Combining Traditional and Molecular Techniques Supports the Discovery of a Novel Species During Environmental Surveillance in a Healthcare Facility.

surveillance plays a significant role not only to prevent the risk of infection but also to study the ecology of isolates, their characteristics, and how their prevalence changes in the environment. The difficulty in isolation, identification, and typing results in a low notification rate; therefore, human infection is still underestimated. In addition, during surveillance, the special attention given to leads to an underestimation of the prevalence and risk of infection for other species. This study describes the workflow performed during environmental surveillance that resulted in the isolation of two strains, named 8cVS16 and 9fVS26, associated with the genus . Traditional and novel approaches such as standard culture technique, MALDI-TOF MS, gene sequencing, and whole-genome sequencing (WGS) analysis were combined to demonstrate that isolates belong to a novel species. The strain characteristics, the differences between macrophage infectivity potential (), RNA polymerase β subunit (), and reference gene sequences, the average nucleotide identity (ANI) of 90.4%, and the DNA-DNA digital hybridization (dDDH) analysis of 43% demonstrate that these isolates belong to a new species. The finding suggests that, during the culture technique, special attention should be paid to the characteristics of the isolates that are less associated with the genus in order to investigate the differences found using more sensitive methods. The characterization of the two newly discovered isolates based on morphological, biochemical, and microscopic characteristics is currently underway and will be described in another future study.