Soil Collected in the Great Smoky Mountains National Park Yielded a Novel Species, .

Soil samples collected in the Great Smoky Mountains National Park yielded a isolate that could not be classified to the species level. Whole-genome sequence-based average nucleotide identity BLAST and DNA-DNA Hybridization analyses confirmed this isolate to be a novel species with the highest similarity to (ANI = 93.9%, isDDH = 55.9%). Additional whole-genome-based analysis using the Genome Taxonomy Database Toolkit further supported delineation as a novel species, as this tool failed to assign a species identification. Phenotypic and genotypic characterization results indicate that this species is nonpathogenic. Specifically, the novel species described here is phenotypically (i) nonhemolytic and (ii) negative for phosphatidylinositol-specific phospholipase C activity; the draft genome lacks all virulence genes found in the pathogenicity islands 1, 2, 3, and 4 as well as the internalin genes and . While the type strain contains an apparently intact catalase gene (), this strain is phenotypically catalase-negative (an unusual characteristic for species). Additional analyses identified a nonsynonymous mutation in a conserved codon of that is likely linked to the catalase-negative phenotype. Rapid species identification systems, including two biochemical and one matrix-assisted laser desorption/ionization, misidentified this novel species as either L. monocytogenes or . We propose the name and the type strain is FSL L7-0020 (=ATCC TSD-239). is a novel species that originated from a US National Park and it will be the first identified to date without official standing in the nomenclature. Validation was impeded by the National Park's requirements for strain access, ultimately deemed too restrictive by the International Committee on Systematics of Prokaryotes. However, lack of valid status should not detract from the significance of adding a novel species to the clade. Notably, detection of non species in a food processing environment indicate conditions that could facilitate the presence of the pathogen L. monocytogenes. If isolated, our data show a potential for to be misidentified as another notably L. monocytogenes. Therefore, developers of spp. detection and identification methods, who historically only include validly published species in their validation studies, should include to ensure accurate results.