Evangeliou, N. | Search bioRxiv

bioRxiv2026

Virus-host interactions on volcanic ash from Mount Etna

Volcanic ash represents an extreme and dynamic habitat, yet it hosts diverse microbial communities with largely unexplored viral diversity. This study investigated bacterial and viral populations in volcanic ash from Mount Etna (Italy) collected during the eruption, focusing on microbial novelty, activity, and virus-host interactions. Taxonomic profiling revealed that Pseudomonas and Telluria were the dominant bacterial genera, both frequently detected in airborne environments. In contrast, enrichment cultures with volcanic ash were dominated by spore-forming members of the phylum Bacillota, highlighting their resilience under harsh conditions. Metagenomic analysis recovered 19 high-quality metagenome-assembled genomes, including four previously undescribed bacterial species. Replication rate estimates showed that certain taxa were metabolically active, particularly at one sampling site. The presence of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) systems with spacers matching viral sequences suggested viral predation pressure on volcanic ash. A total of 1139 viral operational taxonomic units (vOTUs) were identified, of only around half (660 vOTUs) showed similarities to known phages, underscoring the presence of novel viruses. Shared vOTUs across sites revealed the presence of both a core virome and site-specific viral populations. Virus-host predictions indicated frequent interactions with hosts from multiple Gammaproteobacterial genera. Additionally, a 336 kb jumbo phage genome exhibited extensive metabolic capabilities and genetic autonomy. Experimental work identified a unique lytic Bacillus-infecting phage ("Phoenix") with limited propagation capacity. Furthermore, prophage induction experiments revealed active, morphologically diverse temperate phages across multiple bacterial host strains. Overall, these findings highlight volcanic ash as a reservoir of microbial and viral diversity, shaped by environmental extremes and dynamic ecological interactions. HighlightsMetagenomic and cultivation experiments were used to study viruses on volcanic ash Novelty of viral and bacterial species was detected Viral-bacterial interactions in metagenomes from volcanic ash were detected Cultivatable bacteria were mainly spore-forming Bacilli species and harbored inducible prophages

microbiology↗

bioRxiv2026

Virus-host interactions and viral population dynamics across atmospheric cloud events

BackgroundCloud water harbors diverse microbial communities despite its extreme oligotrophic conditions. However, the ecological and evolutionary dynamics of viruses in these transient atmospheric habitats remain poorly understood. Clouds have traditionally been regarded primarily as passive carriers of microorganisms rather than as active ecological environments supporting microbial interactions. In this study, cloud water was sampled at Mount Verde, Cape Verde Islands (744 m a.s.l.). We performed metagenomic analyses of iron-flocculated cloud water alongside genome analyses of a bacterial isolate and metagenome-assembled genomes using established bioinformatic approaches. Viral diversity, virus-host interactions, metabolic functions, genetic adaptations, and viral population dynamics across cloud events were investigated. In addition, UV-B resistance experiments were conducted for a novel cloud-water isolate. ResultsWe isolated 24 cloud water bacteria, including four novel species lineages, and recovered 62 high-quality metagenome-assembled genomes, including 10 novel species lineages. We identified 458 viral operational taxonomic units and 237 virus-host linkages across diverse prokaryotic hosts, revealing active viral predation across diverse bacterial taxa. In addition, CRISPR spacer matches from isolates of novel bacterial lineages such as Deinococcus nubigenus MPC36 were found. Viruses carried genes involved in host adaptation to environmental stressors, including cold-shock response, UV radiation resistance, and osmotic stress. In addition, viral populations exhibited SNP-level microdiversity and shifts in single-nucleotide variant composition across temporally proximate cloud events, indicating rapid population turnover. Experimental characterization of the cloud isolate Curtobacterium nubigenum MPC39 further revealed pronounced resistance to UV-B radiation and the presence of an inducible prophage, Curtobacterium phage vB_CnuS_Cirrus1 assigned to the new viral family Nebulaviridae, which could be validated in transmission electron microscopy. Reconstructed genomes from cloud-associated bacteria encoded carbon monoxide dehydrogenase genes and UV resistance genes, suggesting trace gas metabolism and enhanced UV protection as survival strategies in oligotrophic cloud droplets. In silico replication rates estimated using iRep were consistent with active bacterial replication at the time of sampling. ConclusionsTogether, these findings demonstrate that clouds are not merely passive carriers of microorganisms, but dynamic atmospheric ecosystems in which virus-host interactions shape microbial diversity and contribute to microbial turnover, atmospheric dispersal, and cloud-water biogeochemistry.

microbiology↗