The purpose of this study was to identify candidate novel species performing beneficial ecological functions from soil. The hypothesis was that novel culturable species with antibiotic production potential could be readily found from soil in the Nature Canyon at Los Angeles Pierce College. 137 bacterial isolates from soil were grown on nutrient agar, ISP-6 medium, or LB agar and partial 16S was ascertained using the Sanger platform for regions V1-V4. One of the potentially novel isolates, Pseudomonas sp. 21-C3-ER, was isolated on nutrient agar from fallow soil in an agricultural nature preserve in Los Angeles. The soil conditions were 3.7% moisture, clay texture by touch, 13.37% organic matter, pH=6.63, TDS=638, 150 ppm nitrogen, 114 ppm phosphate, and 42 ppm potash. Whole genome sequencing was performed on the DNBseq platform and species identification was performed with TRUEBacID; the closest relative was Pseudomonas brassicacearum subsp. neoaurantiaca which was 92.05% similar by genomic evidence. 197 virulence factors were identified including flagellar biosynthesis, motor proteins, Beta-lactamase, Fosfomycin, copper, and sulfur drug resistance. Gram stain of strain 21-C3-ER revealed gram negative rods. Rods were visualized with scanning electron microscopy, revealing individuals approximately 1 µm in length. Secondary metabolite analysis using antiSMASH showed a variety of post-translationally modified proteins and a promysalin analog. The qualitative test for pyoverdine siderophore production using UV light was positive. Metabolic analysis using RAST indicated genes were present for pyoverdine production, antimicrobial resistance, and ammonification. The nitrate reduction test was positive. Denitrifying bacteria may prevent leaching of nitrates from soil, since ammonia has a higher affinity to soil and organic matter than nitrate. We propose Pseudomonas fernandeno as a novel species. The type stain is 21-C3-ER.

Groundwater contamination is an ever-growing environmental issue that has attracted much and undiminished attention for the past half century. Groundwater contamination may originate from both anthropogenic (e.g., hydrocarbons) and natural compounds (e.g., nitrate and arsenic); to tackle the removal of these contaminants, different technologies have been developed and implemented. Recently, bioelectrochemical systems (BES) have emerged as a potential treatment for groundwater contamination, with reported in situ applications that showed promising results. Nitrate and hydrocarbons (toluene, phenanthrene, benzene, BTEX and light PAHs) have been successfully removed, due to the interaction of microbial metabolism with poised electrodes, in addition to physical migration due to the electric field generated in a BES. The selection of proper BESs relies on several factors and problems, such as the complexity of groundwater and subsoil environment, scale-up issues, and energy requirements that need to be accounted for. Modeling efforts could help predict case scenarios and select a proper design and approach, while BES-based biosensing could help monitoring remediation processes. In this review, we critically analyze in situ BES applications for groundwater remediation, focusing in particular on different proposed setups, and we identify and discuss the existing research gaps in the field.

Electrochemical methods are used for the treatment of both municipal and industrial wastewater, either independently or in conjunction with biological methods. This conjunction is used for pretreatment or posttreatment of biologically treated wastewater. In our work, we focused on the combination of these processes, where pre-electrolysis was used to produce dissolved iron before the activation process. Electrolysis was also directly introduced into the activation using either iron or carbon electrodes. The surface of one iron electrode was 32.2 cm2, the applied voltage was 21 V, and the supplied current was 270 mA. The surface of one carbon electrode was 7.54 cm2, the supplied current was 82.5 mA, and the applied voltage was 21 V. From laboratory research on the treatment of synthetic municipal wastewater using a combination of electrolysis and activation processes, it resulted that the use of iron electrodes increased the efficiency of phosphorus removal compared to its precipitation with iron salts. The electrolysis had a positive effect on the sedimentation properties of sludge and the destruction of filamentous microorganisms. Electrolysis negatively affected the respiration rates of activated sludge and the efficiency of denitrification. However, it did not have a negative impact on the nitrification activity of the sludge.

Genes coding for enzymes of the denitrification pathway appear randomly distributed among isolates of the ancestral genus Thermus, but only in few strains of the species T. thermophilus the pathway has been studied to a certain detail. Here, we review the enzymes involved in this pathway present in T. thermophilus NAR1, a strain extensively employed as a model for nitrate respiration, on the light of its full sequence recently assembled through a combination of PacBio and Illumina technologies in order to counteract the systematic errors introduced by the former technique. The genome of this strain is divided in four replicons, a chromosome of 2,021,843 pb, two megaplasmids of 370,865 and 77,135 bp and a small plasmid of 9,799 pb. Nitrate respiration is encoded in the largest megaplasmid, pTTHNP4, within a region that includes operons for O2 and nitrate sensory systems, a nitrate reductase, nitrate and nitrite transporters and a nitrate specific NADH dehydrogenase, in addition to multiple insertion sequences (IS), suggesting its mobility-prone nature. Despite nitrite is the final product of nitrate respiration in this strain, the megaplasmid encodes two putative nitrite reductases of the cd1 and Cu-containing types, apparently inactivated by IS. No nitric oxide reductase genes have been found within this region, although the NorR sensory gene, needed for its expression, is found near the inactive nitrite respiration system. These data clearly support that partial denitrification in this strain is the consequence of recent deletions and IS insertions in genes involved in nitrite respiration. Based on these data, the capability of this strain to transfer or acquire denitrification clusters by horizontal gene transfer is discussed.

In this study, a three-dimensional bioelectrochemical reactor system (3D-BERs) with granular activated carbon (GAC) epitomizes a novel treatment technology for treating nitrate-polluted water. The conventional denitrification process faces many challenges, including the huge demand for organic carbon, long-term accumulation of intermediate products, and the adaptation period. Results shown that under the optimal conditions of the COD/NO3--N ratio was 1.5, the denitrification efficiency reached 98.62%, when compared to 81.12% at COD/ NO3--N ratio of 3.5, and the initial pH of 7.5 ± 0.5, NO3--N was entirely removed at 2.2 h without accumulation of nitrite. The high initial ratio of NO2--N/NO3--N is mainly to accelerate the denitrification rate by accelerating the reduction of nitrite. Denitrification process followed by zero-order kinetics linear model for at different concentrations of inlet NO3--N, and achieved higher denitrification rate at greater inlet NO3--N concentration. High-throughput sequencing shows that the community structure and relative abundance of bacteria changed significantly, especially at the genes and the phyla level in immobilized GAC particles. Microbial composition enhanced the removal of nitrogen at the inner surface (IS) and bottom surface (BS) of immobilized GAC carriers. Therefore, this system is expected to be a more efficient and useful supplement or a cost-effective alternative compared to the traditional low carbon to nitrogen wastewater treatment system.

To meet the growing interest in the development of innovative biological nutrient removal (BNR) alternatives for energy neutrality, resource recovery, and decarbonization, the adsorption/bio-oxidation (A/B) process has been widely studied for carbon capture and shortcut nitrogen (N) removal, while limited research has focused on incorporating enhanced biological phosphorus (P) removal (EBPR), mainly due to the differential carbon usage characteristics of functionally relevant microorganisms. Here, a full-scale pilot testing with an integrated system combining A-stage high-rate activated sludge (HRAS) with B-stage partial (de)nitrification/anammox and side-stream EBPR (HRAS-P(D)N/A-S2EBPR) was conducted treating real municipal wastewater. The results demonstrated that, despite the low influent carbon load, the B-stage P(D)N-S2EBPR system achieved effective and stable P removal performance, especially when the volatile fatty acid (VFA) load from A-stage was elevated. Sludge fermentation in both A-stage and B-stage promoted carbon redistribution and likely provided more competitive advantages for ammonium-oxidizing bacteria and polyphosphate accumulating organisms, leading to carbon-efficient shortcut N removal mainly through partial nitrification pathway and influent carbon-independent EBPR simultaneously. Exposure to high VFA levels was considered a potential selection factor for the suppression of nitrite-oxidizing bacteria in the system. The involvement of internal carbon-accumulating organisms would potentially play an important role in endogenous denitrification. This study provided new insights into the effects of incorporating side-stream EBPR into the A/B process on microbial ecology, metabolic activities, and system performance.

Nitrogen (N) is an essential nutrient and plays an important role in plant growth and physiology. In addition, N is also the limiting nutrient in most terrestrial ecosystems. The increasing use of N fertilizers increases agricultural production, but also has negative impacts on biodiversity, water quality and increases emissions of greenhouse gases such as nitrogen oxides (NOx) and nitrous oxide (N2O) into the atmosphere. N2O is a strong greenhouse gas and the product of microbial transformation processes of N introduced into soil and groundwater (nitrification and denitrification). The production of N2O in soils is highly dependent on oxygen and water content, soil temperature and texture and the available amount of reactive nitrogen (NO3- or NH4+). In agricultural soils, N2O emissions are also influenced by the type of fertilizer used, crops grown, soil pH and NO3- concentration. Refined forms of land management, such as the cultivation of legumes, can reduce the use of fertilizers and thus also the emission of N2O. Legumes can use symbiotic nodule bacteria (rhizobia) to bind atmospheric N2 and make it available to the plant. Non-symbiotic soil microorganisms such as cyanobacteria or other heterotrophic and autotrophic prokaryotes are also able to fix N2. Furthermore, the N2-fixing enzyme nitrogenase, which is specific for most organisms, is not specific for other N2 compounds, especially N2O. To study the N2 and N2O fixation potential of soils and legumes, experiments with isotope-enriched N-gases (100 mol% 15N2 and 15N2O) were performed. Three different soil types (forest, meadow and wetland) as well as legume plants inoculated with rhizobia bacteria (Rhizobium leucaneae) (Leucaena leucocephala) were incubated in microcosms with different mixing ratios of N2 and N2O over a period of one week. The measured 15N enrichment was then used to determine the biological uptake rates. Both in plants and in soils admixtures of up to 40% N2O had no influence on the N2 fixation rate. N2 uptake rates were 1.2 ± 0.4 ug N gdw-1 d- 1in forest and grassland soils and 3.9 ± 1.2 ug N gdw-1 d- 1 in wetland soils. In contrast, the N2 fixation rates of leguminous plants were significantly higher (130 ± 40 ug N gdw-1 d- 1), with the highest accumulation not in root nodules but in the roots. In both plants and soil, the N2O fixation of 0.2 ± 0.1 ug N gdw-1 d- 1 could only be measured from a concentration of 4 vol%. The results indicate that both soils and legumes have a high potential to bind atmospheric nitrogen in the form of both N2 but not N2O. The fate of nitrogen in soil needs to be further investigated. N2 fixation seems to play an important role in wetland soils. Legumes in particular have the potential for NH4+ input and N2O reduction, as most of the solid nitrogen has been transferred to the roots, which can contribute significantly to soil nutrient input.

Preventing the rebound of black and odorous water bodies is critical for improving the ecological environment of water bodies. This study investigated the effect and mechanism of in-situ improvement of sediment microenvironment by nitrate in tailwater of wastewater treatment plants combined with aerobic denitrifying bacteria under low-DO regulation (TailN+CFM+LDO). On the 60th day of remediation, the dissolved oxygen and oxidation-reduction potential in the overlying water increased to 5.6 mg/L and 300 mV, respectively, the acid volatile sulfide in the sediment decreased by 70.4%, and the organic matter in the sediment was reduced by 62.7%, in which the HFOM was degraded from 105 g/kg to 56 g/kg, and the potential risk of water reverting to black and odorous conditions significantly decreased. Amplicon sequencing analysis revealed that the relative abundance of electroactive bacteria Thiobacillus and Pseudomonas with denitrification capacity was found to be significantly higher in the TailN+CFM+LDO group than in the other remediation groups. Functional prediction of 16S sequencing results indicated that both the quantity and activity of critical microbial enzymes involved in nitrification and denitrification processes could enhance in the TailN+CFM+LDO group. These results improved our understanding of the improvement of the sediment microenvironment and could thus facilitate its application.

This article deals with the possibility of partial replacement of blast furnace slag with fly ash after denitrification by the Selective Non-Catalytic Reduction (SNCR) method in alkali-activated materials. In the experiment, the basic physical-mechanical properties and durability properties were tested, the hydration reaction was monitored in a calorimeter and infrared spectroscopy was performed. Results were compared between mixtures prepared with fly ash without denitrification and also with reference mixture based only on alkali-activated blast furnace slag. The basic result is the finding, that hybrid alkali-systems with fly ash after denitrification show similar trends as hybrid alkali-systems with fly ash without denitrification. The significant effect of fly ash is manifested especially in terms of resistance to freeze-thaw. The reactions in the calorimeter show a slower development of reactions with increasing replacement of slag by fly ash. In the case of testing resistance to leaching in demineralised water, a decrease of flexural strength was found, which corresponds to the conclusions of strength testing, that long-term deposition of bodies in water causes deterioration of mechanical properties.

Emerging pollutants as pharmaceuticals have been focusing international attention for few decades. Ciprofloxacin (CIP) is a common drug widely found in effluents from hospitals, industrial and different wastewater treatment plants, as well as rivers. In this work, the lab-scale 3D-BER system was established, and more than 90% of the antibiotic CIP removal from the Low C/N wastewater. Best results were obtained with current intensity, and different C/N ratio significantly improve the removal of CIP and nitrates, when the ideal conditions were; C/N = 1.5-3.5, pH =7.0-7.5, and I = 60 mA. The highest removal efficiency of CIP = 94.20 %, NO3--N= 95.53 % and total nitrogen (TN) = 84.27 %, respectively. In this novel system, the autotrophic-heterotrophic denitrifying bacteria played vital role for the removal of CIP and enhanced denitrification process. Thus, autotrophic denitrifying bacteria uses CO2 and H2 as carbon sources to reduce nitrates to N2. This system has the assortment and prosperous community revealed at the current intensity of 60 mA, and the analysis of bacterial community structure in effluent samples fluctuates under different condition of C/N ratios. According to the results of LC-MS/MS analysis, the intermediate products were proposed after efficient biodegradation of CIP. Microbial community on biodegrading was mostly found at phylum, and class level was dominantly responsible for the NO3--N and biodegradation of CIP. This work can provide some new insights towards the biodegradation of CIP and the efficient removal of nitrates from low C/N wastewater treatment by the novel 3D-BER system.

The low-temperature environment significantly inhibits the growth and metabolism of denitrifying bacteria, leading to an excessive concentration of ammonia nitrogen and total nitrogen in sewage treatment plants during the cold season. In this study, an efficient denitrifying strain of heterotrophic nitrification–aerobic denitrification(HN-AD) bacteria named HS2 was isolated and screened from industrial sewage of a chemical factory in Inner Mongolia at 8°C. The strain was confirmed to be Achromobacter spiritinus, a colorless rod-shaped bacterium. When cultured with sodium succinate as the carbon source, a carbon-to-nitrogen ratio of 20-30, a shaking rate of 150-180 r/min, and the initial pH of 6-10, the Strain HS2 exhibited excellent nitrogen removal at 8°C. Through the results of whole-genome sequencing, gene amplification, and gas product detection, the Strain HS2 was determined to possess key enzyme genes in both nitrification and denitrification pathways, suggesting a HN-AD pathway of NH4+-N→NH2OH→NO2--N→NO→N2O→N2. At 8°C, the Strain HS2 could completely remove ammonia nitrogen from industrial sewage with an initial concentration of 127.23 mg/L. Microbial species diversity analysis of the final sewage confirmed Achromobacter sp. as the dominant genus, this indicated that the low-temperature denitrifying strain HS2 plays an important role in nitrogen removal of actual low-temperature sewage.

This study describes the seasonal course of denitrifying and nitrate reducing bacteria in a dimictic mesotrophic lake (Lake Scharmützelsee, Brandenburg, Germany) within a three-year period from 2011 to 2013. The bacterial cell numbers were quantified by fluorescence microscopy, most probable number (MPN) and PCR dependent quantification of the chromosomal 16S rDNA and of the nirS and nirK gene copy number. The highest seasonal differences (up to three orders of magnitudes) has been measured by MPN in the epilimnion. This variation was not reflected by PCR dependent approaches or direct microscopical enumeration. At adverse conditions (low temperature and/or low nitrate concentrations), the differences between MPN and gene copy numbers increased up to five orders of magnitudes and decreased to one magnitude at favorable environmental conditions. These results can be explained best by an increasing ratio of viable but not cultivable (VBNC) cells or dead cells at impairing conditions. In the hypolimnion, the courses of MPN and nir gene copy numbers were similar. This can be explained by higher feeding pressure and therefore smaller amounts of dormant cells. In the pelagial in general, the total cell numbers enumerated by either microscopical or molecular approaches were similar. In the sediment, more than 99 % of the DNA was obviously not related to viable bacteria but was rather “dead DNA” in dead cells or adsorbed to particle surfaces.