Research on halophilic microorganisms is important due to their relation to fundamental questions of survival of living organisms in a hostile environment. Here we introduce a novel method to stain halophiles with MitoTracker fluorescent dyes in their growth medium. The method is based on membrane-potential sensitive dyes, which were originally used to label mitochondria in eukaryotic cells. We demonstrate that these fluorescent dyes provide high staining efficiency and are beneficial for multi-staining purposes due to the spectral range covered (from orange to deep red). In contrast with other fluorescent dyes used so far, MitoTracker does not affect growth rate, and remains in cells after several washing steps and several generations in cell culture. The suggested dyes were tested on three archaeal (Hbt. salinarum, Haloferax sp., Halorubrum sp.) and two bacterial (Salicola sp., Halomonas sp.) strains of halophilic microorganisms. The new staining approach provides new insights into biology of Hbt. salinarum. We demonstrated the interconversion of rod-shaped cells of Hbt. salinarium to spheroplasts and submicron-sized spheres, as well as the cytoplasmic integrity of giant rod Hbt. salinarum species. By expanding the variety of tools available for halophile detection, MitoTracker dyes overcome long-standing limitations in fluorescence microscopy studies of halophiles.
Maintenance of the wastewater treatment plants and increasing the efficiency of existing aerobic biological reactors depend on the stability of activated sludge characteristics under varying wastewater parameters within significant limits and/or influence of some environmental factors. The steady microbial communities observed in biofilms and anaerobic granules of activated sludge can serve as successful samples of formation of the similar aerobic systems. The granular aerobic sludge obtained in the course of our researches is an ideal "plant" on treatment of biogenic pollution at both low and high concentrations. It demonstrates high ability for treatment and stability to adverse factors. To improve aerobic wastewater treatment characteristics, a possibility of using impact of stress conditions upon activated sludge has been studied. Under conditions of fractional hydrogen peroxide addition at diffused lighting, the granular aerobic activated sludge adapted to hydrogen peroxide has been obtained. This sludge has got good sedimentary properties and it differs from the control sample in the species diversity, improved treatment characteristics and also resistance to the stressor. It also endures an impact of one-time hydrogen peroxide addition up to 1.2-1.5 g H2O2/l. The conditions under which the steady aerobic granules of the diameter from 2 to 5 mm were formed with high treatment ability have been chosen. The granules were being stabilized at passages with hydrogen peroxide treatment and they endured up to 2.4-3.0 g/l of one-time H2O2 addition.
Microbial fuel cell (MFC) is a new technology that uses microorganisms to extract energy from complex organic mixtures. On the basis of aerobic granular sludge we have selected a nitrogen-fixing community of microorganisms that was immobilized on a carbon material (graphite foam, carbon nanotubes). The MFC anode has been developed on the basis of selected biological material. A membraneless glucose / oxygen MFC with bioanode and cathode based on non-platinum group metals or laccase enzyme has been developed. A mathematical model describing the processes in the MFC has been developed, on its base the calculations have been carried out.
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