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Spatial and temporal expression of two cytosolic glutamine synthetase genes in Scots pine: functional implications on nitrogen metabolism during early stages of conifer development.

Ammonium assimilation during the initial stages of Scots pine growth involves two cytosolic glutamine synthetase (GS, EC: 6.3.1.2) isoenzymes encoded by separate genes, GS1a and GS1b. GS1a was most exclusively expressed in photosynthetic tissues of the seedling whereas GS1b was expressed ubiquitously showing higher levels in non-photosynthetic tissues such as root and hypocotyl. Temporal expression analysis has shown that when germination starts GS1b is the predominant form in the embryo, however, its relative abundance in the tissue decreased in the postgerminative stages when green cotyledons are developed. In contrast GS1a was present at a low level in the embryo but its abundance increased markedly during germination and seedling growth. These data suggest that GS1a and GS1b genes display different and non-redundant roles in the nitrogen metabolism of conifers. The precise localization of individual transcripts by in situ hybridization strongly supports this possibility. GS1 gene products are mainly expressed in different cellular types: GS1a in chlorophylic parenchyma and GS1b in the vascular bundles of all tissues examined in the seedling. Our data support that glutamine biosynthesis in pine seedlings follows a different pattern related to angiosperms involving two cytosolic GS proteins: one of them a typical cytosolic GS which may be involved in the generation of glutamine for N transport and a second cytosolic GS generating amino donors for the biosynthesis of major N compounds in photosynthetic tissues, a closer role to angiosperm chloroplastic GS. The results are discussed with regard to recent studies on N mobilization and metabolism during the initial stages of conifer development.

Cycadopsida↗

Mouse model for human arginase deficiency.

Deficiency of liver arginase (AI) causes hyperargininemia (OMIM 207800), a disorder characterized by progressive mental impairment, growth retardation, and spasticity and punctuated by sometimes fatal episodes of hyperammonemia. We constructed a knockout mouse strain carrying a nonfunctional AI gene by homologous recombination. Arginase AI knockout mice completely lacked liver arginase (AI) activity, exhibited severe symptoms of hyperammonemia, and died between postnatal days 10 and 14. During hyperammonemic crisis, plasma ammonia levels of these mice increased >10-fold compared to those for normal animals. Livers of AI-deficient animals showed hepatocyte abnormalities, including cell swelling and inclusions. Plasma amino acid analysis showed the mean arginine level in knockouts to be approximately fourfold greater than that for the wild type and threefold greater than that for heterozygotes; the mean proline level was approximately one-third and the ornithine level was one-half of the proline and ornithine levels, respectively, for wild-type or heterozygote mice--understandable biochemical consequences of arginase deficiency. Glutamic acid, citrulline, and histidine levels were about 1.5-fold higher than those seen in the phenotypically normal animals. Concentrations of the branched-chain amino acids valine, isoleucine, and leucine were 0.4 to 0.5 times the concentrations seen in phenotypically normal animals. In summary, the AI-deficient mouse duplicates several pathobiological aspects of the human condition and should prove to be a useful model for further study of the disease mechanism(s) and to explore treatment options, such as pharmaceutical administration of sodium phenylbutyrate and/or ornithine and development of gene therapy protocols.

Animals↗

Biodeterioration of natural stone with special reference to nitrifying bacteria.

An evaluation of field data from historical buildings in Germany showed that chemoorganotrophic bacteria are the most numerous microorganisms in building stones, followed by fungi and nitrifying bacteria. Chemoorganotrophic bacteria and fungi were present in almost every sample. Ammonia and nitrite oxidizers were found in 55 and 62% of the samples, respectively. Within months, natural stone was colonized by chemoorganotrophic microorganisms. The highest cell numbers were usually found near the surface. The colonization of natural stone by nitrifying bacteria took several years. The highest cell numbers were in some cases found underneath the surface. Nitrifying bacteria showed a preference for calcareous material with a medium pore radius between 1 and 10 microns. Cell numbers of nitrifying bacteria did not correlate to the nitrate content of the stone material. We demonstrated that the stone inhabiting microflora can cause significant loss of nitrate by denitrification. Our data strongly suggested that microbial colonization of historical buildings was enhanced by anthropogenic air pollution. Samples taken from stone material with a pore radius < or = 1 micron had significantly higher cell numbers when they were covered with black crusts. A comparison of samples taken between 1990-1995 from buildings throughout Germany showed that in Eastern Germany a significantly stronger colonization with facultatively methylotrophic bacteria and nitrifying bacteria existed. The same was true for natural stone from an urban exposure site when compared to material from a rural exposure site. Data from outdoor exposure and laboratory simulation experiments indicated that the colonization of calcareous stone by nitrifying bacteria was enhanced by chemical weathering.

Biodegradation, Environmental↗

The anammox case-a new experimental manifesto for microbiological eco-physiology.

The anaerobic ammonium oxidation process is a new process for ammonia removal from wastewater. It is also a new microbial physiology that was previously believed to be impossible. The identification of Candidatus Brocadia anammoxidans and its relatives as the responsible bacteria was only possible with the development of a new experimental approach. That approach is the focus of this paper. The approach is a modernisation of the Winogradsky/Beyerinck strategy of selective enrichment and is based on the introduction of the molecular toolbox and modern bioreactor engineering to microbial ecology. It consists of five steps: (1) postulation of an ecological niche based on thermodynamic considerations and macro-ecological field data; (2) engineering of this niche into a laboratory bioreactor for enrichment culture; (3) black-box physiological characterisation of the enrichment culture as a whole; (4) phylogenetic characterisation of the enriched community using molecular tools; (5) physical separation of the dominant members of the enrichment culture using gradient centrifugation and the identification of the species of interest in accordance with Koch's postulates; (6) verification of the in situ importance of these species in the actual ecosystems. The power of this approach is illustrated with a case study: the identification of the planctomycetes responsible for anaerobic ammonium oxidation. We argue that this was impossible using molecular ecology or conventional 'cultivation based techniques' alone. We suggest that the approach might also be used for the microbiological study of many interesting microbes such as anaerobic methane oxidisers.

Bacteria, Anaerobic↗

Human Rhesus B and Rhesus C glycoproteins: properties of facilitated ammonium transport in recombinant kidney cells.

The mammalian Rh (Rhesus) protein family belongs to the Amt/Mep (ammonia transporter/methylammonium permease)/Rh superfamily of ammonium transporters. Whereas RhCE, RhD and RhAG are erythroid specific, RhBG and RhCG are expressed in key organs associated with ammonium transport and metabolism. We have investigated the ammonium transport function of human RhBG and RhCG by comparing intracellular pH variation in wild-type and transfected HEK-293 (human embryonic kidney) cells and MDCK (Madin-Darby canine kidney) cells in the presence of ammonium (NH4+/NH3) gradients. Stopped-flow spectrofluorimetry analysis, using BCECF [2',7'-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein] as a pH-sensitive probe, revealed that all cells submitted to inwardly or outwardly directed ammonium gradients exhibited rapid alkalinization or acidification phases respectively, which account for ammonium movements in transfected and native cells. However, as compared with wild-type cells known to have high NH3 lipid permeability, RhBG- and RhCG-expressing cells exhibited ammonium transport characterized by: (i) a five to six times greater kinetic rate-constant; (ii) a weak temperature-dependence; and (iii) reversible inhibition by mercuric chloride (IC50: 52 microM). Similarly, when subjected to a methylammonium gradient, RhBG- and RhCG-expressing cells exhibited kinetic rate constants greater than those of native cells. However, these constants were five times higher for RhBG as compared with RhCG, suggesting a difference in substrate accessibility. These results, indicating that RhBG and RhCG facilitate rapid and low-energy-dependent bi-directional ammonium movement across the plasma membrane, favour the hypothesis that these Rh glycoproteins, together with their erythroid homologue RhAG [Ripoche, Bertrand, Gane, Birkenmeier, Colin and Cartron (2005) Proc. Natl. Acad. Sci. U.S.A. 101, 17222-17227] constitute a family of NH3 channels in mammalian cells.

Animals↗

Ammonia acquisition in enteric bacteria: physiological role of the ammonium/methylammonium transport B (AmtB) protein.

Homologues of the amtB gene of enteric bacteria exist in all three domains of life. Although their products are required for transport of the ammonium analogue methylammonium in washed cells, only in Saccharomyces cerevisiae have they been shown to be necessary for growth at low NH4+ concentrations. We now demonstrate that an amtB strain of Escherichia coli also grows slowly at low NH4+ concentrations in batch culture, but only at pH values below 7. In addition, we find that the growth defect of an S. cerevisiae triple-mutant strain lacking the function of three homologues of the ammonium/methylammonium transport B (AmtB) protein [called methylammonium/ammonium permeases (MEP)] that was observed at pH 6.1 is relieved at pH 7.1. These results provide direct evidence that AmtB participates in acquisition of NH4+/NH3 in bacteria as well as eucarya. Because NH3 is the species limiting at low pH for a given total concentration of NH4+ + NH3, results with both organisms indicate that AmtB/MEP proteins function in acquisition of the uncharged form. We confirmed that accumulation of [14C]methylammonium depends on its conversion to gamma-N-methylglutamine, an energy-requiring reaction catalyzed by glutamine synthetase, and found that at pH 7, constitutive expression of AmtB did not relieve the growth defects of a mutant strain of Salmonella typhimurium that appears to require a high internal concentration of NH4+/NH3. Hence, contrary to previous views, we propose that AmtB/MEP proteins increase the rate of equilibration of the uncharged species, NH3, across the cytoplasmic membrane rather than actively transporting-that is, concentrating-the charged species, NH4+.

Bacterial Proteins↗

[The influence of aging on cerebral energy metabolism following post-hypoglycemic recovery in the rat. Pharmacologic application].

Influence of aging on cerebral energetic metabolism was evaluated during and after severe hypoglycemia in rats respectively 20 (adults), 60 (matures) or 100 (senescents) week-old. Cerebral content of carbohydrates, amino-acids, ammonia, ATP, ADP, AMP, creatine phosphate and creatine was analysed after 20 min insulin induced hypoglycemia and after 20 min hypoglycemic recovery induced by glucose infusion. In the rats of different ages tested, effect of raubasine (0.85 mg X kg-1 i.p. and i.v.), almitrine (2.68 mg X kg-1 i.p. and i.v.) and association almitrine plus raubasine (at the same doses) on post-hypoglycemic recovery was tested. Aging does not affect the cerebral metabolic disorders occurring in severe hypoglycemia, but rather the metabolic changes during the post-hypoglycemic restitution. In fact there is lower restitution of the concentrations of cerebral cortical metabolites in older rats: the concentrations of many amino-acids and adenylate nucleotides remains largely abnormal. Compared with saline treated post-hypoglycemic rats, raubasine decreases by 15 to 20% cerebral glucose and pyruvate contents in "adults" and "matures" rats and by 10 to 15% glutamate content in rat of different ages tested. Almitrine decreases by 20% cerebral glucose concentration in "matures" and "senescent" rats. In this latter group, almitrine decreases lactate and ammonium contents and increases by 23% glutamine level. In rats of all ages that were submitted to 20 min insulin induced hypoglycemia followed by 20 min glucose induced post-hypoglycemic recovery, the association almitrine plus raubasine decreases by 20 to 30% cerebral glucose, pyruvate and lactate contents and decreases by 15% glutamate. In older brains the association almitrine plus raubasine decreases by 50% cerebral content in ammonium and concomitantly induces an equivalent increase in glutamine content. The effect of the combination almitrine plus raubasine is characterized by an increase in rate of metabolic recovery process in all ages tested.

Aging↗

Ozonation of naphthalenesulphonic acid in the aqueous phase in the presence of basic activated carbons.

The present study aimed to explore the possibility of increasing the purification efficacy of ozone in the removal of high-toxicity contaminants by using carbons of basic character and to analyze the mechanism involved in this process. These carbons were prepared by treating a commercial activated carbon (Witco, W) with ammonia (W-A), ammonium carbonate (W-C), or urea (W-U), under high pressure and temperature. The ammonia and carbonate treatments slightly increased the mesoporosity and, to a greater degree, the macroporosity of carbon W, whereas the urea treatment produced an increase in the porosity across the whole range of pore sizes. In addition, treatment of the activated carbon with these nitrogenating agents produced a marked change in the chemical nature of its surface. Thus, according to the pH of the point of zero charge (pHPZC) values obtained for each sample, carbon W was neutral (pHPZC = 7.12), but the treated carbons were basic, especially carbon W-U (pHPZC = 8.85). This basicity results from an increased concentration of basic oxygenated and nitrogenated surface functional groups, as confirmed by the results of elemental and XPS analyses. An increase in the degradation of 1,3,6-naphthalenetrisulfonic acid was observed when the activated carbon samples were added to the system. This degradation was especially enhanced in the presence of carbon W-U. The increased NTS degradation rate in the presence of the activated carbon is due to an increased concentration of highly reactive radicals in the system. When the catalytic activity of the activated carbon samples was related to their chemical and textural characteristics, it was found that: (i) The catalytic activity increased with an increase in the surface basicity. Interestingly, in the sample with greatest catalytic activity in NTS ozonation, carbon W-U, most of the nitrogenated surface groups introduced were pyrrol groups. These groups increase the electronic density of the basal plane of the activated carbon, thereby enhancing the reduction of ozone on the surface and the generation of highly reactive radicals in the system. (ii) The greater catalytic activity of carbon W-U may also be partly related to its greater surface area and higher volume of mesopores and macropores; these large pores facilitate access of the ozone to the surface active centers of the carbon, increasing its catalytic activity. The presence of the activated carbon samples during NTS ozonation also favored the removal of total organic carbon present in the solution, due to (a) transformation of organic matter into CO2 through the generation of highly reactive species catalyzed by the presence of the activated carbons (catalytic contribution) and (b) adsorption of NTS oxidation byproducts on the activated carbon (adsorptive contribution). The results obtained show that activated carbons treated with nitrogenating agents are very promising catalysts for application in the ozonation of aromatic compounds.

Adsorption↗

The participation of hydrogen peroxide in methyl jasmonate-induced NH(4)(+) accumulation in rice leaves.

Ammonium is a central intermediate in the nitrogen metabolism of plants. We have previously shown that methyl jasmonate (MJ) not only increases the content of H(2)O(2), but also causes NH(4)(+) accumulation in rice leaves. More recently, H(2)O(2) is thought to constitute a general signal molecule participating in the recognition of and the response to stress factors. In this study, we examined the role of H(2)O(2) as a link between MJ and subsequent NH(4)(+) accumulation in detached rice leaves. MJ treatment resulted in an accumulation of NH(4)(+) in detached rice leaves, which was preceded by a decrease in the activity of glutamine synthetase (GS) and an increase in the specific activities of protease and phenylalanine ammonia-lyase (PAL). GS, PAL, and protease appear to be the enzymes responsible for the accumulation of NH(4)(+) in MJ-treated detached rice leaves. Dimethylthiourea (DMTU), a chemical trap for H(2)O(2), was observed to be effective in inhibiting MJ-induced NH(4)(+) accumulation in detached rice leaves. Scavengers of free radicals (sodium benzoate, SB, and glutathione, GSH), nitric oxide donor (N-tert-butyl-alpha-phenylnitrone, PBN), the inhibitors of NADPH oxidase (diphenyleneiodonium chloride, DPI, and imidazole, IMD), and inhibitors of phosphatidylinositol 3-kinase (wortmannin, WM, and LY 294002, LY), which have previously been shown to prevent MJ-induced H(2)O(2) production in detached rice leaves, inhibited MJ-induced NH(4)(+) accumulation. Similarly, changes in enzymes responsible for NH(4)(+) accumulation induced by MJ were observed to be inhibited by DMTU, SB, GSH, PBN DPI, IMD, WM, or LY. Seedlings of rice cultivar Taichung Native 1 (TN1) are jasmonic acid (JA)-sensitive and those of cultivar Tainung 67 (TNG67) are JA-insensitive. On treatment with JA, H(2)O(2) accumulated in the leaves of TN1 seedlings but not in the leaves of TNG67. Ethylene action inhibitor, silver thiosulfate, was observed to inhibit MJ- and abscisic acid-induced accumulation of NH(4)(+) and changes in enzymes responsible for NH(4)(+) accumulation in detached rice leaves, suggesting that the action of MJ and ABA is ethylene dependent.

Abscisic Acid↗

Removal of nutrients in various types of constructed wetlands.

The processes that affect removal and retention of nitrogen during wastewater treatment in constructed wetlands (CWs) are manifold and include NH(3) volatilization, nitrification, denitrification, nitrogen fixation, plant and microbial uptake, mineralization (ammonification), nitrate reduction to ammonium (nitrate-ammonification), anaerobic ammonia oxidation (ANAMMOX), fragmentation, sorption, desorption, burial, and leaching. However, only few processes ultimately remove total nitrogen from the wastewater while most processes just convert nitrogen to its various forms. Removal of total nitrogen in studied types of constructed wetlands varied between 40 and 55% with removed load ranging between 250 and 630 g N m(-2) yr(-1) depending on CWs type and inflow loading. However, the processes responsible for the removal differ in magnitude among systems. Single-stage constructed wetlands cannot achieve high removal of total nitrogen due to their inability to provide both aerobic and anaerobic conditions at the same time. Vertical flow constructed wetlands remove successfully ammonia-N but very limited denitrification takes place in these systems. On the other hand, horizontal-flow constructed wetlands provide good conditions for denitrification but the ability of these system to nitrify ammonia is very limited. Therefore, various types of constructed wetlands may be combined with each other in order to exploit the specific advantages of the individual systems. The soil phosphorus cycle is fundamentally different from the N cycle. There are no valency changes during biotic assimilation of inorganic P or during decomposition of organic P by microorganisms. Phosphorus transformations during wastewater treatment in CWs include adsorption, desorption, precipitation, dissolution, plant and microbial uptake, fragmentation, leaching, mineralization, sedimentation (peat accretion) and burial. The major phosphorus removal processes are sorption, precipitation, plant uptake (with subsequent harvest) and peat/soil accretion. However, the first three processes are saturable and soil accretion occurs only in FWS CWs. Removal of phosphorus in all types of constructed wetlands is low unless special substrates with high sorption capacity are used. Removal of total phosphorus varied between 40 and 60% in all types of constructed wetlands with removed load ranging between 45 and 75 g N m(-2) yr(-1) depending on CWs type and inflow loading. Removal of both nitrogen and phosphorus via harvesting of aboveground biomass of emergent vegetation is low but it could be substantial for lightly loaded systems (cca 100-200 g N m(-2) yr(-1) and 10-20 g P m(-2) yr(-1)). Systems with free-floating plants may achieve higher removal of nitrogen via harvesting due to multiple harvesting schedule.

Biodegradation, Environmental↗

Long-term effects of poultry litter, alum-treated litter, and ammonium nitrate on aluminum availability in soils.

Research has shown that alum [Al(2)(SO(4))(3).14H(2)O] applications to poultry litter can greatly reduce phosphorus (P) runoff, as well as decrease ammonia (NH(3)) volatilization. However, the long-term effects of fertilizing with alum-treated litter are unknown. The objectives of this study were to evaluate the long-term effects of normal poultry litter, alum-treated litter, and ammonium nitrate (NH(4)NO(3)) on aluminum (Al) availability in soils, Al uptake by tall fescue (Festuca arundinacea Schreb.), and tall fescue yields. A long-term study was initiated in April of 1995. There were 13 treatments (unfertilized control, four rates of normal litter, four rates of alum-treated litter, and four rates of NH(4)NO(3)) in a randomized block design. All fertilizers were broadcast applied to 52 small plots (3.05 x 1.52 m) cropped to tall fescue annually in the spring. Litter application rates were 2.24, 4.49, 6.73, and 8.98 Mg ha(-1) (1, 2, 3, and 4 tons acre(-1)); NH(4)NO(3) rates were 65, 130, 195, and 260 kg N ha(-1) and were based on the amount of N applied with alum-treated litter. Soil pH, exchangeable Al (extracted with potassium chloride), Al uptake by fescue, and fescue yields were monitored periodically over time. Ammonium nitrate applications resulted in reductions in soil pH beginning in Year 3, causing exchangeable Al values to increase from less than 1 mg Al kg(-1) soil in Year 2 to over 100 mg Al kg(-1) soil in Year 7 for many of the NH(4)NO(3) plots. In contrast, normal and alum-treated litter resulted in an increase in soil pH, which decreased exchangeable Al when compared to unfertilized controls. Severe yield reductions were observed with NH(4)NO(3) beginning in Year 6, which were due to high levels of acidity and exchangeable Al. Aluminum uptake by forage and Al runoff from the plots were not affected by treatment. Fescue yields were highest with alum-treated litter (annual average = 7.36 Mg ha(-1)), followed by normal litter (6.93 Mg ha(-1)), NH(4)NO(3) (6.16 Mg ha(-1)), and the control (2.89 Mg ha(-1)). These data indicate that poultry litter, particularly alum-treated litter, may be a more sustainable fertilizer than NH(4)NO(3).

Alum Compounds↗

Amounts and sources of protein for dairy calves.

Female Holstein calves (178) were fed rations varying from 10 to 17% crude protein (air dry) from 30 to 102 days of age. Ten percent was inadequate resulting in less body weight, gain, withers height, and heart girth than rations containing 12 to 17% crude protein. In trial 1 addition of urea, biuret, or ammonia-molasses to a 12% diet to make a 16% crude protein diet allowed gains and weights indistinguishable from those on a 12 or 16% crude protein soybean meal ration. In another trial, addition of urea, biuret, or ammonium propionate to a 10% ration to make a 14% ration allowed insignificantly greater gains and body weight. Gains and body weight were greater for the 14% soybean meal ration than the three non-protein-nitrogen rations for calves to 71 days of age but equal for 71 to 102 days. Gain, body weight, dry matter, and protein intakes at most dates and intervals were greater for calves fed the 14% soybean meal ration than the 10% ration. When 10, 12, 14, and 16% soybean meal rations were compared simultaneously, only the 10% crude protein ration gave inferior performance. Gains were reduced when average daily crude protein intake for the 72 day trail was 258 g or less. Calves fed lupine or dried poultry waste did not gain or consume as much as those fed comparable soybean meal rations. Calves fed a 17% crude protein ration from several protein sources plus alfalfa hay gained no much than those fed the 12 to 16% crude protein rations. Regression analysis relating dry matter and protein intake to body weight or gains indicated both dry matter intake and protein percentage were about equally important from day 41 to 72 and that after this age dry matter intake was of much greater importance than protein percentage. Protein intake expressed as g/day was not of importance in this regression. Weaning at day 42 allowed for greater body weight and gains than weaning at day 32. The difference in body weight was 2.5 kg at day 42 and 7 kg at day 102.

Animal Feed↗

Roles of SpoT and FNR in NH4+ assimilation and osmoregulation in GOGAT (glutamate synthase)-deficient mutants of Escherichia coli.

An osmosensitive mutant of Escherichia coli was isolated and shown to harbor two mutations that were together necessary for osmosensitivity. One (ossB) was an insertion mutation in the gltBD operon, which encodes the enzyme glutamate synthase (GOGAT), involved in ammonia assimilation and L-glutamate biosynthesis. The other (ossA) was in the fnr gene, encoding the regulator protein FNR for anaerobic gene expression. Several missense or deletion mutations in fnr and gltBD behaved like ossA and ossB, respectively, in conferring osmosensitivity. A mutation affecting the DNA-binding domain of FNR was recessive to fnr+ with respect to the osmotolerance phenotype but was dominant-negative for its effect on expression of genes in anaerobic respiration. Our results may most simply be interpreted as suggesting the requirement for monomeric FNR during aerobic growth of E. coli in high-osmolarity media, presumably for L-glutamate accumulation via the GOGAT-independent pathway (catalyzed by glutamate dehydrogenase [GDH]), but the mechanism of FNR action is not known. We also found that the spoT gene (encoding guanosine 3',5'-bispyrophosphate [ppGpp] synthetase II/ppGpp-3' pyrophosphohydrolase), in multiple copies, overcomes the defect in NH4+ assimilation associated with GOGAT deficiency and thereby suppresses osmosensitivity in gltBD fnr strains. Enhancement of GDH activity in these derivatives appears to be responsible for the observed suppression. Its likely physiological relevance was established by the demonstration that growth of gltBD mutants (that are haploid for spoT+) on moderately low [NH4+] was restored with the use of C sources poorer than glucose in the medium. Our results raise the possibility that SpoT-mediated accumulation of ppGpp during C-limited growth leads to GDH activation and that the latter enzyme plays an important role in N assimilation in situ hitherto unrecognized from studies on laboratory-grown cultures.

Aerobiosis↗

Comparison of low cost measurement techniques for long-term monitoring of atmospheric ammonia.

An inter-comparison of techniques for long-term sampling of atmospheric ammonia (NH3) was conducted with a view to establishing a national network with > 50 sites. Key requirements were for: a low cost system, simplicity and durability to enable a postal exchange with local site operators, a precision of < +/- 20% for monthly sampling at expected NH3 concentrations of 1-2 micrograms m-3, a detection limit sufficient to resolve the small NH3 concentrations (< 0.2 microgram m-3) expected in remote parts of the UK, and a quantitative means to establish quality control. Five sampling methods were compared: A, a commercially available membrane diffusion tube (exposed in triplicate), with membranes removed immediately after sampling; B, the above method, with the membranes left in place until analysis; C, open-ended diffusion tubes (exposed with 4 replicates); D, a new active sampling diffusion denuder system; and E, an active sampling bubbler system. Method D consisted of two 0.1 m acid coated glass denuders in series with sampling at approximately 0.3 l min-1. These methods were deployed at 6 locations in the UK and the Netherlands and compared against reference estimates. Method D was the most precise and sensitive of the techniques compared, with a detection limit of < 0.1 microgram m-3. The bubbler provided a less precise estimate of NH3 concentration, and also suffered several practical drawbacks. The diffusion tubes were found to correlate with the reference at high concentrations (> 3 micrograms m-3), but were less precise and overestimated NH3 at smaller concentrations. Of the passive methods, A was the most precise and C the least precise. On the basis of the results, method D has been implemented in the national network, together with application of method A to explore spatial variability in regions with expected high NH3 concentrations.

Air Pollutants↗

Effects of atmospheric ammonia (NH3) on terrestrial vegetation: a review.

At the global scale, among all N (nitrogen) species in the atmosphere and their deposition on to terrestrial vegetation and other receptors, NH3 (ammonia) is considered to be the foremost. The major sources for atmospheric NH3 are agricultural activities and animal feedlot operations, followed by biomass burning (including forest fires) and to a lesser extent fossil fuel combustion. Close to its sources, acute exposures to NH3 can result in visible foliar injury on vegetation. NH3 is deposited rapidly within the first 4-5 km from its source. However, NH3 is also converted in the atmosphere to fine particle NH4+ (ammonium) aerosols that are a regional scale problem. Much of our current knowledge of the effects of NH3 on higher plants is predominantly derived from studies conducted in Europe. Adverse effects on vegetation occur when the rate of foliar uptake of NH3 is greater than the rate and capacity for in vivo detoxification by the plants. Most to least sensitive plant species to NH3 are native vegetation > forests > agricultural crops. There are also a number of studies on N deposition and lichens, mosses and green algae. Direct cause and effect relationships in most of those cases (exceptions being those locations very close to point sources) are confounded by other environmental factors, particularly changes in the ambient SO2 (sulfur dioxide) concentrations. In addition to direct foliar injury, adverse effects of NH3 on higher plants include alterations in: growth and productivity, tissue content of nutrients and toxic elements, drought and frost tolerance, responses to insect pests and disease causing microorganisms (pathogens), development of beneficial root symbiotic or mycorrhizal associations and inter species competition or biodiversity. In all these cases, the joint effects of NH3 with other air pollutants such as all-pervasive O3 or increasing CO2 concentrations are poorly understood. While NH3 uptake in higher plants occurs through the shoots, NH4+ uptake occurs through the shoots, roots and through both pathways. However, NH4+ is immobile in the soil and is converted to NO3- (nitrate). In agricultural systems, additions of NO3- to the soil (initially as NH3 or NH4+) and the consequent increases in the emissions of N2O (nitrous oxide, a greenhouse gas) and leaching of NO3- into the ground and surface waters are of major environmental concern. At the ecosystem level NH3 deposition cannot be viewed alone, but in the context of total N deposition. There are a number of forest ecosystems in North America that have been subjected to N saturation and the consequent negative effects. There are also heathlands and other plant communities in Europe that have been subjected to N-induced alterations. Regulatory mitigative approaches to these problems include the use of N saturation data or the concept of critical loads. Current information suggests that a critical load of 5-10 kg ha(-1) year(-1) of total N deposition (both dry and wet deposition combined of all atmospheric N species) would protect the most vulnerable terrestrial ecosystems (heaths, bogs, cryptogams) and values of 10-20 kg ha(-1) year(-1) would protect forests, depending on soil conditions. However, to derive the best analysis, the critical load concept should be coupled to the results and consequences of N saturation.

Air Pollutants↗