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We present the genome organization and molecular characterization of the three  Formica exsecta  viruses, along with ORF predictions, and functional annotation of genes. The  Formica exsecta virus-4  (FeV4; GenBank ID:  MF287670 ) is a newly discovered negative-sense single-stranded RNA virus representing the first identified member of order  Mononegavirales  in ants, whereas the  Formica exsecta virus-1  (FeV1; GenBank ID:  KF500001 ), and the  Formica exsecta virus-2  (FeV2; GenBank ID:  KF500002 ) are positive single-stranded RNA viruses initially identified (but not characterized) in our earlier study. The new virus FeV4 was found by re-analyzing data from a study published earlier. The  Formica exsecta virus-4  genome is 9,866 bp in size, with an overall G + C content of 44.92%, and containing five predicted open reading frames (ORFs). Our bioinformatics analysis indicates that gaps are absent and the ORFs are complete, which based on our comparative genomics analysis suggests that the genomes are complete. Following the characterization, we validate virus infection for FeV1, FeV2 and FeV4 for the first time in field-collected worker ants. Some colonies were infected by multiple viruses, and the viruses were observed to infect all castes, and multiple life stages of workers and queens. Finally, highly similar viruses were expressed in adult workers and queens of six other  Formica  species:  F. fusca ,  F. pressilabris ,  F. pratensis, F. aquilonia, F. truncorum  and  F. cinerea . This research indicates that viruses can be shared between ant species, but further studies on viral transmission are needed to understand viral infection pathways.

Genome organization and molecular characterization of the three Formica exsecta viruses—FeV1, FeV2 and FeV4