Stems of Phragmites australis are buffering methane and carbon dioxide emissions.

Internal concentrations in the culm nodes of Phragmites australis and fluxes of methane (CH4) and carbon dioxide (CO2) were recorded in the treatment bed of constructed wetland (CW) with subsurface wastewater horizontal flow. Fluxes of CH4 and CO2 from the CW treatment bed were in ranges of 0 to 490 µmol m-2 h-1 and from 0 to 4499 µmol m-2 h-1 for CH4 and CO2, respectively. The highest CH4 soil fluxes were recorded in the unvegetated coarse gravel inflow zone of the CW treatment bed. The nearby inflow zone exhibited the highest CO2 fluxes. Internal culm node concentrations of CH4 and CO2 were related to oxygen (O2) stem concentrations and environmental conditions during diurnal courses. The concentrations of CH4 and CO2 gases were significantly correlated and opposing O2 concentrations. Culm node parameters and shoot density of P. australis influenced internal gas concentrations and the buffering of CH4 and CO2 emissions. The effect of buffering CH4 emissions is distinctive in the outflow zone of the treatment bed and is less important in the highly polluted inflow zone of the CW. Buffering of CH4 and partially also CO2 emissions by stems of P. australis is a process which affects the diurnal dynamics of CH4 and CO2 fluxes from common reed wetlands.

Portable Measurement System for in situ Estimation of Oxygen and Carbon Fluxes of Submerged Plants.

The metabolism of submerged plants is commonly characterized by oxygen development. The turnover rates of carbon dioxide and other inorganic carbon species, however, are assessed only at distinct points in time after incubation or calculated through shifts in pH and total alkalinity. A novel three parameter measurement system was developed in order to improve this issue and to gain a better understanding of the metabolism of aquatic plants. It allows the simultaneous and continuous assessment of oxygen concentration, partial pressure of carbon dioxide and pH with optical sensors without the need of taking water samples. Plants or plant parts can be enclosed in a chamber, while the surrounding water is either flushed through or circulated within the system. The method was evaluated in regards to measurement time and possible stress reactions during measurement. Its applicability in situ was confirmed with Elodea nuttallii and Ceratophyllum demersum. The measurement system will enable deeper insights into the metabolism and response of aquatic plants to changing environmental conditions, especially related to carbon fixation.

Beneficial effects of intermittent feedstock management on biogas and methane production.

Intermittent supply of easily degradable carbohydrates can be used for on-demand biogas production. The study tested the effects of splitting feeding portions of sugar beet silage (S) on biogas production rates and total yield, respectively and if methane production rates follow those ones of biogas. Four experimental AD reactors were operated for 117 days at organic loading rates of 2.0 kgVS m-3 d-1 and VS ratios of maize silage (M) to S of 3:1. While M was supplied hourly (h0-h12), reactors differed only regarding the intermittent S supply, provided at once (h0), twice (h0, h1) and three times (h0, h1, h2) per twelve-hour observation period. Biogas and methane production rates rose simultaneously after S supply and lasted depending on S intakes. Biogas and methane yields were significantly increased at S given once and twice per period. Appropriate feedstock management can thus influence production rates and increase biogas and methane yields.

Different response of bacteria, archaea and fungi to process parameters in nine full-scale anaerobic digesters.

Biogas production is a biotechnological process realized by complex bacterial, archaeal and likely fungal communities. Their composition was assessed in nine full-scale biogas plants with distinctly differing feedstock input and process parameters. This study investigated the actually active microbial community members by using a comprehensive sequencing approach based on ribosomal 16S and 28S rRNA fragments. The prevailing taxonomical units of each respective community were subsequently linked to process parameters. Ribosomal rRNA of bacteria, archaea and fungi, respectively, showed different compositions with respect to process parameters and supplied feedstocks: (i) bacterial communities were affected by the key factors temperature and ammonium concentration; (ii) composition of archaea was mainly related to process temperature; and (iii) relative abundance of fungi was linked to feedstocks supplied to the digesters. Anaerobic digesters with a high methane yield showed remarkably similar bacterial communities regarding identified taxonomic families. Although archaeal communities differed strongly on genus level from each other, the respective digesters still showed high methane yields. Functional redundancy of the archaeal communities may explain this effect. 28S rRNA sequences of fungi in all nine full-scale anaerobic digesters were primarily classified as facultative anaerobic Ascomycota and Basidiomycota. Since the presence of ribosomal 28S rRNA indicates that fungi may be active in the biogas digesters, further research should be carried out to examine to which extent they are important players in anaerobic digestion processes.

Characteristics of on-demand biogas production by using sugar beet silage.

On-demand electricity generation can be achieved by just-in-time biogas production instantly utilized in co-generation units. For this goal, easily degradable substrates like sugar beet silage have a high potential. Potential for on-demand biogas production from co-digestion of sugar beet silage (SS) with grass silage (GS) was evaluated in two experiments at organic loading rates (OLRs) of 1.5 kgVS m-3 day-1 and 2.5 kgVS m-3 day-1, respectively. Each experiment was fed with intermittent feeding system at 8 hrs interval at the same feedstock ratios (volatile solids based) of GS:SS-1:0, 3:1 and 1:3, respectively. Modelling by Gaussian equation was performed in order to understand the effects of SS on biogas production. Addition of sugar beet silage led to maximum biogas production within a short time, but it differed significantly depending on feedstock ratios and OLRs, respectively. At OLR 1.5 kgVS m-3 day-1, during mono fermentation of grass silage maximum biogas production rate of 0.27 lN hr-1 was reached at 2.74 hrs. Production rate did not change at feedstock ratio of GS:SS-3:1 but increased to 0.64 lN hr-1 at GS:SS-1:3 within a shorter time span (1.58 hrs). On the contrary, at OLR of 2.5 kgVS m-3 day-1 time span between feedstock input and maximum biogas production did not differ significantly (p > 0.05) among the reactors. Biogas production rates were 0.60 lN hr-1 within 2.27 hrs and 0.82 lN hr-1 within 2.30 hrs at GS:SS-3:1 and GS:SS-1:3, respectively. Surprisingly, there was no time lag between maximum biogas and methane production rates, irrespectively of OLR. This implies that once the whole microbial community is adapted to intermittent substrate input, the metabolic products are instantly utilized through the all steps of anaerobic substrate degradation. Applying this finding opens new perspectives for on-demand biogas energy production.

Co-Digestion of Sugar Beet Silage Increases Biogas Yield from Fibrous Substrates.

This study tested the hypothesis that the easily degradable carbohydrates of the sugar beet silage (S) will improve the anaerobic digestion of grass silage (G) more profoundly compared to co-digestion of sugar beet silage with maize silage (M). M : S and G : S mixtures were tested in two continuous laboratory-scale AD experiments at volatile solid ratios of 1 : 0, 6 : 1, 3 : 1, and 1 : 3 at organic loading rates of 1.5 kgVS m-3 day-1. While the sugar beet effects in mixtures with maize silage were negligible, co-digestion with grass silage showed a beneficial performance. There, the specific methane production rate was 0.27 lN kg-1VS h-1at G : S ratio of 6 : 1 compared to G : S 1 : 0 with 0.14 lN kg-1VS h-1. In comparison to G : S 1 : 0, about 44% and 62% higher biogas yields were obtained at G : S 6 : 1 and 3 : 1, respectively. Also, the highest methane concentration was found in G : S at ratio of 1 : 3. Synergistic increase of methane yield was found in co-digestion in both experiments, but higher effect was realized in G : S, independently of the amount of sugar beet silage. The findings of this study emphasize the improvement of AD of grass silage by even low addition of sugar beet silage.

Diurnal dynamics of oxygen and carbon dioxide concentrations in shoots and rhizomes of a perennial in a constructed wetland indicate down-regulation of below ground oxygen consumption.

Wetland plants actively provide oxygen for aerobic processes in submerged tissues and the rhizosphere. The novel concomitant assessment of diurnal dynamics of oxygen and carbon dioxide concentrations under field conditions tests the whole-system interactions in plant-internal gas exchange and regulation. Oxygen concentrations ([O2]) were monitored in-situ in central culm and rhizome pith cavities of common reed (Phragmites australis) using optical oxygen sensors. The corresponding carbon dioxide concentrations ([CO2]) were assessed via gas samples from the culms. Highly dynamic diurnal courses of [O2] were recorded, which started at 6.5-13 % in the morning, increased rapidly up to 22 % during midday and declined exponentially during the night. Internal [CO2] were high in the morning (1.55-17.5 %) and decreased (0.04-0.94 %) during the rapid increase of [O2] in the culms. The observed negative correlations between [O2] and [CO2] particularly describe the below ground relationship between plant-mediated oxygen supply and oxygen use by respiration and biogeochemical processes in the rhizosphere. Furthermore, the nocturnal declining slopes of [O2] in culms and rhizomes indicated a down-regulation of the demand for oxygen in the complete below ground plant-associated system. These findings emphasize the need for measurements of plant-internal gas exchange processes under field conditions because it considers the complex interactions in the oxic-anoxic interface.

Functionally redundant but dissimilar microbial communities within biogas reactors treating maize silage in co-fermentation with sugar beet silage.

Numerous observations indicate a high flexibility of microbial communities in different biogas reactors during anaerobic digestion. Here, we describe the functional redundancy and structural changes of involved microbial communities in four lab-scale continuously stirred tank reactors (CSTRs, 39°C, 12 L volume) supplied with different mixtures of maize silage (MS) and sugar beet silage (SBS) over 80 days. Continuously stirred tank reactors were fed with mixtures of MS and SBS in volatile solid ratios of 1:0 (Continuous Fermenter (CF) 1), 6:1 (CF2), 3:1 (CF3), 1:3 (CF4) with equal organic loading rates (OLR 1.25 kgVS m(-3) d(-1) ) and showed similar biogas production rates in all reactors. The compositions of bacterial and archaeal communities were analysed by 454 amplicon sequencing approach based on 16S rRNA genes. Both bacterial and archaeal communities shifted with increasing amounts of SBS. Especially pronounced were changes in the archaeal composition towards Methanosarcina with increasing proportion of SBS, while Methanosaeta declined simultaneously. Compositional shifts within the microbial communities did not influence the respective biogas production rates indicating that these communities adapted to environmental conditions induced by different feedstock mixtures. The diverse microbial communities optimized their metabolism in a way that ensured efficient biogas production.

Dynamics of biofilm formation during anaerobic digestion of organic waste.

Biofilm-based reactors are effectively used for wastewater treatment but are not common in biogas production. This study investigated biofilm dynamics on biofilm carriers incubated in batch biogas reactors at high and low organic loading rates for sludge from meat industry dissolved air flotation units. Biofilm formation and dynamics were studied using various microscopic techniques. Resulting micrographs were analysed for total cell numbers, thickness of biofilms, biofilm-covered surface area, and the area covered by extracellular polymeric substances (EPS). Cell numbers within biofilms (10(11) cells ml(-1)) were up to one order of magnitude higher compared to the numbers of cells in the fluid reactor content. Further, biofilm formation and structure mainly correlated with the numbers of microorganisms present in the fluid reactor content and the organic loading. At high organic loading (45 kg VS m(-3)), the thickness of the continuous biofilm layer ranged from 5 to 160 µm with an average of 51 µm and a median of 26 µm. Conversely, at lower organic loading (15 kg VS m(-3)), only microcolonies were detectable. Those microcolonies increased in their frequency of occurrence during ongoing fermentation. Independently from the organic loading rate, biofilms were embedded completely in EPS within seven days. The maturation and maintenance of biofilms changed during the batch fermentation due to decreasing substrate availability. Concomitant, detachment of microorganisms within biofilms was observed simultaneously with the decrease of biogas formation. This study demonstrates that biofilms of high cell densities can enhance digestion of organic waste and have positive effects on biogas production.

Leaf allocation patterns and 13C and 15N natural abundances of tropical lianas (Passiflora sp.) as dependent on external climbing support.

The transformation from self-supporting lianas to host-supported climbing lianas is related to re-allocation of biomass and nutrients among plant organs. Therefore, first, variations in leaf mass per area (LMA), leaf carbon and nitrogen allocation and (13)C and (15)N natural abundances were analysed among three tropical Passiflora species (P. edulis, P. ligularis, and P. tripartita) in a greenhouse study. Second, the influence of a climbing support was considered for each species and parameter. P. ligularis leaves were most enriched in (13)C in both treatments when compared with the other two species. This enrichment was caused by a high LMA, which is related to a high internal resistance to CO(2) diffusion. For P. edulis and P. tripartita, δ(13)C was additionally increasing with nitrogen content per area. Generally, there were no differences when considering carbon and nitrogen allocation to leaves of host-supported and self-supporting lianas. The only hints towards increased investment into leaves after the transition from self-supporting to host-supported stages could be seen by a trend to increased leaf areas and masses. δ(13)C values of supported P. edulis or P. tripartita plants were significantly increasing faster than those of non-supported plants once the interactions of leaf mass or nitrogen content per area were accounted for. Hence, the offer of a climbing support had only a minor impact on δ(13)C or δ(15)N values in vitro, but this could be different with increasing age of lianas in vivo.

Stability of a biogas-producing bacterial, archaeal and fungal community degrading food residues.

The resident microbiota was analyzed in a mesophilic, continuously operating biogas plant predominantly utilizing food residues, stale bread, and other waste cosubstrates together with pig manure and maize silage. The dominating bacterial, archaeal, and eukaryotic community members were characterized by two different 16S/18S rRNA gene culture-independent approaches. Prokaryotic 16S rRNA gene and eukaryotic 18S rRNA gene clone libraries were constructed and further analyzed by restriction fragment length polymorphism (RFLP), 16S/18S rRNA gene sequencing, and phylogenetic tree reconstruction. The most dominant bacteria belonged to the phyla Bacteriodetes, Chloroflexus, and Firmicutes. On the family level, the bacterial composition confirmed high differences among biogas plants studied so fare. In contrast, the methanogenic archaeal community was similar to that of other studied biogas plants. Furthermore, it was possible to identify fungi at the genus level, namely Saccharomyces and Mucor. Both genera, which are important for microbial degradation of complex compounds, were up to now not found in biogas plants. The results revealed their long-term presence as indicated by denaturating gradient gel electrophoresis (DGGE). The DGGE method confirmed that the main members of the microbial community were constantly present over more than one-year period.

Modelling tree roots in mixed forest stands by inhomogeneous marked Gibbs point processes.

The aim of the paper is to apply point processes to root data modelling. We propose a new approach to parametric inference when the data are inhomogeneous replicated marked point patterns. We generalize Geyer's saturation point process to a model, which combines inhomogeneity, marks and interaction between the marked points. Furthermore, the inhomogeneity influences the definition of the neighbourhood of points. Using the maximum pseudolikelihood method, this model is then fitted to root data from mixed stands of Norway spruce (Picea abies (L.) Karst.) and European beech (Fagus sylvatica L.) to quantify the degree of root aggregation in such mixed stands. According to the analysis there is no evidence that the two root systems are not independent.

Fine root dynamics of mature European beech (Fagus sylvatica L.) as influenced by elevated ozone concentrations.

Fine root dynamics (diameter < 1 mm) in mature Fagus sylvatica, with the canopies exposed to ambient or twice-ambient ozone concentrations, were investigated throughout 2004. The focus was on the seasonal timing and extent of fine root dynamics (growth, mortality) in relation to the soil environment (water content, temperature). Under ambient ozone concentrations, a significant relationship was found between fine root turnover and soil environmental changes indicating accelerated fine root turnover under favourable soil conditions. In contrast, under elevated ozone, this relationship vanished as the result of an altered temporal pattern of fine root growth. Fine root survival and turnover rate did not differ significantly between the different ozone regimes, although a delay in current-year fine root shedding was found under the elevated ozone concentrations. The data indicate that increasing tropospheric ozone levels can alter the timing of fine root turnover in mature F. sylvatica but do not affect the turnover rate.

Community assembly of terrestrial testate amoebae: how is the very first beginning characterized?

Testate amoebae play an important role at the very first beginning of succession on land. We used litterbags buried into four different soils to study the early colonization (which occurred within less than 55 days) and establishment of testate amoebae. The litterbag cellulose exposed at the youngest mining site poor in nitrogen and phosphorus was colonized firstly in high abundances, whereas the substrate introduced into the reference sites of undisturbed soil was colonized slowly and in low densities. Besides the (expected) small-sized r-strategists (e.g., Euglypha rotunda, Tracheleuglypha dentata, and Trinema lineare), large-sized K-strategists (e.g., Centropyxis spp., Phryganella acropodia) occurred in remarkably high densities on all sites. Species that colonized the cellulose in high densities (e.g., P. acropodia and T. dentata) were found extremely rarely in the adjacent source substrate and vice versa, stressing the importance of the target substrate quality. In the course of the experiment, the influencing environmental factors became more complex, as shown by redundancy analysis (RDA). Concerning the amoebal community, there was a change from variability to stability, as visualized by cluster analysis. Adjacent litterbags within an investigation site revealed amoebal species and abundances with an increasing similarity during exposition time, whereas the litterbags between the four investigation sites were colonized differently. These observations point to a stochastic (variable) beginning of community assembly, changing to a more deterministic (stable) course. No species replacement has been observed, which is an essential part of most successional theories. Thus, the more flexible concept of "community assembly" should be considered instead of "succession" for protozoa. The stochastic beginning of community assembly and the lack of species replacement are explained by a neutral community model.

Clustered root distribution in mature stands of Fagus sylvatica and Picea abies.

Distribution of small roots (diameter between 2 mm and 5 mm) was studied in 19 pits with a total of 72 m(2) trench profile walls in pure stands of Fagus sylvatica and Picea abies. Root positions within the walls were marked and transformed into x-coordinates and y-coordinates. In a GIS-based evaluation, zones of potential influence around each root were calculated. The total potential influence produced isoline maps of relative root influence zones, thus indicating small root clustering. The questions studied were (1) whether there were marked clusters of small roots in the soil and (2) whether trees surrounding the pit (defined as tree density) correlate with the root abundance and distribution on the trench profile walls. Small roots of both species showed maximum abundance in the top 20 cm of the soil, where pronounced root clusters occurred next to areas with only low root accumulation. The area of root clusters did not differ significantly between the two stands. Weighted clumping, WC, calculated as a product of root class, and its area was used as an index of root clustering, which again did not differ between beech and spruce stands. However, evaluations on a single root level showed that beech achieved the same degree of clustering with lower number of roots. Regardless of soil properties related to root clusters, a significantly higher clustering acquired per root for beech than for spruce suggests beech to be more efficient in belowground acquisition of space. Because none of the parameters describing root clustering were correlated with tree density around the investigated soil profiles, clusters of small roots are inherently present within the tree stands.