Sabine Marie Podmirseg

Going for mainstream deammonification from bench to full scale for maximized resource efficiency

A three-pronged coordinated research effort was undertaken by cooperating utilities at three different experimental scales investigating bioaugmentation, enrichment and performance of anammox organisms in mainstream treatment. Two major technological components were applied: density-based sludge wasting by a selective cyclone to retain anammox granules and intermittent aeration to repress nitrite oxidizers. This paper evaluates process conditions and operation modes to direct more nitrogen to the resource-saving metabolic route of deammonification.

The dynamic bacterial communities of a melting High Arctic glacier snowpack

Snow environments can occupy over a third of land surface area, but little is known about the dynamics of snowpack bacteria. The effect of snow melt on bacterial community structure and diversity of surface environments of a Svalbard glacier was examined using analyses of 16S rRNA genes via T-RFLP, qPCR and 454 pyrosequencing. Distinct community structures were found in different habitat types, with changes over 1 week apparent, in particular for the dominant bacterial class present, Betaproteobacteria. The differences observed were consistent with influences from depositional mode (snowfall vs aeolian dusts), contrasting snow with dust-rich snow layers and near-surface ice. Contrary to that, slush as the decompositional product of snow harboured distinct lineages of bacteria, further implying post-depositional changes in community structure. Taxa affiliated to the betaproteobacterial genus Polaromonas were particularly dynamic, and evidence for the presence of betaproteobacterial ammonia-oxidizing bacteria was uncovered, inviting the prospect that the dynamic bacterial communities associated with snowpacks may be active in supraglacial nitrogen cycling and capable of rapid responses to changes induced by snowmelt. Furthermore the potential of supraglacial snowpack ecosystems to respond to transient yet spatially extensive melting episodes such as that observed across most of Greenland’s ice sheet in 2012 merits further investigation.

Anaerobic Fungi and Their Potential for Biogas Production

Plant biomass is the largest reservoir of environmentally friendly renewable energy on earth. However, the complex and recalcitrant structure of these lignocellulose-rich substrates is a severe limitation for biogas production. Microbial pro-ventricular anaerobic digestion of ruminants can serve as a model for improvement of converting lignocellulosic biomass into energy. Anaerobic fungi are key players in the digestive system of various animals, they produce a plethora of plant carbohydrate hydrolysing enzymes. Combined with the invasive growth of their rhizoid system their contribution to cell wall polysaccharide decomposition may greatly exceed that of bacteria. The cellulolytic arsenal of anaerobic fungi consists of both secreted enzymes, as well as extracellular multi-enzyme complexes called cellulosomes. These complexes are extremely active, can degrade both amorphous and crystalline cellulose and are probably the main reason of cellulolytic efficiency of anaerobic fungi. The synergistic use of mechanical and enzymatic degradation makes anaerobic fungi promising candidates to improve biogas production from recalcitrant biomass. This chapter presents an overview about their biology and their potential for implementation in the biogas process.

Application of denaturing gradient gel electrophoresis for analysing the gut microflora of Lumbricus rubellus Hoffmeister under different feeding conditions

The earthworm, Lumbricus rubellus, plays an essential role in soil ecosystems as it affects organic matter decomposition and nutrient cycling. By ingesting a mixture of organic and mineral material, a variety of bacteria and fungi are carried to the intestinal tract of the earthworm. To get a better understanding of the interactions between L. rubellus and the microorganisms ingested, this study tried to reveal if the diet affects the composition of the gut microflora of L. rubellus or if its intestinal tract hosts an indigenous, species-specific microbiota. A feeding experiment with L. rubellus was set up; individuals were collected in the field, transferred to a climate chamber and fed with food sources of different quality (dwarf shrub litter, grass litter or horse dung) for six weeks. DNA was extracted from the guts of the earthworms, as well as from the food sources and the surrounding soil, and further analysed by a molecular fingerprinting method, PCR-DGGE (Polymerase Chain Reaction -- Denaturing Gradient Gel Electrophoresis). We were able to demonstrate that the gut microbiota was strongly influenced by the food source ingested and was considerably different to that of the surrounding soil. Sequencing of dominant bands of the bacterial DGGE fingerprints revealed a strong occurrence of y-Proteobacteria in all gut samples, independent of the food source. A specific microflora in the intestinal tract of L. rubellus, robust against diet changes, could not be found.

Expanding DEMON Sidestream Deammonification Technology Towards Mainstream Application.

A cross-Atlantic R&D-cooperation involving three large utilities investigated the feasibility of mainstream deammonification-the application of partial nitritation/anammox for full-plant treatment of municipal wastewater at ambient temperatures. Two major process components have been implemented, 1) bioaugmentation of aerobic- and anaerobic ammonia oxidizers (AOB and AMX) from the DEMON-sidestream sludge liquor treatment to the mainstream and 2) implementation of hydrocyclones to select for anammox granules and retain them in the system. Different operation modes have been tested at laboratory- and pilot-scale in order to promote the short-cut (more direct anammox route) in nitrogen removal metabolism. At the full-scale installation at Strass WWTP, stable repression of nitrite oxidizing biomass (NOB) has been achieved for several months. Significant anammox enrichment in the mainstream has been monitored while high efficiency in the sidestream-process has been maintained (96% annual average ammonia removal).

A novel fixed fibre biofilm membrane process for on-site greywater reclamation requiring no fouling control

On‐site greywater treatment and reuse in urban areas bears the potential to reduce huge quantities of wastewater and lower freshwater shortages. Until now dissemination of small, single household applications has been rather limited as simple and high quality water producing, but also cost‐effective treatment units have not been developed so far. This paper proposes a new process, based on a concurrently working hollow‐fibre membrane as fixed biofilm support and filtration device. Bioreactor characteristics, influence of different aeration rates, membrane flux development, as well as structure and composition of biofilm were monitored to evaluate the performance of the tested pilot unit. The introduced process achieved international water reuse guidelines, worked soundly and could, compared to conventional micro MBR, significantly reduce energy demand (<1.4 kWh m−3). Fouling control by air scouring and chemical cleaning was not required once flux had stabilized. The biofilm analysis showed a porous, spongy‐like structure. Microbiological investigation revealed a community of sheathed bacteria and nematodes that could play an important role in the flux stabilisation effect. In general, the study confirmed the suitability of the presented process for greywater treatment and provides valuable design data for future optimization and systematic analysis. Biotechnol. Bioeng. 2015;112: 484–493.

Wood ash amendment to biogas reactors as an alternative to landfilling? A preliminary study on changes in process chemistry and biology

Wood ash addition to biogas plants represents an alternative to commonly used landfilling by improving the reactor performance, raising the pH and alleviating potential limits of trace elements. This study is the first on the effects of wood ash on reactor conditions and microbial communities in cattle slurry-based biogas reactors. General process parameters [temperature, pH, electrical conductivity, ammonia, volatile fatty acids, carbon/nitrogen (C/N), total solids (TS), volatile solids, and gas quantity and quality] were monitored along with molecular analyses of methanogens by polymerase chain reaction- denaturing gradient gel electrophoresis and modern microarrays (archaea and bacteria). A prompt pH rise was observed, as was an increase in C/N ratio and volatile fatty acids. Biogas production was inhibited, but recovered to even higher production rates and methane concentration after single amendment. High sulphur levels in the wood ash generated hydrogen sulphide and potentially hampered methanogenesis. Methanosarcina was the most dominant methanogen in all reactors; however, diversity was higher in ash-amended reactors. Bacterial groups like Firmicutes, Proteobacteria and Acidobacteria were favoured, which could improve the hydrolytic efficiency of the reactors. We recommend constant monitoring of the chemical composition of the used wood ash and suggest that ash amendment is adequate if added to the substrate at a rate low enough to allow adaptation of the microbiota (e.g. 0.25 g g−1 TS). It could further help to enrich digestate with important nutrients, for example phosphorus, calcium and magnesium, but further experiments are required for the evaluation of wood ash concentrations that are tolerable for anaerobic digestion.

Quantitative and qualitative effects of bioaugmentation on ammonia oxidisers at a two-step WWTP

Large waste water treatment plants (WWTP) often operate nitrification in two different process environments: the cold-dilute sewage is treated in the mainstream nitrification/denitrification system, while the high strength ammonia liquors from sludge dewatering are treated in a separate high temperature reactor (SBR). This study investigates transfer from nitrifier biomass into a two-stage WWTP, commonly referred to as bioaugmentation. Besides the quantitation of ammonia oxidising bacteria (AOB), community differences were analysed with two techniques, denaturing gradient gel electrophoresis and real-time PCR melt curve analysis. It was shown that, without bioaugmentation, two distinct AOB communities establish in the mainstream and in the SBR, respectively. A gradual shift of the two AOB communities with increasing pump rates between the systems could be demonstrated. These molecular findings support process engineering experience, that cycling of waste activated sludge improves the ability of AOB to adapt to different process environments.

The use of extracellular DNA as a proxy for specific microbial activity

The ubiquity and relevance of extracellular DNA (exDNA) are well-known and increasingly gaining importance in many fields of application such as medicine and environmental microbiology. Although sources and types of exDNA are manifold, ratios of specific DNA-molecules inside and outside of living cells can give reliable information about the activity of entire systems and of specific microbial groups or species. Here, we introduce a method to discriminate between internal (iDNA), as well as bound and free exDNA, and evaluate various DNA fractions and related ratios (ex:iDNA) regarding their applicability to be used as a fast, convenient, and reliable alternative to more tedious RNA-based activity measurements. In order to deal with microbial consortia that can be regulated regarding their activity, we tested and evaluated the proposed method in comparison to sophisticated dehydrogenase- and RNA-based activity measurements with two anaerobic microbial consortia (anaerobic fungi and syntrophic archaea and a microbial rumen consortium) and three levels of resolution (overall activity, total bacteria, methanogenic archaea). Furthermore, we introduce a 28S rRNA gene-specific primer set and qPCR protocol, targeting anaerobic fungi (Neocallimastigomycota). Our findings show that the amount of actively released free exDNA (fDNA) strongly correlates with different activity measurements and is thus suggested to serve as a proxy for microbial activity.

The effect of maize silage as co-substrate for swine manure on the bacterial community structure in biogas plants

The qualitative and quantitative changes in the bacterial community composition in two mesophilic, commercially used biogas plants were monitored by denaturing gradient gel electrophoresis (DGGE) and real-time PCR. The main objective was to evaluate the influence of the co-substrate maize silage on total bacteria and some selected bacterial groups by comparing full-scale reactors fed solely with pig manure or additionally with maize silage. DGGE fingerprints reflected shifts in the bacterial community structure associated with maize silage as co-substrate and the real-time PCR results showed clear changes in the quantitative composition of the bacterial consortia of each fermenter. A clear dominance of Clostridia in all surveyed fermenters and considerably lower abundance of Bacteroidetes in the biogas plant fed with maize silage was shown.