Abstract

In mid-February 2015, a large number of deaths were observed in the sole extant population of an endangered species of freshwater snapping turtle, Myuchelys georgesi, in a coastal river in New South Wales, Australia. Mortalities continued for approximately 7 weeks and affected mostly adult animals. More than 400 dead or dying animals were observed and population surveys conducted after the outbreak had ceased indicated that only a very small proportion of the population had survived, severely threatening the viability of the wild population. At necropsy, animals were in poor body condition, had bilateral swollen eyelids and some animals had tan foci on the skin of the ventral thighs. Histological examination revealed peri-orbital, splenic and nephric inflammation and necrosis. A virus was isolated in cell culture from a range of tissues. Nucleic acid sequencing of the virus isolate has identified the entire genome and indicates that this is a novel nidovirus that has a low level of nucleotide similarity to recognised nidoviruses. Its closest relatives are nidoviruses that have recently been described in pythons and lizards, usually in association with respiratory disease. In contrast, in the affected turtles, the most significant pathological changes were in the kidneys. Real time PCR assays developed to detect this virus demonstrated very high virus loads in affected tissues. In situ hybridisation studies confirmed the presence of viral nucleic acid in tissues in association with pathological changes. Collectively these data suggest that this virus is the likely cause of the mortalities that now threaten the survival of this species. Bellinger River Virus is the name proposed for this new virus.

Highlights

  • The Bellinger River snapping turtle, Myuchelys georgesi, is a species of freshwater turtle that, prior to this outbreak, was rare and has a very restricted habitat

  • When Buffalo African green monkey kidney (BGM) cells were inoculated with pooled homogenates of spleen and lung tissues from 5 animals, after 2 passages cytopathology consisting of lytic destruction of cells in small foci was observed after approximately 5 days

  • Limited cytopathology was observed in Vero cells while no microscopic evidence of virus replication was detected in hamster lung (HmLu-1), avian (CEF), fish (SB, FHM, SSN-1), reptile (VH2) or mosquito (C6/36) cell lines

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Summary

Introduction

The Bellinger River snapping turtle, Myuchelys georgesi, is a species of freshwater turtle that, prior to this outbreak, was rare and has a very restricted habitat. A multi-agency investigation was undertaken to establish the cause and extent of the outbreak. More than 400 dead or dying animals were observed and population surveys conducted after the outbreak had ceased indicated that only a small proportion of the total population had survived with very few adults remaining. A number of moribund and dead M. georgesi were collected for post mortem examination at the Australian Registry of Wildlife Health, Taronga Conservation Society Australia Mosman, NSW. Tissue samples were referred to a number of different laboratories to test for a wide range of potential pathogens, toxins and water quality assessment. We report the isolation and characterisation of a novel nidovirus and provide evidence for its involvement as the principal pathogen in this disease outbreak

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