A hotspot of diversity: novel Shewanella species isolated from Baltic Sea sediments delineate a sympatric species complex

Two bacterial strains, SP1S1- 4T T and SP2S1- 2T, T , were isolated from sediment samples collected in the Stockholm archipelago in November 2021. Following whole- genome sequencing, these strains were identified as tentatively belonging to two novel Shewanella genospecies, based on digital DNA-DNA hybridization, as implemented in the Type Strain Genome Server. Shewanella septentrionalis, , Shewanella baltica and Shewanella hafniensis were, in this order and within a narrow genomic relatedness range, their closest genotypic relatives. Additional sampling and sequencing efforts led to the retrieval of distinct isolates that were monophyletic with SP1S1- 4T T and SP2S1- 2T, T , respectively, based on phylogenomic analysis of whole- genome sequences. Comparative analyses of genome sequence data, which included BLAST-based average nucleotide identity, core genome- based and core proteome- based phylogenomics, in addition to MALDI- TOF MS- based protein profiling, confirmed the distinctness of the putative novel genospecies with respect to their closest genotypic relatives. A comprehensive phenotypic characterisation of SP1S1- 4T T and SP2S1- 2T T revealed only minor differences with respect to the type strains of S. septentrionalis, , S. baltica and S. hafniensis. . Based on the collective phylogenomic, proteomic, and phenotypic evidence presented here, we describe two novel genospecies within the genus Shewanella, , for which the names Shewanella scandinavica sp. nov. and Shewanella vaxholinensis sp. nov. are proposed. The type strains are, respectively, SP2S1- 2T T (=CCUG 76457T=CECT T =CECT 30688T), T ), with a draft genome sequence of 5 041 805 bp and a G+C content of 46.3 mol%, and SP1S1- 4T T (=CCUG 76453T=CECT T =CECT 30684T), T ), with a draft genome sequence of 4 920147 bp and a G+C content of 46.0 mol%. Our findings suggest the existence of a species complex formed by the species S. baltica, , S. septentrionalis, , S. scandinavica sp. nov., and S. vaxholinensis sp. nov., with S. hafniensis falling in the periphery, where distinct genomic species clusters could be identified. However, this does not exclude the possibility of a continuum of genomic diversity within this sedimental ecosystem, as discussed herein with additional sequenced isolates.