DUPLICATE DO NOT PUBLISH - Coral Reef Viruses in Kane'ohe Bay, Hawai'i: Abundance, Diversity & Environmental Drivers

Viruses are a ubiquitous component of coral reef ecosystems, with several viral types, from at least seven prokaryotic and 20 eukaryotic virus families currently characterised from the surface mucopolysaccharide layer (SML), coral tissue and the water column. However, little is known about the ecology and function of these viruses. For example, what are the environmental drivers of viral abundance and diversity on coral reefs? In this study, the abundance and distribution of virus-like particles (VLPs) associated with the SML and reef water of the coral Montipora capitata were determined using epifluorescence microscopy, while transmission electron microscopy was employed to determine the morphological diversity of VLPs. Sampling was conducted across the Coconut Island Marine Reserve (CIMR) reef system, Kane’ohe Bay, O’ahu, Hawai’i. Viral abundance was correlated with select environmental drivers and prokaryote abundance, while non-metric multidimensional scaling was used to determine the key environmental drivers of the viral community assemblage. The water column contained high concentrations of VLPs (5.98 × 107 ml-1) and prokaryotes (3.11 × 106 ml-1), consistent with the considerable anthropogenic impacts at this location. In comparison, the SML contained lower concentrations of VLPs (2.61 × 107 ml-1) and prokaryotes (2.08 × 106 ml-1); of note, the densities of viruses and prokaryotes in the SML were strongly coupled while those in the reef water were not. VLP density in the water column varied spatially across the reef, with the most sheltered site and the only one not situated on the reef crest having a greater VLP density than the other sites. Temporal variations in the density of microbes (i.e. viruses and prokaryotes) in the reef water were pronounced, while in the SML microbial densities remained constant. However, no specific environmental drivers of this variability could be identified. In contrast, temperature and water quality were correlated with shifts in the morphological diversity of VLPs across the reef. Small ( 100 nm) Podoviridae-like VLPs and elongate Myoviridae-like VLPs, as well as lemon-shaped VLPs of both size classes showed positive associations with turbidity, while large filamentous VLPs, Geminiviridae-like VLPs and rod-shaped VLPs were positively associated with temperature. These results demonstrate that the viral community of Coconut Island’s reef is highly diverse, and subject to spatial and temporal change, especially in the water column. However, while the environmental drivers of viral diversity were partly elucidated, we are still a long way from understanding the drivers of viral abundance. More detailed study, both spatially and temporally, of the CIMR environment is required, as is comprehensive molecular analysis of the viral community of Kane’ohe Bay. Only then can we begin to understand the importance of viruses to the health and function of this, and other reef sites.