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Eubacterium rangiferina, a novel usnic acid-resistant bacterium from the reindeer rumen

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Abstract

Reindeer are able to eat and utilize lichens as an important source of energy and nutrients. In the current study, the activities of antibiotic secondary metabolites including usnic, antranoric, fumarprotocetraric, and lobaric acid commonly found in lichens were tested against a collection of 26 anaerobic rumen bacterial isolates from reindeer (Rangifer tarandus tarandus) using the agar diffusion method. The isolates were identified based on their 16S ribosomal ribonucleic acid (rRNA) gene sequences. Usnic acid had a potent antimicrobial effect against 25 of the isolates, belonging to Clostridiales, Enterococci, and Streptococci. Isolates of Clostridia and Streptococci were also susceptible to atranoric and lobaric acid. However, one isolate (R3_91_1) was found to be resistant to usnic, antranoric, fumarprotocetraric, and lobaric acid. R3_91_1 was also seen invading and adhering to lichen particles when grown in a liquid anaerobic culture as demonstrated by transmission electron microscopy. This was a Gram-negative, nonmotile rod (0.2–0.7 × 2.0–3.5 μm) with a deoxyribonucleic acid G + C content of 47.0 mol% and main cellular fatty acids including 15:0 anteiso-dimethyl acetal (DMA), 16:0 iso-fatty acid methyl ester (FAME), 13:0 iso-3OH FAME, and 17:0 anteiso-FAME, not matching any of the presently known profiles in the MIDI database. Combined, the phenotypic and genotypic traits including the 16S rRNA gene sequence show that R3_91_1 is a novel species inside the order Clostridiales within the family Lachnospiraceae, for which we propose the name Eubacterium rangiferina. This is the first record of a rumen bacterium able to tolerate and grow in the presence of usnic acid, indicating that the rumen microorganisms in these animals have adapted mechanisms to deal with lichen secondary metabolites, well known for their antimicrobial and toxic effects.

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Acknowledgments

This project was funded by The Reindeer Husbandry Research Fund (Project A5299), The University of Tromsø and the Norwegian Research Fund (Project S5258), and the Roald Amundsen Centre for Arctic Research (University of Tromsø). It is linked to the framework of the International Polar Year as part of the IPY consortium IPY no. 399 EALAT: Climate change and reindeer husbandry. Dr. Tove H. Aagnes Utsi is acknowledged for allowing access to her rumen bacterial isolates and Randi Olsen at the Electron Microscopy Laboratory at the Medical Faculty, University of Tromsø, for her help with the TEM. We thank Dr. André-Denis G. Wright for his invaluable help with the maximum-likelihood tree, and we are grateful to Dr. Lee R. Krumholz and Dr. Alexandra H. Smith for their input on E. oxidoreducens. The 16S rRNA gene sequences of our rumen bacterial isolates, the whole-cell fatty acid profile, and the G + C content of strain R3_91_1 were determined at BCCM™/LMG (Belgium).

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Sundset, M.A., Kohn, A., Mathiesen, S.D. et al. Eubacterium rangiferina, a novel usnic acid-resistant bacterium from the reindeer rumen. Naturwissenschaften 95, 741–749 (2008). https://doi.org/10.1007/s00114-008-0381-0

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