A tentative new species Aphanomyces fennicus sp. nov. interferes with molecular diagnostic methods for crayfish...

Several isolates of an unknown oomycete resembling the genus Aphanomyces were obtained into laboratory culture from samples of noble crayfish (Astacus astacus) in 2016-2017. The crayfish were kept in cages in connection with a study on an eventually persistent crayfish plague infection in a small Finnish lake, following an acute episode of the disease in 2010. Despite the close resemblance of the isolates to the causative agent of crayfish plague, Aphanomyces astaci, and the positive results obtained in OIE recommended A. astaci-specific ITS-based conventional PCR and qPCR molecular assays, the isolates can be distinguished from A. astaci by morphological features concerning hyphal structure and chlamydospore formation, as well as using the randomly amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) method, microsatellite-based genotyping, the pathogenicity test and phylogenetic analysis based on ITS sequencing. The name Aphanomyces fennicus sp. novum is proposed for this close relative of A. astaci. The detection of this tentative novel species giving false-positive results in existing diagnostic assays for the crayfish plague highlights the importance of careful interpretation of the results from molecular methods, especially concerning crayfish with low-level infections, excluding the possibility to verify the results from clinical or sequencing data.