Laboratory of Microbial Ecology

Head

Natalia O. Kozyrovska

Ph.D. (Mol. Biol.), Senior Staff Scientist
Tel.: +380 (44) 200-03-60
Fax: +380 (44) 526-07-59
E-mail: kozyrna@ukr.net

Education and Degrees:

1971–1976 Graduate Student, Faculty of Biology, Taras Shevchenko National University of Kyiv, Kyiv, Ukraine, M.Sc. (biology, microbiology)

1984 Ph.D. (molecular biology)

Honours, Prizes, Awards:

1989 DAAD (German Academic Exchange Service) Grant

1993 Royal Society Grant

2003 WIPO (World Intellectual Property Organization) Gold medal for an invention

Research Area:

Scientific and biotechnological aspects of polymicrobial community study

Сurrent Research Activities and Recent Achievements:

Interaction of microorganisms with plants: the mechanism of plant protection against biotic and abiotic factors, involving endophytic microbial populations.

The concept of plant protection under the assistance of communities of resident endophytic bacteria is elaborated. In addition to known plant defence mechanisms (basic and induced resistance), the reserve mechanism is revealed, which involves latent endophytic microorganisms. The effectiveness of this mechanism depends on the plant genotype-associated endophytic microbiome and its ability to resuscitate from the latent state and to respond to external factors. Biocontrol agents used for plant protection may serve as activators of the latent state. The reaction of plants on pathogens after priming by biocontrol agents depends on the endophytic microbiome structure and its quality. The concept may explain, why the same microbial preparations for plant protection act differently on different plant species and even varieties, and provides a tool for selection of plant varieties with beneficial endophytic microbiome composition.

Fig. 1. a – Survival of potato plants under post vitro conditions; b – primed in vitro plants; c – increase of bacterial OTUs inside potato roots after priming with PGPB (on results of 16S rDNA PCR/TRFLP (Hha І)-analysis)
Polymicrobial communities: communal interactions with an emphasis on а biofilm formation. Design of biodegradable composites based on nanocellulose for use in medical and aerospace industries.

Kombucha polymicrobial community (a tea “fungus”) is known within two millennia, and for the first time, the full microbiome of one of the ecotype (Ukrainian) has been revealed by DNA barcoded pyrosequencing. Construction of consortia, using different kombucha community members, for a high throughput synthesis of cellulose showed that the level of kombucha cellulose film synthesis depends on partners-duals in the community. Biodegradable antimicrobial composites based on nanocellulose were designed for a wide application.

Fig. 2. Stages of new composite nanomaterials manufacture: growing biofilm (a), purification from bacterial cells (b), modification of pure cellulose (c) (joint materials of IMBG, IBOPCH NASU).
Communication effects of extracellular membrane vesicles of multimicrobial culture kombucha under normal and stressful conditions.

The influence of diet on human health is a fast progressing background of the modern research. The diverse naturally-occurring microbial communities involved in the fermentation of functional foods and provide with valuable metabolites, essential elements and the cell-free components like extracellular membrane vesicles (EMV) (Figure). EMVs are nanosized membrane-bound spheres and carry cell surface molecular patterns from parental cell membranes. EMVs as carriers of any fragile cargo (a selective set of mRNA, miRNA, DNA, proteins) potentially can play a surrogate role of the cell they derived from. EMVs are involved in communications between cells and function to the benefit of maternal population survival. At the same time, in some cases, EMVs can pose risk of delivery into the macroorganism altered molecular patterns. The purpose of this study is to determine the communication effects of EMVs shed by probiotic microbial communities within the community and in conjunction with macroorganism under normal and stressful conditions.

Fig. 3. Extracellular membrane vesicles from kombucha bacterial-yeast community (scanning electron micrograph) (A) and Komagataeibacter intermedius (transmission electron micrographs) (B,C). Deformations and aggregations of extracellular membrane vesicles after the exposure of kombucha samples in cosmic conditions outboard of the International Space Station (D) (Kharina et al., 2015; Podolich et al., 2017).

National Grants:

Projects of National Academy of Sciences of Ukraine:

  • 2012–2017 N 47 Project: “Participation in international project BIOMEX" (NASU)
  • 2015-2016 N49 Project: Hydrogel dressings based on nanocelulose bacteria for the treatment of burns and wounds
  • 2009 N 44 Project: “System biotechnology of potato protection and molecular tests of potato pathogens”

Projects of Ministry of Education and Science of Ukraine:

  • 2009–2010 M/431 2009–2010 M/431 Ukrainian-German Project: “A study of model organisms for a use on a Lunar base”

International Grants:

  • 5th Framework Programme (FP5) Project: “The use of mycorrhizal fungi in phytoremediation projects”
  • BMBF (German Federal Ministry of Education and Research) Project: “Biological and Mars Experiment (BIOMEX)”
  • COST (European Cooperation in Science and Technology) Action FA1103 Project: “Endophytes in Biotechnology and Agriculture”
  • STCU (Science and Technology Center in Ukraine) N16 Project: “Molecular interactions between bacteria under microgravity”

Collaboration:

with Ukrainian organizations:

  • Institute of Biology, Kyiv National Taras Shevchenko University
  • Institute of Physics, National Academy of Sciences of Ukraine
  • Institute of Biochemistry of the National Academy of Sciences of Ukraine
  • Institute of Bioorganic Chemistry and Petrochemistry of the National Academy of Sciences of Ukraine
  • Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine
  • Institute of Evolutionary Ecology of the National Academy of Sciences of Ukraine
  • Nanomedtech LLC

with foreign organizations:

  • ESA Research and Scientific Support Department, ESTEC/SCI-PD, Noordwijk, The Netherlands
  • Oulu University, Oulu,Finland
  • Institute of Planetary Research, Aerospace Center, Berlin, Germany
  • Vinča Institute of Nuclear Sciences, University of Belgrade, Belgrade, Serbia
  • University of Prétore, Prétore, South Africa

Selected publications:

  1. Orlovska, I. , Zubova, G. , Shatursky, O. , ... Borisova, T. , Kozyrovska, N. Extracellular membrane vesicles derived from Komagataeibacter oboediens exposed on the International Space Station fuse with artificial eukaryotic membranes in contrast to vesicles of reference bacterium. Biochimica et Biophysica Acta Biomembranes, 2024, 1866(3), 184290
  2. Cockell, C.S. , Kozyrovska, N.O. , Kulyk, I.V., ... Yatskiv, Y.S. , Zubova, G.V. Planetary research and the search for life beyond Earth in Kyiv, Ukraine Nature Astronomy, 2024, 8(2), pp. 158–159
  3. O. V. Podolich, G. V. Zubova, I. V. Orlovska, O. E. Kukharenko, T. V. Shyryna, L. G. Palchykovska, L. A. Zaika, I. E. Zaets, N. O. Kozyrovska. Komagataeibacter oboediens changes outer membrane vesicleassociated activitіes after exposure on the International Space Station. Biopolymers and Cell. -2023. № 39(2): 131-145
  4. Mickael Baqué, Theresa Backhaus, Joachim Meeßen, Natalia Kozyrovska. Biosignature stability in space enables their use for life detection on Mars. Science Advances, 2022, 8(36), eabn7412
  5. Daniel Santana de Carvalho, Ana Paula Trovatti Uetanabaro...Olga Kukharenko,Iryna Orlovska, Olga Podolich...Natalia O. Kozyrovska. The Space-Exposed Kombucha Microbial Community Member Komagataeibacter oboediens Showed Only Minor Changes in Its Genome After Reactivation on Earth. Frontiers in Microbiology, 2022, 13, 782175
  6. Lee, I., Podolich, O., Brenig, B., ... Barh, D., Kim, B.-S. Metagenome-Assembled Genomes of Komagataeibacter from Kombucha Exposed to Mars-Like Conditions Reveal the Secrets in Tolerating Extraterrestrial Stresses. Journal of Microbiology and Biotechnology, 2022, 32(8), pp. 967–975
  7. Zmejkoski, D.Z., Marković, Z.M., Mitić, D.D., Kozyrovska, N.O., ... Bugárová, N., Todorović Marković, B.M. Antibacterial composite hydrogels of graphene quantum dots and bacterial cellulose accelerate wound healing. Journal of Biomedical Materials Research - Part B Applied Biomaterials, 2022, 110(8), pp. 1796–1805
  8. Raffaella Sabatino, … Olga Podolich, Natalia Kozyrovska, Andrea Di Cesare. Metagenome Analysis Reveals a Response of the Antibiotic Resistome to Mars-like Extraterrestrial Conditions. ASTROBIOLOGY. 22 (9): 1072-1080. doi: http://doi.org/10.1089/ast.2021.0176
  9. Iryna Orlovska, Olga Podolich, Olga Kukharenko, Iryna Zaets, Oleg Reva, Ludmila Khirunenko, Danica Zmejkoski, Sergiy Rogalsky, Debmalya Barh, Sandeep Tiwari, Madangchanok Imchen, Ranjith Kumavath, Aristóteles Góes-Neto, Vasco Azevedo, Bertram Brenig, Jean-Pierre de Vera, Natalia Kozyrovska. Bacterial cellulose retains robustness but its synthesis declines after exposure to the Mars-like environment simulated outside the International Space Station//Astrobiology. – 2021. http://doi.org/10.1089/ast.2020.2332
  10. Aristóteles Góes‐Neto, Olga Kukharenko, Iryna Orlovska, Olga Podolich, Madangchanok Imchen, Ranjith Kumavath, Rodrigo Bentes Kato, Daniel Santana de Carvalho, Sandeep Tiwari, Bertram Brenig, Vasco Azevedo, Oleg Reva, Jean‐Pierre P. De Vera, Natalia Kozyrovska, Debmalya Barh. Shotgun metagenomic analysis of kombucha mutualistic community exposed to Mars‐like environment outside the International Space Station// Environ. Microbiol. -2021. https://doi.org/10.1111/1462-2920.15405
  11. D.Z. Zmejkoski, Z.M. Marković, M.D. Budimir, N.M. Zdravković, D.D. Trišić, N. Bugárová, M.Danko, N.O. Kozyrovska, Z.Špitalský, A.Kleinová, S.B. Kuzman, V.B. Pavlović, B.M. Todorović Marković. Photoactive and antioxidant nanochitosan dots/biocellulose hydrogels for wound healing treatment//Materials Science and Engineering: C. Volume 122, 2021, 111925, ISSN 0928-4931. https://doi.org/10.1016/j.msec.2021.111925
  12. D.Z. Zmejkoski, Z.M. Marković, N.M. Zdravković, D.D. Trišić, M.D. Budimir, S.B. Kuzman, N.O. Kozyrovska, I.V. Orlovska, N. Bugárová, Đ.Ž. Petrović, M. Kováčová, A.Kleinová, Z.Špitalský, V.B. Pavlović, B.M. Todorović Marković.//Bactericidal and antioxidant bacterial cellulose hydrogels doped with chitosan as potential urinary tract infection biomedical agent.//RSC Adv. -2021. -11:8559-8568. https://doi.org/10.1039/D0RA10782D
  13. Olga Podolich, Olga Kukharenko, Iryna Zaets, Iryna Orlovska, Larysa Palchykovska, Leonid Zaika, Serhii Sysoliatin, Ganna Zubova, Oleg Reva, Maxym Galkin, Tetyana Horid’ko, Halyna Kosiakova, Tatiana Borisova, Volodymyr Kravchenko, Mykola Skoryk, Maxym Kremenskoy, Preetam Ghosh, Debmalya Barh, Aristóteles Góes-Neto, Vasco Azevedo, Jean-Pierre de Vera and Natalia Kozyrovska. Fitness of outer membrane vesicles from Komagataeibacter intermedius is altered under the impact of simulated Mars-like stressors outside the International Space Station.//Frontiers in Microbiology. -2020. 11, 1-14. doi.org/10.3389/fmicb.2020.01268
  14. Kukharenko O.Ye., Terzova V.O., Zubova G.V. Protein aggregates carry non-genetic memory in bacteria after stresses. //Biopolym. Cell.– 2020. -36(6):409-422. http://dx.doi.org/10.7124/bc.000A3F
  15. Kozyrovska N. O.,Vidmachenko A. P., Steklov A. F. Determination of biosafety of nano-vesicles and viruses in astromaterials. International scientific conference Astronomical School of Young Scientists. December 11–12, Kyiv, Ukraine.- 2020. p. 65-68
  16. Imchen, M., Kumavath, R., Vaz, A.B.M., (...), Podolich, O., Azevedo, V. 16S rRNA Gene Amplicon Based Metagenomic Signatures of Rhizobiome Community in Rice Field During Various Growth Stages. Frontiers in Microbiology 10,2103, 2019
  17. Podolich, O., Kukharenko, O., Haidak, A., (...), Kozyrovska, N., De Vera, J.-P.P. Multimicrobial Kombucha Culture Tolerates Mars-Like Conditions Simulated on Low-Earth Orbit. Astrobiology 19(2), pp. 183-196, 2019
  18. De Vera, J.-P., Alawi, M., Backhaus, T., (...), Wolter, D., Zucconi, L. Limits of Life and the Habitability of Mars: The ESA Space Experiment BIOMEX on the ISS. Astrobiology 19(2), pp. 145-157, 2019
  19. Orlovska I.V., Yakovenko I.O., Haidak A.H., Zmejkoski D.Z., Kozyrovska N.O. Regenerative medicine advanced therapy for the non-healing cutaneous wound. Biopolymers and Cell. – 2018. – 34 ( 3): P 171–195 doi:http://dx.doi.org/10.7124/bc.000979
  20. Zmejkoski D., Spasojević D., Orlovska I., Kozyrovska N., Soković M., Glamočlija J., Dmitrović S., Matović B., Tasić N., Maksimović V., Sosnin M., Radotić K. Bacterial cellulose-lignin composite hydrogel as a promising agent in chronic wound healing. Int J Biol Macromol. – 2018. – 18(Pt A):494-503. doi:10.1016/j.ijbiomac.2018.06.067
  21. S.Rogalsky, Jean-François Bardeaub, Stanislav Makhno, Natalia Babkina, Oksana Tarasyuk, Tetiana Cherniavska, Iryna Orlovska, Natalia Kozyrovska, Olexander Brovko. New proton conducting membrane based on bacterial cellulose/polyaniline nanocomposite film impregnated with guanidinium-based ionic liquid. Polymer, 2018,142:183-195.
  22. Podolich O, Zaets I, Kukharenko O, Orlovska I, Reva O, Khirunenko L, Sosnin M, Haidak A, Shpylova S, Rohutskyy I, Kharina A, Skoryk М, Kremenskoy M, Klymchuk D, Demets R, de Vera JP, Kozyrovska N. The First Space-Related Study of a Kombucha Multimicrobial Cellulose-Forming Community: Preparatory Laboratory Experiments. Orig Life Evol Biosph, 2017, 47(2):169-185.
  23. Podolich O, Zaets I, Kukharenko O, Orlovska I, Reva O, Khirunenko L, Sosnin M, Haidak A, Shpylova S, Rabbow E, Skoryk M, Kremenskoy M, Demets R, Kozyrovska N, de Vera JP. Kombucha Multimicrobial Community under Simulated Spaceflight and Martian Conditions. Astrobiology, 2017, 17(5):459-469.
  24. O. Podolich, I. Zaets, O. Kukharenko, I. Orlovska, O. Reva, J.-P. deVera, N. Kozyrovska et al. The First Space-Related Study of a KombuchaMultimicrobial Cellulose-Forming Community: Preparatory Laboratory Experiments. Origins of Life and Evolution of Biospheres. March 2016. DOI: 10.1007/s11084-016-9483-4.
  25. I. Ye. Zaets, O. V. Podolich, O. N. Reva, N. O. Kozyrovska. DNA metabarcoding of microbial communities for healthcare. Biopolymers and Cell. 2016, 32(1): 3–8.
  26. Janne Koskimäki, Marena Kajula, Juho Hokkanen, Emmi-LeenaIhantola, Jong HKim, Heidi Hautajärvi, Elina Hankala, Marko Suokas, Johanna Pohjanen, Olga Podolich, Natalia Kozyrovska, et al. Methyl-esterified 3-hydroxybutyrateoligomersprotectbacteria from hydroxyl radicals. Nat Chem Biol. 2016 May;12(5):332-338.
  27. Podolich O, Ardanov P, Zaets I, Pirttilä AM, Kozyrovska N. Reviving of the endophytic bacterial community as a putative mechanism of plant resistance. Plant and Soil. 2015, 388(1-2):367-377.
  28. Reva ON, Zaets IЕ, Ovcharenko LP, Kukharenko OE, Shpylova SP, Podolich OV, de Vera J-P, Kozyrovska NO. Metabarcoding of the kombucha microbial community grown in different microenvironments. AMB Express. 2015, 5(1):35. doi: 10.1186/s13568-015-0124-5
  29. Kharina A, Podolich O, Faidiuk Iu, Zaika S, Haidak A, Kukharenko O, Zaets I, Tovkach F, Reva O, Kremenskoy M, Kozyrovska N. Temperate bacteriophages collected by outer membrane vesicles in Komagataeibacter intermediu. J Basic Microbiol. 2015, 55(4):509-513.
  30. Ardanov P, Lyastchenko S, Karppinen K, Häggman H, Kozyrovska N, Pirttilä AM. Effects of Methylobacterium sp. on emergence, yield, and disease prevalence in three cultivars of potato (Solanum tuberosum L.) were associated with the shift in endophytic microbial community. Plant and Soil. 2015, 5:1-12.
  31. Kukharenko O, Jean-François Bardeau, Zaets I, et al. Promising low cost antimicrobial composite material based on bacterial cellulose and polyhexamethylene guanidine hydrochloride. European Polymer Journal. 2014;60:247-254.
  32. Zaets I, Kukharenko O, Podolich O, et al. Bacterial cellulose may provide the microbial-life biosignature in the rock records. Adv Space Res. 2014;53(5):828–835.
  33. Podolich O, Ardanov P, Zaets I, et al. Reviving of the endophytic bacterial community as a putative mechanism of plant resistance. Plant Soil. 2014; DOI 10.1007/s11104-014-2235-1
  34. Kozyrovska NO. Crosstalk between endophytes and the plant host within informationprocessing networks. Biopolym. Cell. 2013; 29(3):234–43. doi: 10.7124/bc.00081D
  35. Ardanov P, Sessitsch A, Huggman H, Kozyrovska N, Pirttila AM. Methylobacteriuminduced endophyte community changes correspond with protection of plants against pathogen attack. PLoS ONE. 2012; 7:e46802. doi:10.1371/journal.pone.0046802
  36. Kozyrovska N, Reva O, Goginyan V, de Vera JP. Kombucha microbiome as a probiotic: a view from the perspective of postgenomics and synthetic ecology. Biopolym. Cell. 2012; 28(2):103–10. doi:10.7124/bc.000034
  37. Zaets I, Kozyrovska N, Heavy metal resistance in Plants: a putative role of endophytic Bacteria In: A. Zaidi et al. (eds.), Toxicity of heavy metals to legumes and bioremediation. SpringerVerlag, Wien 2012. doi: 10.1007/978-3-7091-0730-0_12
  38. Zaetz I, Burlak O, Rogutskyy I. et al. Bioaugmentation strategy of growing plants for lunar bases. Adv. Space Res. 2011; 47: 1071–8. doi: 10.1016/j.asr.2010.11.014
  39. Ardanov P, Ovcharenko L, Zaets I, Kozyrovska N, Pirttila AM Endophytic bacteria enhancing growth and disease resistance of potato (Solanum tuberosum L.). Biocontrol. 2011; 56 (1):43–9.
  40. Burlak O, Lar O, Rogutskyy IS, et al. A bacterial consortium alleviating a lowdose gamma irradiation effect in kalanchoe plantlets. Space Sci. Technol. 2010; 1:32–40.
  41. Podolich O, Laschevskyy V, Ovcharenko L, Kozyrovska N, Pirttila AM. Methylobacterium sp. resides in unculturable state in potato tissues in vitro and becomes culturable after induction by Pseudomonas fluorescens IMGB163. J. Appl. Microbiol. 2009; 116: 728–32. doi:10.1111/j.1365-2672.2008.03951.x