MANITOBA'S MESOSALINE MICROBES' METAGENOMIC MAPPING

Water has been described as the most precious resource in Manitoba, Canada, yet little is known about the microbial inhabitants of these waters. This is particularly true for the saline springs that are > 600 km from the closest marine ecosystem, where microbial communities are largely uncharacterized. No information on the composition, longevity or resilience after disruptions is available for these systems, which means that no framework exists for biodiversity conservation at the microbial level. With this work, we aim to use eDNA technologies to guide cultivation efforts to enrich for bacteria and archaea from these springs. To target anaerobic diversity, enrichment cultures were established in anoxic, moderately saline media (14.5 ppt) supplemented with several carbon sources for heterotrophs, and carbon dioxide for autotrophs, with a N₂/CO₂/H₂ (88:7.5:4.25 vol/vol) headspace. Subsurface sediment from the springs was added as 10% v/v inoculum, and enrichments were incubated at room temperature, under circumneutral pH. After 13 days of incubation, metagenomic sequencing of cultures showed selection for new Desulfobacterota, Campylobacterota, Chloroflexota and Nanobdellota. This work established the first enrichment cultures for these organisms, with some described only at the order or family level, highlighting their unique genomic diversity. Future work entails characterizing these organisms to serve as anchor points for monitoring of the microbial communities in these springs, as the high degree of novelty may be indicative of endemic species in need of conservation.

Presentation Preference: Poster

Primary Presenter: Ayush Sharma, University of Manitoba (sharm86@myumanitoba.ca)

Authors:

Ayush Sharma, University of Manitoba  (sharm86@myumanitoba.ca)

Ben Kilbrei, University of Manitoba  (kilbreib@myumanitoba.ca)

Marike Palmer, University of Manitoba  (Marike.Palmer@umanitoba.ca)