respectively. Only 0.91 mg/L of NO2--N was accumulated with the elimination of 1000 mg/L NH4+-N. A concentration of 5 mg/L exogenous hydroxylamine was toxic and further inhibited heterotrophic nitrification and aerobic denitrification (HN-AD). The NH4+-N and NO2--N elimination capacities of strain EM-A1 were specifically inhibited by 2-Octyne (OCT) over 4 µmol/L and diethyldithiocarbamate (DDC) over 0.5 mmol/L, respectively. Above 25 mg/L procyanidin (PCY) inhibited the bioconversion of NO3--N and NO2--N. The results demonstrated that strain EM-A1 had HN-AD capacity under halophilic conditions, and has great potential for use in the treatment of nitrogen pollution wastewater; this study also provides new insights into this strain's nitrogen elimination mechanism, helping advance environmental biotechnology.

Efficient detoxication of hydroxylamine and nitrite through heterotrophic nitrification and aerobic denitrification by Acinetobacter johnsonii EN-J1.

The co-existence of hydroxylamine (NH2OH) and nitrite (NO2 --N) can aggravate the difficulty of wastewater treatment. The roles of hydroxylamine (NH2OH) and nitrite (NO2 --N) in accelerating the elimination of multiple nitrogen sources by a novel isolated strain of Acinetobacter johnsonii EN-J1 were investigated in this study. The results demonstrated that strain EN-J1 could eliminate 100.00% of NH2OH (22.73 mg/L) and 90.09% of NO2 --N (55.32 mg/L), with maximum consumption rates of 1.22 and 6.75 mg/L/h, respectively. Prominently, the toxic substances NH2OH and NO2 --N could both facilitate nitrogen removal rates. Compared with the control treatment, the elimination rates of nitrate (NO3 --N) and NO2 --N were enhanced by 3.44 and 2.36 mg/L/h after supplementation with 10.00 mg/L NH2OH, and those of ammonium (NH4 +-N) and NO3 --N were improved by 0.65 and 1.00 mg/L/h after the addition of 50.00 mg/L NO2 --N. Furthermore, the nitrogen balance results indicated that over 55.00% of the initial total nitrogen was transformed into gaseous nitrogen by heterotrophic nitrification and aerobic denitrification (HN-AD). Ammonia monooxygenase (AMO), hydroxylamine oxidoreductase (HAO), nitrate reductase (NR), and nitrite reductase (NIR), which are essential for HN-AD, were detected at levels of 0.54, 0.15, 0.14, and 0.01 U/mg protein, respectively. All findings confirmed that strain EN-J1 could efficiently execute HN-AD, detoxify NH2OH and NO2 --N, and ultimately promote nitrogen removal rates.

Enhanced heterotrophic nitrification and aerobic denitrification performance of Glutamicibacter arilaitensis EM-H8 with different carbon sources.

Different carbon sources for Glutamicibacter arilaitensis EM-H8 were evaluated for ammonium nitrogen (NH4+-N), nitrate nitrogen (NO3--N) and nitrite nitrogen (NO2--N) removal. Strain EM-H8 could rapidly remove NH4+-N, NO3--N and NO2--N. The highest removal rates measured for different forms of nitrogen with different carbon sources were 5.94 mg/L/h for NH4+-N with sodium citrate, 4.25 mg/L/h for NO3--N with sodium succinate, and 3.88 mg/L/h for NO2--N with sucrose. The Nitrogen balance showed that strain EM-H8 could convert 77.88% of the initial nitrogen into nitrogenous gas when NO2--N was selected as the sole nitrogen source. The presence of NH4+-N increased the removal rate of NO2--N from 3.88 to 4.02 mg/L/h. In an enzyme assay, ammonia monooxygenase, nitrate reductase and nitrite oxidoreductase were detected at 0.209, 0.314, and 0.025 U/mg protein, respectively. These results demonstrate that strain EM-H8 performs well for nitrogen removal, and shows excellent potential for simple and efficient removal of NO2--N from wastewater.

[Denitrifying phosphate accumulating organisms and its mechanism of nitrogen and phosphorus removal].

Water eutrophication poses great threats to protection of water environment. Microbial remediation of water eutrophication has shown high efficiency, low consumption and no secondary pollution, thus becoming an important approach for ecological remediation. In recent years, researches on denitrifying phosphate accumulating organisms and their application in wastewater treatment processes have received increasing attention. Different from the traditional nitrogen and phosphorus removal process conducted by denitrifying bacteria and phosphate accumulating organisms, the denitrifying phosphate accumulating organisms can simultaneously remove nitrogen and phosphorus under alternated anaerobic and anoxic/aerobic conditions. It is worth noting that microorganisms capable of simultaneously removing nitrogen and phosphorus absolutely under aerobic conditions have been reported in recent years, but the mechanisms remain unclear. This review summarizes the species and characteristics of denitrifying phosphate accumulating organisms and the microorganisms capable of performing simultaneous nitrification-denitrification and phosphorous removal. Moreover, this review analyzes the relationship between nitrogen removal and phosphorus removal and the underlying mechanisms, discusses the challenges of denitrifying phosphorus removal, and prospects future research directions, with the aim to facilitate process improvement of denitrifying phosphate accumulating organisms.

Klebsiella oxytoca (EN-B2): A novel type of simultaneous nitrification and denitrification strain for excellent total nitrogen removal during multiple nitrogen pollution wastewater treatment.

The poor total nitrogen (TN) removal rate achieved using microorganisms to treat wastewater polluted with multiple types of nitrogen was improved using a novel simultaneous nitrification and denitrification strain (Klebsiella oxytoca EN-B2). Strain EN-B2 rapidly eliminated ammonium, nitrate, and nitrite, giving maximum elimination rates of 4.58, 7.46, and 7.83 mg/(L h), respectively, equivalent to TN elimination rates of 4.35, 6.92, and 7.11 mg/(L h), respectively. The simultaneous nitrification and denitrification system gave ammonium and nitrite elimination rates of 7.14 and 9.17 mg/(L h), respectively, and a TN elimination rate ≥ 9.0 mg/(L h). Nitrogen balance calculations indicated that 51.22 %, 31.62 % and 46.82 % of TN in systems containing only ammonium, nitrite, and nitrate, respectively, were lost as nitrogenous gases. The ammonia monooxygenase, hydroxylamine oxidoreductase, nitrate reductase and nitrite reductase enzyme activities were determined. The results indicated that strain EN-B2 can be used to treat wastewater polluted with multiple types of nitrogen.

Nitrogen removal characterization and functional enzymes identification of a hypothermia bacterium Pseudomonas fragi EH-H1.

A novel hypothermic strain, Pseudomonas fragi EH-H1, was found to effectively perform heterotrophic nitrification and aerobic denitrification at 15 °C. This strain could consume 100 %, 100 % and 99.95 % of ammonium (54.90 mg∙L-1), nitrate (56.12 mg∙L-1) and nitrite (54.15 mg∙L-1), accompanied by peak removal rates of 5.51, 3.63 and 3.14 mg/L/h, respectively. The ammonium was removed preferentially during simultaneous nitrification and denitrification. Notably, the elimination rate of the toxic nitrite nitrogen remained approximately 3.14 mg/L/h, whether supplemented with ammonium or not. Stepwise inhibition experiments revealed that the key enzymes of ammonia monooxygenase (AMO) and nitrite oxidoreductase (NiR) for nitrification and denitrification coexisted in strain EH-H1. AMO, nitrate reductase and NiR were successfully expressed and detected at 0.637, 0.239 and 0.018 U/mg proteins, respectively. Overall, strain EH-H1 had an outstanding ability to remove nitrogen at low temperatures and could provide guidance for cryogenic wastewater treatment.

Isolation of Bioactive Compounds, Antibacterial Activity, and Action Mechanism of Spore Powder From Aspergillus niger xj.

Aspergillus fungi can produce a wide range of secondary metabolites, and they have represented a potential resource of novel bioactive compounds. Bacterial plant diseases have a serious impact on the sustainable development of agriculture worldwide, so it is necessary to use natural antibacterial compounds in microorganisms to control plant pathogens. This study was conducted to investigate the bioactive compounds of Aspergillus niger xj, three plant pathogens (Agrobacterium tumefaciens T-37, Erwinia carotovora EC-1, and Ralstonia solanacearum RS-2) were used as indicator bacteria, according to the biological activity tracking, five compounds were isolated from A. niger xj spore powder, and characterization of compounds was done by NMR (1H-NMR and 13C-NMR) and EI-MS and was identified as ergosterol (1), ß-sitosterol (2), 5-pentadecylresorcinol (3), 5-hydroxymethyl-2-furancarboxylic acid (4), and succinimide (5). Compounds 3 and 5 were isolated from A. niger xj for the first time. The minimum inhibitory concentration (MIC) of five compounds against three plant pathogens was evaluated, the results showed that compound 4 exhibited the strongest antibacterial activity against tested bacteria, and RS-2 was the most sensitive to compound 4, showing the lowest MIC of 15.56 µg/ml. We concluded that the mechanism of action of the compound 4 against RS-2 might be described as compound 4 acting on bacterial protein synthesis and intracellular metabolism according to the results of the scanning electron microscopy observation, permeability of cell membrane and SDS-PAGE. These results indicated that compound 4 has good potential to be as a biocontrol agent. In conclusion, the results from this study demonstrated that the compounds with antibacterial activity are of great significance of the prevention and control of plant phytopathogenic bacteria, and they may be applicable to exploring alternative approaches to integrated control of phytopathogens.

Mutation breeding of Aspergillus niger by atmospheric room temperature plasma to enhance phosphorus solubilization ability.

Background: Phosphorus (P) is abundant in soils, including organic and inorganic forms. Nevertheless, most of P compounds cannot be absorbed and used by plants. Aspergillus niger v. Tiegh is a strain that can efficiently degrade P compounds in soils. Methods: In this study, A. niger xj strain was mutated using Atmospheric Room Temperature Plasma (ARTP) technology and the strains were screened by Mo-Sb Colorimetry with strong P-solubilizing abilities. Results: Compared with the A. niger xj strain, setting the treatment time of mutagenesis to 120 s, four positive mutant strains marked as xj 90-32, xj120-12, xj120-31, and xj180-22 had higher P-solubilizing rates by 50.3%, 57.5%, 55.9%, and 61.4%, respectively. Among them, the xj120-12 is a highly efficient P solubilizing and growth-promoting strain with good application prospects. The growth characteristics such as plant height, root length, and dry and fresh biomass of peanut (Arachis hypogaea L.) increased by 33.5%, 43.8%, 43.4%, and 33.6%, respectively. Besides available P, the chlorophyll and soluble protein contents also vary degrees of increase in the P-solubilizing mutant strains. Conclusions: The results showed that the ARTP mutagenesis technology can improve the P solubilization abilities of the A. niger mutant strains and make the biomass of peanut plants was enhanced of mutant strains.

Ammonium and hydroxylamine can be preferentially removed during simultaneous nitrification and denitrification by Pseudomonas taiwanensis EN-F2.

To overcome a large amount of nitrite accumulation and poor removal rate for hydroxylamine, a simultaneous nitrification and denitrification (SND) bacterium was isolated and identified as Pseudomonas taiwanensis EN-F2 by DNA sequencing. Strain EN-F2 could remove 100% of ammonium (52.90 mg/L), 100% of hydroxylamine (23.32 mg/L), 86.99% of nitrite (56.32 mg/L) and 89.21% of nitrate (56.18 mg/L) with a maximum removal rate of 8.72, 2.12, 4.55 and 5.80 mg/L/h, respectively. Ammonium and hydroxylamine could be preferentially removed during the SND process. The nitrite removal rate and cell growth were substantially enhanced by 2.10 mg/L/h and 0.45 after supplementation of hydroxylamine. The specific activities of ammonia monooxygenase (AMO), hydroxylamine oxidoreductase (HAO), nitrate reductase (NR), nitrite reductase (NIR) were successfully detected as 0.95, 0.31, 0.42 and 0.03 U/mg protein, respectively. All results demonstrated that strain EN-F2 could perform SND to remove multiple nitrogen sources from wastewater.

New insight into the nitrogen removal capacity and mechanism of Streptomyces mediolani EM-B2.

The utilization of actinomycetes as the bioresources for heterotrophic nitrification and aerobic denitrification is rarely reported due to the lack of work to explore their nitrogen biodegradation capabilities. Streptomyces mediolani EM-B2 belonging to actinomycetes could effectively remove high concentration of multiple nitrogen forms, and the maximum removal rates of ammonium, nitrate and nitrite reached 3.46 mg/(L·h), 1.71 mg/(L·h) and 1.73 mg/(L·h), respectively. Nitrite was preferentially consumed from the simultaneous nitrification and denitrification reaction system. Nitrogen balance analysis uncovered that more than 37% of the initial total nitrogen was converted to nitrogenous gas by aerobic denitrification. Experiments with specific inhibitors of nitrification and denitrification revealed that strain EM-B2 contained ammonia monooxygenase, hydroxylamine oxidoreductase, nitrate reductase and nitrite oxidoreductase, which were successfully expressed and detected as 0.43, 0.59, 0.12 and 0.005 U/mg proteins, respectively. These findings may provide new insights into the actinomycetes for bioremediation of nitrogen pollution wastewater.

Efficient hydroxylamine removal through heterotrophic nitrification by novel bacterium Glutamicibacter arilaitensis EM-H8.

Hydroxylamine, an intermediate product in the nitrification process, is widely found in nature. However, hydroxylamine accumulation can decrease the biological nitrogen removal efficiency by reducing the activities, inhibiting the reproductions, and even causing the death of microorganisms. In this study, a novel heterotrophic nitrification bacterium was separated from biogas digester and identified as Glutamicibacter arilaitensis EM-H8. Strain EM-H8 exhibited efficient hydroxylamine removal (93.75%). The optimal conditions for hydroxylamine removal were as follows: Carbon source, glucose; C/N ratio, 25; temperature, 20 °C; inoculum size, 0.53 × 108 CFU; and shaking speed, 150 rpm. The nitrogen balance results using strain EM-H8 showed that about 26.86% of the initial nitrogen was removed as nitrogenous gas, while 4.6% was converted into biomass under aerobic conditions, confirming that strain EM-H8 possessed the capacity for heterotrophic nitrification. Furthermore, the successful expression of hydroxylamine oxidase (0.065 U/mg protein) showed that strain EM-H8 had the ability to transform hydroxylamine from wastewater.

Hydroxylamine and nitrite are removed effectively by Streptomyces mediolani strain EM-B2.

Biological nitrogen removal is primarily conducted by bacteria and fungi rather than actinomycetes. However, accumulations of nitrite and hydroxylamine could significantly impair the biological nitrogen removal process. A strain of Streptomyces mediolani, termed EM-B2, was isolated from a cow dung fermentation biogas digester. The strain removed more than 99% of ammonium and 78% of total nitrogen in the presence of glucose and under environmental conditions of 30 °C, a carbon/nitrogen ratio of 15, 7.4 mg/L dissolved oxygen and a pH range of 7.5-9.0. Maximal removal rates were 2.29 mg/L/h for ammonium, 1.90 mg/L/h for nitrate and 2.01 mg/L/h for nitrite. The removal efficiencies of hydroxylamine and total nitrogen peaked at 81.48% and 60.38%, respectively. Notably, hydroxylamine and nitrite were never detected during the heterotrophic nitrification and aerobic denitrification. Nitrate rather than nitrite was accumulated from the process of hydroxylamine oxidation. These findings indicate that S. mediolani strain EM-B2 performs heterotrophic nitrification and aerobic denitrification, and can be used to remove hydroxylamine and nitrite from wastewater.

Hypothermia Pseudomonas taiwanensis J488 exhibited strong tolerance capacity to high dosages of divalent metal ions during nitrogen removal process.

The nitrogen metabolic pathways of Pseudomonas taiwanensis J488 have not been confirmed from genomic function analysis and its divalent metal ion resistance remains poorly understood. In this study, the key denitrifying gene of Pseudomonas taiwanensis J488, nirB, was determined by draft genome sequencing. The nitrification of ammonium was insensitive to high concentrations of Ca(II), Mn(II), Zn(II), and Cd(II). Similarly, complete nitrite removal was achieved despite Mn(II) and Zn(II) reaching concentrations up to 30 mg/L. Furthermore, the efficiency of nitrate removal was significantly enhanced by 1.33%, 3.33%, 5.99%, and 1.53% with the addition of 0.5 mg/L Ca(II), 20 mg/L Mn(II), 5 mg/L Zn(II), and 2 mg/L Cd(II), respectively, comparison with the control. The bacterial growth in both nitrifying and denitrifying processes was substantially promoted by various dosages of divalent metal ions. These results indicate that divalent metal ions would not severely limit the capacity of strain J488 to purify nitrogen-polluted wastewater.

Isolation and Identification of Antibacterial Bioactive Compounds From Bacillus megaterium L2.

Bacterial metabolites exhibit a variety of biologically active compounds including antibacterial and antifungal activities. It is well known that Bacillus is considered to be a promising source of bioactive secondary metabolites. Most plant pathogens have an incredible ability to mutate and acquire resistance, causing major economic losses in the agricultural field. Therefore, it is necessary to use the natural antibacterial compounds in microbes to control plant pathogens. This study was conducted to investigate the bio-active compounds of Bacillus megaterium L2. According to the activity guidance of Agrobacterium tumefaciens T-37, Erwinia carotovora EC-1 and Ralstonia solanacearum RS-2, five monomeric compounds, including erucamide (1), behenic acid (2), palmitic acid (3), phenylacetic acid (4), and ß-sitosterol (5), were fractionated and purified from the crude ethyl acetate extract of B. megaterium. To our knowledge, all compounds were isolated from the bacterium for the first time. To understand the antimicrobial activity of these compounds, and their minimum inhibitory concentrations (MICs) (range: 0.98∼500 µg/mL) were determined by the broth microdilution method. For the three tested pathogens, palmitic acid exhibited almost no antibacterial activity (>500 µg/mL), while erucamide had moderate antibacterial activity (MIC = 500 µg/mL). Behenic acid showed MICs of 250 µg/mL against T-37 and RS-2 strains with an antibacterial activity. ß-sitosterol showed significant antimicrobial activity against RS-2. ß-sitosterol showed remarkable antimicrobial activity against RS-2 with an MIC of 15.6 µg/mL. In addition, with the antimicrobial activity, against T-37 (62.5 µg/mL) and against EC-1 (125 µg/mL) and RS-2 (15.6 µg/mL) strains notably, phenylacetic acid may be interesting for the prevention and control of phytopathogenic bacteria. Our findings suggest that isolated compounds such as behenic acid, ß-sitosterol, and phenylacetic acid may be promising candidates for natural antimicrobial agents.

Nitrous oxide produced directly from ammonium, nitrate and nitrite during nitrification and denitrification.

A hypothermia aerobic denitrifying bacterium, Pseudomonas taiwanensis strain J488, can effectively remove multiple nitrogen sources from wastewater at 15 °C. The ammonium, nitrate and nitrite removal efficiencies were 100 %, 92.61 % and 92.49 %, respectively. Strain J488 could survive with hydroxylamine as sole nitrogen source and its removal efficiency was 97.71 %. The removal efficiency of ammonium was 100 % even in the presence of the classical inhibitors of nitrification allylthiourea and diethyldithiocarbamate. These findings fundamentally changed the picture that the ammonia monooxygenase could be inhibited by the copper chelators of allylthiourea or diethyldithiocarbamate. Similarly, the nitrite removal capacity of strain J488 was not sensitive to inhibition by Pb2+, and its removal efficiency was also 100 %. Additionally, by identifying the intermediates accumulation of nitrification and denitrification, using nitrification and denitrification inhibitors, measuring enzyme activities and determining N2O concentrations, it was demonstrated that N2O could be produced directly from ammonium, nitrate and nitrite.