Effects of dry-deposited sulphur dioxide on fungal decomposition of angiosperm tree leaf litter II. Chemical content of leaf litters.

Ash (Fraxinus excelsior L.), birch (Betula spp.), hazel (Corylus avellana L.), sessile oak [Quercus petraea (Mattuschka) Liebl.] and sycamore (Acer pseudoplatanus L.) leaf litters from a non-polluted and a heavily sulphur dioxide (SO2 )-polluted woodland were fumigated with environmentally-realistic concentrations (0.010-0.030 µl l-1 ) of SO2 for 16-68 wk in an open-air field-fumigation experiment. Fumigation markedly increased sulphate and protons in leachates from the litters and decreased calcium and magnesium contents of the leaves. However, there were few differences in the responses between leaf litters from the two woodlands. This was attributed to rapid sulphate wash-out from the litters from the heavily polluted woodland, so that the litters from the two sites quickly reached the same sulphate status.

Effects of dry-deposited sulphur dioxide on fungal decomposition of angiosperm tree leaf litter III. Decomposition rates and fungal respiration.

Ash (Fraxinus excelsior L.), birch (Betula spp.), hazel (Corylus avellana L.), sessile oak [Quercus petraea (Mattuschka) Liebl.] and sycamore (Acer pseudoplatanus L.) leaf litters from a virtually non-polluted and a heavily sulphur dioxide (SO2 )-polluted woodland were fumigated with environmentally-realistic concentrations (0.010-0.030µl l-1 ) of SO2 for 16-68 wk in an open-air field-fumigation experiment. Fumigation inhibited the respiration (CO2 evolution) and decomposition rates of the leaf litters. However, there were few differences in the responses between leaf litters from the two woodlands. In addition, pure cultures of four saprotrophic fungi were grown individually on irradiated hazel litter and exposed to c. 0.030µl l-1 of gaseous SO2 , for 28 d in the laboratory. The gas inhibited the respiration of Phoma exigua Desm. and Phoma macrostoma Mont, but not the respiration of Cladosporium cladosporioides (Fres.) de Vries or Coniothyrium quercinum Sacc. var. glandicola Grove. These results in part substantiated findings of previous experiments examining the effects of SO2 on the structures of saprotrophic fungal communities. The effects of SO2 on fungal decomposition of angiosperm tree leaf litter as possible causes of forest decline are discussed.

Effects of dry-deposited sulphur dioxide on fungal decomposition of angiosperm tree leaf litter I. Changes in communities of fungal saprotrophs.

Comparisons of the saprotrophic fungi isolated from ash (Fraxinus excelsior L.), birch (Betula spp.), hazel (Corylus avellana L.), pedunculate oak (Quercus robur L.), sessile oak [Q. petraea (Mattuschka) Liebl.] and sycamore (Acer pseudoplatanus L.) leaf litters from three woodlands exposed to low, medium and high levels (c. 0.0→ 0.060µl l-1 ) of sulphur dioxide (SO2 ) showed that the composition of the fungal communities differed between sites. Fumigation of angiosperm tree leaf litters from the least and the most polluted site with environmentally-realistic concentrations (0.010-0.030 µl1 ) of SO2 for 16-68 wk in an open-air field-fumigation experiment resulted in marked changes in the composition of the fungal communities in the leaf litters, comparable with differences found between the woodland sites. Cladosporium spp., Epicoccutn nigrum Link, Fitsarium spp. and Phoma exigua Desm. were less commonly isolated from leaf litters exposed to SO2 , whereas Coniothyrium quercinum Sacc. var. glandicola Grove, Cylindrocarpon orthosporum (Sacc.) Wollenw. and Penicillium spp. were more frequently isolated from fumigated litters. However, few differences could be detected in the response to SO2 of the mycofloras of leaf litters originating from different woodland sites. In general, SO2 did not affect the total extent of fungal occupancy of the microsites in the litter, as fungal species which decreased in abundance on exposure to the gas appeared to be replaced in the litter by other species more tolerant to the gas. SO2 therefore appeared to be selectively toxic to saprotrophic fungi isolated from these litters.

Elevated CO2 reduces field decomposition rates of Betula pendula (Roth.) leaf litter.

The effect of elevated atmospheric CO2 and nutrient supply on elemental composition and decomposition rates of tree leaf litter was studied using litters derived from birch (Betula pendula Roth.) plants grown under two levels of atmospheric CO2 (ambient and ambient +250 ppm) and two nutrient regimes in solar domes. CO2 and nutrient treatments affected the chemical composition of leaves, both independently and interactively. The elevated CO2 and unfertilized soil regime significantly enhanced lignin/N and C/N ratios of birch leaves. Decomposition was studied using field litter-bags, and marked differences were observed in the decomposition rates of litters derived from the two treatments, with the highest weight remaining being associated with litter derived from the enhanced CO2 and unfertilized regime. Highly significant correlations were shown between birch litter decomposition rates and lignin/N and C/N ratios. It can be concluded, from this study, that at levels of atmospheric CO2 predicted for the middle of the next century a deterioration of litter quality will result in decreased decomposition rates, leading to reduction of nutrient mineralization and increased C storage in forest ecosystems. However, such conclusions are difficult to generalize, since tree responses to elevated CO2 depend on soil nutritional status.

Root production and turnover in an upland grassland subjected to artificial soil warming respond to radiation flux and nutrients, not temperature.

Root demographic processes (birth and death) were measured using minirhizotrons in the soil warming experiments at the summit of Great Dun Fell, United Kingdom (845 m). The soil warming treatment raised soil temperature at 2 cm depth by nearly 3°C. The first experiment ran for 6 months (1994), the second for 18 (1995-1996). In both experiments, heating increased death rates for roots, but birth rates were not significantly increased in the first experiment. The lack of stimulation of death rate in 1996 is probably an artefact, caused by completion of measurements in late summer of 1996, before the seasonal demography was concluded: root death continued over the winter of 1995-1996. Measurements of instantaneous death rates confirmed this: they were accelerated by warming in the second experiment. In the one complete year (1995-1996) in which measurements were taken, net root numbers by the end of the year were not affected by soil warming. The best explanatory environmental variable for root birth rate in both experiments was photosynthetically active radiation (PAR) flux, averaged over the previous 5 (first experiment) or 10 days (second experiment). In the second experiment, the relationship between birth rate and PAR flux was steeper and stronger in heated than in unheated plots. Death rate was best explained by vegetation temperature. These results provide further evidence that root production acclimates to temperature and is driven by the availability of photosynthate. The stimulation of root growth due to soil warming was almost certainly the result of changes in nutrient availability following enhanced decomposition.

Combined use of open-air and indoor fumigation systems to study effects of SO2 on leaching processes in Scots pine litter.

Both an open-air fumigation system and a laboratory-based system were used to expose decomposing Scots pine (Pinus sylvestris L.) needles to controlled concentrations of SO(2) (arithmetic mean

PAHs associated with the leaves of three deciduous tree species. I--Concentrations and profiles.

Results for the concentrations of total polycyclic aromatic hydrocarbons (Sigma PAH) and the PAH profile in leaves from three deciduous tree species from the same woodland are presented, and discussed with reference to environmental and leaf-related variables. There were significant differences between oak, ash and hazel leaves in their Sigma PAH concentrations (sum of 23 PAHs), and in the relative contribution of individual PAHs to the sum. Leaves exhibiting pubescence (hairiness) were found to have significantly higher Sigma PAH concentrations than hairless leaves, regardless of their position in the vegetation strata of the wood. Hazel leaves from the understorey had a PAH profile consisting of a greater proportion of the 4-, 5- and 6-ring PAHs than oak or ash from the canopy. This was concluded to be the result of the filtering effect of the main canopy on the air passing over and through it, with subsequent transfer of particles and attendant PAHs to the understorey below. The proportion of Sigma PAH contributed by the 6-ring PAH in hazel leaves was negatively correlated with distance from the southern edge of the canopy. It is proposed that the predominantly windward edges of the woodland, where atmospheric turbulence is likely to be greatest, favoured the deposition of particle-bound PAHs to leaves.

Dynamics of PAH deposition, cycling and storage in a mixed-deciduous (Quercus-Fraxinus) woodland ecosystem.

Estimates of standing biomass and fluxes of biomass in a mixed-deciduous woodland were derived, and used with results for concentrations of seven polycyclic aromatic hydrocarbons (PAHs) in different compartments of the woodland system to quantitatively assess some of the key fluxes and burdens of PAHs in this complex system. We quantified PAH burdens in air, in leaves of three deciduous tree species, in leaf litter and in soil, and uptake of PAHs by the tree leaves; PAH fluxes in litterfall, and deposition to the litter layer on the woodland floor during winter were calculated from these data. Air burdens exhibited marked seasonal variations for all compounds, with lowest values in summer when combustion-related emissions were low. Leaves did not accumulate large burdens of PAHs while on the trees and consequently, litterfall-associated fluxes of PAHs were small, representing only a fraction of the burdens in the litter layer to which they were deposited. Higher PAH burdens in air in winter, combined with the organic-matter-rich nature of the litter layer, are thought to be responsible for fluxes of PAHs to the litter layer in winter being 20-170 times the peak litterfall fluxes. The soil compartment was calculated to contain 25 years' worth of deposition of benzo[ghi]perylene, the most recalcitrant PAH in this study. Storage quotients for fluoranthene, pyrene, benzo[k]fluoranthene and benzo[a]pyrene burdens in soil represented 7-10 years' worth of deposition, while fluorene and phenanthrene storage in soil approached unity with inputs (1 and 3 years' worth of deposition, respectively). The relative importance of storage and loss processes was therefore closely related to the physico-chemical properties of the PAH, and is discussed in relation to the cycling of carbon in the woodland.

Soil solution nitrogen and cations influenced by (NH4)2SO4 deposition in a coniferous forest.

The effects of chronically enhanced (NH(4))(2)SO(4) deposition on ion concentrations in soil solution and ionic fluxes were investigated in a Picea abies plot at Grizedale forest, NW England. Soil cores closed at the base and containing a ceramic suction cup sampler were 'roofed' and watered every 2 weeks with bulk throughfall collected in the field. Treatments consisted of the inclusion of living roots from mature trees in the lysimeters and increasing (NH(4))(2)SO(4) deposition (NS treatment) to ambient + 75 kg N ha(-1) a(-1). Rainfall, throughfall and soil solutions were collected every 2 weeks during 18 months, and analysed for major cations and anions. NO(3)(-) fluxes significantly increased following NS treatment, and were balanced by increased Al(3+) losses. Increased SO(4)(2-) concentrations played a minor role in controlling soil solution cation concentrations. The soil exchange complex was dominated by Al and, during the experimental period, cores of all treatments 'switched' from Ca(2+) to Al(3+) leaching, leading to mean [Formula: see text] molar ratios in soil solution of NS treated cores of 0.24. The experiment confirmed that the most sensitive soils to acidification (through deposition or changing environmental conditions) are those with low base saturation, and with a pH in the lower Ca, or Al buffer ranges.

Aluminium and fluoride in the water supply and their removal for haemodialysis.

Aluminium and fluoride in the water supply and their removal for haemodialysis have been investigated in the Trent Region, U.K., and wide variations noted. The efficiency of removal of these elements from the mains water supplying home haemodialysis units by different water treatment systems currently installed has been assessed and a follow-up study performed 8 months later. Some improvement in water treatment was noted, with the percentage of home dialysis units receiving waters within the proposed European Economic Community guidelines for aluminium rising from 61.7 to 72.1%. Removal of fluoride by the different treatment mechanisms is analogous to that for aluminium. A comparison of new and older water treatment systems has shown that there is a deterioration in performance with use. However, some cases of poor removal may be due to the installation of unsuitable equipment, or, more probably, due to a change in the waters used to supply the different homes. Thus, adequate maintenance of equipment and frequent sampling of both untreated and treated waters are required in order to maintain the provision of waters suitable for the preparation of dialysate.

PAHs associated with the leaves of three deciduous tree species. II: uptake during a growing season.

Leaves from three species of deciduous tree (oak, ash and hazel) were sampled at intervals through a growing season in a mature, mixed-deciduous woodland. Polycyclic aromatic hydrocarbon (PAH) concentrations remained within a small range for all species between May and September, deviating significantly only when increases in atmospheric concentrations of PAHs (notably from the 'Bonfire night' festival in early autumn) have been shown. We concluded that the influence of air concentrations was more important than meteorological conditions (temperature, humidity and rainfall) in determining plant concentrations of PAHs over a growing season. Concentrations of 4-, 5- and 6-ring PAHs were positively correlated with time for all species, but there were significant differences in the PAH profile between species sampled from the canopy (oak and ash) compared with the understorey (hazel). Oak and ash had similar PAH profiles, while hazel leaves had proportionally greater concentrations of the heavier molecular weight (4-, 5- and 6-ring) PAHs, and the ratios of these compounds to 3-ring PAHs was positively correlated with time. This affirms earlier work conducted on the same species in the same woodland, where we concluded that the canopy was filtering particles and attendant PAHs from air passing over or through it, and that these particles were transferred to the understorey and the woodland floor.

Respiration of the external mycelium in the arbuscular mycorrhizal symbiosis shows strong dependence on recent photosynthates and acclimation to temperature.

* Although arbuscular mycorrhizal (AM) fungi are a major pathway in the global carbon cycle, their basic biology and, in particular, their respiratory response to temperature remain obscure. * A pulse label of the stable isotope (13)C was applied to Plantago lanceolata, either uninoculated or inoculated with the AM fungus Glomus mosseae. The extra-radical mycelium (ERM) of the fungus was allowed to grow into a separate hyphal compartment excluding roots. We determined the carbon costs of the ERM and tested for a direct temperature effect on its respiration by measuring total carbon and the (13)C:(12)C ratio of respired CO(2). With a second pulse we tested for acclimation of ERM respiration after 2 wk of soil warming. * Root colonization remained unchanged between the two pulses but warming the hyphal compartment increased ERM length. delta(13)C signals peaked within the first 10 h and were higher in mycorrhizal treatments. The concentration of CO(2) in the gas samples fluctuated diurnally and was highest in the mycorrhizal treatments but was unaffected by temperature. Heating increased ERM respiration only after the first pulse and reduced specific ERM respiration rates after the second pulse; however, both pulses strongly depended on radiation flux. * The results indicate a fast ERM acclimation to temperature, and that light is the key factor controlling carbon allocation to the fungus.

Using delta15N values to characterise the nitrogen nutrient pathways from intensive animal units.

Previous studies on foliar delta15N values, in certain bryophytes, have indicated signature similarities to source pollutants. The object of this study was to investigate the effect further, by examining the mechanisms whereby isotopic fractionation occurs in systems such as atmospheric ammonia (NH3), throughfall, vegetation and soil. Measurements taken in and around point emission sources will then be used to characterise the various fractionation effects associated with these N transformations, as well as to demonstrate some of the issues associated with using delta15N values as pollution indicators. The atmospheric dispersion model UK-ADMS has also been used to model atmospheric delta15NH3 emissions, with signatures exhibiting marked negative shifts immediately downwind of an agricultural NH3 source. Similar dispersion patterns were mapped for NH3 concentration data illustrating the link between these two forms of measurement.

Hydrogeochemical studies for Al, F and Fe in waters supplying haemodialysis units in the Trent region, U.K.

Geographical and seasonal variations in the concentrations of Al, F and Fe in tap water supplying home haemodialysis units within the administrative area of the Trent Regional Health Authority, have been examined. Aluminium and fluoride were considered for their implications in bone disorders and iron for its competitive binding with aluminium to transferrin in serum. Four different water treatment systems are currently used and the extractive performance of these have been assessed. In general, the current Department of Health and Social Security's (DHSS) guidelines for aluminium in water used to prepare dialysate are met (30 µg/l), but the proposed European Economic Community's (EEC) levels of 10 µg/l, are not. The removal of fluoride and iron from water by the different treatments was comparable to the removal of aluminium. A comparison between old and new equipment highlights the importance of regular monitoring and maintenance.

Carbon assimilation and turnover in grassland vegetation using an in situ (13)CO(2) pulse labelling system.

A mobile laboratory was developed to administer a controlled flow of (13)C labelled CO(2) at ambient concentrations ( approximately 350 ppm) in the field. The stable isotope delivery (SID) system consists of an isotope-mixing unit with flow control to a series of 12 independent labelling chambers. In-line CPU controlled infrared gas analysers allow automated measurement of chamber CO(2) concentrations and gas flow management. A preliminary experiment was established on an upland pasture located at the NERC Soil Biodiversity experimental site, Sourhope, UK, in August 1999. The objective of this investigation was to determine the magnitude of pulse-derived C incorporation into a typical upland plant community. To achieve this, the SID system was deployed to pulse-label vegetation with CO(2) enriched with (13)C (50 atom %) at ambient concentrations ( approximately 350 ppm) on two consecutive days in August 1999. Samples of headspace CO(2), shoot and root were taken on four occasions over a period of 28 days after (13)C labelling. These materials were then prepared for (13)C/(12)C ratio determination by continuous-flow/combustion/isotope ratio mass spectrometry (CF-C-IRMS). Results showed that pulse derived CO(2)-C was assimilated at a rate of 128 +/- 32 microg g shoot-C hour(-1). Dynamic samplings showed that pulse-derived (13)C concentrations in the labelled plant tissues declined by 77.4 +/- 6% after 48 hours. The rapid decline in (13)C concentrations in plant matter was the result of C loss from the plant in the form of respired CO(2) and root exudates, and dilution by subsequent unlabelled C assimilates. This novel system offers considerable potential for in situ tracer investigations.

A review of, and improvements to, the techniques of trace element analysis of raw water for home dialysis.

The optimum operating conditions for the measurement of aluminium and iron concentrations in water by atomic absorption spectrometry have been determined experimentally. The characteristics of an ion-selective electrode for the measurement of fluoride have also been investigated. Contrary to other work, the addition of acid to samples did not improve their storage stability, although the acid-washing of polyethylene containers is recommended for the storage of water.

Detection and classification of atmospheric methane oxidizing bacteria in soil.

Well-drained non-agricultural soils mediate the oxidation of methane directly from the atmosphere, contributing 5 to 10% towards the global methane sink. Studies of methane oxidation kinetics in soil infer the activity of two methanotrophic populations: one that is only active at high methane concentrations (low affinity) and another that tolerates atmospheric levels of methane (high affinity). The activity of the latter has not been demonstrated by cultured laboratory strains of methanotrophs, leaving the microbiology of methane oxidation at atmospheric concentrations unclear. Here we describe a new pulse-chase experiment using long-term enrichment with 12CH4 followed by short-term exposure to 13CH4 to isotopically label methanotrophs in a soil from a temperate forest. Analysis of labelled phospholipid fatty acids (PLFAs) provided unambiguous evidence of methane assimilation at true atmospheric concentrations (1.8-3.6 p.p.m.v.). High proportions of 13C-labelled C18 fatty acids and the co-occurrence of a labelled, branched C17 fatty acid indicated that a new methanotroph, similar at the PLFA level to known type II methanotrophs, was the predominant soil micro-organism responsible for atmospheric methane oxidation.

Stable-isotope probing as a tool in microbial ecology.

Microorganisms are responsible for driving the biogeochemical cycling of elements on Earth. Despite their importance and vast diversity, the taxonomic identity of the microorganisms involved in any specific process has usually been confined to that small fraction of the microbiota that has been isolated and cultivated. The recent coupling of molecular biological methods with stable-isotope abundance in biomarkers has provided a cultivation-independent means of linking the identity of bacteria with their function in the environment. Here we show that 13C-DNA, produced during the growth of metabolically distinct microbial groups on a 13C-enriched carbon source, can be resolved from 12C-DNA by density-gradient centrifugation. DNA isolated from the target group of microorganisms can be characterized taxonomically and functionally by gene probing and sequence analysis. Application of this technique to investigate methanol-utilizing microorganisms in soil demonstrated the involvement of members of two phylogenetically distinct groups of eubacteria; the alpha-proteobacterial and Acidobacterium lineages. Stable-isotope probing thus offers a powerful new technique for identifying microorganisms that are actively involved in specific metabolic processes under conditions which approach those occurring in situ.

(13)C incorporation into DNA as a means of identifying the active components of ammonia-oxidizer populations.

AIMS: To identify active CO2-assimilating species of ammonia-oxidizing bacteria in fresh water sediment. METHODS AND RESULTS: Enrichment cultures were incubated in the presence of 13C labelled CO2, and 13C-DNA successfully resolved from 12C-DNA by caesium chloride density gradient ultracentrifugation of DNA extracts. Ammonia-oxidizer DNA recovered from these gradients was amplified and characterised by Temporal Temperature Gradient Gel Electrophoresis (TTGE), with confirmatory sequence analysis to identify the metabolically active components of the population. CONCLUSION: The 12C-DNA fraction was dominated by nitrosospiras, in contrast to the 13C-DNA fraction which was largely nitrosomonad DNA, in support of the hypothesis that nitrosomonads out-compete nitrosospiras in laboratory culture. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of stable isotype incorporation into ammonia-oxidizer DNA could therefore circumvent the problems associated with RNA detection to identify metabolically active species in situ.

Aluminium release from the Sorbsystem D-3160 and D-3260 cartridges.

Recent clinical information indicates that the aluminium level of patients' serum can rise while they are being treated with the Sorbsystem haemodialysis system. Further evidence suggests that the quality of aluminium removed from the dialysing fluid by the sorbent cartridge utilized in this equipment is time-dependent. Evaluation of the sorbent cartridges D-3160 and D-3260 has shown that ultimately aluminium is satisfactorily removed from the dialysing fluid. However, the time taken to achieve the safe condition of less than 30 micrograms/litre of aluminium is dependent on the quantity of aluminium washed out from a new cartridge before being reabsorbed, and the highly variable quantity contained in the diluting water. The results have also shown that the level of aluminium recommended by the DHSS in April 1982 (30 micrograms/litre) is not reliably attained following the manufacturer's recommended pretreatment, and a further 60 min of operation in the 'dialyse' mode is necessary. It should be noted that, as a result of evaluation of the Sorbsystem, the DHSS have issued guidelines on the use of this equipment.