AbstractThe intracellular endosymbiotic proteobacteriaWolbachiahave evolved across the phyla nematoda and arthropoda. InWolbachiaphylogeny, supergroup F is the only clade known so far with members from both arthropod and filarial nematode hosts and therefore can provide unique insights into their evolution and biology. In this study, four new supergroup FWolbachiagenomes have been assembled using a metagenomic assembly and binning approach,wMoz andwMpe from the human filarial parasitesMansonella ozzardiandM. perstans, andwOcae andwMoviF from the blue mason beeOsmia caerulescensand the sheep kedMelophagus ovinus, respectively. A comprehensive phylogenomic analysis revealed two distinct lineages of filarialWolbachiain supergroup F, indicating multiple horizontal transfer events between arthropod and nematode hosts. The analysis also reveals that the evolution ofWolbachia-filaria symbioses is accompanied by a convergent pseudogenization and loss of the bacterioferritin gene, a phenomenon found to be shared by all filarialWolbachia, even those outside supergroup F. These observations indicate that differences in heme metabolism might be a key feature distinguishing filarial and arthropodWolbachia. The new genomes provide a valuable resource for further studies on symbiosis, evolution, and the discovery of new antibiotics to treat mansonellosis.