DAO 95:9-17 (2011)  -  DOI: https://doi.org/10.3354/dao02334

Detection and quantification of the crayfish plague agent in natural waters: direct monitoring approach for aquatic environments

David A. Strand1,2, Arne Holst-Jensen1, Hildegunn Viljugrein3, Bente Edvardsen4, Dag Klaveness2, Japo Jussila5, Trude Vrålstad1,2,*

1Section of Mycology, Norwegian Veterinary Institute, Pb 750, Sentrum, 0106 Oslo, Norway
2Microbial Evolution Research Group (MERG), Department of Biology, University of Oslo, Pb 1066, Blindern, 0316 Oslo, Norway
3Section of Epidemiology, Norwegian Veterinary Institute, Pb 750, Sentrum, 0106 Oslo, Norway
4Marine Biology, Department of Biology, University of Oslo, Pb 1066, Blindern, 0316 Oslo, Norway
5Department of Biosciences, University of Eastern Finland, Kuopio campus, PL 1627, 70211 Kuopio, Finland
*Corresponding author. Email:

ABSTRACT: Aphanomyces astaci, a specialised parasite of North American freshwater crayfish, is the disease agent of crayfish plague that is lethal to European freshwater crayfish. The life cycle of A. astaci has been inferred from experimental laboratory studies, but less is known about its natural sustainability and ecology. To address such questions, tools for monitoring of A. astaci directly in aquatic environments are needed. Here, we present an approach for detecting and quantifying A. astaci directly from water samples using species-specific TaqMan® minor groove binder real-time PCR. Samples of a 10-fold dilution series from ~104 to ~1 spore of A. astaci were repeatedly tested, and reliable detection down to 1 spore was demonstrated. Further, to simulate real-life samples from natural water bodies, water samples from lakes of various water qualities were spiked with spores. The results demonstrated that co-extracted humic acids inhibit detection significantly. However, use of bovine serum albumin or the TaqMan® Environmental Master Mix largely removes this problem. The practical application of the approach was successfully demonstrated on real-life water samples from crayfish farms in Finland hosting infected North American signal crayfish Pacifastacus leniusculus. Direct monitoring of A. astaci from aquatic environments may find application in the management of wild noble crayfish Astacus astacus stocks, improved aquaculture practices and more targeted conservation actions. The approach will further facilitate studies of A. astaci spore dynamics during plague outbreaks and in carrier crayfish populations, which will broaden our knowledge of the biology of this devastating crayfish pathogen.


KEY WORDS: Aphanomyces astaci · Crayfish plague · Filtration · Molecular detection · Aquatic ­environments · Quantitative real-time PCR


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Cite this article as: Strand DA, Holst-Jensen A, Viljugrein H, Edvardsen B, Klaveness D, Jussila J, Vrålstad T (2011) Detection and quantification of the crayfish plague agent in natural waters: direct monitoring approach for aquatic environments. Dis Aquat Org 95:9-17. https://doi.org/10.3354/dao02334

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