Assessing the reliability of peatland GPP measurements by remote sensing: From plot to landscape scale.

Estimates of peatland carbon fluxes based on remote sensing data are a useful addition to monitoring methods in these remote and precious ecosystems, but there are questions as to whether large-scale estimates are reliable given the small-scale heterogeneity of many peatlands. Our objective was to consider the reliability of models based on Earth Observations for estimating ecosystem photosynthesis at different scales using the Forsinard Flows RSPB reserve in Northern Scotland as our study site. Three sites across the reserve were monitored during the growing season of 2017. One site is near-natural blanket bog, and the other two are at different stages of the restoration process after removal of commercial conifer forestry. At each site we measured small (flux chamber) and landscape scale (eddy covariance) CO2 fluxes, small scale spectral data using a handheld spectrometer, and obtained corresponding satellite data from MODIS. The variables influencing GPP at small scale, including microforms and dominant vegetation species, were assessed using exploratory factor analysis. A GPP model using land surface temperature and a measure of greenness from remote sensing data was tested and compared to chamber and eddy covariance CO2 fluxes; this model returned good results at all scales (Pearson's correlations of 0.57 to 0.71 at small scale, 0.76 to 0.86 at large scale). We found that the effect of microtopography on GPP fluxes at the study sites was spatially and temporally inconsistent, although connected to water content and vegetation species. The GPP fluxes measured using EC were larger than those using chambers at all sites, and the reliability of the TG model at different scales was dependent on the measurement methods used for calibration and validation. This suggests that GPP measurements from remote sensing are robust at all scales, but that the methods used for calibration and validation will impact accuracy.

Towards a microbial process-based understanding of the resilience of peatland ecosystem service provisioning - A research agenda.

Peatlands are wetland ecosystems with great significance as natural habitats and as major global carbon stores. They have been subject to widespread exploitation and degradation with resulting losses in characteristic biota and ecosystem functions such as climate regulation. More recently, large-scale programmes have been established to restore peatland ecosystems and the various services they provide to society. Despite significant progress in peatland science and restoration practice, we lack a process-based understanding of how soil microbiota influence peatland functioning and mediate the resilience and recovery of ecosystem services, to perturbations associated with land use and climate change. We argue that there is a need to: in the short-term, characterise peatland microbial communities across a range of spatial and temporal scales and develop an improved understanding of the links between peatland habitat, ecological functions and microbial processes; in the medium term, define what a successfully restored 'target' peatland microbiome looks like for key carbon cycle related ecosystem services and develop microbial-based monitoring tools for assessing restoration needs; and in the longer term, to use this knowledge to influence restoration practices and assess progress on the trajectory towards 'intact' peatland status. Rapid advances in genetic characterisation of the structure and functions of microbial communities offer the potential for transformative progress in these areas, but the scale and speed of methodological and conceptual advances in studying ecosystem functions is a challenge for peatland scientists. Advances in this area require multidisciplinary collaborations between peatland scientists, data scientists and microbiologists and ultimately, collaboration with the modelling community. Developing a process-based understanding of the resilience and recovery of peatlands to perturbations, such as climate extremes, fires, and drainage, will be key to meeting climate targets and delivering ecosystem services cost effectively.

The potential for modelling peatland habitat condition in Scotland using long-term MODIS data.

Globally, peatlands provide an important sink of carbon in their near natural state but potentially act as a source of gaseous and dissolved carbon emission if not in good condition. There is a pressing need to remotely identify peatland sites requiring improvement and to monitor progress following restoration. A medium resolution model was developed based on a training dataset of peatland habitat condition and environmental covariates, such as morphological features, against information derived from the Moderate Resolution Imaging Spectroradiometer (MODIS), covering Scotland (UK). The initial, unrestricted, model provided the probability of a site being in favourable condition. Receiver operator characteristics (ROC) curves for restricted training data, limited to those located on a peat soil map, resulted in an accuracy of 0.915. The kappa statistic was 0.8151, suggesting good model fit. The derived map of predicted peatland condition at the suggested 0.56 threshold was corroborated by data from other sources, including known restoration sites, areas under known non-peatland land cover and previous vegetation survey data mapped onto inferred condition categories. The resulting locations of the areas of peatland modelled to be in favourable ecological condition were largely confined to the North and West of the country, which not only coincides with prior land use intensity but with published predictions of future retraction of the bioclimatic space for peatlands. The model is limited by a lack of spatially appropriate ground observations, and a lack of verification of peat depth at training site locations, hence future efforts to remotely assess peatland condition will require more appropriate ground-based monitoring. If appropriate ground-based observations could be collected, using remote sensing could be considered a cost-efficient means to provide data on changes in peatland habitat condition.

Interactions among fungal community structure, litter decomposition and depth of water table in a cutover peatland.

Peatlands are important reservoirs of carbon (C) but our understanding of C cycling on cutover peatlands is limited. We investigated the decomposition over 18 months of five types of plant litter (Calluna vulgaris, Eriophorum angustifolium, Eriophorum vaginatum, Picea sitchensis and Sphagnum auriculatum) at a cutover peatland in Scotland, at three water tables. We measured changes in C, nitrogen (N) and phosphorus (P) in the litter and used denaturing gradient gel electrophoresis to investigate changes in fungal community composition. The C content of S. auriculatum litter did not change throughout the incubation period whereas vascular plant litters lost 30-40% of their initial C. There were no differences in C losses between low and medium water tables, but losses were always significantly less at the high water table. Most litters accumulated N and E. angustifolium accumulated significant quantities of P. C, N and P were significant explanatory variables in determining changes in fungal community composition but explained <25% of the variation. Litter type was always a stronger factor than water table in determining either fungal community composition or turnover of C, N and P in litter. The results have implications for the ways restoration programmes and global climate change may impact upon nutrient cycling in cutover peatlands.

Summer drought decreases soil fungal diversity and associated phenol oxidase activity in upland Calluna heathland soil.

Natural moisture limitation during summer drought can constitute a stress for microbial communities in soil. Given globally predicted increases in drought frequency, there is an urgent need for a greater understanding of the effects of drought events on soil microbial processes. Using a long-term field-scale drought manipulation experiment at Clocaenog, Wales, UK, we analysed fungal community dynamics, using internal transcribed spacer-denaturing gradient gel electrophoresis (DGGE), over a 1-year period in the 6th year of drought manipulation. Ambient seasonality was found to be the dominant factor driving variation in fungal community dynamics. The summer drought manipulation resulted in a significant decline in the abundance of dominant fungal species, both independently of, and in interaction with, this seasonal variation. Furthermore, soil moisture was significantly correlated with the changes in fungal diversity over the drought manipulation period. While the relationship between species diversity and functional diversity remains equivocal, phenol oxidase activity was decreased by the summer drought conditions and there was a significant correlation with the decline of DGGE band richness among the most dominant fungal species during the drought season. Climatically driven events such as droughts may have significant implications for fungal community diversity and therefore, have the potential to interfere with crucial ecosystem processes, such as organic matter decomposition.

Calluna vulgaris-dominated upland heathland sequesters more CO2 annually than grass-dominated upland heathland.

It has been shown in many habitats worldwide, that a shift in vegetation composition between woody shrub and graminoid dominance can influence carbon (C) cycling. Due to land management practices and environmental change, UK upland heath vegetation has historically undergone shifts in dominance from the woody dwarf shrub Calluna vulgaris (Calluna) to species poor graminoid swards. The consequences of this for C sequestration are unknown. We compared annual net ecosystem exchange (NEE) of carbon dioxide (CO2) between building phase Calluna- and grass-dominated communities within three upland heaths in Scotland, measuring c. monthly over a year. Light and temperature response curves were generated, and the parameters derived were applied to continuous light and temperature data to extrapolate CO2 fluxes over the full year and generate estimates of annual CO2 sequestration for each vegetation type. Grass-dominated communities had higher ecosystem respiration rates than Calluna-dominated communities, attributed to graminoids having greater metabolic demands and producing more labile litter which decomposes readily. Both communities had similar gross primary productivity over the year; the net result being higher NEE within the Calluna-dominated than the grass-dominated community (-2.36 ± 0.23 and -1.78 ± 0.18 µmol CO2m(-2)s(-1) respectively). Modelled CO2 fluxes over a year showed both communities to be CO2 sinks. The Calluna-dominated community sequesters -3.45 ± 0.96 t C ha(-1)yr(-1), double that sequestered by the grass-dominated community at 1.61 ± 0.57 t C ha(-1)yr(-1). Potential rate of C sequestration by upland heath is comparable to that of woodland and the increase in total sequestration that could be gained from habitat restoration may equate to c. 60% of the annual UK C sink attributed to forest land management. National C sequestration by heathlands is also more than double that by peatlands. Management of graminoid-dominated upland heath should promote Calluna re-establishment, thus providing a C benefit in addition to benefits to biodiversity, grazing and sporting interests.

Survival of Escherichia coli O157:H7 in private drinking water wells: influences of protozoan grazing and elevated copper concentrations.

The survival characteristics of Escherichia coli O157:H7 in private drinking water wells were investigated to assess the potential for human exposure. A non-toxigenic, chromosomally lux-marked strain of E. coli O157:H7 was inoculated into well water from four different sites in the North East of Scotland. These waters differed significantly in their heavy metal contents as well as nutrient and bacterial grazer concentrations. Grazing and other biological factors were studied using filtered (3 and 0.2 microm) and autoclaved water. The survival of E. coli O157:H7 was primarily decreased by elevated copper concentrations. This hypothesis was supported by acute toxicity assay data. In addition, significant protozoan predation effects were observed in untreated water when compared with survival rates in filtered water. The combination of these two factors in particular determines the survival time of the pathogen in a private water well. It therefore appears that wells with higher water quality as assessed using the European Union Drinking Water Directive standards will also allow survival of E. coli O157:H7 for much longer periods.

Long term repeated prescribed burning increases evenness in the basidiomycete laccase gene pool in forest soils.

Repeated prescribed burning alters the biologically labile fraction of nutrients and carbon of soil organic matter (SOM). Using a long-term (30 years) repeated burning experiment where burning has been carried out at a 2- or 4-year frequency, we analysed the effect of prescribed burning on gross potential C turnover rates and phenol oxidase activity in relation to shifts in SOM composition as observed using Fourier-transform infrared spectroscopy. In tandem, we assessed the genetic diversity of basidiomycete laccases. While the overall effect of burning was a decline in phenol oxidase activity, Shannon diversity and evenness of laccases was significantly higher in burned sites. Co-correspondence analysis of SOM composition and laccase operational taxonomic unit frequency data also suggested a strong correlation. While this correlation could indicate that the observed increase in laccase genetic diversity due to burning is due to increased resource diversity, a temporal replacement of the most abundant members of the assembly by an otherwise dormant pool of fungi cannot be excluded. As such, our results fit the intermediate disturbance hypothesis. Effects were stronger in plots burned in 2-year rotations, suggesting that the 4-year burn frequency may be a more sustainable practice to ensure the long-term stability of C cycling in such ecosystems.

Changes in fungal community composition in response to vegetational succession during the natural regeneration of cutover peatlands.

Despite the importance of peatlands as a major store of sequestered carbon and the role of fungi in releasing sequestered C, we know little about the community structure of fungi in peatlands. We investigated these across a gradient of naturally regenerating peatland vegetation using denaturing gradient gel electrophoresis (DGGE) and clone libraries of fragments of the fungal rRNA internal transcribed spacer (ITS) region. Significant changes in the fungal community structure of peat samples at different stages of regeneration were observed, which relate to the composition of the vegetation recolonizing these sites. Cloning and sequence analysis also demonstrated a potential shift in the relative abundance of the main fungal phyla. Some of the clones identified to genus level were highly related to fungi known to play a role in the degradation of plant litter or wood in similar ecosystems and/or form mycorrhizal associations. In addition, several fungal isolates highly related to peat clones were obtained, and their enzymic capacity to degrade structural plant tissues was assessed. Together, these results suggest that the fungal community composition of peat may be an important indicator of the status of regeneration and potential carbon sequestration of cutover peatlands.

Potential pitfalls in the quantitative molecular detection of Escherichia coli O157:H7 in environmental matrices.

The detection sensitivity and potential interference factors of a commonly used assay based on real-time polymerase chain reaction (PCR) for Escherichia coli O157:H7 using eae gene-specific primers were assessed. Animal wastes and soil samples were spiked with known replicate quantities of a nontoxigenic strain of E. coli O157:H7 in a viable or dead state and as unprotected DNA. The detection sensitivity and accuracy of real-time PCR for E. coli O157:H7 in animal wastes and soil is low compared to enrichment culturing. Nonviable cells and unprotected DNA were shown to produce positive results in several of the environmental samples tested, leading to potential overestimates of cell numbers due to prolonged detection of nonviable cells. This demonstrates the necessity for the specific calibration of real-time PCR assays in environmental samples. The accuracy of the eae gene-based detection method was further evaluated over time in a soil system against an activity measurement, using the bioluminescent properties of an E. coli O157:H7 Tn5luxCDABE construct. The detection of significant numbers of viable but nonculturable (VBNC) as well as nonviable and possibly physically protected cells as shown over a period of 90 days further complicates the use of real-time PCR assays for quick diagnostics in environmental samples and infers that enrichment culturing is still required for the final verification of samples found positive by real-time PCR methods.

Soil macropores and compaction control the leaching potential of Escherichia coli O157:H7.

The influence of soil structure in controlling leaching of Escherichia coli O157:H7 through soil was investigated under controlled conditions using both intact and repacked soil cores. Leaching rates of E. coli O157:H7 decreased with increasing dry bulk density and were significantly increased by the presence of earthworm (Lumbricus terrestris) burrows in repacked cores. For intact cores, the percentage of E. coli O157:H7 that leached through replicate cores within 72 h varied from 0.01% to 24%. In contrast, the dry bulk densities of intact cores varied only slightly and were not significantly correlated with leaching. Differences in the numbers of E. coli O157:H7 cells in the leachates were not related to variability in the flow volumes, which were relatively constant as a result of the experimental design, but were strongly correlated with the variations in concentrations of E. coli O157:H7 in the leachates. Relatively small variations in the internal structure of soil cores can therefore significantly affect the pathway that cells can take through soil. Factors such as compaction and the occurrence of pores providing preferential flow are prime determinants in the degree of leaching of E. coli O157:H7 through soil.

Effect of nematodes on rhizosphere colonization by seed-applied bacteria.

There is much interest in the use of seed-applied bacteria for biocontrol and biofertilization, and several commercial products are available. However, many attempts to use this strategy fail because the seed-applied bacteria do not colonize the rhizosphere. Mechanisms of rhizosphere colonization may involve active bacterial movement or passive transport by percolating water or plant roots. Transport by other soil biota is likely to occur, but this area has not been well studied. We hypothesized that interactions with soil nematodes may enhance colonization. To test this hypothesis, a series of microcosm experiments was carried out using two contrasting soils maintained under well-defined physical conditions where transport by mass water flow could not occur. Seed-applied Pseudomonas fluorescens SBW25 was capable of rhizosphere colonization at matric potentials of -10 and -40 kPa in soil without nematodes, but colonization levels were substantially increased by the presence of nematodes. Our results suggest that nematodes can have an important role in rhizosphere colonization by bacteria in soil.

The potential for anaerobic mineralisation of hydrocarbon constituents of oily drill cuttings from the North Sea seabed.

The potential for natural attenuation of hydrocarbons in oily drill cuttings from the seabed beneath a North Sea oil platform was investigated. The study focused on the anaerobic degradation of n-hexadecane, n-octacosane and naphthalene using additions of 14C-labelled analogues to drill cuttings samples and was conducted under realistic seabed conditions (except pressure) over an 11-month period. No mineralisation of naphthalene was detected over this time period and mineralisation of octacosane represented only 0.5-1.5% of the added label. In contrast, mineralisation of hexadecane was 10-49% after 11 months of incubation. Selective inhibition of key functional bacterial groups revealed the key role of both sulfate reduction and methanogenesis in the degradation process. This study demonstrates the potential for natural attenuation of at least some hydrocarbon constituents of oily drill cuttings under realistic environmental conditions and highlights the involvement of a wide functional consortium in the natural attenuation process.

A stable bioluminescent construct of Escherichia coli O157:H7 for hazard assessments of long-term survival in the environment.

A chromosomally lux-marked (Tn5 luxCDABE) strain of nontoxigenic Escherichia coli O157:H7 was constructed by transposon mutagenesis and shown to have retained the O157, H7, and intimin phenotypes. The survival characteristics of this strain in the experiments performed (soil at -5, -100, and -1,500 kPa matric potential and artificial groundwater) were indistinguishable from the wild-type strain. Evaluation of potential luminescence was found to be a rapid, cheap, and quantitative measure of viable E. coli O157:H7 Tn5 luxCDABE populations in environmental samples. In the survival studies, bioluminescence of the starved populations of E. coli O157:H7 Tn5 luxCDABE could be reactivated to the original levels of light emission, suggesting that these populations remain viable and potentially infective to humans. The attributes of the construct offer a cheap and low-risk substitute to the use of verocytotoxin-producing E. coli O157:H7 in long-term survival studies.