Aviaja Anna Hansen

A conceptual ecosystem model of microbial communities in enhanced biological phosphorus removal plants

The microbial populations in 25 full-scale activated sludge wastewater treatment plants with enhanced biological phosphorus removal (EBPR plants) have been intensively studied over several years. Most of the important bacterial groups involved in nitrification, denitrification, biological P removal, fermentation, and hydrolysis have been identified and quantified using quantitative culture-independent molecular methods. Surprisingly, a limited number of core species was present in all plants, constituting on average approx. 80% of the entire communities in the plants, showing that the microbial populations in EBPR plants are rather similar and not very diverse, as sometimes suggested. By focusing on these organisms it is possible to make a comprehensive ecosystem model, where many important aspects in relation to microbial ecosystems and wastewater treatment can be investigated. We have reviewed the current knowledge about these microorganisms with focus on key ecophysiological factors and combined this into a conceptual ecosystem model for EBPR plants. It includes the major pathways of carbon flow with specific organic substances, the dominant populations involved in the transformations, interspecies interactions, and the key factors controlling their presence and activity. We believe that the EBPR process is a perfect model system for studies of microbial ecology in water engineering systems and that this conceptual model can be used for proposing and testing theories based on microbial ecosystem theories, for the development of new and improved quantitative ecosystem models and is beneficial for future design and management of wastewater treatment systems.

MiDAS: the field guide to the microbes of activated sludge.

The Microbial Database for Activated Sludge (MiDAS) field guide is a freely available online resource linking the identity of abundant and process critical microorganisms in activated sludge wastewater treatment systems to available data related to their functional importance. Phenotypic properties of some of these genera are described, but most are known only from sequence data. The MiDAS taxonomy is a manual curation of the SILVA taxonomy that proposes a name for all genus-level taxa observed to be abundant by large-scale 16 S rRNA gene amplicon sequencing of full-scale activated sludge communities. The taxonomy can be used to classify unknown sequences, and the online MiDAS field guide links the identity to the available information about their morphology, diversity, physiology and distribution. The use of a common taxonomy across the field will provide a solid foundation for the study of microbial ecology of the activated sludge process and related treatment processes. The online MiDAS field guide is a collaborative workspace intended to facilitate a better understanding of the ecology of activated sludge and related treatment processes—knowledge that will be an invaluable resource for the optimal design and operation of these systems. Database URL: http://www.midasfieldguide.org

Microbial communities involved in enhanced biological phosphorus removal from wastewater: A model system in environmental biotechnology

Enhanced biological phosphorus removal (EBPR) is one of the most advanced and complicated wastewater treatment processes applied today, and it is becoming increasingly popular worldwide as a sustainable way to remove and potentially reuse P. It is carried out by complex microbial communities consisting primarily of uncultured microorganisms. The EBPR process is a well-studied system with clearly defined boundaries which makes it very suitable as a model ecosystem in microbial ecology. Of particular importance are the transformations of C, N, and P, the solid-liquid separation properties and the functional and structural stability. A range of modern molecular methods has been used to study these communities in great detail including single cell microbiology, various -omics methods, flux analyses, and modeling making this one of the best studied microbial ecosystems so far. Recently, an EBPR core microbiome has been described and we present in this article some highlights and show how this complex microbial community can be used as model ecosystem in environmental biotechnology.

A Facility for Long-Term Mars Simulation Experiments: The Mars Environmental Simulation Chamber (MESCH)

We describe the design, construction, and pilot operation of a Mars simulation facility comprised of a cryogenic environmental chamber, an atmospheric gas analyzer, and a xenon/mercury discharge source for UV generation. The Mars Environmental Simulation Chamber (MESCH) consists of a double-walled cylindrical chamber. The double wall provides a cooling mantle through which liquid N(2) can be circulated. A load-lock system that consists of a small pressure-exchange chamber, which can be evacuated, allows for the exchange of samples without changing the chamber environment. Fitted within the MESCH is a carousel, which holds up to 10 steel sample tubes. Rotation of the carousel is controlled by an external motor. Each sample in the carousel can be placed at any desired position. Environmental data, such as temperature, pressure, and UV exposure time, are computer logged and used in automated feedback mechanisms, enabling a wide variety of experiments that include time series. Tests of the simulation facility have successfully demonstrated its ability to produce temperature cycles and maintain low temperature (down to -140 degrees C), low atmospheric pressure (5-10 mbar), and a gas composition like that of Mars during long-term experiments.

Activity and stability of a complex bacterial soil community under simulated Martian conditions

A simulation experiment with a complex bacterial soil community in a Mars simulation chamber was performed to determine the effect of Martian conditions on community activity, stability and survival. At three different depths in the soil core short-term effects of Martian conditions with and without ultraviolet (UV) exposure corresponding to 8 Martian Sol were compared. Community metabolic activities and functional diversity, measured as glucose respiration and versatility in substrate utilization, respectively, decreased after UV exposure, whereas they remained unaffected by Martian conditions without UV exposure. In contrast, the numbers of culturable bacteria and the genetic diversity were unaffected by the simulated Martian conditions both with and without UV exposure. The genetic diversity of the soil community and of the colonies grown on agar plates were evaluated by denaturant gradient gel electrophoresis (DGGE) on DNA extracts. Desiccation of the soil prior to experimentation affected the functional diversity by decreasing the versatility in substrate utilization. The natural dominance of endospores and Gram-positive bacteria in the investigated Mars-analogue soil may explain the limited effect of the Mars incubations on the survival and community structure. Our results suggest that UV radiation and desiccation are major selecting factors on bacterial functional diversity in terrestrial bacterial communities incubated under simulated Martian conditions. Furthermore, these results suggest that forward contamination of Mars is a matter of great concern in future space missions.

Effects of Long-Term Simulated Martian Conditions on a Freeze-Dried and Homogenized Bacterial Permafrost Community

Indigenous bacteria and biomolecules (DNA and proteins) in a freeze-dried and homogenized Arctic permafrost were exposed to simulated martian conditions that correspond to about 80 days on the surface of Mars with respect to the accumulated UV dose. The simulation conditions included UV radiation, freeze-thaw cycles, the atmospheric gas composition, and pressure. The homogenized permafrost cores were subjected to repeated cycles of UV radiation for 3 h followed by 27 h without irradiation. The effects of the simulation conditions on the concentrations of biomolecules; numbers of viable, dead, and cultured bacteria; as well as the community structure were determined. Simulated martian conditions resulted in a significant reduction of the concentrations of DNA and amino acids in the uppermost 1.5 mm of the soil core. The total number of bacterial cells was reduced in the upper 9 mm of the soil core, while the number of viable cells was reduced in the upper 15 mm. The number of cultured aerobic bacteria was reduced in the upper 6 mm of the soil core, whereas the community structure of cultured anaerobic bacteria was relatively unaffected by the exposure conditions. As explanations for the observed changes, we propose three causes that might have been working on the biological material either individually or synergistically: (i) UV radiation, (ii) UV-generated reactive oxygen species, and (iii) freeze-thaw cycles. Currently, the production and action of reactive gases is only hypothetical and will be a central subject in future investigations. Overall, we conclude that in a stable environment (no wind-/pressure-induced mixing) biological material is efficiently shielded by a 2 cm thick layer of dust, while it is relatively rapidly destroyed in the surface layer, and that biomolecules like proteins and polynucleotides are more resistant to destruction than living biota.

Dynamics of the Fouling Layer Microbial Community in a Membrane Bioreactor.

Membrane fouling presents the greatest challenge to the application of membrane bioreactor (MBR) technology. Formation of biofilms on the membrane surface is the suggested cause, yet little is known of the composition or dynamics of the microbial community responsible. To gain an insight into this important question, we applied 16S rRNA gene amplicon sequencing with a curated taxonomy and fluorescent in situ hybridization to monitor the community of a pilot-scale MBR carrying out enhanced biological nitrogen and phosphorus removal with municipal wastewater. In order to track the dynamics of the fouling process, we concurrently investigated the communities of the biofilm, MBR bulk sludge, and the conventional activated sludge system used to seed the MBR system over several weeks from start-up. As the biofilm matured the initially abundant betaproteobacterial genera Limnohabitans, Hydrogenophaga and Malikia were succeeded by filamentous Chloroflexi and Gordonia as the abundant species. This study indicates that, although putative pioneer species appear, the biofilm became increasingly similar to the bulk community with time. This suggests that the microbial population in bulk water will largely determine the community structure of the mature biofilm.

An ultraviolet simulator for the incident Martian surface radiation and its applications

Ultraviolet (UV) radiation can act on putative organic/biological matter at the Martian surface in several ways. Only absorbed, but not transmitted or reflected, radiation energy can be photo-chemically effective. The most important biological UV effects are due to photochemical reactions in nucleic acids, DNA or RNA, which constitute the genetic material of all cellular organisms and viruses. Protein or lipid effects generally play a minor role, but they are also relevant in some cases. UV radiation can induce wavelengths-specific types of DNA damage. At the same time it can also induce the photo-reversion reaction of a UV induced DNA photoproduct of nucleic acid bases, the pyrimidine dimers. Intense UVB and UVC radiation, experienced on early Earth and present-day Mars, has been revealed to be harmful to all organisms, including extremophile bacteria and spores. Moreover, the formation of oxidants, catalytically produced in the Martian environment through UV irradiation, may be responsible for the destruction of organic matter on Mars. Following this, more laboratory simulations are vital in order to investigate and understand UV effects on organic matter in the case of Mars. We have designed a radiation apparatus that simulates the anticipated Martian UV surface spectrum between 200 and 400 nm (UVC-UVA). The system comprises a UV enhanced xenon arc lamp, special filter-sets and mirrors to simulate the effects of the Martian atmospheric column and dust loading. We describe the technical setup and performance of the system and discuss its uses for different applications. The design is focused on portability, therefore, the Mars-UV simulator represents a device for several different Mars simulation facilities with specific emphasis on Mars research topics.

The use of complex microbial soil communities in Mars simulation experiments

Mars simulation studies have in the past mainly investigated the effect of the simulation conditions such as UV radiation, low pressure and temperature on pure cultures and much has been learnt about the survival potential of sporeformers such as Bacillus subtilis. However, this approach has limitations as the studies only investigate the properties of a very limited number of microorganisms. In this paper we propose that Mars simulations should be carried out with complex microbial communities of Martian analogues such as permafrost or the deep biosphere. We also propose that samples from these environments should be studied by a number of complementary methods and claim that these methods in combination can provide a comprehensive picture of how imposed Martian conditions affect the microbial community and in particular the survival of its constituents - microbes as well as biological material in general. As an interesting consequence this approach can lead to the isolation of bacteria, which are more recalcitrant to the imposed Martian conditions than the pure cultures that have previously been studied.