Evolution of an alternative genetic code in the Providencia symbiont of the haematophagous leech Haementeria acuecueyetzin.

Strict blood-feeding animals are confronted with a strong B vitamin de_ciency. Blood-feeding leeches from the Glossiphoniidae family, similarly to haematophagous insects, have evolved specialised organs called bacteriomes to harbour symbiotic bacteria. Leeches of the Haementeria genus have two pairs of globular bacteriomes attached to the oesophagus which house intracellular 'Candidatus Providencia siddallii' bacteria. Previous work analysing a draft genome of the Providencia symbiont of the Mexican leech Haementeria officinalis showed that, in this species, the bacteria hold a reduced genome capable of synthesising B vitamins. In this work, we aimed to expand our knowledge on the diversity and evolution of Providencia symbionts of Haementeria. For this purpose, we sequenced the symbiont genomes of three selected leech species. We found that all genomes are highly syntenic and have kept a stable genetic repertoire, mirroring ancient insect endosymbionts. Additionally, we found B vitamin pathways to be conserved among these symbionts, pointing to a conserved symbiotic role. Lastly and most notably, we found that the symbiont of Haementeria acuecueyetzin has evolved an alternative genetic code, affecting a portion of its proteome and showing evidence of a lineage-specific and likely intermediate stage of genetic code reassignment.