Lactic acid bacteria were isolated from freshwater puddles in a meadow–shrub habitat near Skálholt, Iceland. Two Gram-positive, catalase-negative isolates were recovered and initially identified as Latilactobacillus sakei by MALDI-TOF MS. Whole-genome sequencing confirmed one isolate as L. sakei , whereas the second isolate represented a distinct lineage closely related to L. sakei . Genome sizes of both isolates were larger than previously reported for the genus, necessitating an emended description of Latilactobacillus . Comparative analyses showed that average nucleotide identity and digital DNA–DNA hybridization values between the novel lineage and recognized Latilactobacillus species were ≤95.57% and ≤63.0%, respectively, supporting its recognition as a separate species. On the basis of genotypic, phylogenetic and phenotypic evidence, the name Latilactobacillus parasakei sp. nov. is proposed, with strain 5-91 T (=LMG 34054 T =DSM 120140 T ) as the type strain.
