Metagenomic Analyses Reveal a Constrained Network of Nutritional Symbionts in Kissing Bugs

Triatominae bugs are the main vectors of Chagas disease in Latin America and rely on microbial nutritional symbiosis to complement their haematophagous diet with B-vitamins. While Rhodococcus bacteria have been identified as key symbionts, diverse metabarcoding analyses have suggested additional candidates. However, symbiont genomic data and metabolic capabilities remain largely uncharacterized. To address this gap, we generated metagenomic assemblies for 14 Triatominae and captured 15 bacterial genomes belonging to 4 genera ( Rhodococcus, Wolbachia, Symbiopectobacterium and Arsenophonus ) across 9 triatominae species. We identified five co-infection cases, including one involving two distinct Arsenophonus symbionts, one exhibiting hallmarks of massive genome degradation. Phylogenetic analyses revealed that Triatominae-associated symbionts form monophyletic groups within each genus, suggesting common origins followed by co-evolution with their hosts. Annotation of vitamin B metabolic genes indicates that most symbionts harbour incomplete pathways, with evidence of metabolic complementation between co-infecting symbionts. Additionally, we identified bacterial genes laterally transferred into host insect genomes, interpreted as footprints of present or past symbiotic associations. Nearly all Triatominae genomes displayed transferred genes from all four bacterial genera, including hosts with no detectable symbiont in genome assemblies. Taken together with these discoveries support the existence of a stable and limited network of four possible nutritional symbiont lineages with rare evidence of symbiont turn-overs.