Christian Schurig

Microbial cell-envelope fragments and the formation of soil organic matter: a case study from a glacier forefield

Genesis of soil organic matter (SOM) during pedogenesis is still a matter of controversy in soil science. Recently, it was hypothesized that microbial cell-envelope fragments contribute significantly to SOM formation. We tested the relevance of this process during pedogenesis by evaluating the development of SOM along a chronosequence of a glacier forefield (Damma glacier). Samples of increasing soil age collected along the forefield were analyzed for C and N contents, phospholipid and total fatty acids (PLFA and tFA), water contact angle, micro-hydrophobicity and surface coverage by microbial cell-envelope residues. The surface coverage was visualized and quantified by analysis of representative, equally-scaled scanning electron micrographs (SEM). Increasing SOM contents were accompanied by increasing coverage and overall abundance of microbial cell-envelope fragments as evaluated on the basis of scanning electron microscopy; this is also reflected in the amounts of tFA and PLFA, the trend of C/N ratios, and the increasing hydrophobicity and water contact angles of the soil samples. Using SEM and the image analysis approach, we can provide a process-based description of the development of SOM in the newly developing ecosystem of the glacier forefield. The majority of small-sized SOM visible with scanning electron microscopy appears to consist of bacterial cell envelope fragments that remain stable after cell death, such that their shape does not change with soil age. Our results show the importance of microbial processing of SOM, and highlight the existence of microbial necromass as a significant part of the fine-particulate SOM even in later stages of soil development.

Methods for visualising active microbial benzene degraders in in situ microcosms

Natural attenuation maybe a cost-efficient option for bioremediation of contaminated sites but requires knowledge about the activity of degrading microbes under in situ conditions. In order to link microbial activity to the spatial distribution of contaminant degraders, we combined the recently improved in situ microcosm approach, so-called ‘direct-push bacterial trap’ (DP-BACTRAP), with nano-scale secondary ion mass spectrometry (NanoSIMS) analysis on samples from contaminated constructed wetlands. This approach is based on initially sterile microcosms amended with 13C-labelled benzene as a source of carbon and energy for microorganisms. The microcosms were introduced directly in the constructed wetland, where they were colonised by indigenous microorganisms from the sediment. After incubation in the field, the samples were analysed by NanoSIMS, scanning electron microscopy (SEM) and fluorescence microscopy in order to visualise 13C-labelled microbial biomass on undisturbed samples from the microcosms. With the approach developed, we successfully visualised benzene-degrading microbes on solid materials with high surface area by means of NanoSIMS. Moreover, we could demonstrate the feasibility of NanoSIMS analysis of unembedded porous media with a highly complex topography, which was frequently reasoned to not lead to sufficient results.

Characterisation of microbial activity in the framework of natural attenuation without groundwater monitoring wells?: a new Direct-Push probe

At many contaminated field sites in Europe, monitored natural attenuation is a feasible site remediation option. Natural attenuation includes several processes but only the microbial degradation leads to real contaminant removal and very few methods are accepted by the authorities providing real evidence of microbial contaminant degradation activity. One of those methods is the recently developed in situ microcosm approach (BACTRAP®). These in situ microcosms consist of perforated stainless steel cages or PTFE tubes filled with an activated carbon matrix that is amended with 13C-labelled contaminants; the microcosms are then exposed within groundwater monitoring wells. Based on this approach, natural attenuation was accepted by authorities as a site remediation option for the BTEX-polluted site Zeitz in Germany. Currently, the in situ microcosms are restricted to the use inside groundwater monitoring wells at the level of the aquifer. The (classical) system therefore is only applicable on field sites with a network of monitoring wells, and only microbial activity inside the monitoring wells at the level of the aquifer can be assessed. In order to overcome these limitations, a new Direct-Push BACTRAP probe was developed on the basis of the Geoprobe® equipment. With respect to the mechanical boundary conditions of the DP technique, these new probes were constructed in a rugged and segmented manner and are adaptable to various sampling concepts. With this new probe, the approach can be extended to field sites without existing monitoring wells, and microbial activity was demonstrated to be measureable even under very dry conditions inside the vadose zone above the aquifer. In a field test, classical and Direct-Push BACTRAPs were applied in the BTEX-contaminated aquifer at the ModelPROBE reference site Zeitz (Germany). Both types of BACTRAPs were incubated in the centre and at the fringe of the BTEX plume. Analysis of phospholipid fatty acid (PLFA) patterns showed that the bacterial communities on DP-BACTRAPs were more similar to the soil than those found on classical BACTRAPs. During microbial degradation of the 13C-labelled substrate on the carrier material of the microcosms, the label was only slightly incorporated into bacterial biomass, as determined by PLFA analysis. This provides clear indication for decreased in situ natural attenuation potential in comparison to earlier sampling campaigns, which is presumably caused by a large-scale source remediation measure in the meantime. In conclusion, Direct-Push-based BACTRAPs offer a promising way to monitor natural attenuation or remediation success at field sites which are currently inaccessible by the technique due to the lack of monitoring wells or due to a main contamination present within the vadose zone.

Soil wettability can be explained by the chemical composition of particle interfaces - An XPS study

Soil wettability (quantified in terms of contact angle, CA) is crucial for physical, chemical, and biological soil functioning. As the CA is determined by components present within the outmost nanometer of particles, this study applied X-ray photoelectron spectroscopy (XPS) with a maximum analysis depth of 10 nm to test the relationship between CA and surface elemental composition, using soil samples from a chronosequence where CA increased from 0° (0 yrs) to about 98° (120 yrs). Concurrently, as seen by XPS, C and N content increased and the content of O and the mineral-derived cations (Si, Al, K, Na, Ca, Mg, Fe) decreased. The C content was positively correlated with CA and least squares fitting indicated increasing amounts of non-polar C species with soil age. The contents of O and the mineral-derived cations were negatively correlated with CA, suggesting an increasing organic coating of the minerals that progressively masked the underlying mineral phase. The atomic O/C ratio was found to show a close negative relationship with CA, which applied as well to further sample sets of different texture and origin. This suggests the surface O/C ratio to be a general parameter linking surface wettability and surface elemental composition.

Spatially Resolved Quantification by NanoSIMS of Organic Matter Sorbed to (Clay) Minerals

Soils are highly heterogeneous entities in which organic and inorganic as well as living and non-living building blocks interact to form biogeochemical interfaces. While processes at these interfaces occur at the micrometer or submicrometer scale, they are thought to influence the behavior of soils at the global scale, e.g. soils as carbon sinks. Analytical methodologies with a high resolution are, therefore, required in order to investigate these processes with the final goal to understand biogeochemical-interface formation mechanistically. In the present study, sorption experiments of water-extractable organic matter on model minerals, such as boehmite and illite, were performed. Adsorption of organic matter on the minerals was quantified by conventional bulk-scale methods and compared with data from nanoSIMS measurements. From the data obtained, scaling factors have been developed which permit the quantification of organic matter in the secondary ion images provided by nanoSIMS.