Thermospores, the dormant resting stages of thermophilic bacteria, have been shown to be frequent but enigmatic components of cold marine sediments around the world. Multiple hypotheses have been proposed to explain their distribution, emphasizing their potential as model organisms for studying microbial dispersal via ocean currents. In the Arctic Ocean, the abundance and diversity of thermospores have previously been assumed to be low. However, this assessment has been based on data mainly from the western fjords of Svalbard, thus leaving most of the Arctic unexplored. Here, we expand the knowledge about the distribution of thermospores in the Arctic Ocean by investigating the abundance and diversity of thermospores in heated shelf sediments from three sites in the outer Laptev Sea. Two of the sites are located in an area with methane-emitting cold seeps with a thermogenic source signature suggestive of an origin in a deep hydrocarbon reservoir, while the third site is a reference site not known to be impacted by seepage. We found that activity of viable thermospore populations was more prominent at one of the investigated seep sites. This finding is supported by both radiotracer growth experiments showing thermophilic, sulfate-reducing activity triggered by heating, as well as 16S gene sequence analyses showing significantly enriched ASVs affiliated to the phylum Firmicutes following high-temperature incubations. An enrichment of the sulfate-reducing, endospore-forming class Desulfotomaculia in heated samples compared to unheated samples was also observed. Furthermore, several ASVs identified at the seep site are closely related to thermospore-producing bacteria associated with the deep biosphere, including hydrocarbon and hydrothermal systems. Based on the combined information from induced activity, estimated abundance, and phylogenetic composition using 16S rRNA gene sequencing, we propose likely source environments and dispersal vectors for thermospores in the Arctic Ocean.
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