{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T00:02:59Z","timestamp":1790294579348,"version":"4.1.0"},"reference-count":56,"publisher":"Cambridge University Press (CUP)","issue":"8","license":[{"start":{"date-parts":[[2019,4,12]],"date-time":"2019-04-12T00:00:00Z","timestamp":1555027200000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/www.cambridge.org\/core\/terms"}],"content-domain":{"domain":["cambridge.org"],"crossmark-restriction":true},"short-container-title":["Parasitology"],"published-print":{"date-parts":[[2019,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p><jats:italic>Aphanomyces astaci<\/jats:italic>causes crayfish plague, which is a devastating disease of European freshwater crayfish. The likely first introduction of<jats:italic>A. astaci<\/jats:italic>into Europe was in the mid-19th century in Italy, presumably with the introduction of North American crayfish. These crayfish can carry<jats:italic>A. astaci<\/jats:italic>in their cuticle as a benign infection.<jats:italic>A<\/jats:italic><jats:italic>phanomyces astaci<\/jats:italic>rapidly spread across Europe causing the decline of the highly susceptible indigenous crayfish species. Random amplified polymorphic DNA-PCR analysis of<jats:italic>A. astaci<\/jats:italic>pure cultures characterized five genotype groups (A, B, C, D and E). Current<jats:italic>A. astaci<\/jats:italic>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<jats:italic>A. astaci<\/jats:italic>genotyping molecular markers that target the mitochondrial DNA, increasing the sensitivity of the genotyping tools. The assays were validated on DNA extracts of<jats:italic>A. astaci<\/jats:italic>pure cultures, crayfish tissue extractions from crayfish plague outbreaks and tissue extractions from North American carriers. We demonstrate the presence of<jats:italic>A. astaci<\/jats:italic>genotype groups A and B in UK waters.<\/jats:p>","DOI":"10.1017\/s0031182019000283","type":"journal-article","created":{"date-parts":[[2019,4,12]],"date-time":"2019-04-12T07:17:05Z","timestamp":1555053425000},"page":"1022-1029","update-policy":"https:\/\/doi.org\/10.1017\/policypage","source":"Crossref","is-referenced-by-count":14,"title":["Improved method for genotyping the causative agent of crayfish plague (<i>Aphanomyces astaci<\/i>) based on mitochondrial DNA"],"prefix":"10.1017","volume":"146","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9426-8627","authenticated-orcid":false,"given":"Diana","family":"Minardi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3010-6637","authenticated-orcid":false,"given":"David J.","family":"Studholme","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Birgit","family":"Oidtmann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tobia","family":"Pretto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1033-1335","authenticated-orcid":false,"given":"Mark","family":"van der Giezen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"56","published-online":{"date-parts":[[2019,4,12]]},"reference":[{"key":"S0031182019000283_ref50","first-page":"192","article-title":"On the host range and origin of the crayfish plague fungus","volume":"52","author":"Unestam","year":"1972","journal-title":"Report of the Institute of Freshwater Research, Drottningholm"},{"key":"S0031182019000283_ref29","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2164-12-84"},{"key":"S0031182019000283_ref13","doi-asserted-by":"publisher","DOI":"10.3391\/ai.2017.12.4.10"},{"key":"S0031182019000283_ref4","doi-asserted-by":"publisher","DOI":"10.1089\/cmb.2012.0021"},{"key":"S0031182019000283_ref51","doi-asserted-by":"publisher","DOI":"10.1099\/00221287-60-1-77"},{"key":"S0031182019000283_ref6","doi-asserted-by":"publisher","DOI":"10.1038\/sj.tpj.6500167"},{"key":"S0031182019000283_ref47","doi-asserted-by":"publisher","DOI":"10.3354\/dao02334"},{"key":"S0031182019000283_ref49","doi-asserted-by":"publisher","DOI":"10.1093\/bib\/bbs017"},{"key":"S0031182019000283_ref38","first-page":"303","article-title":"An improved isolation method for the cultivation of the crayfish plague fungus, Aphanomyces astaci","volume":"12","author":"Oidtmann","year":"1999","journal-title":"Freshwater Crayfish"},{"key":"S0031182019000283_ref11","doi-asserted-by":"publisher","DOI":"10.1016\/S0953-7562(09)80716-8"},{"key":"S0031182019000283_ref54","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2014.07.008"},{"key":"S0031182019000283_ref23","doi-asserted-by":"publisher","DOI":"10.1016\/j.jip.2011.08.002"},{"key":"S0031182019000283_ref52","doi-asserted-by":"publisher","DOI":"10.3354\/dao02575"},{"key":"S0031182019000283_ref7","doi-asserted-by":"publisher","DOI":"10.1094\/PHYTO-12-14-0373-R"},{"key":"S0031182019000283_ref33","first-page":"215","article-title":"Nota sulla malattia dei gamberi che ammorb\u00f2 le acque del veronese nell'anno 1861","volume":"XLI","author":"Martinati","year":"1862","journal-title":"Memorie Dell'accademia D'agricoltura Commercio Ed Arti Di Verona"},{"key":"S0031182019000283_ref26","unstructured":"Li H (2013) Aligning sequence reads, clone sequences and assembly contigs with BWA-MEM. eprint arXiv:1303.3997v1 [q-GN]."},{"key":"S0031182019000283_ref1","first-page":"603","article-title":"Geographical spread of bacterial and fungal diseases of crustaceans","volume":"15","author":"Alderman","year":"1996","journal-title":"Revue Scientifique et Technique (International Office of Epizootics)"},{"key":"S0031182019000283_ref34","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0036518"},{"key":"S0031182019000283_ref39","doi-asserted-by":"publisher","DOI":"10.3354\/dao072053"},{"key":"S0031182019000283_ref30","first-page":"171","article-title":"Co-invaders: the effects of alien parasites on native hosts","volume":"3","author":"Lymbery","year":"2014","journal-title":"International Journal for Parasitology: Parasites and Wildlife"},{"key":"S0031182019000283_ref31","doi-asserted-by":"publisher","DOI":"10.3354\/dao02955"},{"key":"S0031182019000283_ref35","doi-asserted-by":"publisher","DOI":"10.1016\/j.jip.2018.06.002"},{"key":"S0031182019000283_ref18","doi-asserted-by":"publisher","DOI":"10.1016\/0044-8486(94)90243-7"},{"key":"S0031182019000283_ref22","doi-asserted-by":"publisher","DOI":"10.1051\/kmae\/2018001"},{"key":"S0031182019000283_ref28","doi-asserted-by":"publisher","DOI":"10.1016\/S0044-8486(97)00153-1"},{"key":"S0031182019000283_ref43","first-page":"16","article-title":"Crayfish plague threatens UK stock","volume":"7","author":"Polglase","year":"1984","journal-title":"Fish Farmer"},{"key":"S0031182019000283_ref16","first-page":"394","article-title":"A review of the ever increasing threat to European crayfish from non-indigenous crayfish species","volume":"11","author":"Holdich","year":"2009","journal-title":"Knowledge and Management of Aquatic Ecosystems"},{"key":"S0031182019000283_ref42","doi-asserted-by":"publisher","DOI":"10.1007\/s10530-010-9890-9"},{"key":"S0031182019000283_ref48","doi-asserted-by":"publisher","DOI":"10.1111\/jfd.12472"},{"key":"S0031182019000283_ref25","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkq1019"},{"key":"S0031182019000283_ref24","doi-asserted-by":"publisher","DOI":"10.1080\/03949370.2014.897652"},{"key":"S0031182019000283_ref2","unstructured":"Alderman DJ (2003) Aphanomycosis of crayfish: crayfish plague. Research and Development Technical Report W2-064. Cefas, Weymouth."},{"key":"S0031182019000283_ref3","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2761.1984.tb01205.x"},{"key":"S0031182019000283_ref5","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms6030058"},{"key":"S0031182019000283_ref8","first-page":"361","article-title":"Pallasea quadrispinosa G.O. Sars specimens as vectors of aquatic zoosporic fungi parasiting on fish","volume":"13","author":"Czeczuga","year":"2004","journal-title":"Polish Journal of Environmental Studies"},{"key":"S0031182019000283_ref9","doi-asserted-by":"publisher","DOI":"10.1127\/nova_hedwigia\/2015\/0230"},{"key":"S0031182019000283_ref10","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2109.1993.tb00655.x"},{"key":"S0031182019000283_ref12","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2014.02.020"},{"key":"S0031182019000283_ref14","first-page":"95","article-title":"Bioedit: a user-friendly biological sequence alignment editor and analysis program for Windows 95\/98\/NT","volume":"41","author":"Hall","year":"1999","journal-title":"Nucleic Acids Symposium Series"},{"key":"S0031182019000283_ref15","doi-asserted-by":"publisher","DOI":"10.1002\/aqc.3270010204"},{"key":"S0031182019000283_ref19","doi-asserted-by":"publisher","DOI":"10.1017\/S0031182016002419"},{"key":"S0031182019000283_ref20","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1515\/9783110438666-013","volume-title":"Biological Invasions in Changing Ecosystems, Vectors, Ecological Impacts, Management and Predictions","author":"Jussila","year":"2015"},{"key":"S0031182019000283_ref21","doi-asserted-by":"publisher","DOI":"10.1007\/s10530-017-1496-z"},{"key":"S0031182019000283_ref17","doi-asserted-by":"publisher","DOI":"10.1080\/03949370.2014.903380"},{"key":"S0031182019000283_ref27","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btp352"},{"key":"S0031182019000283_ref32","doi-asserted-by":"publisher","DOI":"10.1017\/S0031182018000227"},{"key":"S0031182019000283_ref36","doi-asserted-by":"publisher","DOI":"10.1002\/aqc.2674"},{"key":"S0031182019000283_ref37","first-page":"1203","article-title":"Sulla mortalita dei gamberi (Astacus fluviatilis L.) nel Veneto e pi\u00f9 particolarmente nella provincia trevigiana","volume":"10","author":"Ninni","year":"1865","journal-title":"Atti Instituto Veneto"},{"key":"S0031182019000283_ref40","volume-title":"Manual of Diagnostic Tests for Aquatic Animals","year":"2018"},{"key":"S0031182019000283_ref41","doi-asserted-by":"publisher","DOI":"10.1016\/j.jip.2018.05.006"},{"key":"S0031182019000283_ref44","unstructured":"Pretto T , Tosi F , Sandoval-Sierra JV , Grandjean F , Manfrin A and Di\u00e9guez-Uribeondo J (2014) Characterization of Aphanomyces astaci in white-clawed crayfish Austropotamobius pallipes from Northern Italy: considerations regarding a crayfish plague outbreak., in: IAA & CSJ Joint International Conference on Crustacea. Sapporo, Japan."},{"key":"S0031182019000283_ref45","doi-asserted-by":"publisher","DOI":"10.1016\/j.funbio.2013.10.007"},{"key":"S0031182019000283_ref46","doi-asserted-by":"publisher","DOI":"10.1017\/S0950268805005303"},{"key":"S0031182019000283_ref53","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2008.12.022"},{"key":"S0031182019000283_ref55","doi-asserted-by":"publisher","DOI":"10.1002\/bies.20529"},{"key":"S0031182019000283_ref56","doi-asserted-by":"crossref","first-page":"3741","DOI":"10.1128\/AEM.63.10.3741-3751.1997","article-title":"Inhibition and facilitation of nucleic acid amplification","volume":"63","author":"Wilson","year":"1997","journal-title":"Applied and Environmental Microbiology"}],"container-title":["Parasitology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.cambridge.org\/core\/services\/aop-cambridge-core\/content\/view\/S0031182019000283","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,9,15]],"date-time":"2022-09-15T20:44:54Z","timestamp":1663274694000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.cambridge.org\/core\/product\/identifier\/S0031182019000283\/type\/journal_article"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,4,12]]},"references-count":56,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2019,7]]}},"alternative-id":["S0031182019000283"],"URL":"https:\/\/doi.org\/10.1017\/s0031182019000283","relation":{},"ISSN":["0031-1820","1469-8161"],"issn-type":[{"value":"0031-1820","type":"print"},{"value":"1469-8161","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,4,12]]},"assertion":[{"value":"Copyright \u00a9 Cambridge University Press 2019\u00a0","name":"copyright","label":"Copyright","group":{"name":"copyright_and_licensing","label":"Copyright and Licensing"}}]}}