BACKGROUND: Inflammatory bowel diseases (IBDs) are multifactorial illnesses of the intestine, to which microorganisms are contributing. Among the contributing microorganisms, sulfate-reducing bacteria (SRB) are suggested to be involved in the process of bowel inflammation due to the production of hydrogen sulfide (H2S) by dissimilatory sulfate reduction. The aims of our research were to physiologically examine SRB in fecal samples of patients with IBD and a control group, their identification, the study of the process of dissimilatory sulfate reduction (sulfate consumption and H2S production) and biomass accumulation. Determination of biogenic elements of the SRB and evaluation of obtained parameters by using statistical methods were also included in the research. The material for the research consisted of 14 fecal samples, which was obtained from patients and control subjects. METHODS: Microscopic techniques, microbiological, biochemical, biophysical methods and statistical analysis were included. RESULTS: Colonies of SRB were isolated from all the fecal samples, and subsequently, 35 strains were obtained. Vibrio-shaped cells stained Gram-negative were dominant in all purified studied strains. All strains had a high percentage of similarity by the 16S rRNA gene with deposited sequences in GenBank of Desulfovibrio vulgaris. Cluster analysis of sulfate reduction parameters allowed the grouping of SRB strains. Significant (p < 0.05) differences were not observed between healthy individuals and patients with IBD with regard to sulfate reduction parameters (sulfate consumption, H2S and biomass accumulation). Moreover, we found that manganese and iron contents in the cell extracts are higher among healthy individuals in comparison to unhealthy individuals that have an intestinal bowel disease, especially ulcerative colitis. CONCLUSIONS: The observations obtained from studying SRB emphasize differences in the intestinal microbial processes of healthy and unhealthy people.

Archaea Physiology and Biotechnology Group Department of Functional and Evolutionary Ecology Universität Wien Althanstraße 14 1090 Vienna Austria

Centre of Region Hana for Biotechnological an Agricultural Research Central Laboratories and Research Support Faculty of Science Palacky University Olomouc 77900 Olomouc Czech Republic

Department of Experimental Biology Faculty of Science Masaryk University Kamenice 753 5 62500 Brno Czech Republic

Department of Medical Biophysics Danylo Halytsky Lviv National Medical University 69 Pekarska St 79010 Lviv Ukraine

Department of Plant Origin Foodstuffs Hygiene and Technology Faculty of Veterinary Hygiene and Ecology University of Veterinary and Pharmaceutical Sciences 61242 Brno Czech Republic

Faculty of Biology University of Salamanca Campus Miguel de Unamuno C Donantes de sangre s n 37007 Salamanca Spain

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Katz J.A., Itoh J., Fiocchi C. Pathogenesis of inflammatory bowel disease. Curr. Opin. Gastroenterol. 1999;15:291. doi: 10.1097/00001574-199907000-00003. PubMed DOI

Danese S., De La Motte C., Fiocchi C. Platelets in inflammatory bowel disease: Clinical, pathogenic, and therapeutic implications. Am. J. Gastroenterol. 2004;99:938–945. doi: 10.1111/j.1572-0241.2004.04129.x. PubMed DOI

Danese S., Fiocchi C. Etiopathogenesis of inflammatory bowel diseases. World J. Gastroenterol. 2006;12:4807. doi: 10.3748/wjg.v12.i30.4807. PubMed DOI PMC

Scaldaferri F., Fiocchi C. Inflammatory bowel disease: Progress and current concepts of etiopathogenesis. J. Dig. Dis. 2007;8:171–178. doi: 10.1111/j.1751-2980.2007.00310.x. PubMed DOI

Fiocchi C. Inflammatory bowel disease: Etiology and pathogenesis. Gastroenterology. 1998;115:182–205. doi: 10.1016/S0016-5085(98)70381-6. PubMed DOI

Loubinoux J., Bronowicji J.P., Pereira I.A. Sulphate-reducing bacteria in human feces and their association with inflammatory diseases. FEMS Microbiol. Ecol. 2002;40:107–112. doi: 10.1111/j.1574-6941.2002.tb00942.x. PubMed DOI

Rowan F.E., Docherty N.G., Coffey J.C., O’Connell P.R. Sulphate-reducing bacteria and hydrogen sulphide in the aetiology of ulcerative colitis. Br. J. Surg. 2009;96:151–158. doi: 10.1002/bjs.6454. PubMed DOI

Kushkevych I., Dordević D., Vítězová M. Toxicity of hydrogen sulfide toward sulfate-reducing bacteria Desulfovibrio piger Vib-7. Arch. Microbiol. 2019;201:389–397. doi: 10.1007/s00203-019-01625-z. PubMed DOI

Kushkevych I., Dordević D., Kollar P., Vítězová M., Drago L. Hydrogen Sulfide as a Toxic Product in the Small–Large Intestine Axis and its Role in IBD Development. J. Clin. Med. 2019;8:1054. doi: 10.3390/jcm8071054. PubMed DOI PMC

Kushkevych I., Kotrsová V., Dordević D., Buňková L., Vítězová M., Amedei A. Hydrogen Sulfide Effects on the Survival of Lactobacilli with Emphasis on the Development of Inflammatory Bowel Diseases. Biomolecules. 2019;9:752. doi: 10.3390/biom9120752. PubMed DOI PMC

Cummings J.H., Macfarlane G.T., Macfarlane S. Intestinal Bacteria and Ulcerative Colitis. Curr. Issues Intest. Microbiol. 2003;4:9–20. PubMed

Gibson G.R., Cummings J.H., Macfarlane G.T. Growth and activities of sulphate-reducing bacteria in gut contents of health subjects and patients with ulcerative colitis. FEMS Microbiol. Ecol. 1991;86:103–112. doi: 10.1111/j.1574-6968.1991.tb04799.x. DOI

Coutinho C.M.L.M., Coutinho-Silva R., Zinkevich V., Pearce C.B., Ojcius D.M., Beech I. Sulphate-reducing bacteria from ulcerative colitis patients induce apoptosis of gastrointestinal epithelial cells. Microb. Pathog. 2017;112:126–134. doi: 10.1016/j.micpath.2017.09.054. PubMed DOI

Kushkevych I., Vítězová M., Kos J., Kollár P., Jampilek J. Effect of selected 8-hydroxyquinoline-2-carboxanilides on viability and sulfate metabolism of Desulfovibrio piger. J. Appl. Biomed. 2018;16:241–246. doi: 10.1016/j.jab.2018.01.004. DOI

Kushkevych I., Kollar P., Suchy P., Parak T., Pauk K., Imramovsky A. Activity of selected salicylamides against intestinal sulfate-reducing bacteria. Neuro Endocrinol. Lett. 2015;36:106–113. PubMed

Kushkevych I.V. Kinetic properties of pyruvate ferredoxin oxidoreductase of intestinal sulfate-reducing bacteria Desulfovibrio piger Vib-7 and Desulfomicrobium sp. Rod-9. Pol. J. Microbiol. 2015;64:107–114. doi: 10.33073/pjm-2015-016. PubMed DOI

Barton L.L., Hamilton W.A. Sulphate-Reducing Bacteria: Environmental and Engineered Systems. Cambridge University Press; Cambridge, UK: 2010. p. 553.

Kotrsová V., Kushkevych I. Possible methods for evaluation of hydrogen sulfide toxicity against lactic acid bacteria. Biointerface Res. Appl. Chem. 2019;9:4066–4069.

Pitcher M.C., Cummings J.H. Hydrogen sulphide: A bacterial toxin in ulcerative colitis? Gut. 1996;39:1–4. doi: 10.1136/gut.39.1.1. PubMed DOI PMC

Kushkevych I., Leščanová O., Dordević D., Jančíková S., Hošek J., Vítězová M., Buňková L., Drago L. The sulfate-reducing microbial communities and meta-analysis of their occurrence during diseases of small-large intestine axis. J. Clin. Med. 2019;8:1656. doi: 10.3390/jcm8101656. PubMed DOI PMC

Kushkevych I., Fafula R., Parak T., Bartoš M. Activity of Na+/K+-activated Mg2+-dependent ATP hydrolase in the cell-free extracts of the sulfate-reducing bacteria Desulfovibrio piger Vib-7 and Desulfomicrobium sp. Rod-9. Acta Vet. Brno. 2015;84:3–12. doi: 10.2754/avb201585010003. DOI

Kushkevych I.V. Activity and kinetic properties of phosphotransacetylase from intestinal sulfate-reducing bacteria. Acta Biochem. Pol. 2015;62:1037–1108. doi: 10.18388/abp.2014_845. PubMed DOI

Kushkevych I., Dordević D., Vítězová M., Kollár P. Cross-correlation analysis of the Desulfovibrio growth parameters of intestinal species isolated from people with colitis. Biologia. 2018;73:1137–1143. doi: 10.2478/s11756-018-0118-2. DOI

Kushkevych I., Dordević D., Vítězová M. Analysis of pH dose-dependent growth of sulfate-reducing bacteria. Open Med. 2019;14:66–74. doi: 10.1515/med-2019-0010. PubMed DOI PMC

Tomasova L., Konopelski P., Ufnal M. Gut bacteria and hydrogen sulfide: The new old players in circulatory system homeostasis. Molecules. 2016;21:1558. doi: 10.3390/molecules21111558. PubMed DOI PMC

Kováč J., Vítězová M., Kushkevych I. Metabolic activity of sulfate-reducing bacteria from rodents with colitis. Open Med. 2018;13:344–349. doi: 10.1515/med-2018-0052. PubMed DOI PMC

Kushkevych I., Vítězová M., Fedrová P., Vochyanová Z., Paráková L., Hošek J. Kinetic properties of growth of intestinal sulphate-reducing bacteria isolated from healthy mice and mice with ulcerative colitis. Acta Vet. Brno. 2017;86:405–411. doi: 10.2754/avb201786040405. DOI

Gibson G.R., Macfarlane G.T., Cummings J.H. Occurrence of sulphate-reducing bacteria in human faeces and the relationship of dissimilatory sulphate reduction to methanogenesis in the large gut. J. Appl. Bacteriol. 1988;65:103–111. doi: 10.1111/j.1365-2672.1988.tb01498.x. PubMed DOI

Černý M., Vítězová M., Vítěz T., Bartoš M., Kushkevych I. Variation in the distribution of hydrogen producers from the clostridiales order in biogas reactors depending on different input substrates. Energies. 2018;11:3270. doi: 10.3390/en11123270. DOI

Kushkevych I., Vítězová M., Vítěz T., Bartoš M. Production of biogas: Relationship between methanogenic and sulfate-reducing microorganisms. Open Life Sci. 2017;12:82–91. doi: 10.1515/biol-2017-0009. DOI

Kushkevych I., Vítězová M., Vítěz T., Kováč J., Kaucká P., Jesionek W., Bartoš M., Barton L. A new combination of substrates: Biogas production and diversity of the methanogenic microorganisms. Open Life Sci. 2018;13:119–128. doi: 10.1515/biol-2018-0017. PubMed DOI PMC

Peck H.D., Legall J., Vanbeeumen J. Biochemistry of dissimilatory sulfate reduction. Philos. Trans. R. Soc. Lond. B Biol. Sci. 1982;298:443–466. PubMed

Kushkevych I., Dordević D., Kollar P. Analysis of physiological parameters of Desulfovibrio strains from individuals with colitis. Open Life Sci. 2018;13:481–488. doi: 10.1515/biol-2018-0057. PubMed DOI PMC

Loubinoux J., Valente F.M.A., Pereira I.A.C. Reclassification of the only species of the genus Desulfomonas, Desulfomonas pigra, as Desulfovibrio piger comb. nov. Int. J. Syst. Evol. Microbiol. 2002;52:1305–1308. PubMed

Postgate J.R. The Sulfate-Reducing Bacteria. 2nd ed. Cambridge University Press; Cambridge, UK: 1984. p. 208.

Abdulina D., Kováč J., Iutynska G., Kushkevych I. ATP sulfurylase activity of sulfate-reducing bacteria from various ecotopes. 3Biotech. 2020;10:55. doi: 10.1007/s13205-019-2041-9. PubMed DOI PMC

Kushkevych I., Kollar P., Ferreira A.L., Palma D., Duarte A., Lopes M.M., Bartos M., Pauk K., Imramovsky A., Jampilek J. Antimicrobial effect of salicylamide derivatives against intestinal sulfate-reducing bacteria. J. Appl. Biomed. 2016;14:125–130. doi: 10.1016/j.jab.2016.01.005. DOI

Kushkevych I., Kos J., Kollar P., Kralova K., Jampilek J. Activity of ring-substituted 8-hydroxyquinoline-2-carboxanilides against intestinal sulfate-reducing bacteria Desulfovibrio piger. Med. Chem. Res. 2018;27:278–284. doi: 10.1007/s00044-017-2067-7. DOI

Dordević D., Jančíková S., Vítězová M., Kushkevych I. Hydrogen sulfide toxicity in the gut environment: Meta-analysis of sulfate-reducing and lactic acid bacteria in inflammatory processes. J. Adv. Res. 2020;25:1–15. doi: 10.1016/j.jare.2020.03.003. PubMed DOI PMC

Kushkevych I., Dordević D., Vítězová M. Possible synergy effect of hydrogen sulfide and acetate produced by sulfate-reducing bacteria on inflammatory bowel disease development. J. Adv. Res. 2020;25:1–8. doi: 10.1016/j.jare.2020.03.007. PubMed DOI PMC

Moore W.E., Johnson J.L., Holdeman L.V. Emendation of Bacteroidaceae and Butyrivibrio and descriptions of Desulfomonas gen. nov. and ten new species of the genera Desulfomonas, Butyrivibrio, Eubacterium, Clostridium and Ruminococcus. Int. J. Syst. Bact. 1976;26:238–252. doi: 10.1099/00207713-26-2-238. DOI

Brenner D.J., Krieg N.R., Staley J.T., Garrity G.M. Bergey’s Manual of Systematic Bacteriology. 2nd ed. The United States of America Springer; Boston, MA, USA: 2005. Volume two: The proteobacteria, part C: The alpha-, beta-, delta-, and epsilonproteobacteria; p. 1388.

Kováč J., Kushkevych I. New modification of cultivation medium for isolation and growth of intestinal sulfate-reducing bacteria; Proceedings of the International PhD Students Conference Mendel Net; Brno, Czech Republic. 6–7 November 2019; pp. 702–707.

Weisburg W.G., Barns S.M., Pelletier D.A., Lane D.J. 16S ribosomal DNA amplification for phylogenetic study. J. Bacteriol. 1991;173:697–703. doi: 10.1128/JB.173.2.697-703.1991. PubMed DOI PMC

Persing D.H. Molecular Microbiology: Diagnostic Principles and Practice. 2nd ed. ASM Press; Washington, DC, USA: 2011. p. 960.

Kolmert A., Wikstrom P., Hallberg K.B. A fast and simple turbidimetric method for the determination of sulfate in sulfate-reducing bacterial cultures. J. Microbiol. Methods. 2000;41:179–184. doi: 10.1016/S0167-7012(00)00154-8. PubMed DOI

Sugiyama M. Reagent Composition for Measuring Hydrogen Sulfide and Method for Measuring Hydrogen. 6,340,596. U.S. Patent. 2002 Jan 22

Bailey N.T.J. Statistical Methods in Biology. Cambridge University Press; Cambridge, UK: 1995.

Florin T.H., Neale G., Goretski S. Sulfate in food and beverages. J. Food Compos. Anal. 1993;6:140–151. doi: 10.1006/jfca.1993.1016. DOI

Kushkevych I., Kováč J., Vítězová M., Vítěz T., Bartoš M. The diversity of sulfate-reducing bacteria in the seven bioreactors. Arch. Microbiol. 2018;200:945–950. doi: 10.1007/s00203-018-1510-6. PubMed DOI

Kushkevych I., Cejnar J., Treml J., Dordević D., Kollar P., Vítězová M. Recent Advances in Metabolic Pathways of Sulfate Reduction in Intestinal Bacteria. Cells. 2020;9:698. doi: 10.3390/cells9030698. PubMed DOI PMC

Beauchamp R.O., Bus J.S., Popp J.A., Boreiko C.J., Andjelkovich D.A., Leber P. A critical review of the literature on hydrogen sulfide toxicity. CRC Crit. Rev. Toxicol. 1984;13:25–97. doi: 10.3109/10408448409029321. PubMed DOI

Blachier F., Davila A.M., Mimoun S. Luminal sulfide and large intestine mucosa: Friend or foe? Amino Acids. 2010;39:335–347. doi: 10.1007/s00726-009-0445-2. PubMed DOI

Attene-Ramos M.S., Wagner E.D., Plewa M.J., Gaskins H.R. Evidence that hydrogen sulfide is a genotoxic agent. Mol. Cancer Res. 2006;4:9–14. doi: 10.1158/1541-7786.MCR-05-0126. PubMed DOI

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