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Improved method for genotyping the causative agent of crayfish plague (Aphanomyces astaci) based on mitochondrial DNA

Published online by Cambridge University Press:  12 April 2019

Diana Minardi
Biosciences, University of Exeter, Stocker Road, EX4 4QD, Exeter, UK Centre for Environment, Fisheries and Aquaculture Science (Cefas), Barrack Road, DT4 8UB, Weymouth, UK The Centre for Sustainable Aquaculture Futures, UK
David J. Studholme
Biosciences, University of Exeter, Stocker Road, EX4 4QD, Exeter, UK
Birgit Oidtmann
Centre for Environment, Fisheries and Aquaculture Science (Cefas), Barrack Road, DT4 8UB, Weymouth, UK
Tobia Pretto
National Reference Laboratory for Fish, Crustacean and Mollusc Pathologies, Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe), Viale dell'Università, 10, 35020, Legnaro, Padova, Veneto, Italy
Mark van der Giezen
Biosciences, University of Exeter, Stocker Road, EX4 4QD, Exeter, UK The Centre for Sustainable Aquaculture Futures, UK
E-mail address:


Aphanomyces astaci causes crayfish plague, which is a devastating disease of European freshwater crayfish. The likely first introduction of A. astaci into Europe was in the mid-19th century in Italy, presumably with the introduction of North American crayfish. These crayfish can carry A. astaci in their cuticle as a benign infection. Aphanomyces astaci rapidly spread across Europe causing the decline of the highly susceptible indigenous crayfish species. Random amplified polymorphic DNA-PCR analysis of A. astaci pure cultures characterized five genotype groups (A, B, C, D and E). Current A. astaci genotyping techniques (microsatellites and genotype-specific regions, both targeting nuclear DNA) can be applied directly to DNA extracted from infected cuticles but require high infection levels. Therefore, they are not suitable for genotyping benign infections in North American crayfish (carriers). In the present study, we combine bioinformatics and molecular biology techniques to develop A. astaci genotyping molecular markers that target the mitochondrial DNA, increasing the sensitivity of the genotyping tools. The assays were validated on DNA extracts of A. astaci pure cultures, crayfish tissue extractions from crayfish plague outbreaks and tissue extractions from North American carriers. We demonstrate the presence of A. astaci genotype groups A and B in UK waters.

Research Article
Copyright © Cambridge University Press 2019 

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Current Address: Future Animal & Public Health, Endemics & Traceability, Animal & Plant Health, Department for Environment, Food and Rural Affairs (Defra), Nobel House, 17 Smith Square, SW1P 3JR, London, UK.


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