In vitro Biofilm Formation by Bioluminescent Bacteria Isolated from the Marine Fish Gut
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ENVIRONMENTAL MICROBIOLOGY
In vitro Biofilm Formation by Bioluminescent Bacteria Isolated from the Marine Fish Gut Olga Burtseva 1 & Olga Baulina 1 & Anna Zaytseva 1 & Tatiana Fedorenko 1 & Konstantin Chekanov 1,2
&
Elena Lobakova 1
Received: 11 September 2020 / Accepted: 24 November 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract The internal surface of the animal gastrointestinal tract is covered by microbial biofilms. They play an important role in the development and functioning of the host organism and protect it against pathogens. Microbial communities of gastrointestinal biofilms are less elucidated than luminal microbiota. Therefore, the studies of biofilm formation by gastrointestinal microorganisms are a topical issue. For the first time, we report the formation of a biofilm in vitro by the strains of bioluminescent bacteria isolated from the intestines of marine fish. These bacteria exhibit co-aggregation and tend to attach to solid surfaces. The attachment of cells is accompanied by appearance of the pili. Then, we observed the formation of microcolonies and the production of extracellular polymer substances (EPSs) connecting bacterial cells into an integrated system. The presence of acidic polysaccharides is shown in the EPS when using the ruthenium red staining. Acidic polysaccharides in this matrix is a biochemical evidence of microbial biofilms. On the fibers of the polymer matrix, these bacteria form the “mushroom body”-type structures. Matured biofilms exhibit a specific three-dimensional architecture with pores and channels formed by cells and EPS. We also demonstrated the formation of a biofilm by binary culture of the luminous enterobacterium Kosakonia cowanii and a Gram-positive Macrococcus sp. The data obtained help to understand the role of these bacteria in the intestines of fish. They lead to a new study in the field of investigation of the intestinal microbiome of fish. Keywords Biofilm . Fish gut microbiota . Bioluminescent bacteria . Photobacterium . Aliivibrio . Schewanella
Abbreviations EPS Extracellular polymeric substances TEM Transmission electron microscopy SEM Scanning electron microscopy VPS Vibrio exopolysaccharide
Introduction Gut microbiota plays a key role in the development and functioning of a host macroorganism and protects it against pathogens [1, 2]. Microorganisms produce vitamins and essential
* Konstantin Chekanov [email protected] 1
Department of Bioengineering, Faculty of Biology, Lomonosov Moscow State University, Moscow, Russia
2
National Research Nuclear University MEPhi, Centre for Humanities Research and Technology, Moscow, Russia
fatty acids and promote the hydrolysis of complex organic compounds [1–3]. Luminal microbial communities are most studied in the gut [1, 2]. At the same time, a significant part of gut microbes is organized into the biofilms, multicellular supraorganismal structures formed by microorganisms immersed into polymer matrix (glycocalyx) co-aggregated between each other and/or adhered t
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