Ute Risse-Buhl

Protists with different feeding modes change biofilm morphology.

The effect of Dexiostoma (filter feeder), Vannella, Chilodonella (raptorial feeders), Spumella, and Neobodo (direct interception feeders) on the morphology of multispecies bacterial biofilms was investigated in small flow cells. The filter feeder Dexiostoma campylum did not alter biofilm volume and porosity but stimulated the formation of larger microcolonies compared with ungrazed biofilms. In contrast, the raptorial feeder Vannella sp. efficiently grazed bacteria from the biofilm surface, leading to smaller microcolonies and lower maximal and basal layer thickness compared with ungrazed biofilms. Microcolony formation was not stimulated in the presence of the sessile Spumella sp. Chilodonella uncinata rasped bacteria from the outer surface leading to mushroom-shaped microcolonies. In the presence of C. uncinata and Spumella sp., the biofilm volume was 2.5-6.3 times lower compared with ungrazed biofilms. However, the biofilm porosity and the ratio of biofilm surface area to biofilm volume were 1.5-3.7 and 1.2-1.8 times higher, respectively. Thus, exchange of nutrients and gases between the biofilm and its surrounding fluid should also be improved in deeper biofilm layers, hence accelerating microbial growth.

Colonization dynamics of biofilm-associated ciliate morphotypes at different flow velocities.

The impact of flow velocity on initial ciliate colonization dynamics on surfaces were studied in the third order Ilm stream (Thuringia, Germany) at a slow flowing site (0.09ms(-1)) and two faster flowing sites (0.31ms(-1)) and in flow channels at 0.05, 0.4, and 0.8ms(-1). At the slow flowing stream site, surfaces were rapidly colonized by ciliates with up to 60 cells cm(-2) after 24h. In flow channels, the majority of suspended ciliates and inorganic matter accumulated at the surface within 4.5h at 0.05ms(-1). At 0.4ms(-1) the increase in ciliate abundance in the biofilm was highest between 72 and 168h at about 3 cells cm(-2)h(-1). Faster flow velocities were tolerated by vagile flattened ciliates that live in close contact to the surface. Vagile flattened and round filter feeders preferred biofilms at slow flow velocities. Addition of inorganic particles (0, 0.6, and 7.3mgcm(-2)) did not affect ciliate abundance in flow channel biofilms, but small ciliate species dominated and number of species was lowest (16 species cm(-2)) in biofilms at high sediment content. Although different morphotypes dominated the communities at contrasting flow velocities, all functional groups contributed to initial biofilm communities implementing all trophic links within the microbial loop.

Tracking the autochthonous carbon transfer in stream biofilm food webs

Food webs in the rhithral zone rely mainly on allochthonous carbon from the riparian vegetation. However, autochthonous carbon might be more important in open canopy streams. In streams, most of the microbial activity occurs in biofilms, associated with the streambed. We followed the autochthonous carbon transfer toward bacteria and grazing protozoa within a stream biofilm food web. Biofilms that developed in a second-order stream (Thuringia, Germany) were incubated in flow channels under climate-controlled conditions. Six-week-old biofilms received either ¹³C- or ¹²C-labeled CO₂, and uptake into phospholipid fatty acids was followed. The dissolved inorganic carbon of the flow channel water became immediately labeled. In biofilms grown under 8-h light/16-h dark conditions, more than 50% of the labeled carbon was incorporated in biofilm algae, mainly filamentous cyanobacteria, pennate diatoms, and nonfilamentous green algae. A mean of 29% of the labeled carbon reached protozoan grazer. The testate amoeba Pseudodifflugia horrida was highly abundant in biofilms and seemed to be the most important grazer on biofilm bacteria and algae. Hence, stream biofilms dominated by cyanobacteria and algae seem to play an important role in the uptake of CO₂ and transfer of autochthonous carbon through the microbial food web.

Phagotrophic Protist Diversity in the Groundwater of a Karstified Aquifer – Morphological and Molecular Analysis

To clarify the structure of microbial food webs in groundwater, knowledge about the protist diversity and feeding strategies is essential. We applied cultivation‐dependent approaches and molecular methods for further understanding of protist diversity in groundwater. Groundwater was sampled from a karstified aquifer located in the Thuringian Basin (Thuringia, Germany). Cultivable protist abundance estimated up to 8,000 cells/L. Eleven flagellates, 10 naked amoebae, and one ciliate morpho‐species were detected in groundwater enrichment cultures. Most of the flagellates morpho‐species, typically < 10 μm, were sessile or free swimming suspension feeders, e.g., Spumella spp., Monosiga spp., and mobile, surface‐associated forms that grasp biofilms, e.g., Bodo spp. Naked amoebae, typically < 35 μm, that grasp biofilms were represented by, e.g., Vahlkampfia spp., Vannella spp., and Hartmanella spp. The largest fraction of the 18S rRNA gene sequences was affiliated with Spumella‐like Stramenopiles. Besides, also sequences affiliated with fungi and metazoan grazers were detected in clone libraries of the groundwater. We hypothesize that small sized protist species take refuge in the structured surface of the fractures and fissures of the karstified aquifer and mainly feed on biofilm‐associated or suspended bacteria.

Colonization dynamics of ciliate morphotypes modified by shifting sandy sediments.

Sandy stream-bed sediments colonized by a diverse ciliate community are subject to various disturbance regimes. In microcosms, we investigated the effect of sediment shifting on the colonization dynamics of 3 ciliate morphotypes differing in morphology, behavior and feeding strategy. The dynamics of the ciliate morphotypes inhabiting sediment pore water and overlying water were observed at 3 sediment shifting frequencies: (1) stable sediments, (2) periodically shifting sediments such as migrating ripples, and (3) continuously shifting sediments as occurring during scour events of the uppermost sediment. Sediment shifting significantly affected the abundance and growth rate of the ciliate morphotypes. The free-swimming filter feeder Dexiostoma campylum was vulnerable to washout by sediment shifting since significantly higher numbers occurred in the overlying water than in pore water. Abundance of D. campylum only increased in pore water of stable sediments. On the contrary, the vagile grasper feeder Chilodonella uncinata and the sessile filter feeder Vorticella convallaria had positive growth rates and successfully colonized sediments that shifted periodically and continuously. Thus, the spatio-temporal pattern of sediment dynamics acts as an essential factor of impact on the structure, distribution and function of ciliate communities in sand-bed streams.

The role of hydrodynamics in shaping the composition and architecture of epilithic biofilms in fluvial ecosystems

Previous laboratory and on-site experiments have highlighted the importance of hydrodynamics in shaping biofilm composition and architecture. In how far responses to hydrodynamics can be found in natural flows under the complex interplay of environmental factors is still unknown. In this study we investigated the effect of near streambed turbulence in terms of turbulent kinetic energy (TKE) on the composition and architecture of biofilms matured in two mountainous streams differing in dissolved nutrient concentrations. Over both streams, TKE significantly explained 7% and 8% of the variability in biofilm composition and architecture, respectively. However, effects were more pronounced in the nutrient richer stream, where TKE significantly explained 12% and 3% of the variability in biofilm composition and architecture, respectively. While at lower nutrient concentrations seasonally varying factors such as stoichiometry of dissolved nutrients (N/P ratio) and light were more important and explained 41% and 6% of the variability in biofilm composition and architecture, respectively. Specific biofilm features such as elongated ripples and streamers, which were observed in response to the uniform and unidirectional flow in experimental settings, were not observed. Microbial biovolume and surface area covered by the biofilm canopy increased with TKE, while biofilm thickness and porosity where not affected or decreased. These findings indicate that under natural flows where near bed flow velocities and turbulence intensities fluctuate with time and space, biofilms became more compact. They spread uniformly on the mineral surface as a film of densely packed coccoid cells appearing like cobblestone pavement. The compact growth of biofilms seemed to be advantageous for resisting hydrodynamic shear forces in order to avoid displacement. Thus, near streambed turbulence can be considered as important factor shaping the composition and architecture of biofilms grown under natural flows.

Contrasting habitats but comparable microbial decomposition in the benthic and hyporheic zone

Input of allochthonous leaf litter is the main carbon source for heterotrophic metabolism in low-order forested streams. A major part of this leaf litter is accumulated at benthic retention structures or buried in the hyporheic zone. As a result of hyporheic sediment characteristics, hyporheic physicochemistry differs from that of the benthic zone selecting the microbial community. The present study aimed at understanding the influence of the hydrological and physiochemical differences between the benthic and hyporheic zone on microbial leaf litter decomposition and on the structure and function of the associated microbial community. Leached leaves of Alnus glutinosa were exposed for 62days in 250-μm mesh bags in the benthic zone and buried in the hyporheic zone at a depth of 2-3cm. Decomposition rates were comparable for both zones. In contrast, respiration, bacterial abundance, ergosterol content, fungal spore production and richness of fungal morphotypes were lower associated with hyporheic than with benthic leaves. Microbial community structure displayed zone-dependent temporal dynamics. Thus, the microbial community carried out leaf litter decomposition independently of its structure. These results suggest that carbon processing is not necessarily impaired by environmental constraints because the community structure may compensate those constraints (i.e. functional redundancy).

The Biota of Intermittent Rivers and Ephemeral Streams: Prokaryotes, Fungi, and Protozoans

Microbial diversity and function in intermittent rivers and ephemeral streams (IRES) are tightly linked to specific habitat availability and hydrological phases. The intensity and frequency of the different phases (especially drying and rewetting) affect community composition and key functions, mainly linked to biogeochemical processes. Resistance and resilience strategies are distinct among microorganism groups—bacteria, archaea, fungi, protozoans—and strongly depend on different types of microhabitat or refuge available. The biodiversity of prokaryotes in IRES is strongly affected by hydrology but microhabitat conditions and type of benthic substrate significantly affect their community composition. Fungi are very sensitive to drying but use several refuges, including the terrestrial habitat, and resistance strategies. Protozoans show a wide range of survival strategies and several species can resist harsh conditions such as anoxia in drying pools. Thus, they become especially relevant for ecosystem functions when other organisms are inhibited. This sensitivity causes “waves” of microbial functions and biodiversity to covary with hydrological phases, potentially affecting ecosystem functioning and higher trophic levels. Microbially mediated functions in IRES are perhaps the most critical to freshwater ecosystem services such as nitrogen and carbon cycling. Therefore, efforts to manage and restore IRES will depend on improved understanding of hydrological controls on microbial communities and functions across space and time.

Molecular and Morphological Snapshot Characterisation of the Protist Communities in Contrasting Alpine Glacier Forefields

Phagotrophic protist diversity in oligotrophic soils such as alpine glacier forefields is still poorly studied. Combining morphologic observations with molecular-based analyses, we assessed the diversity of major phagotrophic protist groups in two contrasting glacier fore- fields in the Swiss Alps (Tiefen glacier forefield, siliceous bedrock, and Wildstrubel glacier forefield, calcareous bedrock), at sites differing in soil development. Ciliates and heterotrophic flagellates could be detected with both approaches, while amoebae could be observed only microscopically. Soils from Tiefen and Wildstrubel glacier forefields harboured distinctly different ciliate, flagellate and amoebae commu- nities. The ciliate clone libraries from the Tiefen glacier forefield were dominated by Oligohymenophorea-related sequences while those from the Wildstrubel glacier forefield were dominated by Spirotrichea-related sequences. Testate amoebae morphospecies of the genera Corythion, Cryptodifflugia, Euglypha and Tracheleuglypha were restricted to the Tiefen glacier forefield, while Centropyxis and Trinema to the Wildstrubel one. No ciliate sequences and only a few ciliate and testate amoebae morphospecies could be retrieved from unvegetated soils of both glacier forefields. The ciliate and testate amoebae community detected at unvegetated sites were a subset of the community developed at vegetated sites. Overall, our results suggest that alpine glacier forefields are colonised by a diverse community of phagotrophic protists which seems to be shaped by bedrock geology and vegetation cover.