Photon costs of shoot and root NO3-, and root NH4+, assimilation in terrestrial vascular plants considering associated pH regulation, osmotic and ontogenetic effects.

The photon costs of photoreduction/assimilation of nitrate (NO3-) into organic nitrogen in shoots and respiratory driven NO3- and NH4+ assimilation in roots are compared for terrestrial vascular plants, considering associated pH regulation, osmotic and ontogenetic effects. Different mechanisms of neutralisation of the hydroxyl (OH-) ion necessarily generated in shoot NO3- assimilation are considered. Photoreduction/assimilation of NO3- in shoots with malic acid synthesis and either accumulation of malate in leaf vacuoles or transport of malate to roots and catabolism there have a similar cost which is around 35% less than that for root NO3- assimilation and around 20% less than that for photoreduction/assimilation of NO3-, oxalate production and storage of Ca oxalate in leaf vacuoles. The photon cost of root NH4+ assimilation with H+ efflux to the root medium is around 70% less than that of root NO3- assimilation. These differences in photon cost must be considered in the context of the use of a combination of locations of NO3- assimilation and mechanisms of acid-base regulation, and a maximum of 4.9-9.1% of total photon absorption needed for growth and maintenance that is devoted to NO3- assimilation and acid-base regulation.

Ecological risk assessment of glyphosate and its possible effect on bacterial community in surface sediments of a typical shallow Lake, northern China.

Glyphosate is a widely used herbicide worldwide and its prevalent presence in aquatic ecosystems poses a threat to living organisms. This study evaluated potential ecological risk of glyphosate to sediment-dwelling organisms and assessed the probable effect of glyphosate on structure and predicated function of sediment-attached bacterial communities from a large shallow lake in northern China based on 16S rRNA high-throughput sequencing. Results suggested that glyphosate showed a medium to high concentration (up to 8.63 mg/kg) and chronic risk to sediment-dwelling organisms (10% samples exhibiting medium to high risk quotient), especially in sites nearby farmland and residential areas in August. Bacterial community identification based on 16S rRNA sequence indicated some species of dominant phylum Proteobacteria and Campilobacterota (e.g., Steroidobacteraceae, Thiobacillus, Gallionellaceae, Sulfurimonadaceae) were stimulated while some species of dominant phylum Actinobacteriota, Acidobacteriota and Firmicutes (e.g., Nocardioidaceae, Microtrichales, Vicinamibacteraceae, Paenisporosarcina) were inhibited by glyphosate accumulation. The stimulating species were related to sulfur-oxidizing, sulfate-, iron-, or nitrate-reducing bacteria; The inhibiting species were related to plant bacterial endophytes, polyphosphate-accumulating organisms (PAOs) and denitrifers. Correspondingly, promoted bacterial metabolic functions of "sulfite respiration", "nitrogen respiration", "aromatic compound degradation" and "nitrification" but suppressed "cellulolysis", "manganese oxidation", "anoxygenic photoautotrophy S oxidizing" and "nitrate denitrification" were predicated on functional annotation of prokaryotic taxa. Although these results could only partly suggest the impacts of glyphosate on the bacterial communities due to the lack of actual results from control experiments, the identified Steroidobacteraceae could be thought as a bioindicator in the future mechanism study for the ecological effect and bioremediation of glyphosate. This work intends to arise the concern about the depletion of biodiversity and bacterial metabolic functions with contribution of glyphosate in part in eutrophic lakes.

Nitrite lowers the oxygen cost of ATP supply in cultured skeletal muscle cells by stimulating the rate of glycolytic ATP synthesis.

Dietary nitrate lowers the oxygen cost of human exercise. This effect has been suggested to result from stimulation of coupling efficiency of skeletal muscle oxidative phosphorylation by reduced nitrate derivatives. In this paper, we report the acute effects of sodium nitrite on the bioenergetic behaviour of cultured rat (L6) myocytes. At odds with improved efficiency of mitochondrial ATP synthesis, extracellular flux analysis reveals that a ½-hour exposure to NaNO2 (0.1-5 µM) does not affect mitochondrial coupling efficiency in static myoblasts or in spontaneously contracting myotubes. Unexpectedly, NaNO2 stimulates the rate of glycolytic ATP production in both myoblasts and myotubes. Increased ATP supply through glycolysis does not emerge at the expense of oxidative phosphorylation, which means that NaNO2 acutely increases the rate of overall myocellular ATP synthesis, significantly so in myoblasts and tending towards significance in contractile myotubes. Notably, NaNO2 exposure shifts myocytes to a more glycolytic bioenergetic phenotype. Mitochondrial oxygen consumption does not decrease after NaNO2 exposure, and non-mitochondrial respiration tends to drop. When total ATP synthesis rates are expressed in relation to total cellular oxygen consumption rates, it thus transpires that NaNO2 lowers the oxygen cost of ATP supply in cultured L6 myocytes.

Does energy cost constitute the primary cause of ammonium toxicity in plants?

Nitrate (NO3-) and ammonium (NH4+) are the main nitrogen (N) sources and key determinants for plant growth and development. In recent decades, NH4+, which is a double-sided N compound, has attracted considerable amounts of attention from researchers. Elucidating the mechanisms of NH4+ toxicity and exploring the means to overcome this toxicity are necessary to improve agricultural sustainability. In this review, we discuss the current knowledge concerning the energy consumption and production underlying NH4+ metabolism and toxicity in plants, such as N uptake; assimilation; cellular pH homeostasis; and functions of the plasma membrane (PM), vacuolar H+-ATPase and H+-pyrophosphatase (H+-PPase). We also discuss whether the overconsumption of energy is the primary cause of NH4+ toxicity or constitutes a fundamental strategy for plants to adapt to high-NH4+ stress. In addition, the effects of regulators on energy production and consumption and other physiological processes are listed for evaluating the possibility of high energy costs associated with NH4+ toxicity. This review is helpful for exploring the tolerance mechanisms and for developing NH4+-tolerant varieties as well as agronomic techniques to alleviate the effects of NH4+ stress in the field.

Differential responses of bloom-forming Ulva intestinalis and economically important Gracilariopsis lemaneiformis to marine heatwaves under changing nitrate conditions.

Marine heatwaves (MHWs) are affecting the survival of macroalgae. However, little is known regarding how the impacts of MHWs are regulated by nitrogen availability. In this study, we investigated the physiological and genetic responses of a green-tide macroalga Ulva intestinalis Linnaeus and a commercially cultivated macroalga Gracilariopsis lemaneiformis (Bory) E.Y. Dawson, Acleto & Foldvik under different nitrate conditions to simulated MHWs. Under nitrogen limited conditions (LN), heatwaves did not significantly affect biomass or Fv/Fm of U. intestinalis although it led to an earlier biomass decline due to more reproduction events, and meanwhile an upregulation in genes related to TCA cycle and oxidative phosphorylation was detected, supporting sporulation. Under nitrogen replete conditions (HN), heatwaves did not change biomass, Fv/Fm or photosynthetic pigments but reduced reproduction rate along with insignificant change of oxidative phosphorylation and TCA cycle related genes. Meanwhile, genes related to photosynthesis and glutathione metabolism were upregulated. Regarding G. lemaneiformis, heatwaves reduced its Fv/Fm and photosynthetic pigments content, leading to bleaching and death, and photosynthesis-related genes were also downregulated at LN. Fv/Fm was improved and photosynthesis-related genes were up-regulated by the combination of nitrogen enrichment and heatwaves, whereas G. lemaneiformis remained bleached and died by day 12. Therefore, U. intestinalis could survive heatwaves through shifting to micropropagules at LN and protecting its photosynthesis at HN. In contrast, G. lemaneiformis died of bleaching when suffering heatwaves regardless of nitrogen availability. These findings suggest that in future oceans with eutrophication and MHWs, the harmful alga U. intestinalis may have more advantages over the economic alga G. lemaneiformis.

Maximizing US nitrate removal through wetland protection and restoration.

Growing populations and agricultural intensification have led to raised riverine nitrogen (N) loads, widespread oxygen depletion in coastal zones (coastal hypoxia)1 and increases in the incidence of algal blooms.Although recent work has suggested that individual wetlands have the potential to improve water quality2-9, little is known about the current magnitude of wetland N removal at the landscape scale. Here we use National Wetland Inventory data and 5-kilometre grid-scale estimates of N inputs and outputs to demonstrate that current N removal by US wetlands (about 860 ± 160 kilotonnes of nitrogen per year) is limited by a spatial disconnect between high-density wetland areas and N hotspots. Our model simulations suggest that a spatially targeted increase in US wetland area by 10 per cent (5.1 million hectares) would double wetland N removal. This increase would provide an estimated 54 per cent decrease in N loading in nitrate-affected watersheds such as the Mississippi River Basin. The costs of this increase in area would be approximately 3.3 billion US dollars annually across the USA-nearly twice the cost of wetland restoration on non-agricultural, undeveloped land-but would provide approximately 40 times more N removal. These results suggest that water quality improvements, as well as other types of ecosystem services such as flood control and fish and wildlife habitat, should be considered when creating policy regarding wetland restoration and protection.

Probabilistic risk assessment of nitrates for Austrian adults and estimation of the magnitude of their conversion into nitrites.

Nitrates occur in food naturally, as contaminants or additives. The health implications attributed to ingested nitrates result primarily from their conversion into nitrites and subsequent methemoglobinemia, carcinogenicity induced by N-nitroso-compounds and cardiovascular, endocrine, metabolic, reproductive and developmental effects. The present study comprises a probabilistic tiered risk assessment of nitrates for Austrian adults through the diet with the application of the Monte Carlo simulation method in alternative optimistic and pessimistic scenarios. Risk estimates are of concern regarding the upper exposures, which exceed the Acceptable Daily Intake (ADI) in almost all scenarios and population groups. Exposure is elevated when all dietary sources are considered and the ADI is exceeded by already the mean intake for vegetarians. Leafy vegetables are major contributors to the intake. Contribution of cured meat is very low. Estimates of the conversion of nitrates into nitrites were used to assess the combined exposure to both species. When the average intake of nitrates and nitrites is considered, the mean exposure to nitrites is lower or close to the ADI for individuals with average conversion capacity. However, upper tail combined intake can lead to a multifold exceedance of the ADI of nitrites for individuals with both high and average conversion capacity.

Investigation into the Concentrations and Sources of Nitrates and Nitrites in Milk and Plant-Based Powders.

Milk powders in the United States (US) may contain nitrates and nitrites from several potential sources. These sources include the ingestion of nitrates and nitrites by dairy cows during grazing and drinking, nitric acid used during the sanitization of dairy equipment, and the production of nitrous oxides in directly heated spray dryers. Recently, milk powders manufactured in the US have been rejected during import to other countries because nitrite concentrations were greater than 2 mg/kg (ppm). To date, the concentrations of nitrates and nitrites in milk and plant-based powders in the US are unknown. In this study the nitrate and nitrite concentrations present in diverse milk powders were investigated including 81 milk powders from local and online retailers from 2015 to 2018. In addition, 71 commercial milk powders were obtained from blinded production facilities. Nitrate and nitrite concentrations were determined using ion chromatography with conductivity and UV detection. A subset of samples was analyzed for N-nitrosodimethylamine using gas chromatography-mass spectrometry. Carbon and nitrogen bulk isotope ratios analyzed using isotope ratio mass spectrometry were used to obtain some insights into the production method (organic vs conventional) and geographic source of the milk powder samples. Background nitrate concentrations in US-produced milk powder samples averaged 17 ± 12 mg/kg. Nitrite was detected at concentrations greater than 2 mg/kg in 5 out of 39 different brands of retail milk and plant-based powders. Of these brands, two were plant-based (soy and coconut) powders and the other three had consistently high nitrites. The analysis of milk powders using stable isotope analysis revealed further information about the cow's diet.

Incorporating Exogenous and Endogenous Exposures into Dietary Risk Assessment of Nitrates and Nitrites in Vegetables: A Probabilistic Integrated Toxicokinetic Modeling Approach.

This study aimed to estimate the dietary risk of nitrates and nitrites in vegetables based on internal dose in a probabilistic manner by integrating exogenous exposure based on measured concentrations in vegetables with endogenous exposure using a toxicokinetic (TK) model. We optimized and validated a previous TK model and incorporated Monte Carlo simulations to account for variability across different age populations for predicting internal dose. High levels of nitrates were detected in leafy vegetables (from 545 ± 274 to 1641 ± 873 mg/kg). Nitrite contents of vegetables were generally low (from 1.26 ± 1.40 to 8.20 ± 14.1 mg/kg). The dietary risk was found to be different based on internal versus external dose, suggesting that it is critical to include endogenous nitrite formation into risk assessment. Nitrate and nitrite exposure from vegetables is unlikely to result in appreciable risks for most populations but may be a potential risk for preschoolers.

Oxidative stress assessment in breath-hold diving.

PURPOSE: Breath-hold diving results in significant changes in blood gases' levels. Challenging variations in oxygen partial pressures may induce reactive oxygen species (ROS) production that exacerbate oxidative stress and, consequently, affect endothelial function. The aim of this study was to investigate the effects of breath-hold diving on oxidative stress damage, assessing ROS production. Nitric oxide metabolites, inducible nitric oxide synthase (iNOS), aminothiols, and renal function were evaluated too as markers of redox status and renal damage. METHODS: ROS production was assessed with electron paramagnetic resonance. Oxidative status values were measured at pre- and post-40 m dive in a deep swimming pool (Y-40) from six divers (mean age 46.6 ± 9.3 years; height 176 ± 4 cm; BMI 25 ± 2.9 kg/m2). RESULTS: Significant (p < 0.05) increases at post-dive of ROS production rate (0.158 ± 0.003 vs 0.195 ± 0.006 µmol min-1), lipid peroxidation (8-isoprostane: 375.67 ± 195.62 vs 420.49 ± 232.31 pg mg-1 creatinine), nitrate (27.91 ± 19.71 vs 30.80 ± 20.44 µM), iNOS (31.30 ± 4.52 vs 35.68 ± 6.72 IU mL-1) and neopterin concentration (96.20 ± 40.41 vs 118.76 ± 27.84 µmol mol-1 creatinine) were recorded. Conversely, the antioxidant capacity significantly decreased (3.423 ± 0.089 vs 3.015 ± 0.284 mM) after immersion. CONCLUSION: Overproduction of ROS and consequent oxidative damage to lipids of membrane and antioxidant capacity decreasing reflect also a hypoxic condition, which in the breath-hold diving typically occurs in the last few meters below the surface. iNOS produces NO in large quantities under the examined extreme conditions. Neopterin and creatinine concentration level increased, suggesting an "impairment of renal function" as a likely physiological response to PaO2 variations during dive activity.

High-efficient nitrogen removal from municipal wastewater via two-stage nitritation/anammox process: Long-term stability assessment and mechanism analysis.

This study focused on the long-term stability of a novel two-stage partial-nitritation/anammox (PN/A) process treating municipal wastewater with fluctuated water quality. Specifically, two parallel sequencing batch reactors (SBRs) were used for removing organic matters and achieving complete nitritation, while the expanded granular sludge bed (ANA-EGSB) was used for anammox. With the influent ammonium concentration varying from 32 to 79 mg/L and the average hydraulic retention time of 3.39 h in this system, more than 93% of ammonium was removed and the effluent TIN was 4.8-11.8 mg/L. The partial denitrifying occurring in the anammox reactor could reduce nitrate to nitrite that was reused by anammox bacterium, enhancing the TIN removal efficiency. Further, the "overconsumption of ammonium" under anaerobic conditions was observed in ANA-EGSB. Microbial community analysis showed that Nitrosomonas (AOB) were the dominant nitrifying bacteria in the nitritation SBR and Candidatus_Brocadia with the relative abundance of 6-13% dominated in ANA-EGSB.

Interstitial water microbial communities as an indicator of microbial denitrifying capacity in wood-chip bioreactors.

The discharge from food production greenhouses (greenhouse effluent) contains high nutrient and salt concentrations, which, if directly released, can have adverse effects on the environment. Wood-chip bioreactors are increasingly popular passive water treatment systems favoured for their economical denitrification in treating agricultural field tile drainage. Microbial communities are central to denitrification; however little is known about the maturation of microbial communities in wood-chip bioreactors treating greenhouse effluents. In this study, multiple subsurface flow wood-chip bioreactors, each vegetated with a different plant species, together with an unplanted unit, received synthetic greenhouse effluent with elevated nitrate concentrations. The hybrid bioreactors were operated for over 2 years, during which time water samples were collected from the inlet, outlet and within the reactors. The increasing denitrification rate in the bioreactor planted with Typha angustifolia (narrowleaf cattail) correlated with increasing microbial activity and metabolic richness, measured by the carbon utilization patterns in Biolog® EcoPlates. Increased denitrifying gene (nirS) copies (determined by quantitative polymerase chain reaction, qPCR), and near-complete nitrate removal were observed in the T. angustifolia and unplanted reactors after 16 and 23 months of operation respectively. The findings suggested that an acclimation period of at least one year can be expected in unseeded bioreactors planted with T. angustifolia, while bioreactors without vegetation may require a longer time to maximize their denitrification capacity. These results are important for the design and operation of wood-chip bioreactors, which are expected to be more commonly applied in the future.

A metatranscriptomics-based assessment of small-scale mixing of sulfidic and oxic waters on redoxcline prokaryotic communities.

Lateral intrusions of oxygen caused by small-scale mixing are thought to shape microbial activity in marine redoxclines. To examine the response of prokaryotes to such mixing events we employed a shipboard mixing experiment in the euxinic central Baltic Sea: oxic, nitrate containing and sulfidic water samples without detectable oxygenized substances were incubated directly or after mixing. While nitrate, nitrite and ammonium concentrations stayed approximately constant in all incubations, we observed a decrease of sulfide after the contact with oxygen in the sulfide containing incubations. The transcription of marker genes from chemolithoauthotrophic key players including archaeal nitrifiers as well as gammaproteobacterial and campylobacterial autotrophic organisms that couple denitrification with sulfur-oxidation were followed at four time points within 8.5 h. The temporally contrasting transcriptional profiles of gammaproteobacterial and campylobacterial denitrifiers that depend on the same inorganic substrates pointed to a niche separation. Particular archaeal and campylobacterial marker genes involved in nitrification, denitrification and sulfur oxidation, which depend on oxidized substrates, were highly upregulated in the anaerobic sulfidic samples. We suggest that, despite the absence of measurable oxygenated compounds in the sulfidic water, frequent intermittent small-scale intrusions stimulate the permanent upregulation of genes involved in nitrification, denitrification and sulfur oxidation.

Understanding the response of Desulfovibrio desulfuricans ATCC 27774 to the electron acceptors nitrate and sulfate - biosynthetic costs modulate substrate selection.

Sulfate-reducing bacteria (SRB) are a diverse group of anaerobic microorganisms that obtain their energy from dissimilatory sulfate reduction. Some SRB species have high respiratory versatility due to the possible use of alternative electron acceptors. A good example is Desulfovibrio desulfuricans ATCC 27774, which grows in the presence of nitrate (end product: ammonium) with higher rates and yields to those observed in sulfate containing medium (end product: sulfide). In this work, the mechanisms supporting the respiratory versatility of D. desulfuricans were unraveled through the analysis of the proteome of the bacterium under different experimental conditions. The most remarkable difference in the two-dimensional gel electrophoresis maps is the high number of spots exclusively represented in the nitrate medium. Most of the proteins with increase abundance are involved in the energy metabolism and the biosynthesis of amino acids (or proteins), especially those participating in ammonium assimilation processes. qPCR analysis performed during different stages of the bacterium's growth showed that the genes involved in nitrate and nitrite reduction (napA and nrfA, respectively) have different expressions profiles: while napA did not vary significantly, nrfA was highly expressed at a 6h time point. Nitrite levels measured along the growth curve revealed a peak at 3h. Thus, the initial consumption of nitrate and concomitant production of nitrite must induce nrfA expression. The activation of alternative mechanisms for energy production, aside several N-assimilation metabolisms and detoxification processes, solves potential survival problems in adapting to different environments and contributes to higher bacterial growth rates.

Metabolic Compensation of Fitness Costs Is a General Outcome for Antibiotic-Resistant Pseudomonas aeruginosa Mutants Overexpressing Efflux Pumps.

It is generally assumed that the acquisition of antibiotic resistance is associated with a fitness cost. We have shown that overexpression of the MexEF-OprN efflux pump does not decrease the fitness of a resistant Pseudomonas aeruginosa strain compared to its wild-type counterpart. This lack of fitness cost was associated with a metabolic rewiring that includes increased expression of the anaerobic nitrate respiratory chain when cells are growing under fully aerobic conditions. It was not clear whether this metabolic compensation was exclusive to strains overexpressing MexEF-OprN or if it extended to other resistant strains that overexpress similar systems. To answer this question, we studied a set of P. aeruginosa mutants that independently overexpress the MexAB-OprM, MexCD-OprJ, or MexXY efflux pumps. We observed increased expression of the anaerobic nitrate respiratory chain in all cases, with a concomitant increase in NO3 consumption and NO production. These efflux pumps are proton/substrate antiporters, and their overexpression may lead to intracellular H+ accumulation, which may in turn offset the pH homeostasis. Indeed, all studied mutants showed a decrease in intracellular pH under anaerobic conditions. The fastest way to eliminate the excess of protons is by increasing oxygen consumption, a feature also displayed by all analyzed mutants. Taken together, our results support metabolic rewiring as a general mechanism to avoid the fitness costs derived from overexpression of P. aeruginosa multidrug efflux pumps. The development of drugs that block this metabolic "reaccommodation" might help in reducing the persistence and spread of antibiotic resistance elements among bacterial populations.IMPORTANCE It is widely accepted that the acquisition of resistance confers a fitness cost in such a way that in the absence of antibiotics, resistant populations will be outcompeted by susceptible ones. Based on this assumption, antibiotic cycling regimes have been proposed in the belief that they will reduce the persistence and spread of resistance among bacterial pathogens. Unfortunately, trials testing this possibility have frequently failed, indicating that resistant microorganisms are not always outcompeted by susceptible ones. Indeed, some mutations do not result in a fitness cost, and in case they do, the cost may be compensated for by a secondary mutation. Here we describe an alternative nonmutational mechanism for compensating for fitness costs, which consists of the metabolic rewiring of resistant mutants. Deciphering the mechanisms involved in the compensation of fitness costs of antibiotic-resistant mutants may help in the development of drugs that will reduce the persistence of resistance by increasing said costs.

The cost of surviving nitrogen excess: energy and protein demand in the lichen Cladonia portentosa as revealed by proteomic analysis.

MAIN CONCLUSION: Different nitrogen forms affect different metabolic pathways in lichens. In particular, the most relevant changes in protein expression were observed in the fungal partner, with NO 3- mostly affecting the energetic metabolism and NH 4+ affecting transport and regulation of proteins and the energetic metabolism much more than NO 3- did. Excess deposition of reactive nitrogen is a well-known agent of stress for lichens, but which symbiont is most affected and how, remains a mystery. Using proteomics can expand our understanding of stress effects on lichens. We investigated the effects of different doses and forms of reactive nitrogen, with and without supplementary phosphorus and potassium, on the proteome of the lichen Cladonia portentosa growing in a 'real-world' simulation of nitrogen deposition. Protein expression changed with the nitrogen treatments but mostly in the fungal partner, with NO3- mainly affecting the energetic metabolism and NH4+ also affecting the protein synthesis machinery. The photobiont mainly responded overexpressing proteins involved in energy production. This suggests that in response to nitrogen stress, the photobiont mainly supports the defensive mechanisms initiated by the mycobiont with an increased energy production. Such surplus energy is then used by the cell to maintain functionality in the presence of NO3-, while a futile cycle of protein production can be hypothesized to be induced by NH4+ excess. External supply of potassium and phosphorus influenced differently the responses of particular enzymes, likely reflecting the many processes in which potassium exerts a regulatory function.

Feasibility of two low-cost organic substrates for inducing denitrification in artificial recharge ponds: Batch and flow-through experiments.

Anaerobic batch and flow-through experiments were performed to assess the capacity of two organic substrates to promote denitrification of nitrate-contaminated groundwater within managed artificial recharge systems (MAR) in arid or semi-arid regions. Denitrification in MAR systems can be achieved through artificial recharge ponds coupled with a permeable reactive barrier in the form of a reactive organic layer. In arid or semi-arid regions, short-term efficient organic substrates are required due to the short recharge periods. We examined the effectiveness of two low-cost, easily available and easily handled organic substrates, commercial plant-based compost and crushed palm tree leaves, to determine the feasibility of using them in these systems. Chemical and multi-isotopic monitoring (δ15NNO3, δ18ONO3, δ34SSO4, δ18OSO4) of the laboratory experiments confirmed that both organic substrates induced denitrification. Complete nitrate removal was achieved in all the experiments with a slight transient nitrite accumulation. In the flow-through experiments, ammonium release was observed at the beginning of both experiments and lasted longer for the experiment with palm tree leaves. Isotopic characterisation of the released ammonium suggested ammonium leaching from both organic substrates at the beginning of the experiments and pointed to ammonium production by DNRA for the palm tree leaves experiment, which would only account for a maximum of 15% of the nitrate attenuation. Sulphate reduction was achieved in both column experiments. The amount of organic carbon consumed during denitrification and sulphate reduction was 0.8‰ of the total organic carbon present in commercial compost and 4.4% for the palm tree leaves. The N and O isotopic fractionation values obtained (εN and εO) were -10.4‰ and -9.0‰ for the commercial compost (combining data from both batch and column experiments), and -9.9‰ and -8.6‰ for the palm tree column, respectively. Both materials showed a satisfactory capacity for denitrification, but the palm tree leaves gave a higher denitrification rate and yield (amount of nitrate consumed per amount of available C) than commercial compost.

Resource stoichiometry and the biogeochemical consequences of nitrogen deposition in a mixed deciduous forest.

Ecosystems often show differential sensitivity to chronic nitrogen (N) deposition; hence, a critical challenge is to improve our understanding of how and why site-specific factors mediate biogeochemical responses to N enrichment. We examined the extent to which N impacts on soil carbon (C) and N dynamics depend on microbial resource stoichiometry. We added N to forest plots dominated by ectomycorrhizal (ECM) trees, which have litter and soil pools rich in organic N and relatively wide C:N ratios, and adjacent forest plots dominated by arbuscular mycorrhizal (AM) trees, which have litter and soil pools rich in inorganic N and relatively narrow C:N ratios. While microbes in both plot types exhibited fairly strict biomass homeostasis, microbes in AM- and ECM-dominated plots differed in their physiological responses to N addition. Microbes in ECM plots responded to N enrichment by decreasing their investment in N-acquisition enzymes (relative to C-acquisition enzymes) and increasing N mineralization rates (relative to C mineralization rates), suggesting that N addition alleviated microbial N demand. In contrast, heterotrophic microbial activities in AM plots were unaffected by N addition, most likely as a result of N-induced increases in net nitrification (60% increase relative to control plots) and nitrate mobilization (e.g., sixfold increases in mobilization relative to control plots). Combined, our findings suggest the stoichiometric differences between AM and ECM soils are the primary drivers of the observed responses. Plant and microbial communities characterized by wide C:N are more susceptible to N-induced changes in decomposition and soil C dynamics, whereas communities characterized by narrow C:N are more susceptible to N-induced nitrate leaching losses. Hence, the biogeochemical consequences of N deposition in temperate forests may be driven by the stoichiometry of the dominant trees and their associated microbes.

Rapid drug-susceptibility testing of Mycobacterium tuberculosis clinical isolates to first-line antitubercular drugs by nitrate reductase assay: A comparison with proportion method.

OBJECTIVE/BACKGROUND: Early initiation of therapy in patients with tuberculosis is imperative for its control. Conventional methods of susceptibility testing such as the proportion method (PM) require visual detection and counting of colonies that takes up to 6weeks. Rapid and simple phenotypic methods that have been endorsed by the World Health Organization can serve as alternatives. METHODS: In this study, we evaluated the colorimetric nitrate reductase assay, which utilizes the detection of nitrate reduction as an indicator of growth much earlier compared with PM (within 7-14days). The susceptibility of 75 clinical isolates of Mycobacterium tuberculosis to four first-line antitubercular drugs was tested by nitrate reductase assay and compared with the standard PM. In this assay, inoculation was done on both drug-free and drug-containing Löwenstein-Jensen medium containing sodium nitrate. After incubation for 7-14days, reduction to nitrite was taken as an indicator of growth, which was detected by color change on addition of Griess reagent. RESULTS: Agreement between nitrate reductase assay and PM was 100% for rifampicin, 97.30% for isoniazid, 93.30% for streptomycin, and 98.60% for ethambutol. Cost/isolate with this assay was found to be approximately two times lesser than that of PM. All results were obtained in 7-14days by nitrate reductase assay, which was significantly rapid compared with 42days taken for obtaining results by PM. CONCLUSION: Nitrate reductase assay can be used as a rapid and inexpensive method for drug-susceptibility testing of M. tuberculosis for first-line antitubercular drugs without compromising accuracy of standard methods.

Dynamics of δ(15)N isotopic signatures of different intertidal macroalgal species: Assessment of bioindicators of N sources in coastal areas.

δ(15)N of annual (Ulva sp., Porphyra sp.) and perennial intertidal seaweed species (Chondrus crispus, Fucus sp.) collected on 17 sampling points along the French coast of the English Channel in 2012 and 2013 were assessed on their suitability as bioindicators of N pollution in coastal areas. A sine function applied for δ(15)N time series data showed for all the species the same seasonal trend with lowest δ(15)N values in April and highest in summer but with no significant interspecific differences of amplitude (α) and phase angle (ϕ). This model provides a useful tool for monitoring the inter-annual changes of N pollution. An interspecific variability of δ(15)N values was observed, probably due to their tolerance to emersion. An in vitro study for comparing the kinetic acquisition of the isotopic signal and N uptake mechanisms of each species underlined the influence of algal physiology on the δ(15)N interspecific variability.