Abstract

The koala population in northern Australia has become increasingly fragmented due to natural and man-made barriers and interventions. This situation has created a unique opportunity to study both endogenous and exogenous koala retrovirus (KoRV). To determine the impact that population isolation has had on KoRV diversity in Queensland, 272 koalas from six fragmented koala populations were profiled for their KoRV provirus across two natural biogeographical barriers (the St Lawrence Gap and the Brisbane Valley Barrier), one man-made geographical barrier (the city of Brisbane) and two translocation events (the single movement of koalas to an island and the repeated movement of koalas into a koala sanctuary). Analysis revealed that all koalas tested were KoRV-A positive, with 90 - 96% of the detected KoRV provirus from each koala representing a single, likely endogenous, KoRV-A strain. The next most abundant proviral sequence was a defective variant of the dominant KoRV-A strain, accounting for 3 - 10% of detected provirus. The remaining KoRV provirus represented expected exogenous strains of KoRV and included geographically localized patterns of KoRV-B, -C, -D, -F, -G, and -I. These results indicate that lineage diversification of exogenous KoRV is actively ongoing. In addition, comparison of KoRV provirus within known dam-sire-joey family groups from the koala sanctuary revealed that joeys consistently had KoRV proviral patterns more similar to their dams than their sires in KoRV-B, -C and -D provirus composition. Collectively, this study highlights both the consistency of endogenous KoRV and the diversity of exogenous KoRV across the fragmented koala populations in northern Australia.IMPORTANCE KoRV infection has become a permanent part of koalas in northern Australia. With KoRV presence and abundance linked to more severe chlamydial disease and neoplasia in these koalas, understanding how KoRV exists throughout an increasingly fragmented koala population is a key first step in designing conservation and management strategies. This survey of KoRV provirus in Queensland koalas indicates that endogenous KoRV provirus is ubiquitous and consistent throughout the state while exogenous KoRV provirus is diverse and distinct in fragmented koala populations. Understanding the prevalence and impact of both endogenous and exogenous KoRV will be needed to ensure a future for all koala populations.

Highlights

  • IMPORTANCE Koala retrovirus (KoRV) infection has become a permanent part of koalas in northern Australia

  • To assess KoRV diversity across central east to southeast Queensland, samples from a total of 272 koalas were collected from six study sites (Fig. 1A)

  • Previous study has determined that KoRV proviral profiles generated from multiple sample types can be combined for analysis [12], so all samples were DNA extracted, PCR amplified to obtain the KoRV envelope gene receptor binding region from all subtypes simultaneously, deep amplicon sequenced, and processed to obtain representative operational taxonomic units (OTUs) or strains present in each koala

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Summary

Introduction

IMPORTANCE KoRV infection has become a permanent part of koalas in northern Australia. Within Queensland koalas, full koala genome sequencing, historical koala skin examination, and field studies of KoRV provirus in wild koala populations have strongly suggested that the KoRV-A subtype contains both endogenous (vertically transmitted, based on high-copy [apparently germ line] proviral insertion) and exogenous (horizontally transmitted, based on various strain types) strains, while KoRV-B to -I subtypes appear to contain only exogenous strains (based on low-copy-number [apparently somatic] insertions) [11,12,13,14]. This study compares the diversity of KoRV provirus detected in 272 koalas from six distinct Queensland koala populations, spanning two natural biogeographical barriers, one isolated island population and one long-term captive koala colony. This detailed family comparison revealed that exogenous KoRV patterns detected in joeys were much more related to their dam’s KoRV profile

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