A composite of bacterial cellulose and lignin for a healing of incurable wounds

Biofilm-forming multi-drug resistant pathogens may account for 10% of all hospital acquired infections and can occur in burns, post-surgical wounds, pressure injuries and diabetic foot wounds. Hydrogels – natural polymers or composite materials – represent both a bandage and a matrix for therapeutics in a wound healing. Novel composite based on bacterial cellulose and synthetic analog of lignin - DHP (dehydrogenate polymer) has been designed with the aim to suppress pathogenic agents in incurable wounds. The FT-IR spectra of the BC/DHP membrane established an appearance of characteristic for DHP absorption bands in the composite in the 1600–1510 cm−1 spectral range. A clear position of these bands in BC/DHP showed that DHP had been incorporated into the BC polymeric matrix. In addition, disappearance of the bands at the 3630–3650 cm−1 spectral range assigned to the cellulose free OH vibration, strengthening the bands assigned to intermolecular interactions and an appearance of a new band at 3293 cm−1 could serve as a proof of the interaction of BC molecules and DHP. Micrographs obtained from a scanning electron microscopy proved the dose-dependent interaction of DHP with BC, which resulted in a decrease of the pore number in the cellulose membrane. The antimicrobial activity of composites BC/DHP were tested the in vitro experiments, using Pseudomonas aeruginosa (УМВ-100) (MTT-test) and showed that BC/DHP inhibited P. aeruginosa. BC/DHP composite will be promising hydrogel for a healing of infected incurable wounds.Integrative Biology and Medicine : October 2-7, 2017, Kyiv, Ukraine

Kombucha microbiome as a probiotic: a view from the perspective of post-genomics and synthetic ecology

Probiotics are essential for establishing and maintaining optimal immune health. The probiotic therapy is known from alternative medicine for ages; however, the recent demonstration of the normal microflora to induce innate immunity has introduced the science-based concept of therapeutic application of potentially beneficial probiotic microorganisms for a treatment of functional disorders. Traditionally, probiotics are associated with dairy products, however, novel formulations are needed, first of all, originated from naturally occurring symbiotic microbial communities as the most robust assemblages. Especially, safe and robust probiotics are needed for long-term expeditions, outposts, extraterrestrial permanently-manned bases, where humans are exposed to adverse environmental factors. Kombucha beverage is Symbiotic Culture of Bacteria and Yeast (SCOBY) and associated with health-promoting effects. Kombucha tea/mat is being in use in human livings within millennia as a probiotic drink for healing and health prophylaxis effects, however, new research opportunities promise its «renaissance», going to be used pharmacologically

Enhancing the internal plant colonization rate with endophytic nitrogen-fixing bacteria

Several diazotrophic strains of Klebsiella oxytoca and K. terrigena that colonize the plant-host interior were able to produce the plant cell wall depolymerising enzyme pectate lyase (Pel). The activity of the K. oxytoca enzyme was weaker than that of phytopathogenic bacteria, and it was located mainly inside the cells. A small fraction of the cells (10⁻⁶ to 10⁻⁵) in populations grown in nonselective media was able to grow in a selective medium with polygalactorunate (PC) as sole carbon source. After passage through selective medium cells were converted to the Pet+ -phenotype, and total Pet-activity in population of K. oxytoca increased. The increased level of Pelactivity of K. oxytoca and K. terrigena correlated with a 10-fold higher rate of internal colonization of wheat roots. Cultures of K. oxytoca VN13 grown in selective medium with PG also showed increased stimulation of wheat growth. Seedlings inoculated with such cultures exhibited better development resulting in higher biomass.Декілька штамів Klebsiella oxytoca та K. terrigena, здатних колонізувати рослини зсередини, виділяли пектат ліазу (ПЛ) – фермент, який деполімеризус клітинну стінку. Ак­ тивність ПЛ була нижчою, ніж у фітопатогенних бактерій, і зосереджувалась всередині клітин. Невелика кількість клітин популяції (10⁻⁶ –10⁻⁵ ) спроможна рости на селективному се­редовищі з полігалактуронатом натрію (ПГ), який викори­стовували як джерело вуглецю. Після пасажу через селективне середовище всі клітини набували Реl+ -фенотипу, і загальна ПЛ-активність К. oxytoca зростала. Підвищена ПЛ-активність бактерій K. oxytoca та K. terrigena корелювала з посилен­ням у 10 разів внутрішньої колонізації коренів пшениці. Куль­тура бактерій, яка виростала в селективному середовищі з ПГ, краще стимулювала розвиток пшениці, що проявлялося у збільшенні її біомаси, ніж культура, зрощена без селекціїУ нескольких изученных штаммов Klebsiella oxytoca и K. terrigena, способных колонизировать растения изнутри, обна­ружена активность пектат лиазы (ПЛ) –фермента, кото­рый деполимеризует клеточную стенку. Активность ПЛ была ниже, чем у фитопатогенных бактерий, и сосредоточена внутри клетки. Только небольшая часть популяции клеток (10⁻⁶ –10⁻⁵ ) способна расти на селективной среде с полигалактуронатом, который использовали в качестве источника уг­лерода. После пассажа через селективную среду все клетки приобретают Pel+ фенотип, и при этом общая ПЛ-активность K. oxytoca и K. terrigena возрастает Повышенная ПЛ-активность бактерий коррелировала с уусилением в 10 раз внутренней колонизации корней пшеницы. Культура бактерий K. oxytoca VN13, выросшая в селективных условиях, лучше стимулировала развитие пшеницы, что проявлялось в увеличении ее биомассы

Kombucha microbiome as a probiotic : a view from the perspective of post-genomics and synthetic ecology

Probiotics are essential for establishing and maintaining optimal immune health. The probiotic therapy is known from alternative medicine for ages; however, the recent demonstration of the normal microflora to induce innate immunity has introduced the science-based concept of therapeutic application of potentially beneficial probiotic microorganisms for a treatment of functional disorders. Traditionally, probiotics are associated with dairy products, however, novel formulations are needed, first of all, originated from naturally occurring symbiotic microbial communities as the most robust assemblages. Especially, safe and robust probiotics are needed for longterm expeditions, outposts, extraterrestrial permanently-manned bases, where humans are exposed to adverse environmental factors. Kombucha beverage is Symbiotic Culture of Bacteria and Yeast (SCOBY) and associated with health-promoting effects. Kombucha tea/mat is being in use in human livings within millennia as a probiotic drink for healing and health prophylaxis effects, however, new research opportunities promise its «renaissance », going to be used pharmacologically.http://www.biopolymers.org.uanf201

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

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.National Academy of Sciences of Ukraine (N47/2013)http://www.amb-express.comhb201

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

Metagenome-assembled genome sequences obtained from a reactivated kombucha microbial community exposed to a Mars-like environment outside the International Space Station

Kombucha is a traditional tea fermented by symbiotic microbiota, and it has been known as a functional fermented product. Here, we report four microbial metagenome-assembled genome sequences (MAGs) reconstructed from the microbiomes in kombucha exposed to a Mars-like environment outside the International Space Station.The Bio & Medical Technology Development Program of the National Research Foundation of Korea (NRF), funded by the South Korean government (MSIT), and the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare.https://mra.asm.orgam2022BiochemistryGeneticsMicrobiology 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