The isolation, characterization, and identification of a novel species of bacterium in the Enterobacteriaceae family from Kephart Prong, Great Smoky Mountains National Park

The purpose of this study was to examine a single bacterial species isolated from Great Smoky Mountains Nation Park (GSMNP), characterize its growth requirements, and identify it down to the species level. A polyphasic approach that examined phenotypic, genotypic, and phylogenetic characteristics was used. Phenotypic analysis revealed that the isolate is Gram-negative, rod-shaped, non-motile, oxidase negative, catalase positive, and grows in the presence and absence of oxygen. Growth was observed at temperatures ranging from 4ºC to 37ºC, with optimum growth at 30ºC based on visual observation of colony mass. The pH range for growth was pH7-9, with optimum growth at pH9 based on visual observation of colony mass. The isolate can tolerate up to 1% NaCl in the nutrient media. Genotypic analysis utilizing 16S rDNA sequences and whole genome sequencing (WGS) identified the isolate as a member of the order “Enterobacteriales” and the family Enterobacteriaceae. Phylogenetic analysis supported the isolate’s position in both taxa, but did not cluster the isolate with any specific genera. On the basis of phenotypic, genotypic, and phylogenetic properties, the isolate LD2 represents a novel species of a new genus