Characterization of Gut Symbionts from Wild-caught and Other Diptera: Description of Gen. Nov., Sp. Nov., Sp. Nov., Sp. Nov, and Sp. Nov

Non-culture based surveys show that the bacterial family is widespread in guts of insects, including wild . Relatively few isolates have been described, and none has been described from . We present the isolation and characterization of five strains of from wild-caught flies of the genera (Diptera: Drosophilidae) and (Diptera: Lauxaniidae). Cells are generally rod-shaped, mesophilic, and measure 0.8-2.0 µm long by 0.3-0.5 µm wide. Optimal growth was observed under ambient atmosphere. Reconstruction of phylogenies from the 16S rRNA gene and from single-copy orthologs verify placement of these strains within . Cells exhibited similar fatty acid profiles to those of other . Strain lpD01 shared 74% average nucleotide identity (ANI) with its closest relatives Schmidhempelia bombi Bimp and IPMB12. Results from multiple genome-wide similarity comparisons indicate lpD01 should be classified as a novel species within a novel genus. The major respiratory quinone for lpD01 is ubiquinone Q-8. lpD02, lpD03, lpD04, and BiB are more closely related to CN3 (76, 77, 76, and 77% ANI, respectively) than to other described . Genomic and phylogenetic analyses suggest that lpD03 and lpD04 belong to the same species and that lpD02, lpD03/lpD04, and BiB are each novel species of the genus . The proposed names of these strains are gen. nov., sp. nov. (type strain lpD01 =NCIMB 15517 =ATCC TSD-399), sp. nov (type strain lpD02 =NCIMB 15518 =ATCC TSD-400), sp. nov. (type strain lpD04 =NCIMB 15520 =ATCC TSD-401), and sp. nov (type strain BiB=NCIMB 15516 =ATCC TSD-402). The isolation and characterization of these strains expands the repertoire of culturable bacteria naturally associated with insects, including the model organism .