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Results of an experiment named "Test" on survival and variability of microorganisms in open space near the International Space Station are presented. It was found after two-years exposure, spore-forming bacteria of the species Bacillus subtilis, fungi of the species Aureobasidium pullulans and archaea of the species Methanosarcina mazei S-6 T , deposited on cotton wool, are able to survive, and their numbers decreased equally, regardless of whether the microorganisms belong to different taxonomic groups. The main factors for the long-term survival could be the result of their dehydration and partial lyophilization in the vacuum of near-Earth space. For the first time, after being in outer space, cyst-like cells of the archaea strain M. mazei S-6 T and a 14-day delay in their growth were detected when cultured on a nutrient medium compared to the ground-based control strain. In 30% of fungi species strains A. pullulans, isolated after a two-year stay in outer space, the resistance to γ-radiation increased compared to the control strain. It was found that the reaction to the action of various concentrations of hydrogen peroxide and 1% chlorine in the surviving strains of the fungus A. pullulans on the ISS is less pronounced than in the control strain., (© 2024. The Author(s).)

Spore-forming bacteria have a unique resistance to negative environmental conditions, including aggressive space factors, and are an excellent model for studying adaptation mechanisms and survival strategies at the molecular level. The study analyzed the genome of Bacillus velezensis, which remained viable after a 2-year exposure in outer space on the outer surface of the ISS as part of the Test space experiment. A comparative analysis of the draft genomes of the exhibit strain and the ground control did not reveal significant changes; the average nucleotide identity was 99.98%, which indicates the ability of microorganisms to maintain genome stability in space conditions, due to both increased stress resistance of bacterial spores and efficient operation of the system of repair of accumulated changes. The study of a single nucleotide polymorphism in the genome of B. velezensis revealed nine point substitutions, three of which are in intergenic regions, six in protein-coding genes, three of them are missense mutations, two nucleotide deletions leading to a shift in the reading frame, and one synonymous substitution. The profiles of the housekeeping genes were determined during MLST typing and it was found that the allelic profiles obtained for B. velezensis T15.2 and 924 strains do not correspond to any of the previously described sequence types. The presented results indicate the ability of B. velezensis bacteria to maintain the viability of spores and the integrity of the genome for a long time under extreme conditions of outer space, which is important for the problem of planetary protection, as well as the potential possibility of performing biotechnological processes based on B. velezensis during space exploration.

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.

The effort was made to assess the feasibility to remove microorganisms from surface of materials with the help of high-intensity ultrasound generator <> and further air decontamination using unit <>. The test data guided construction of a system capable to eliminate completely a mature association of microorganisms from aluminum surfaces. The combined use of 2 units resulted in total decontamination of atmosphere.