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DAO 129:85-98 (2018)  -  DOI: https://doi.org/10.3354/dao03239

Novel quantitative PCR assay specific for the emerging Perkinsea amphibian pathogen reveals seasonal infection dynamics

Emily E. Karwacki1, Matthew S. Atkinson1, Robert J. Ossiboff2, Anna E. Savage1,*

1University of Central Florida, Department of Biology, 4110 Libra Dr., Orlando, Florida 32816, USA
2University of Florida, Department of Comparative, Diagnostic, and Population Medicine, 2015 SW 16th Ave., Gainesville, Florida 32610, USA
*Corresponding author:

ABSTRACT: Amphibians are suffering from large-scale population declines worldwide, and infectious diseases are a central driving force. Most pathogen-mediated declines are attributed to 2 pathogens, the fungus Batrachochytrium dendrobatidis and iridoviruses in the genus Ranavirus. However, another emerging pathogen within Perkinsea is associated with mass mortality events in anurans throughout the southeastern USA. Molecular resources for detecting amphibian Perkinsea have been limited to general protistan primers that amplify a range of organisms, not all of which are disease agents. Moreover, the only quantitative method available involves histopathology, which is labor intensive, requires destructive sampling, and lacks sensitivity. Here, we developed a novel quantitative (q)PCR assay that is sensitive and specific for amphibian Perkinsea, providing a resource for rapid and reliable pathogen diagnosis. We used histopathology to confirm that qPCR burdens track the severity of Perkinsea infections across multiple anuran tissues. We also sampled 3 natural amphibian communities in Florida, USA, to assess the prevalence and intensity of amphibian Perkinsea infections across species, seasons, tissues, and life stages. Anurans from 2 of 3 sampling locations were infected, totaling 25.1% of all individuals. Infection prevalence varied significantly among locations, seasons, species, and life stages. Infection intensity was significantly higher in larval tissues than adult tissues, and was significantly different across locations, seasons, and species. Understanding relationships between amphibian Perkinsea infection, other pathogens, and biotic and abiotic cofactors will allow us to assess what drives population declines, improving our ability to develop conservation strategies for susceptible species to reduce global amphibian biodiversity loss.


KEY WORDS: Emerging disease · Amphibian decline · Protozoan · Parasite · Frog · Rana · Hyla · Acris · Anuran · Mortality · Florida


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Cite this article as: Karwacki EE, Atkinson MS, Ossiboff RJ, Savage AE (2018) Novel quantitative PCR assay specific for the emerging Perkinsea amphibian pathogen reveals seasonal infection dynamics. Dis Aquat Org 129:85-98. https://doi.org/10.3354/dao03239

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