Diversity of ssDNA Phages Related to the Family \u3cem\u3eMicroviridae\u3c/em\u3e within the \u3cem\u3eCiona robusta\u3c/em\u3e Gut

The gut microbiome is a complex ecosystem of bacteria, viruses, and fungi that strongly influences animal health. The bacterial component, for example, contributes orders of magnitude more gene products to host physiology than the host genome; thus, changes to the composition of these bacterial communities can have profound influences on the health of the animal. By infecting and lysing their hosts, viruses (particularly viruses infecting bacteria or phages) can affect critical functions in these environments, yet the consequences of these infections remain to be fully described. Most studies investigating gut viromes to date have focused on double-stranded DNA (dsDNA) phages and, consequently, little is known about the smaller single-stranded DNA (ssDNA) phages, which also inhabit gut environments. In this study, we investigated ssDNA phages of the Microviridae family within the gut of an invertebrate organism, Ciona robusta, used as a model system to better understand gut microbial interactions. As a filter feeder, Ciona concentrates dissolved organics and microbes as part of its diet, yet maintains a microbiome distinct from the surrounding water column. We identified 258 unique ssDNA phage genomes representing a diversity of Microviridae subgroups including novel members of the established Gokushovirinae subfamily and several proposed phylogenetic groups (Alpavirinae, Aravirinae, Group D, Parabacteroides prophages, and Pequeñovirus). Over 70% of the genomes belonged to the Gokushovirinae; however, 155 of these genomes did not group with previously described sequences. Our results highlight an unprecedented diversity of ssDNA phages from an animal gut. Furthermore, comparative analysis between samples collected from Ciona specimens with full and cleared guts as well as the surrounding water indicated that Ciona retains a unique and highly diverse community of ssDNA phages. The present study significantly expands the known diversity within the Microviridae family and suggests that Ciona is a promising system for studying the role of ssDNA phages within animal guts