Temperate bacteriophages collected by outer membrane vesicles in Komagataeibacter intermedius

The acetic acid bacteria have mainly relevance for bacterial cellulose production and fermented bio-products manufacture. The purpose of this study was to identify temperate bacteriophages in a cellulose-producing bacterial strain Komagataeibacter intermedius IMBG180. Prophages from K. intermedius IMBG180 were induced with mitomycin C and nalidixic acid. Transmission electron microscopy analysis exhibited tailed bacteriophages belonging to Myoviridae. A PCR assay targeting the capsid gene of the myoviruses proved phylogenetic position of induced phages. Nalidixic acid was poor inducer of prophages, however, it induced the OMV-like particles release. Size of OMVs depended on an antibiotic applied for phage induction and varied in the range of 30–80 and 120–200 nm. Inside some of them, tails of phages have been visible. Under conditions, inducing prophages, OMVs acted as the collectors of formed phage particles, using outer membrane receptors for phage detection (in this case, outer membrane siderophore receptor), and fulfilled therefore ―a cleaning,‖ as well as defensive functions, preventing bacteriophage spread outside population. This is the first description of myoviruses affiliated to K. intermedius, as well as outer membrane vesicles interaction with phages within this host.This research was supported by grant 47/2014 from National Academy of Sciences, Ukraine.http://onlinelibrary.wiley.com/journal/10.1002/(ISSN)1521-4028hb2016Biochemistr

To other planets with upgraded millennial kombucha in rhythms of sustainability and health support

Humankind has entered a new era of space exploration: settlements on other planetary bodies are foreseen in the near future. Advanced technologies are being developed to support the adaptation to extraterrestrial environments and, with a view on the longer term, to support the viability of an independent economy. Biological processes will likely play a key role and lead to the production of life-support consumables, and other commodities, in a way that is cheaper and more sustainable than exclusively abiotic processes. Microbial communities could be used to sustain the crews’ health as well as for the production of consumables, for waste recycling, and for biomining. They can self-renew with little resources from Earth, be highly productive on a per-volume basis, and be highly versatile—all of which will be critical in planetary outposts. Well-de!ned, semi-open, and stress-resistant microecosystems are particularly promising. An instance of it is kombucha, known worldwide as a microbial association that produces an eponymous, widespread soft drink that could be valuable for sustaining crews’ health or as a synbiotic (i.e., probiotic and prebiotic) after a rational assemblage of de!ned probiotic bacteria and yeasts with endemic or engineered cellulose producers. Bacterial cellulose products offer a wide spectrum of possible functions, from leather-like to innovative smart materials during long-term missions and future activities in extraterrestrial settlements. Cellulose production by kombucha is zero-waste and could be linked to bioregenerative life support system (BLSS) loops. Another advantage of kombucha lies in its ability to mobilize inorganic ions from rocks, which may help feed BLSS from local resources. Besides outlining those applications and others, we discuss needs for knowledge and other obstacles, among which is the biosafety of microbial producers

The conceptual approach to the use of postbiotics based on bacterial membrane nanovesicles for prophylaxis of astronauts' health disorders

The functional fermented foods containing live microorganisms and their components are necessary for the normal functioning of the human body as normal gut microbiota needs fuel from external microbial organisms and their nanostructures — membrane vesicles (MVs), excreting outside. The сoncept that MVs may contribute to astronauts’ health probably to the same extent as their parental microbial cells do and be a temporary substitute for living microbial cells until we know more about the behavior of microbes in the space environment. The advantage of MVs is that they are not alive and cannot be changed under unfavorable conditions as microbial organisms may be. As the model, we selected MVs of a robust to environmental factors kombucha multimicrobial culture (KMC), known for its health-promoting characteristics for humans. We exposed KMC on the International Space Station in a hybrid space/Mars-like environment for an initial proof-of-concept stage. In the exposure study, KMC has survived a long-term period in harsh conditions, and the MVs generated by post-flight kombucha community members did not acquire toxicity, despite the changed membrane composition in the environment imitated conditions on the Mars surface. This observation, together with our KMC metagenomic and comparative genomic analyses of the dominant KMC bacterium Komagataeibacter oboediens, showed that the ground reference sample and spaceexposed ones were similar in topology and maintained their stability. In the next stage, we assessed the fitness, safety, and biodistribution of MVs of post-flight K. oboediens and showed that they were altered, but the modifications in membrane structure did not result in toxicity acquisition. Our proof-of-concept strategy is discussed in this review in line with the literature.The National Academy of Sciences of Ukraine Space Research Programme National Academy of Sciences of Ukraine Space Research Program.https://spj.science.org/journal/spaceam2023BiochemistryGeneticsMicrobiology and Plant Patholog

Kombucha multimicrobial community under simulated spaceflight and martian conditions

Kombucha microbial community (KMC) produces a cellulose-based biopolymer of industrial importance and a probiotic beverage. KMC-derived cellulose-based pellicle film is known as a highly adaptive microbial macrocolony - a stratified community of prokaryotes and eukaryotes. In the framework of the multipurpose international astrobiological project "BIOlogy and Mars Experiment (BIOMEX)," which aims to study the vitality of prokaryotic and eukaryotic organisms and the stability of selected biomarkers in low Earth orbit and in a Mars-like environment, a cellulose polymer structural integrity will be assessed as a biomarker and biotechnological nanomaterial. In a preflight assessment program for BIOMEX, the mineralized bacterial cellulose did not exhibit significant changes in the structure under all types of tests. KMC members that inhabit the cellulose-based pellicle exhibited a high survival rate; however, the survival capacity depended on a variety of stressors such as the vacuum of space, a Mars-like atmosphere, UVC radiation, and temperature fluctuations. The critical limiting factor for microbial survival was high-dose UV irradiation. In the tests that simulated a 1-year mission of exposure outside the International Space Station, the core populations of bacteria and yeasts survived and provided protection against UV; however, the microbial density of the populations overall was reduced, which was revealed by implementation of culture-dependent and culture-independent methods. Reduction of microbial richness was also associated with a lower accumulation of chemical elements in the cellulose-based pellicle film, produced by microbiota that survived in the post-test experiments, as compared to untreated cultures that populated the film.This study was supported by National Academy of Sciences of Ukraine (grant 47/2012-15). The pre-flight programs EVTs and SVTs for the EXPOSE-R2 mission were supported by the European Space Agency.http://www.liebertpub.com/overview/astrobiology/992018-05-30hj2017Biochemistr

The first space-related study of a kombucha multimicrobial cellulose-forming community : preparatory laboratory experiments

Biofilm-forming microbial communities are known as the most robust assemblages that can survive in harsh environments. Biofilm-associated microorganisms display greatly increased resistance to physical and chemical adverse conditions, and they are expected to be the first form of life on Earth or anywhere else. Biological molecules synthesized by biofilm -protected microbiomes may serve as markers of the nucleoprotein life. We offer a new experimental model, a kombucha multimicrobial culture (KMC), to assess a structural integrity of a widespread microbial polymer - cellulose - as a biosignature of bacteria-producers for the multipurpose international project "BIOlogical and Mars Experiment (BIOMEX)", which aims to study the vitality of pro- and eukaryotic organisms and the stability of organic biomolecules in contact with minerals to analyze the detectability of life markers in the context of a planetary background. In this study, we aimed to substantiate the detectability of mineralized cellulose with spectroscopy and to study the KMC macrocolony phenotype stability under adverse conditions (UV, excess of inorganics etc.). Cellulose matrix of the KMC macrocolony has been mineralized in the mineral-water interface under assistance of KMC-members. Effect of bioleached ions on the cellulose matrix has been visible, and the FT-IR spectrum proved changes in cellulose structure. However, the specific cellulose band vibration, confirming the presence of beta(1,4)-linkages between monomers, has not been quenched by secondary minerals formed on the surface of pellicle. The cellulose-based KMC macrocolony phenotype was in a dependence on extracellular matrix components (ionome, viriome, extracellular membrane vesicles), which provided its integrity and rigidness in a certain extent under impact of stressful factors.https://link.springer.com/journal/110842018-06-30hj2017Business Managemen

Limits of Life and the Habitability of Mars: The ESA Space Experiment BIOMEX on the ISS

BIOMEX (BIOlogy and Mars EXperiment) is an ESA/Roscosmos space exposure experiment housed within the exposure facility EXPOSE-R2 outside the Zvezda module on the International Space Station (ISS). The design of the multiuser facility supports—among others—the BIOMEX investigations into the stability and level of degradation of space-exposed biosignatures such as pigments, secondary metabolites, and cell surfaces in contact with a terrestrial and Mars analog mineral environment. In parallel, analysis on the viability of the investigated organisms has provided relevant data for evaluation of the habitability of Mars, for the limits of life, and for the likelihood of an interplanetary transfer of life (theory of lithopanspermia). In this project, lichens, archaea, bacteria, cyanobacteria, snow/permafrost algae, meristematic black fungi, and bryophytes from alpine and polar habitats were embedded, grown, and cultured on a mixture of martian and lunar regolith analogs or other terrestrial minerals. The organisms and regolith analogs and terrestrial mineral mixtures were then exposed to space and to simulated Mars-like conditions by way of the EXPOSE-R2 facility. In this special issue, we present the first set of data obtained in reference to our investigation into the habitability of Mars and limits of life. This project was initiated and implemented by the BIOMEX group, an international and interdisciplinary consortium of 30 institutes in 12 countries on 3 continents. Preflight tests for sample selection, results from ground-based simulation experiments, and the space experiments themselves are presented and include a complete overview of the scientific processes required for this space experiment and postflight analysis. The presented BIOMEX concept could be scaled up to future exposure experiments on the Moon and will serve as a pretest in low Earth orbit

Metabarcoding of the kombucha microbial community grown in different microenvironments

Introducing of the DNA metabarcoding analysis of probiotic microbial communities allowed getting insight into their functioning and establishing a better control on safety and efficacy of the probiotic communities. In this work the kombucha poly-microbial probiotic community was analysed to study its flexibility under different growth conditions. Environmental DNA sequencing revealed a complex and flexible composition of the kombucha microbial culture (KMC) constituting more bacterial and fungal organisms in addition to those found by cultural method. The community comprised bacterial and yeast components including cultured and uncultivable microorganisms. Culturing the KMC under different conditions revealed the core part of the community which included acetobacteria of two genera Komagataeibacter (former Gluconacetobacter) and Gluconobacter, and representatives of several yeast genera among which Brettanomyces/Dekkera and Pichia (including former Issatchenkia) were dominant. Herbaspirillum spp. and Halomonas spp., which previously had not been described in KMC, were found to be minor but permanent members of the community. The community composition was dependent on the growth conditions. The bacterial component of KMC was relatively stable, but may include additional member—lactobacilli. The yeast species composition was significantly variable. High-throughput sequencing showed complexity and variability of KMC that may affect the quality of the probiotic drink. It was hypothesized that the kombucha core community might recruit some environmental bacteria, particularly lactobacilli, which potentially may contribute to the fermentative capacity of the probiotic drink. As many KMC-associated microorganisms cannot be cultured out of the community, a robust control for community composition should be provided by using DNA metabarcoding

Bacterial cellulose may provide the microbial-life biosignature in the rock records

Bacterial cellulose (BC) is a matrix for a biofilm formation, which is critical for survival and persistence of microbes in harsh environments. BC could play a significant role in the formation of microbial mats in pristine ecosystems on Earth. The prime objective of this study was to measure to what extent spectral and other characteristics of BC were changed under the performance of BC interaction with the earthly rock – anorthosite – via microorganisms. The spectral analyses (Fourier Transform Infrared FT-IR, spectroscopy, and atomic absorption spectroscopy) showed unprecedented accumulation of chemical elements in the BC-based biofilm. The absorption capacity of IR by BC was shielded a little by mineral crust formed by microorganisms on the BC based biofilm surface, especially clearly seen in the range of 1200-900 cm−1 in FT-IR spectra. Confocal scanning laser microscopy analysis revealed that elements bioleached from anorthosite created surface coats on the BC nanofibril web. At the same time, the vibrational spectra bands showed the presence of the characteristic region of anomeric carbons (960 – 730 cm-1), wherein a band at 897 cm-1 confirmed the presence of β-1,4- linkages, which may serve as the cellulose fingerprint region. Results show that BC may be a biosignature for search signs of living organisms in rock records

Robust symbiotic microbial communities in space research

Naturally occurring symbiotic microbial communities (SMK) are the most robust assemblages for a multipurpose use in keeping humans healthy and soil fertile. Especially, safe and reliable SMK are needed for producing probiotics and ferments valuable for health problems prophylaxis. This is true for long-term expeditions, outposts, extraterrestrial permanently-manned bases where humans are exposed to adverse environmental factors, weakening the immune system. The kombucha beverage has been used in human society within millennia as a probiotic drink which is produced by naturally occurring mixed populations of living microorganisms. Here, we discuss the potential of the kombucha culture for outposts in far future missions