alignment generated by the CLUSTAL algorithm were improved by removing hypervariable position using the program Gblock. a phylogenetic tree reconstructed based on 16S rRNA gene sequences of species of the genus lactobacillus present in the digestive tract of animals revealed that stain BTLCH M1/2T BTLCH M3/2 and M250 3MRA are situlated together with uncharacterized strians of lactobacillus from the digestive tract of honeybee in separate phylogenetic cluster. These lactobacilli from the digestive tract of pollinators represent a novel phylogenetic lineage within the genus lactobacillus. Thus, the novel lactobacilli are phylogenetically distant from the lactobacillus alimentarius cluster which inclued the species l. tucceti, the most closely related taxon with a validly published name based on 16s rRNA gene sequence similarity. Significant differences between this species and the isolates from the digestive tract of bumblebees were demonstrated by analysis of biochemistry characteristics. For these reason, the type strain of L. tucceti was not used for comparative analyses of other phenotypic characteristics.