[Identification of Clinical Isolates of the Bacillus cereus Group and Their Characterization by Mass Spectrometry and Electron Microscopy].

Bacillus cereus is a spore-forming bacterium found in the environment mainly in soil. Bacillus spores are known to be extremely resistant not only to environmental factors, but also to various sanitation regimes. This leads to spore contamination of toxin-producing strains in hospital and food equipment and, therefore, poses a great threat to human health. Two clinical isolates identified as B. cereus and B. cytotoxicus were used in the present work. It was shown that their calcium ion content was significantly lower than that of the reference strains. According to electron microscopy, one of the SRCC 19/16 isolates has an enlarged exosporium, and the SRCC 1208 isolate has large electron-dense inclusions of an unclear nature during sporulation. We can assume that these contain a biologically active component with a cytotoxic effect and possibly play a role in pathogenesis. Comparative chemical, biochemical, physiological, and ultrastructural analysis of spores of clinical isolates and reference strains of B. cereus was performed. The results we obtained deepen our understanding of the properties of spores that contribute to the increased pathogenicity of B. cereus group species.

[Hyper-Resistance of the Bacillus licheniformis 24 Strain to Oxidative Stress Is Associated with Overexpression of Enzymatic Antioxidant System Genes].

At the International Space Station (ISS), artificial living conditions are created and maintained to satisfy human needs, these conditions are also favorable for the growth of numerous microorganisms, molds and bacteria. Among the microorganisms detected on the ISS are those from the automicroflora of crew members, and a significant number of spore-forming bacteria. In most cases, this group of microorganisms gives rise to strains that are able to colonize, grow and reproduce on interior materials and equipment of stations, and may be involved in biodestructive processes. These bacteria show increased resistance to various stress factors, for example, DNA-damaging and oxidizing agents. The molecular mechanisms of this resistance to stress are poorly understood. As part of the sanitary-microbiological monitoring of the ISS habitat, the Bacillus licheniformis 24 strain was isolated. Here, we demonstrated that this strain has increased resistance to hydrogen peroxide and Paraquat when compared to the "terrestrial" B. licheniformis B-10956 strain. B. licheniformis 24 overexpressed genes encoding enzymes that neutralize reactive oxygen species, such as KatX catalase and the superoxide dismutases SodA and SodF. Apart from this, in comparison with B. licheniformis B-10956, of B. licheniformis 24 cells had lower hydrogen sulfide production that was associated with sharply reduced expression of the cysIJ operon that encodes sulfite reductase. The results indicate that enzymatic antioxidant protective systems make a more significant contribution to the hyper-resistance of Bacillus strains to oxidizing agents than components of non-enzymatic systems, such as hydrogen sulfide.

The Biological Threat: The Threat of Planetary Quarantine Failure as a Result of Outer Space Exploration by Humans.

The paper presents the results of experiments with spore-forming bacteria and microscopic fungi performed in the framework of the Russian Research Program outside the International Space Station. It has been found that microorganisms not only survive in this extreme environment, but also retain reproductive ability. Moreover, most microorganisms exhibit an increase in biochemical activity and resistance to antimicrobial agents, specifically antibiotics. These findings are of obvious interest to the developers of both planetary quarantine methods and biomedical safety systems for manned space exploration missions. In addition, they demonstrate the necessity of experiments on the exposure of bio-objects to simulated environmental factors beyond Earth's magnetosphere.

[Biotechnological Potential of the Bacillus subtilis 20 Strain].

Bacillus subtilis bacteria play an important role in veterinary medicine, medicine, and biotechnology, and the permanently growing demand for biotechnological products fuels the improvement of the properties of biotechnological strains. B. subtilis strains with improved characteristics maybe obtained by rational design and the directed evolution technologies, or be found among newly described strains. In the course of the long-term microbiome composition studies in the Russian segment of the International Space Station, the B. subtilis 20 strain was isolated, this strain shows the capacity for rapid growth and considerable biomass accumulation, as well as increased resistance to acidification of the environment in comparison to the "terrestrial" B. subtilis 168 strain. What is more, B. subtilis 20 is hyperresistant to the DNA and protein damaging factors that are linked to the overexpression of the genes controlling DNA repair, hydrogen sulfide production, and reactive oxygen species neutralization. The described properties of B. subtilis 20 are indicative of its considerable potential as a promising producer of biologically active compounds.

[Results of salad machine experiment within the MARS-500 project].

The salad machine experiment was aimed to fulfill performance testing of a prototype of space conveyor-type cylindrical greenhouse PHYTOCYCLE-SL, to study growth and development of plants, and to evaluate microbial contamination of equipment in the closed manned environment. Crops of leaf cabbage Brassica chinensis L., cultivar Vesnianka were raised in the time interval between MARS-500 days 417 and 515. The greenhouse proved it serviceability demanding 17 min/(man x d) in the normal mode. Most likely that the slow growth rate and deviations in plant morphology were caused by the presence of volatile pollutants in the greenhouse compartment Accumulation of micromycetes was observed at the sites of humid surfaces contact with ambient air; reduction of the artificial soil area contacting with air decreased population of micromycetes in 40 times. Cabbage leafs were free of pathogenic microflora. These results of the experiment helped develop recommendations on how to work out some units and systems in projectable greenhouse VITACYCLE-T

[Main results of the Biorisk experiment on the International Space Station].

To get better appreciation of the margins of phenotypic adaptation and genotypic changes in bacteria-fungi associations within the typical microbiota residing on structural materials of space-flown equipment, developed were a program and hardware for a series of experiments under the general name BIORISK. Protocol of each experimental cycle is based on the well-proven method of exposure of "passive" samples of materials (Biorisk-KM), microorganisms-materials systems inside the ISS service module (Biorisk-MSV), and microorganisms-materials systems on the outside of the ISS SM (Biorisk-MSN). Each six months the samples are returned to the laboratory in conjunction with crew rotation. Already the first in-hand data from the experiment point to the dramatic effect of space flight on growth, reproduction, and biological properties of test microbes and fungi. Thus, the activity of enzymes that characterize the pathogenic potential (RNA-ase and DNA-ase), and resistance of microorganisms to aseptic agents were found increased.

[Air purification aboard space vehicles]. / Obezzarazhivanie vozdushnoi sredy na bortu kosmicheskikh letatel'nykh apparatov.

Air purification unit POTOK 150MK has been designed to clean air of space vehicles from microbial contaminant. Comparison of the data of analysis of air samples collected in different periods of operation of the space station MIR and ISS reconfirmed efficiency of the unit.

[Comparative evaluation of microorganisms biological characteristics isolated in the orbital complex "Mir" on different phases of its operation]. / Sravnitel'naia otsenka biologicheskikh svoistv mikroorganizmov, vydelennykh v orbital'nom komplekse "Mir" v razlichnye sroki ékspluatatsii.

Colonizing and bio-destructive activities and pathogenetic potential of microorganisms isolated from the MIR environment at different phases of its operations were tested and evaluated in comparison with standard "museum" cultures of analogous species. Comparison of bacterial and fungal species revealed certain cultural, biochemical, and morphological distinctions of the "flight" strains. However, to assess the character and the risk associated with peculiar evolution of microflora in long-term space missions, more careful investigations should be followed with the use of the strain genetic labeling and other modern techniques.