Abstract  The photochemical degradation of two azo and two anthraquinonic dyes was performed using potassium peroxymonosulphate (Oxone®) activated by UV radiation. The fast decolourization of all dyes was observed within 6 min of UV irradiation, with corresponding dye decays higher than 80%. The kinetic rate constants of the dyes' decay were determined, along with the energetic efficiency of the photochemical treatment, taking into account the influence of a few anions commonly present in real wastewaters (i.e., chloride, nitrate, carbonate/bicarbonate and phosphate ions). Chloride and carbonate/bicarbonate ions enhanced dye degradation, whereas phosphate ions exerted an inhibitory effect, and nitrates did not have a predictable influence. The dye decolourization was not associated with efficient mineralization, as suggested by the lack of a significant total organic carbon (TOC) decrease, as well as by the low concentrations of a few detected low molecular weight by-products, including nitrate ions, formaldehyde and organic acids. High molecular weight by-products were also detected by mass spectrometry analysis. The investigated process may be proposed as a convenient pre-treatment to help dye degradation in wastewater during combined treatment methods.

Abstract  Biostimulation to induce reduction of soluble U(VI) to relatively immobile U(IV) is an effective strategy for decreasing aqueous U(VI) concentrations in contaminated groundwater systems. If oxidation of U(IV) occurs following the biostimulation phase, U(VI) concentrations increase, challenging the long-term effectiveness of this technique. However, detecting U(IV) oxidation through dissolved U concentrations alone can prove difficult in locations with few groundwater wells to track the addition of U to a mass of groundwater. We propose the 238U/235U ratio of aqueous U as an independent, reliable tracer of U(IV) remobilization via oxidation or mobilization of colloids. Reduction of U(VI) produces 238U-enriched U(IV), whereas remobilization of solid U(IV) should not induce isotopic fractionation. The incorporation of remobilized U(IV) with a high 238U/235U ratio into the aqueous U(VI) pool produces an increase in 238U/235U of aqueous U(VI). During several injections of nitrate to induce U(IV) oxidation, 238U/235U consistently increased, suggesting 238U/235U is broadly applicable for detecting mobilization of U(IV).

Abstract  The geographical variation of denitrifying bacterial communities and water quality parameters in urban lakes distributed across nine provinces in China were determined. The Illumina sequencing data of the denitrifying encoding gene nirS was examined in the samples collected from nine localities (pairwise geographical distance: 200-2600 km). The results showed that fundamental differences in water quality were observed among different urban lakes. The highest nitrate (2.02 mg L-1) and total nitrogen (3.82 mg L-1) concentrations were observed in Pingzhuang (P < 0.01). The algal cell concentration ranged from 1.29 x 10(8) to 3.0 x 10(9) cell per L. The sequencing data generated a total of 421058 high quality nirS gene reads that resulted in 6369 OTUs (97% cutoff), with Proteobacteria and Firmicutes being the dominant taxa. A co-occurrence network analysis indicated that the top five genera identified as keystone taxa were Dechlorospirillum sp., Alicycliphilus sp., Dechloromonas sp., Pseudogulbenkiania sp., and Paracoccus sp. A redundancy analysis (RDA) further revealed that distinct denitrifying bacterial communities inhabited the different urban lakes, and influenced by urban lake water ammonia nitrogen, manganese and algal cell concentrations. A variance partitioning analysis (VPA) also showed that geographic location was more important than water quality factors in structuring the denitrifying bacterial communities. Together, these results provide new insight into understanding of denitrifying bacterial communities associated with geographically distributed urban lakes on a larger scale, and these results also expand our exploration of aquatic microbial ecology in freshwater bodies.

Abstract  Improving fertility of marginal soils for the sustainable production of biomass is a strategy for reducing land use conflicts between food and energy crops. Digestates can be used as fertilizer and for soil amelioration. In order to promote plant growth and reduce potential adverse effects on roots because of broadcast digestate fertilization, we propose to apply local digestate depots placed into the rhizosphere. We grew Sida hermaphrodita in large mesocosms outdoors for three growing seasons and in rhizotrons in the greenhouse for 3 months both filled with marginal substrate, including multiple sampling dates. We compared digestate broadcast application with digestate depot fertilization and a mineral fertilizer control. We show that depot fertilization promotes a deep reaching root system of S. hermaphrodita seedlings followed by the formation of a dense root cluster around the depot-fertilized zone, resulting in a fivefold increased biomass yield. Temporal adverse effects on root growth were linked to high initial concentrations of ammonium and nitrite in the rhizosphere in either fertilizer application, followed by a high biomass increase after its microbial conversion to nitrate. We conclude that digestate depot fertilization can contribute to an improved cultivation of perennial energy-crops on marginal soils.

Abstract  Ingestion of geophagic materials might affect human health and induce diseases by different ways. The purpose of this study is to determine the geochemical composition of geophagic material consumed especially by pregnant women in Onangama Village, Northern Namibia and to assess its possible health effects. X-ray fluorescence and inductively coupled plasma mass spectrometry were used in order to determine the major, and trace elements as well as anions concentrations of the consumed material. The geochemical analysis revealed high concentrations of aluminium (Al), calcium (Ca), iron (Fe), magnesium (Mg), manganese (Mn), potassium (K), sodium (Na), and silica (Si); and trace elements including arsenic (As), chromium (Cr), mercury (Hg), nickel (Ni) and vanadium (V) as well as sulphate (SO42-), nitrate (NO3-), and nitrite (NO2-) anions comparing to the recommended daily allowance for pregnant women. The pH for some of the studied samples is alkaline, which might increase the gastrointestinal tract pH (pH < 2) and cause a decrease in the bioavailability of elements. The calculated health risk index (HRI > 1) revealed that Al and Mn might be a potential risk for human consumption. Based on the results obtained from the geochemical analysis, the consumption of the studied material might present a potential health risk to pregnant women including concomitant detrimental maternal and foetal effects.

Abstract  To understand the physiological responses of the brown macroalga Macrocystis integrifolia during the marine tidal cycle, two RNA libraries were prepared from algal frond samples collected in the intertidal zone (0 m depth) and subtidal zone (10 m depth). Samples collected from intertidal zone during low tide was considered as abiotic stressed (MI0), while samples collected from subtidal zone was considered as control (MI10). Both RNA libraries were sequenced on Illumina NextSeq 500 which generated approx. 46.9 million and 47.7 million raw paired-end reads for MI0 and MI10, respectively. Among the representative transcripts (RTs), a total of 16,398 RTs (39.20%) from MI0 and 21,646 RTs (39.24%) from MI10 were successfully annotated. A total of 535 unigenes (271 upregulated and 264 downregulated) showed significantly altered expression between MI0 and MI10. In abiotic-stressed condition (MI0), the relative expression levels of genes associated with antioxidant defenses (vanadium-dependent bromoperoxidase, glutathione S-transferase, lipoxygenase, serine/threonine-protein kinase, aspartate Aminotransferase, HSPs), water transport (aquaporin), photosynthesis (light-harvesting complex) protein were significantly upregulated, while in control condition (MI10) most of the genes predominantly involved in energy metabolism (NADH-ubiquinone oxidoreductase/NADH dehydrogenase, NAD(P)H-Nitrate reductase, long-chain acyl-CoA synthetase, udp-n-acetylglucosamine pyrophosphorylase) were overexpressed.

Abstract  Organohalide-respiring bacteria are key players for the turnover of organohalogens. At sites impacted with chlorinated ethenes, bioremediation promotes reductive dechlorination; however, stoichiometric conversion to environmentally benign ethene is not always achieved. We demonstrate that nitrous oxide (N2O), a compound commonly present in groundwater, inhibits organohalide respiration. N2O concentrations in the low micromolar range decreased dechlorination rates and resulted in incomplete dechlorination of tetrachloroethene (PCE) in Geobacter lovleyi strain SZ and of cis-1,2-dichloroethene ( cDCE) and vinyl chloride (VC) in Dehalococcoides mccartyi strain BAV1 axenic cultures. Presumably, N2O interferes with reductive dechlorination by reacting with super-reduced Co(I)-corrinoids of reductive dehalogenases, which is supported by the finding that N2O did not inhibit corrinoid-independent fumarate-to-succinate reduction in strain SZ. Kinetic analyses revealed a best fit to the noncompetitive Michaelis-Menten inhibition model, and determined N2O inhibitory constants, KI, for PCE and cDCE dechlorination of 40.8 ± 3.8 µM and 21.2 ± 3.5 µM in strain SZ and strain BAV1, respectively. The lowest KI value of 9.6 ± 0.4 µM was determined for VC to ethene reductive dechlorination in strain BAV1, suggesting that this crucial dechlorination step for achieving detoxification is most susceptible to N2O inhibition. Groundwater N2O concentrations exceeding 100 µM are not uncommon, especially in watersheds impacted by nitrate runoff from agricultural sources. Thus, dissolved N2O measurements can inform about cDCE and VC stalls at sites impacted with chlorinated ethenes.

Abstract  Breeding for pollution-safe cultivars (PSCs) can reduce pollutant accumulation in crops. However, the PSC breeding would face the risk of nutritional quality reduction, which is usually ignored in conventional breeding programs targeting to increase crop yield or nutritional quality. Thus, the doubt whether the risk would exist has to be clarified for supporting the PSC breeding. In the present study, a newly bred Cd/Pb-PSC of water spinach (Ipomoea aquatic Forsk.) and its parents (QLQ with low-Cd/Pb accumulation ability and T308 with high yield) of water spinach were employed to clarify the above-mentioned issue. Yields, and concentrations of Cd, Pb, nitrite, and organic and inorganic nutrients in shoots of the three experimental lines were determined. There were no significant differences in Cd/Pb concentration between the new PSC and QLQ, in nitrite content between the new PSC and its two parents and in yield between the new PSC and T308. It is decisively significant that shoot concentrations of organic and inorganic nutrients in the Cd/Pb-PSC were as high as those in one of its parents. It is affirmed that the breeding operations (crossing and consequently continuous selfing) for lowering Cd/Pb accumulation capacity of water spinach would not lower the nutritional values of the obtained Cd/Pb-PSCs from the breeding, which should be a pillar that supports the feasibility to minimize Cd/Pb pollution in vegetables using PSC-breeding method.

Abstract  MAIN CONCLUSION: An SPL-type transcription factor, LeSPL-CNR, is negatively involved in NO production by modulating SlNR expression and nitrate reductase activity, which contributes to Cd tolerance. Cadmium (Cd) is a highly toxic pollutant. Identifying factors affecting Cd accumulation in plants is a prerequisite for minimizing dietary uptake of Cd from crops grown with contaminated soil. Here, we report the involvement of a SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE (SPL) transcription factor LeSPL-CNR in Cd tolerance in tomato (Solanum lycopersicum). In comparison with the wild-type Ailsa Craig (AC) plants, the Colourless non-ripening (Cnr) epimutant displayed increased Cd accumulation and enhanced sensitivity to Cd, which was in well accordance with the repression of LeSPL-CNR expression. Cd stress-induced NO production was inhibited by nitrate reductase (NR) inhibitor, but not NO synthase-like enzyme inhibitor. Expression of LeSPL-CNR was negatively correlated with SlNR expression and the NR activity. We also demonstrated that LeSPL-CNR inhibited the SlNR promoter activity in vivo and bound to SlNR promoter sequence that does not contain a known SBP-binding motif. In addition, expression of an IRON-REGULATED TRANSPORTER1, SlIRT1, was more abundant in Cnr roots than AC roots under Cd stress. LeSPL-CNR may thus provide a molecular mechanism linking Cd stress response to regulation of NR-dependent NO production, which then contributes to Cd uptake via SlIRT1 expression in tomato.

Abstract  The implementation of hydrogenotrophic denitrification is limited due to safety concerns, poor H2 utilization and low solubility of H2 gas with the resulting low transfer rate. The current paper presents the main research work conducted on a pressurized hydrogenotrophic reactor for denitrification that was recently developed. The reactor is based on a new concept suggesting that a gas-liquid equilibrium is achieved in the closed headspace of denitrifying reactor, further produced N2 gas is carried out by the effluent and gas purging is not required. The feasibility of the proposed reactor was shown for two effluent concentrations of 10 and 1 mg NO3(-)-N/L. Hydrogen gas utilization efficiencies of 92.8% and 96.9% were measured for the two effluent concentrations, respectively. Reactor modeling predicted high denitrification rates above 4 g NO3(-)-N/(Lreactor·d) at reasonable operational conditions. Hydrogen utilization efficiency was improved up to almost 100% by combining the pressurized reactor with a following open-to-atmosphere polishing unit. Also, the potential of the reactor to remove ClO4(-) was shown.

Abstract  Waste tire rubber (WTR) has been introduced as an alternative, novel media for biofilm development in several experimental systems including attached growth bioreactors. In this context, four laboratory-scale static batch bioreactors containing WTR as a support material for biofilm development were run under anoxic condition for 90 days using waste activated sludge as an inoculum under the influence of different concentrations (2.5, 6.5, 8.5 mg/l) of trivalent ferric iron (Fe(3+)). The data revealed that activated sludge with a Fe(3+) concentration of 8.5 mg/l supported the maximum bacterial biomass [4.73E + 10 CFU/ml cm(2)]; besides, it removed 38% more Chemical oxygen demand compared to Fe(3+) free condition from the reactor. Biochemical testing and 16S rDNA phylogenetic analysis of WTR-derived biofilm communities further suggested the role of varying concentrations of Fe(3+) on the density and diversity of members of Enterobacteria(ceae), ammonium (AOB) and nitrite oxidizing bacteria. Furthermore, Fluorescent in situ hybridization with phylogenetic oligonucleotide probes and confocal laser scanning microscopy of WTR biofilms indicated a significant increase in density of eubacteria (3.00E + 01 to.05E + 02 cells/cm(2)) and beta proteobacteria (8.10E + 01 to 1.42E + 02 cells/cm(2)), respectively, with an increase in Fe(3+) concentration in the reactors, whereas, the cell density of gamma proteobacteria in biofilms decreased.

Abstract  The effects of florfenicol (FF) on the performance, microbial activity and microbial community of a sequencing batch biofilm reactor (SBBR) were evaluated in treating mariculture wastewater. The chemical oxygen demand (COD) and nitrogen removal were inhibited at high FF concentrations. The specific oxygen utilization rate (SOUR), specific ammonium oxidation rate (SAOR), specific nitrite oxidation rate (SNOR) and specific nitrate reduction rate (SNRR) were decreased with an increase in the FF concentration from 0 to 35 mg/L. The chemical compositions of loosely bound extracellular polymeric substances (LB-EPS) and tightly bound EPS (TB-EPS) could be affected with an increase in the FF concentration. The high-throughput sequencing indicated some obvious variations in the microbial community at different FF concentrations. The relative abundance of Nitrosomonas and Nitrospira showed a decreasing tendency with an increase in the FF concentration, suggesting that FF could affect the nitrification process of SBBR. Some genera capable of reducing nitrate to nitrogen gas could be inhibited by the addition of FF in the influent, such as Azospirillum and Hyphomicrobium.

Abstract  The aim of this study was to evaluate the effect of light intensity and temperature on nutrient removal and biomass productivity in a microalgae-bacteria culture and their effects on the microalgae-bacteria competition. Three experiments were carried out at constant temperature and various light intensities: 40, 85 and 125 µE m(-2) s(-1). Other two experiments were carried out at variable temperatures: 23 ± 2°C and 28 ± 2°C at light intensity of 85 and 125 µE m(-2) s(-1), respectively. The photobioreactor was fed by the effluent from an anaerobic membrane bioreactor. High nitrogen and phosphorus removal efficiencies (about 99%) were achieved under the following operating conditions: 85-125 µE m(-2) s(-1) and 22 ± 1°C. In the microalgae-bacteria culture studied, increasing light intensity favoured microalgae growth and limited the nitrification process. However, a non-graduated temperature increase (up to 32°C) under the light intensities studied caused the proliferation of nitrifying bacteria and the nitrite and nitrate accumulation. Hence, light intensity and temperature are key parameters in the control of the microalgae-bacteria competition. Biomass productivity significantly increased with light intensity, reaching 50.5 ± 9.6, 80.3 ± 6.5 and 94.3 ± 7.9 mgVSS L(-1) d(-1) for a light intensity of 40, 85 and 125 µE m(-2) s(-1), respectively.

Abstract  The purpose of this study was to investigate the marginal adaptation and microleakage of SonicFill composite with different photopolymerization techniques. Class II cavities were prepared in 40 premolars and divided into groups according to the photopolymerization technique (n = 10): G1:1200 mW/cm(2) for 20 s; G2:1200 mW/cm(2) for 40 s; G3:Soft-start with 650 mW/cm(2) for 5 s and 1200 mW/cm(2) for 15 s; G4:Soft-start with 650 mW/cm(2) for 10 s and 1200 mW/cm(2) for 30 s. The cavities were restored with OptiBond FL adhesive and SonicFill. Epoxy resin replicas were obtained before and after thermocycling. The occlusal and cervical margins were analyzed with scanning electron microcopy and expressed as the percentage of continuous margins (%CM). The specimens were submitted to microleakage with silver nitrate. ANOVA and Tukey's test revealed that the %CM at the linguo-occlusal margin for group 1 (83.19%) was significantly different from groups 2, 3 and 4, which had values over 95%. At the cervical margin, there was no statistically significant difference between the groups. After thermocycling, there was a significant decrease in %CM. The predominant score of microleakage was 1 in enamel and 3 in dentin. The SonicFill composite led to gap formation and microleakage, especially in the dentin at the cervical margin, regardless of the photopolymerization technique employed.

Abstract  High CO2 is able to ameliorate some negative effects due to climate change and intensify others. This study involves the sweet cherry (Prunus avium) cultivar 'Burlat' grafted on the 'Mariana 2624', 'Adara' and 'LC 52' rootstocks. In a climate chamber at two CO2 concentrations, ambient (400 µmol mol(-1) ) and elevated (800 µmol mol(-1) ), the plants were submitted to waterlogging for 7 d, followed by 7 d of recovery after drainage. Waterlogging drastically decreased the rate of photosynthesis, significantly endangering plant survival, particularly for the 'LC 52' and 'Adara' rootstocks. 'Mariana 2624' was also clearly affected by waterlogging that increased lipid peroxidation and the Cl(-) and SO4(2-) concentrations in all the studied plants. Nevertheless, CO2 was able to overcome this reduction in photosynthesis, augmenting growth, increasing soluble sugars and starch, raising turgor and regulating the concentrations of Cl(-) and SO4(2-) , while lowering the NO3(-) concentration in leaves of all the studied rootstocks. In concordance with these results, the proline levels indicated a more intense stress at control CO2 than at high CO2 for waterlogged plants. 'Mariana 2624' was more resistant to waterlogging than 'Adara', and both were more resistant than 'LC 52' in control CO2 conditions; this clearly enhanced the chance of survival under hypoxia.

Abstract  5,10,15,20-Tetrakis(4-sulfonatophenyl)porphyrinato iron(III) chloride (FeTPPS) is a water-soluble heme analog, which has been used as a scavenger of peroxynitrite in many studies. Similar to heme, it may also possess pseudo-peroxidase activity that could cause protein tyrosine nitration through the peroxidase-H2O2-NO2- pathway. In this paper, we used western blotting and spectrophotometry analysis to study the capability of FeTPPS in catalyzing protein tyrosine nitration. Furthermore, the capability of FeTPPS in catalyzing protein nitration in tissue homogenate and cultured cells was also investigated. Our results showed that FeTPPS induced bovine serum albumin (BSA) nitration in the presence of H2O2 and NaNO2, and the reaction was dose-, time- and pH-dependent. In acidic condition, more protein was nitrated by FeTPPS than heme, which corresponded to their peroxidase activities. Meanwhile, our results also confirmed the catalytic effect of FeTPPS on protein tyrosine nitration in rat brain homogenate and human hepatocellular carcinoma (HepG2) cells. At the end of this study, we used liquid chromatography (LC)-tandem mass spectrometry (MS/MS) to investigate differences of site selectivity between heme and FeTPPS catalyzed protein tyrosine nitration. The result indicated that FeTPPS tended to catalyze tyrosine residues locating in more hydrophilic sites, whereas heme was more likely to induce nitration of tyrosine residues locating in relatively hydrophobic environment. Taken together, this is the first report that FeTPPS is an effective and convenient nitration catalyzer in vitro, and this study confirms that the hydrophilicity of the nitrating agents would play an important role in nitration site selection.

Abstract  Accurate estimation of constituent loads is important for studies of ecosystem mass balance or total maximum daily loads. In response, there has been an effort to develop methods to increase both accuracy and precision of constituent load estimates. The relationship between constituent concentration and stream discharge is often complicated, potentially leading to high uncertainty in load estimates for certain constituents, especially at longer-term (annual) scales. We used the loadflex R package to compare uncertainty in annual load estimates from concentration vs. discharge relationships in constituents of interest in agricultural systems, including ammonium as nitrogen (NH4-N), nitrate as nitrogen (NO3-N), soluble reactive phosphorus (SRP), and suspended sediments (SS). We predicted that uncertainty would be greatest in NO3-N and SS due to complex relationships between constituent concentration and discharge. We also predicted lower uncertainty with a composite method compared to regression or interpolation methods. Contrary to predictions, we observed the lowest uncertainty in annual NO3-N load estimates (relative error 1.5-23%); however, uncertainty was greatest in SS load estimates, consistent with predictions (relative error 19-96%). For all constituents, we also generally observed reductions in uncertainty by up to 34% using the composite method compared to regression and interpolation approaches, as predicted. These results highlight differences in uncertainty among different constituents and will aid in model selection for future studies requiring accurate and precise estimates of constituent load.

Abstract  Free nitrous acid (FNA) exerts a broad range of antimicrobial effects on bacteria, although susceptibility varies considerably among microorganisms. Among nitrifiers found in activated sludge of wastewater treatment processes (WWTPs), nitrite-oxidizing bacteria (NOB) are more susceptible to FNA compared to ammonia-oxidizing bacteria (AOB). This selective inhibition of NOB over AOB in WWTPs bypasses nitrate production and improves the efficiency and costs of the nitrogen removal process in both the activated sludge and anaerobic ammonium oxidation (Anammox) system. However, the molecular mechanisms governing this atypical tolerance of AOB to FNA have yet to be understood. Herein we investigate the varying effects of the antimicrobial FNA on activated sludge containing AOB and NOB using an integrated metagenomics and label-free quantitative sequential windowed acquisition of all theoretical fragment ion mass spectra (SWATH-MS) metaproteomic approach. The Nitrosomonas genus of AOB, on exposure to FNA, maintains internal homeostasis by upregulating a number of known oxidative stress enzymes, such as pteridine reductase and dihydrolipoyl dehydrogenase. Denitrifying enzymes were upregulated on exposure to FNA, suggesting the detoxification of nitrite to nitric oxide. Interestingly, proteins involved in stress response mechanisms, such as DNA and protein repair enzymes, phage prevention proteins, and iron transport proteins, were upregulated on exposure to FNA. In addition enzymes involved in energy generation were also upregulated on exposure to FNA. The total proteins specifically derived from the NOB genus Nitrobacter was low and, as such, did not allow for the elucidation of the response mechanism to FNA exposure. These findings give us an understanding of the adaptive mechanisms of tolerance within the AOB Nitrosomonas to the biocidal agent FNA.

Abstract  Bioaugmentation was used to upgrade the nitrification process in a full-scale municipal WWTP with an A2/O system. A mixture of nitrifying bacteria was inoculated into the bioreactor for a final concentration of 1% (v/v). The upgrade process took 25 days, and the NH4+-N removals reached 94.6% (increased at least by 75%). The effluent concentrations of COD and NH4+-N stabilized at <30 mg/L and <4 mg/L even when the corresponding influent concentrations were over 300 mg/L and 60 mg/L, which met the first-class requirement of the National Municipal Wastewater Discharge Standards of China (COD ≤ 50 mg/L, NH4+-N ≤ 5 mg/L). The succession of the microbial community showed the enhanced NH4+-N removal efficiency mainly resulted from the persistence of introduced ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB), which increased from 0% to 0.4% and from 0.01% to 2.1%, respectively. This bioaugmentation was shown as an effective technology for upgrading or retrofitting conventional systems to tertiary-level.

Abstract  RATIONALE: The isotopic tracer technique is widely used to identify the sources and fate of nitrogen (N) in order to understand the N cycle and contamination in water environments. The stable isotope ratio of ammonium is expected to greatly enhance the tracing analysis by combining it with the traditional nitrate isotope ratio. Diffusion followed by gas-phase trapping is the most commonly applied method for ammonium isotope ratio measurement. Although dissolved organic nitrogen (DON) is abundant in natural water and its breakdown in the diffusion procedure has been reported, the interference of DON with the measurement of ammonium isotope ratios has not been fully examined.

METHODS: This study aims to test the effect of DON contamination by using organic N compounds, viz. humic acid and alanine. A series of diffusion experiments was conducted at a temperature of 80°C for a maximum of 7 days. Ammonia was transferred from alkaline solution and trapped with an acidic filter. This method was applied for samples with ammonium concentrations between 0.5 and 2.0 mg-N/L.

RESULTS: There was no difference between the ammonium N stable isotope ratios for samples with and without added DON compounds; the fractionation between the observed value and the actual value was negligible, in the range of 0.2 to 1.0‰. The modifications of previous studies, i.e. shorter diffusion period, no vigorous shaking and using gas-phase trapping, successfully avoided any breakdown of DON in fresh water samples.

CONCLUSIONS: The modified method provides high precision and accuracy and it is recommended for the analysis of anthropogenically influenced water samples, such as paddy fields, ground water, rivers and lakes.

Abstract  Although Nitrobacter winogradskyi is an important chemoorganotrophic organism for the study of nitrite-oxidizing bacteria physiology as well as nitrification, until now, the mixotrophic medium for this organism growth has not been optimized, comprehensively. In this study, we aimed to improve the growth medium of N. winogradskyi using the one-factor-at-a-time (NaNO2 , glycerol, pH) method. In addition, a further experimental design was carried out based on central composite design with response surface methodology. Different combinations of the three cultural parameters were fitted by multiple regression analysis to calculate the predicted response. Our results suggest that optimal culture condition for the growth of N. winogradskyi was a modified DSMZ 756a medium containing NaNO2 (5·74 g l-1 ) and glycerol (37·88 mmol l-1 ), pH 7·83, a temperature of 28°C and agitation at 120 rev min-1 . The results from a validation experiment (bacterial growth: OD600 1·0293) were close to the value predicted by the quadratic model (OD600 1·0994). In addition, we uncovered the potential mechanism at the cellular and ultrastructural levels. The results indicated that glycerol in the media enhanced the rate of cell division and cell growth by increasing the accumulation of polyphosphates and phosphorus, and high concentrations of NaNO2 provided sufficient energy for growth and contributed to the generation of carboxysomes in cells for CO2 fixation.

SIGNIFICANCE AND IMPACT OF THE STUDY: Due to the extremely slow growth rate and the low growth yield of ammonia-oxidizing bacteria and NOB (nitrite-oxidizing bacteria), nitrification is still the rate-limiting step of nitrogen cycle in the current research. Nitrobacter winogradskyi, an important NOB, participates in the second step of nitrification in water and soil. This study reported an optimized culture condition for N. winogradskyi, which increased the growth yield by 5·06 times than that in the basal medium and uncovered the potential mechanism. We expect our study will contribute to the research on water and soil nitrogen cycle. In addition, the optimized culture conditions have the potential to be suitable for the chemoorganotrophic growth of other nitrifiers.

Abstract  Poly-γ-glutamic acid (γ-PGA) is an important multifunctional biopolymer with various applications, for which adenosine triphosphate (ATP) supply plays a vital role in biosynthesis. In this study, the enhancement of γ-PGA production was attempted through various approaches of improving ATP supply in the engineered strains of Bacillus licheniformis. The first approach is to engineer respiration chain branches of B. licheniformis, elimination of cytochrome bd oxidase branch reduced the maintenance coefficient, leading to a 19.27% increase of γ-PGA yield. The second approach is to introduce Vitreoscilla hemoglobin (VHB) into recombinant B. licheniformis, led to a 13.32% increase of γ-PGA yield. In the third approach, the genes purB and adK in ATP-biosynthetic pathway were respectively overexpressed, with the AdK overexpressed strain increased γ-PGA yield by 14.69%. Our study also confirmed that the respiratory nitrate reductase, NarGHIJ, is responsible for the conversion of nitrate to nitrite, and assimilatory nitrate reductase NasBC is for conversion of nitrite to ammonia. Both NarGHIJ and NasBC were positively regulated by the two-component system ResD-ResE, and overexpression of NarG, NasC, and ResD also improved the ATP supply and the consequent γ-PGA yield. Based on the above individual methods, a method of combining the deletion of cydBC gene and overexpression of genes vgB, adK, and resD were used to enhance ATP content of the cells to 3.53 μmol/g of DCW, the mutant WX-BCVAR with this enhancement produced 43.81 g/L of γ-PGA, a 38.64% improvement compared to wild-type strain WX-02. Collectively, our results demonstrate that improving ATP content in B. licheniformis is an efficient strategy to improve γ-PGA production.

Abstract  Water-soluble organic matter (WSOM) represents a critical fraction of fine particles (PM2.5) in the air, but its changing behaviors and formation mechanisms are not well understood yet, partly due to the lack of fast techniques for the ambient measurements. In this study, a novel system for the on-line measurement of water-soluble components in PM2.5, the particle-into-liquid sampler (PILS)-Nebulizer-aerosol chemical speciation monitor (ACSM), was developed by combining a PILS, a nebulizer, and an ACSM. High time resolution concentrations of WSOM, sulfate, nitrate, ammonium, and chloride, as well as mass spectra, can be obtained with satisfied quality control results. The system was firstly applied in China for field measurement of WSOM. The mass spectrum of WSOM was found to resemble that of oxygenated organic aerosol, and WSOM agreed well with secondary inorganic ions. All evidence collected in the field campaign demonstrated that WSOM could be a good surrogate of secondary organic aerosol (SOA). The PILS-Nebulizer-ACSM system can thus be a useful tool for intensive study of WSOM and SOA in PM2.5.

Abstract  Nitrification plays a crucial role in global nitrogen cycling and treatment processes. However, the relationships between the nitrifier guilds of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) are still poorly understood, especially in freshwater habitats. This study examined the physiological interactions between the AOB and NOB present in a freshwater aquarium biofilter by culturing them, either together or separately, in a synthetic medium. Metagenomic and 16S rRNA gene sequencing revealed the presence and the draft genomes of Nitrosomonas-like AOB as well as Nitrobacter-like NOB in the cultures, including the first draft genome of Nitrobacter vulgaris. The nitrifiers exhibited different growth rates with different ammonium (NH4+) or nitrite concentrations (50-1,500 μM) and the growth rates were elevated under a high bicarbonate (HCO3-) concentration. The half-saturation constant (Ks for NH4+), the maximum growth rate (μmax), and the lag duration indicated a strong dependence on the synergistic relationships between the two guilds. Overall, the ecophysiological and metagenomic results in this study provided insights into the phylogeny of the key nitrifying players in a freshwater biofilter and showed that interactions between the two nitrifying guilds in a microbial community enhanced nitrification.

Abstract  Two lab-scale nitrifying sequencing batch biofilm reactors, with (SBBR_CN) or without the addition of organics (SBBR_N), were operated to investigate potential roles of acyl homoserine lactone (AHL) based quorum sensing. AHLs of N-[(RS)-3-Hydroxybutyryl]-L-homoserine lactone, N-hexanoyl-L-homoserine lactone (C6-HSL) and N-octanoyl-L-homoserine lactone (C8-HSL) were detected in both reactors. C6-HSL and C8-HSL were also detected in batch experiments, especially with stimulated nitrite oxidizing bacteria activities. Quorum sensing affected biofilm formation mainly through the regulation of extracellular protein production. By the metagenomics analysis, many identified genera and species could participate in quorum sensing, quorum quenching and extracellular polymeric substances (EPS) production. A high quorum quenching activity was obtained in SBBR_CN, whereas a high quorum sensing activity in SBBR_N. Nitrosomonas-like ammonia oxidizing bacteria, Nitrospira-like nitrite oxidizing bacteria and Comammox harbored genes for AHL synthesis and EPS production. Possible relationships among AHLs synthesis, biofilm formation and nitrifiers activity were proposed.