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Diseases of Aquatic Organisms

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DAO 135:33-41 (2019)  -  DOI: https://doi.org/10.3354/dao03369

Multi-tool diagnosis of an outbreak of ranavirosis in amphibian tadpoles in the Canadian boreal forest

M. J. Forzán1,*, J. Bienentreu2, D. M. Schock2,3, D. Lesbarrères2

1Cornell Wildlife Health Lab, Department of Population Medicine, Animal Health Diagnostic Center, Cornell University College of Veterinary Medicine, Ithaca, NY 14853, USA
2Department of Biology, Laurentian University, Sudbury, ON, Canada, P3E 2C6
3University Studies and Environmental Technology, Keyano College, Fort McMurray, AB, Canada, T9H 2H7
*Corresponding author:

ABSTRACT: Investigation of mortalities in isolated wild amphibian populations presents diagnostic difficulties that can hinder reaching a definitive diagnosis for the cause of death. Disease can only be diagnosed when pathogen presence (e.g. detection by PCR) is linked to tissue lesions (histopathology) in the host. We report a 2-site outbreak of ranavirosis in wild anuran tadpoles in the boreal forest of Wood Buffalo National Park, Canada, diagnosed by histologic and molecular techniques. Mortalities occurred in wood frog Rana sylvatica tadpoles and boreal chorus frog Pseudacris maculata tadpoles. Lack of mortality in sympatric Canadian toad Bufo (Anaxyrus) hemiophrys tadpoles suggested lower disease susceptibility in this species. In the former 2 species, ranavirosis was diagnosed based on consistent histopathology, immunohistochemistry (IHC), in situ hybridization (ISH), and quantitative PCR results. The most common histopathologic lesion present in wood and boreal chorus frog tadpoles was necrosis of the skin, oral mucosa, renal tubular epithelium, renal hematopoietic tissue, and branchial epithelium. Mild hepatic and pancreatic necrosis and rare intracytoplasmic inclusion bodies in hepatocytes were less common. Skeletal and connective tissues in budding limbs often had multifocal to coalescing necrosis and were intensely positive for ranavirus, with IHC staining even in areas where no obvious necrosis could be observed. Abundant IHC and ISH staining in actively growing tissues support a link between disease emergence and amphibian developmental stage. Our findings provide a definitive diagnosis of ranavirosis in free-living amphibians and highlight the effectiveness of multi-tool approaches to mortality investigation and elucidation of pathogenesis of ranavirosis in wild amphibians.


KEY WORDS: Ranavirus · Hylidae · Wild population · Diagnostic assays · qPCR · Immunohistochemistry · In situ hybridization · Boreal forest


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Cite this article as: Forzán MJ, Bienentreu J, Schock DM, Lesbarrères D (2019) Multi-tool diagnosis of an outbreak of ranavirosis in amphibian tadpoles in the Canadian boreal forest. Dis Aquat Org 135:33-41. https://doi.org/10.3354/dao03369

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