Testate Amoebae Examined by Confocal and Two-Photon Microscopy: Implications for Taxonomy and Ecophysiology

Testate amoebae (TA) are a group of free-living protozoa, important in ecology and paleoecology. Testate amoebae taxonomy is mainly based on the morphological features of the shell, as examined by means of light microscopy or (environmental) scanning electron microscopy (SEM/ESEM). We explored the potential applications of confocal laser scanning microscopy (CLSM), two photon excitation microscopy (TPEM), phase contrast, differential interference contrast (DIC Nomarski), and polarization microscopy to visualize TA shells and inner structures of living cells, which is not possible by SEM or environmental SEM. Images captured by CLSM and TPEM were utilized to create three-dimensional (3D) visualizations and to evaluate biovolume inside the shell by stereological methods, to assess the function of TA in ecosystems. This approach broadens the understanding of TA cell and shell morphology, and inner structures including organelles and endosymbionts, with potential implications in taxonomy and ecophysiolog

Actuoecology of testate amoebae in fresh water and soil environment in enteraction with fungi and their analysis with new microscopic techniques

4 Abstract The present thesis focuses on testate amoebae (TA) and their relationship to their natural environment, as well as on relevant microscopic imaging methods. The bulk of the data has been published in original scientific papers and is compiled into three separate chapters (Pt I, Pt II and Pt III), each annotated by a brief introduction. (Pt I) The methods section is devoted to specialized microscopic techniques employed to broaden the scope of the ecological analyses. In particular, precise discrimination between live and dead individuals, biomass determination inside individual tests and a multi-modal visualization of the cytoplasm and organelles enhance the data. Laser scanning confocal microscopy and two-photon microscopy are the main imaging modalities employed to study TA morphology in detail. The data have implications for taxonomy and ecophysiology, including the use of TA as bioindicators of pollution. (Pt II) An actuoecological analysis focuses on the seasonal variability of TA species composition in a freshwater ecosystem, namely the Komo any ponds in Prague, during the course of the year. The species composition variation is correlated to simultaneously recorded limnological parameters such as temperature, pH, contamination by (heavy) metals (As, Cd, Mn, Ni, Fe, Pb), polycyclic aromatic..

Testate Amoebae Communities in the Rhizosphere of Rhododendron ponticum (Ericaceae) in an Evergreen Broadleaf Forest in Southern Spain

Testate amoebae (TA) are an important part of soil microbial communities and in certain ecosystems they may represent a substantial proportion of total microbial biomass. Their distribution and abundance is driven by various abiotic factors (e.g. pH, organic matter, soil moisture, soil/water chemistry) but comparatively less is known about the role of biotic interactions. TA often co-occur with Ericaceae, a ubiquitous plant family inhabiting acidic soils with poor nutrient status. Ericaceae can significantly change soil properties through production of recalcitrant litter and possibly also due to root exudates and activities of root-inhabiting fungi; this may result in profound modifications of microbial communities. A recent study from northwest England shows that the invasive ericaceous shrub Rhododendron ponticum may significantly modify communities of soil TA. Here, we investigate the effect of pH, organic matter, soil moisture and R. ponticum presence on TA communities within the native range of the ericaceous shrub at two sites in south Spain and compare our results with the previous study from NW England. At the Spanish sites, organic matter content, R. ponticum presence and pH affected occurrence and abundance of several TA species; R. ponticum presence and organic matter content were highly correlated and explained most of the observed variability in TA communities (= the effect of the R. ponticum rhizosphere). R. ponticum rhizosphere affected especially TA with relatively large tests, i.e. Cyclopyxis eurystoma, Phryganella acropodia and Trigonopyxis arcula. Interestingly, T. arcula was also positively associated with R. ponticum in the previously studied British sites. The rhizosphere of the ericaceous shrub appears to have a positive effect on testate amoebae taxon richness at the two studied autochthonous Spanish sites but may reduce taxon richness in the sites in Britain where R. ponticum is an introduced species. Such possible positive/negative effects of native/invasive species, as well as other plant guilds, on TA communities clearly deserve further investigation