Komplikevych S. Relationships between pro-oxidant and antioxidant indicators of metal-resistant bacteria isolated from different biotopes.

Українська версія

Thesis for the degree of Doctor of Philosophy (PhD)

State registration number

0824U003330

Thesis Registration Form

0824U003330.pdf

Applicant for

Solomiia Y. Komplikevych

Specialization

091 - Біологія

Date of defense

27-11-2024

Specialized Academic Board

PhD 6994

Ivan Franko National University of Lviv

Essay

In the dissertation, the relationships between the content of products of the prooxidants action (lipid hydroperoxides, diene conjugates, thiobarbituric acid reactive substances, carbonyl groups in proteins) and antioxidant indicators (activity of antioxidant defense system enzymes, thiols and extracellular polymers content) was established for metal-resistant microorganisms belonging to distinct genera, that were isolated from different biotopes with extreme conditions of existence, under conditions of increasing damaging effect on bacterial cells caused by exposure to heavy metal compounds. The study of changes occurring in the bacteria Rhodopseudomonas yavorovii IMV B-7620, isolated from the Yavorivske Lake (Lviv region, Ukraine), under the influence of ferric(III) citrate, cobalt(II) chloride, copper(II) chloride, and potassium dichromate; Ochrobactrum rhizosphaerae IMV B-7956, isolated from filtrates of Lviv solid waste landfill, and Paenibacillus tundrae IMV B-7915, isolated from a sample containing soil, moss, Deschampsia antarctica (Berthelot Island, Maritime Antarctica), under the influence of copper(II) chloride and potassium dichromate were conducted. It was found that the biomass accumulation of R. yavorovii IMV B-7620 was most inhibited by CoCl2 and CuCl2. P. tundrae IMV B-7915 and O. rhizosphaerae IMV B-7956 are more sensitive to the effects of K2Cr2O7 than CuCl2. Metal salts affect the specific growth rate in the exponential phase and the duration of the lag phase of R. yavorovii IMV B-7620, P. tundrae IMV B-7915 and O. rhizosphaerae IMV B-7956, and patterns of these changes were strain-specific. FeC6H5O7, CoCl2, CuCl2, and K2Cr2O7 cause free radical damage in R. yavorovii IMV B-7620 cells, CuCl2 and K2Cr2O7 – in P. tundrae IMV B-7915 and O. rhizosphaerae IMV B-7956 cells, which was detected by the increase in the content of lipid peroxidation products and carbonyl groups in proteins. In the protection of bacteria R. yavorovii IMV B-7620 under the influence of FeC6H5O7, CoCl2, CuCl2 and K2Cr2O7, P. tundrae IMV B-7915 and O. rhizosphaerae IMV B-7956 under the influence of CuCl2 and K2Cr2O7, the enzymes of the antioxidant defense system are involved (catalase, superoxide dismutase, glutathione peroxidase, glutathione S-transferase and glutathione reductase activities were changed). For the first time, it was found that heavy metals stimulate the formation of extracellular polymers in the bacteria R. yavorovii IMV B-7620, P. tundrae IMV B-7915, and O. rhizosphaerae IMV B-7956, and also affect the qualitative and quantitative composition of pigments in R. yavorovii IMV B-7620, which is important for the survival of these bacteria. Using factor analysis and considering the deviations from the control of the studied parameters, the relationships between the content of products of prooxidants action and antioxidant indicators of bacteria under the influence of various metal salts were revealed. It was found that under the influence of ferric(III) citrate, cobalt(II) chloride, and copper(II) chloride in R. yavorovii IMV B-7620 cells, lipids were damaged. Catalase, superoxide dismutase, glutathione peroxidase, glutathione S-transferase, glutathione reductase activities, as well as lycopene, anhydrorhodovibrin, and bacteriochlorophyll a were involved in the defense of R. yavorovii IMV B-7620 cells. For the survival of P. tundrae IMV B-7915 bacteria under the influence of potassium dichromate, exopolysaccharides and the activity of the glutathione system enzymes are important. Enzymes with peroxidase activity are involved in the protection of O. rhizosphaerae IMV B-7956 cells from direct or indirect effects of lipid peroxidation products formed as a result of the action of copper(II) chloride or potassium dichromate. The positive effect of the studied bacteria in the laboratory conditions on wheat growth was demonstrated: increased seed germination, the length of roots and shoots, and the chlorophyll content in the leaves of seedlings.

Thesis supervisor

Svitlana O. Hnatush

Official opponents

Oksana B. Stoliar
Maryna V. Kryvtsova

Reviewers

Nataliia O. Sybirna
Halyna V. Yavorska

Research papers

1. Komplikevych, S., Maslovska, O., Moravska, T., Yarmoliuk, I., Kulishko, N., Biront, N., Zaritska, Y., & Hnatush, S. (2024). Adaptation of Ochrobactrum rhizosphaerae IMV B-7956 bacteria to the effect of copper (II) chloride. Mikrobiolohichnyi Zhurnal, 86(3), 58–69.

2. Komplikevych, S., Maslovska, O., Halushka, A., & Hnatush, S. (2024). Changes in the pigment composition of Rhodopseudomonas yavorovii IMV B-7620 under the influence of heavy metal salts. Мікробіологія і біотехнологія, 1, 6–21.

3. Komplikevych, S. Ya., Maslovska, O. D., Moravska, T. I., Yarmoliuk, I. V., Biront, N. V., Zaritska, Y. H., & Hnatush, S. O. (2023). Adaptations of the antarctic bacterium Paenibacillus tundrae IMV B-7915 to copper (II) chloride exposure. Ukrainian Antarctic Journal, 21(1), 66–78.

4. Maslovska, O. D., Komplikevych, S. Ya., & Hnatush S. O. (2023). Oxidative stress and protection against it in bacteria. Studia Biologica, 17(2), 153–172.

5. Hnatush, S. O., Maslovska, O. D., Komplikevych, S. Y., & Kovbasa, I. V. (2022). Influence of cobalt chloride and ferric citrate on purple non-sulfur bacteria Rhodopseudomonas yavorovii. Biosyst. Divers., 30(1), 38–45.

6. Hnatush, S. O., Maslovska, O. D., Komplikevych, S. Y., Segin, T. B., & Diakiv, S. V. (2022). Relationships between prooxidant and antioxidant parameters of bacteria of sulfur cycle under the influence of heavy metal compounds. Publishing House “Baltija Publishing”. 100–125.

7. Гнатуш, С. О., Масловська, О. Д., Мороз, О. М., & Комплікевич, С. Я. Свідоцтво про депонування штаму бактерій Ochrobactrum rhizosphaerae K 3 у Депозитарії Інституту мікробіології і вірусології ім. Д. К. Заболотного НАН України з наданням реєстраційного номеру Ochrobactrum rhizosphaerae ІМВ В-7956 від 23 липня 2021 року.

8. Гнатуш, С. О., Перетятко, Т. Б., Мороз, О. М., Масловська, О. Д., & Комплікевич, С. Я. Свідоцтво про депонування штаму бактерій Paenibacillus tundrae 5A–101 у Депозитарії Інституту мікробіології і вірусології ім. Д. К. Заболотного НАН України з наданням реєстраційного номеру Paenibacillus tundrae ІМВ В-7915 від 02 грудня 2020 року.

9. Yarmoliuk, I., Komplikevych, S., Maslovska, O. D., & Hnatush, S. O. (2023). Nonenzymatic adaptations of Ochrobactrum rhizosphaerae IMV B-7956 bacteria under the influence of potassium bichromate. Book of abstracts of the International Scientific and Practical Conference “Modern aspects of microbiology, virology and biotechnology in war and post-war period”, Kyiv, Ukraine, November 15–16, 2023. (P. 277–278). Kyiv: D. K. Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine.

10. Melnyk V., Komplikevych, S., Maslovska, O. D., & Hnatush, S. O. (2023). The effect of potassium bichromate on superoxide dismutase and catalase activities of Paenibacillus tundrae IMB B-7915. Book of abstracts of the International Scientific and Practical Conference “Modern aspects of microbiology, virology and biotechnology in war and post-war period”, Kyiv, Ukraine, November 15–16, 2023. (P. 297–299). Kyiv: D. K. Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine.

11. Moravska, T., Mashchak, S., Komplikevych, S., Maslovska, O. D., & Hnatush, S. O. (2023). Activity of enzymes of the glutathione system of Paenibacillus tundrae IMB B- 7915 under the influence of potassium bichromate. Book of abstracts of the International Scientific and Practical Conference “Modern aspects of microbiology, virology and biotechnology in war and post-war period”, Kyiv, Ukraine, November 15–16, 2023. (P. 153–155). Kyiv: D. K. Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine.

12. Mashchak, S., Moravska, T., Komplikevych, S., Maslovska, O. D., & Hnatush, S. O. (2023). Activity of enzymes of the glutathione system of Ochrobactrum rhizosphaerae IMV B-7956 under the influence of copper (II) chloride. Book of abstracts of the International Scientific and Practical Conference “Modern aspects of microbiology, virology and biotechnology in war and post-war period”, Kyiv, Ukraine, November 15– 16, 2023. (P. 142–143). Kyiv: D. K. Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine.

13. Komplikevych, S. Y., Maslovska, O. D., & Hnatush, S. O. (2023). Adaptations of Antarctic bacteria Paenibacillus tundrae IMB B-7915 to the influence of copper (II) chloride. Book of abstracts of XI International Antarctic Conference dedicated to the 160th anniversary of the birth of Volodymyr Vernadsky, Kyiv, Ukraine, May 10‒12, 2023. (P. 44–45). Kyiv: State Institution National Antarctic Scientific Center.

14. Kanetska, A., Komplikevych, S., Maslovska, O., & Hnatush, S. (2023). Influence of Cu(II) and Cr(VI) ions on the activity of enzymes of glutathione antioxidant system of Rhodopseudomonas yavorovii bacteria. Abstract book of XIX International Scientific Conference for Students and PhD Students “Youth and Progress of Biology”, Lviv, April 26–28, 2023. (P. 144–145). Lviv: Ivan Franko National University of Lviv.

15. Mashchak, S., Koval, I., Komplikevych, S., Maslovska, O., & Hnatush, S. (2023). Lipid peroxidation of Ochrobactrum rhizosphaerae IMB B-7956 under potassium bichromate exposure. Abstract book of XIX International Scientific Conference for Students and PhD Students “Youth and Progress of Biology”, Lviv, April 26–28, 2023. (P. 147). Lviv: Ivan Franko National University of Lviv.

16. Moravska, T., Melnyk, V., Komplikevych, S., Maslovska, O., & Hnatush, S. (2023). Lipid peroxidation of Paenibacillus tundrae IMB B-7915 under the influence of potassium bichromate. Abstract book of XIX International Scientific Conference for Students and PhD Students “Youth and Progress of Biology”, Lviv, April 26–28, 2023. (P. 148). Lviv: Ivan Franko National University of Lviv.

17. Yarmoliuk, I., Komplikevych, S., Maslovska, O., & Hnatush, S. (2023). Non-enzymatic adaptations of Paenibacillus tundrae bacteria under the influence of copper (II) chloride. Abstract book of XIX International Scientific Conference for Students and PhD Students “Youth and Progress of Biology”, Lviv, April 26–28, 2023. (P. 159–160). Lviv: Ivan Franko National University of Lviv.

18. Maslovska, O., Hnatush, S., Komplikevych, S., & Kovbasa, I. (2022). Indicators of free radical damage and activity of enzymes of antioxidant system of bacteria Rhodopseudomonas yavorovii IMV В-7620 under the influence of ferric citrate and cobalt (II) chloride. Abstract book of IV International Scientific Conference Microbiology and Immunology – the development outlook in the 21st century, Kyiv, September 22–23, 2022. (P. 72). Kyiv: Taras Shevchenko National University of Kyiv.

19. Komplikevych, S., Kanetska, A., Melnyk, V., Maslovska, O., & Hnatush, S. (2022). Cu2+ and Cr (VI) induced free radical damages to cells of Rhodopseudomonas yavorovii IMV B-7620. Abstract book of ХVIІI International Scientific Conference for Students and PhD Students “Youth and Progress of Biology”, Lviv, October 6–7, 2022. (P. 114– 115). Lviv: Ivan Franko National University of Lviv.

20. Kulishko, N., Komplikevych, S., Maslovska, O. & Hnatush, S. (2022). Free radical processes in Ochrobactrum rhizosphaerae IMV B-7956 cells under the influence of copper (II) chloride. Abstract book of ХVIІI International Scientific Conference for Students and PhD Students “Youth and Progress of Biology”, Lviv, October 6–7, 2022. (P. 115–116). Lviv: Ivan Franko National University of Lviv.

21. Komplikevych, S., Maslovska, O., & Hnatush, S. (2021). Metal-resistant bacteria Ochrobactrum rhizosphaerae K 3-1, isolated from the lake of infiltrates of Lviv solid waste landfill. Materials of The 1st Ukrainian-Polish Scientific forum AGROBIOPERSPECTIVES, September 29–30, 2021. (P. 56). Lviv: The Animal Biology, vol. 23, no. 3.

22. Komplikevych, S. Y., Maslovska, O. D., Novik, H., Hnatush, S. O., & Fahmi, A. (2021). Properties of biotechnologically promising strain Rhodopseudomonas yavorovii ІМV В-7620. Book of abstracts of 8-th International Conference “Human – Nutrition – Environment”, Rzeszow, Poland, October 13–14, 2021. (P. 67). Rzeszow: University of Rzeszow.

23. Komplikevych, S., Hruna, I., & Maslovska, O. (2020). Physiological and biochemical characteristics of metal-resistant strains of microorganisms from Antarctic biotopes. Book of abstracts of Young Scientists Conference “Youth and modern problems of microbiology and virology”, Kyiv, Ukraine, November 23−26, 2020 (P. 21). Kyiv: Danylo Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine.

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