frigidophilus SAP472: [30] Austropotamobius pallipes (2008, Spain) CHI A. invadans WIC: [6] Brevoortia tyrannus (2004, USA) CHI, MCA, TaqMan A. laevis CBS 107.52 unknown (1952, unknown) CHI, MCA, TaqMan A. helicoides CBS 210.82 unknown (1982, former USSR) CHI, MCA, TaqMan A. repetans LK29 P. leniusculus (2004, Leithakanal, Austria) CHI,

PHYLO A. irregularis CBS 278.81 pond (1981, The Netherlands) CHI, MCA, TaqMan Achlya racemosa CBS 578.67 unknown (1967, Great Britain) CHI Leptolegnia caudata CBS 680.69 unknown (1969, Canada) CHI, MCA, TaqMan Saprolegnia parasitica CBS 540.67 fish hatchery (1967, Great Britian) CHI Aspergillus sp. not assigned horse food (2004, Vienna, Austria) MCA Fusarium solani CBS 181.29 unknown (1929, Germany) CHI Trichosporon cutaneum DSM 70675 sulfite liquor waste CHI Western: western-blot analysis, CHI: partial sequencing of homologous eFT508 concentration chitinase gene(s), RACE: rapid amplification selleck chemical of cDNA ends, PHYLO: determination of ITS nrDNA sequences for phylogenetic analysis, GX: temporal gene expression of Chi2 and Chi3, MCA: qPCR/MCA for qualitative detection of A. astaci, TaqMan: TaqMan qPCR The Aphanomyces strain LK29 was isolated from a healthy signal crayfish (Pacifastacus leniusculus). Physiological and genetic evidence showed that the strain does not fit into any previously identified group of A. astaci. It exhibited properties like repeated zoospore

emergence and lack of sexual reproduction commonly associated with parasitic species. In contrast to A. astaci, the strain LK29 does not express chitinase constitutively during growth or sporulation. Phylogenetic analysis of ITS sequences (Additional file 1A) demonstrated clustering within the A. laevis-repetans clade [29]. In addition, a Blastn search with the 28SrDNA

sequence of LK29 (GenBank:GQ152606, this work) showed close homology to A. laevis (99%, GenBank:AF320584), but clear difference (97% identity) to the A. astaci strains Hö, FDL, GB04 and AZD9291 clinical trial Z12 (AF320583, AF320582, GQ374534, GQ374535, respectively). Until their taxonomic status is fully elucidated the new isolate was assigned to A. repetans. This species is not capable of killing crayfish following standardised experimental infection and is characterised by a high growth rate, and germination in response to nutrients [30]. Sequence determination of the novel A. astaci genes CHI2 and CHI3 Fungal species contain one to twenty GH18 chitinase family genes [28]. In order to develop a robust diagnostic assay for A. astaci, we asked IACS-10759 in vivo whether the chitinolytic system of the pathogen would contain multiple genes of this ancient gene family widely expressed in archea, prokaryotes and eukaryotes [31]. As indicated by the two cross-reacting bands detected in western-blot analysis with antibodies raised against the catalytic GH18 domain, A. astaci contains more than one chitinase-like protein (Figure 1).