Controlled Infection Experiment With Aphanomyces astaci Provides Additional Evidence for Latent Infections and Resistance in Freshwater Crayfish

For 150 years the crayfish plague disease agent Aphanomyces astaci has been the cause of mass mortalities among native European crayfish populations. However, recently several studies have highlighted the great variability of A. astaci virulence and crayfish resistance toward the disease. The main aim of this study was to compare the response of two crayfish species, the European native noble crayfish (Astacus astacus) and the invasive alien marbled crayfish (Procambarus virginalis), to an A. astaci challenge with a highly virulent strain from haplogroup B and a lowly virulent strain from haplogroup A. In a controlled infection experiment we showed a high resistance of marbled crayfish against an A. astaci infection, with zoospores from the highly virulent haplogroup B strain being able to infect the crayfish, but unable to cause signs of disease. Furthermore, we demonstrated a reduced virulence in the A. astaci strain belonging to haplogroup A, as shown by the light symptoms and the lack of mortality in the generally susceptible noble crayfish. Interestingly, in both marbled crayfish and noble crayfish challenged with this strain, we observed a significant decrease of the detected amount of pathogen’s DNA during the experiment, suggesting that this A. astaci haplogroup A strain has a decreased ability of penetrating into the cuticle of the crayfish. Our results provide additional evidence of how drastically strains belonging to A. astaci haplogroup B and haplogroup A differ in their virulence. This study confirmed the adaptation of one specific A. astaci haplogroup A strain to their novel European hosts, supposedly due to reduced virulence. This feature might be the consequence of A. astaci’s reduced ability to penetrate into the crayfish. Finally, we experimentally showed that marbled crayfish are remarkably resistant against the crayfish plague disease and could potentially be latently infected, acting as carriers of highly virulent A. astaci strains.

[1]  Birgit Oidtmann Aphanomyces astaci , 2022, CABI Compendium.

[2]  L. Edsman,et al.  Money Kills Native Ecosystems: European Crayfish as an Example , 2021, Frontiers in Ecology and Evolution.

[3]  C. Jackson,et al.  Tracing the origin of the crayfish plague pathogen, Aphanomyces astaci, to the Southeastern United States , 2021, Scientific Reports.

[4]  L. Pârvulescu,et al.  The spatial distribution ofAphanomyces astacigenotypes across Europe: introducing the first data from Ukraine , 2020 .

[5]  R. Kortet,et al.  The signal crayfish (Pacifastacus leniusculus) in Lake Tahoe (USA) hosts multiple Aphanomyces species. , 2019, Journal of invertebrate pathology.

[6]  A. Schrimpf,et al.  The dimension of biological change caused by autotriploidy: A meta-analysis with triploid crayfish Procambarus virginalis and its diploid parent Procambarus fallax , 2019, Zoologischer Anzeiger.

[7]  A. Schrimpf,et al.  Invasive rusty crayfish (Faxonius rusticus) populations in North America are infected with the crayfish plague disease agent (Aphanomyces astaci) , 2019, Freshwater Science.

[8]  S. Viljamaa-Dirks,et al.  A tentative new species Aphanomyces fennicus sp. nov. interferes with molecular diagnostic methods for crayfish plague. , 2019, Journal of fish diseases.

[9]  F. Lyko,et al.  Ecological plasticity and commercial impact of invasive marbled crayfish populations in Madagascar , 2019, BMC Ecology.

[10]  A. Schrimpf,et al.  Morphological characterization and genotyping of the marbled crayfish and new evidence on its origin. , 2018, Zootaxa.

[11]  A. Schrimpf,et al.  Hidden sites in the distribution of the crayfish plague pathogen Aphanomyces astaci in Eastern Europe: Relicts of genetic groups from older outbreaks? , 2018, Journal of invertebrate pathology.

[12]  A. Schrimpf,et al.  MtDNA allows the sensitive detection and haplotyping of the crayfish plague disease agent Aphanomyces astaci showing clues about its origin and migration , 2018, Parasitology.

[13]  G. Vogt Investigating the genetic and epigenetic basis of big biological questions with the parthenogenetic marbled crayfish: A review and perspectives , 2018, Journal of Biosciences.

[14]  F. Lyko The marbled crayfish (Decapoda: Cambaridae) represents an independent new species. , 2017, Zootaxa.

[15]  H. Kokko,et al.  Aphanomyces astaci isolate from latently infected stone crayfish (Austropotamobius torrentium) population is virulent. , 2017, Journal of invertebrate pathology.

[16]  J. Diéguez-Uribeondo,et al.  Resistance to the crayfish plague, Aphanomyces astaci (Oomycota) in the endangered freshwater crayfish species, Austropotamobius pallipes , 2017, PloS one.

[17]  A. Petrusek,et al.  Predictions of marbled crayfish establishment in conurbations fulfilled: Evidences from the Czech Republic , 2016, Biologia.

[18]  A. Virtala,et al.  Variation in the hyphal growth rate and the virulence of two genotypes of the crayfish plague organism Aphanomyces astaci. , 2016, Journal of fish diseases.

[19]  G. Klobučar,et al.  Prevalence of the pathogen Aphanomyces astaci in freshwater crayfish populations in Croatia. , 2016, Diseases of aquatic organisms.

[20]  A. Mrugała,et al.  Expansion of the marbled crayfish in Slovakia: beginning of an invasion in the Danube catchment? , 2016 .

[21]  A. Kouba,et al.  Hardy exotics species in temperate zone: can “warm water” crayfish invaders establish regardless of low temperatures? , 2015, Scientific Reports.

[22]  A. Mrugała,et al.  Effect of experimental exposure to differently virulent Aphanomyces astaci strains on the immune response of the noble crayfish Astacus astacus. , 2015, Journal of invertebrate pathology.

[23]  A. Mrugała,et al.  Hosts and transmission of the crayfish plague pathogen Aphanomyces astaci: a review. , 2015, Journal of fish diseases.

[24]  A. Mrugała,et al.  Trade of ornamental crayfish in Europe as a possible introduction pathway for important crustacean diseases: crayfish plague and white spot syndrome , 2015, Biological Invasions.

[25]  M. Rantala,et al.  Variation in Resistance to the Invasive Crayfish Plague and Immune Defence in the Native Noble Crayfish , 2014 .

[26]  David A. Strand,et al.  The crayfish plague pathogen can infect freshwater-inhabiting crabs , 2014 .

[27]  L. Edsman,et al.  Population collapses in introduced non-indigenous crayfish , 2014, Biological Invasions.

[28]  V. Jenčič,et al.  Aphanomyces astaci in wild crayfish populations in Slovenia: first report of persistent infection in a stone crayfish Austropotamobius torrentium population. , 2013, Diseases of aquatic organisms.

[29]  A. Vainikka,et al.  Differing virulence of Aphanomyces astaci isolates and elevated resistance of noble crayfish Astacus astacus against crayfish plague. , 2012, Diseases of aquatic organisms.

[30]  C. Chucholl,et al.  The clones are coming - strong increase in marmorkrebs [Procambarus fallax (Hagen, 1870) f. virginalis] records from Europe. , 2012 .

[31]  A. Schrimpf,et al.  Crayfish plague pathogen detected in the Danube Delta - a potential threat to freshwater biodiversity in southeastern Europe. , 2012 .

[32]  H. Kokko,et al.  The diversity of the pathogenic Oomycete (Aphanomyces astaci) chitinase genes within the genotypes indicate adaptation to its hosts. , 2012, Fungal genetics and biology : FG & B.

[33]  C. Hauton The scope of the crustacean immune system for disease control. , 2012, Journal of invertebrate pathology.

[34]  A. Vainikka,et al.  Latent crayfish plague (Aphanomyces astaci) infection in a robust wild noble crayfish (Astacus astacus) population , 2011 .

[35]  A. Petrusek,et al.  Spiny-cheek crayfish Orconectes limosus carry a novel genotype of the crayfish plague pathogen Aphanomyces astaci. , 2011, Journal of invertebrate pathology.

[36]  H. Kokko,et al.  Peracetic acid (PAA) treatment is an effective disinfectant against crayfish plague (Aphanomyces astaci) spores in aquaculture , 2011 .

[37]  G. Scholtz,et al.  The first record of the parthenogenetic Marmorkrebs (Decapoda, Astacida, Cambaridae) in the wild in Saxony (Germany) raises the question of its actual threat to European freshwater ecosystems. , 2010 .

[38]  T. Tengs,et al.  A quantitative TaqMan MGB real-time polymerase chain reaction based assay for detection of the causative agent of crayfish plague Aphanomyces astaci. , 2009, Veterinary microbiology.

[39]  L. Cerenius,et al.  Aphanomyces astaci and Crustaceans , 2008 .

[40]  U. Hopp,et al.  Ontogeny of the Marmorkrebs (marbled crayfish): a parthenogenetic crayfish with unknown origin and phylogenetic position. , 2005, Journal of experimental zoology. Part A, Comparative experimental biology.

[41]  D. Alderman,et al.  Geographical spread of bacterial and fungal diseases of crustaceans. , 1996, Revue scientifique et technique.

[42]  L. Cerenius,et al.  Physiological adaptation of an Aphanomyces astaci strain isolated from the freshwater crayfish Procambarus clarkii , 1995 .

[43]  L. Cerenius,et al.  Analysis of genetic diversity in the crayfish plague fungus, Aphanomyces astaci, by random amplification of polymorphic DNA , 1994 .

[44]  K. Söderhäll,et al.  Psorospermium haeckeli and its interaction with the crayfish defence system , 1993 .

[45]  E. Soylu,et al.  Identification of the pathogenic fungus causing destruction to Turkish crayfish stocks () , 1989 .

[46]  T. Unestam,et al.  On the Adaptation of Aphanomyces astaci as a Parasite , 1969 .

[47]  T. Unestam Studies on the Crayfish Plague Fungus Aphanomyces astaci , 1966 .

[48]  H. Kokko,et al.  Narrow-clawed crayfish in Finland: Aphanomyces astaci resistance and genetic relationship to other selected European and Asian populations , 2020, Knowledge & Management of Aquatic Ecosystems.

[49]  A. Weiperth,et al.  Hungary: a European hotspot of non-native crayfish biodiversity , 2020, Knowledge & Management of Aquatic Ecosystems.

[50]  R. Gross,et al.  First record of an established marbled crayfish Procambarus virginalis (Lyko, 2017) population in Estonia , 2019, BioInvasions Records.

[51]  H. Kokko,et al.  Observations of crayfish plague infections in commercially important narrow-clawed crayfish populations in Turkey , 2018 .

[52]  A. Petrusek,et al.  Slovak section of the Danube has its well-established breeding ground of marbled crayfish Procambarus fallax f. virginalis , 2017 .

[53]  L. Pârvulescu,et al.  First established population of marbled crayfish Procambarus fallax (Hagen, 1870) f. virginalis (Decapoda, Cambaridae) in Romania , 2017 .

[54]  A. Mrugała,et al.  May Cherax destructor contribute to Aphanomyces astaci spread in Central Europe , 2016 .

[55]  R. Kortet,et al.  Invasive Crayfish and Their Invasive Diseases in Europe with the Focus on the Virulence Evolution of the Crayfish Plague 8 , 2015 .

[56]  R. Kortet,et al.  The signal crayfish is vulnerable to both the As and the PsI-isolates of the crayfish plague , 2014 .

[57]  A. Vainikka,et al.  Crayfish plague dilemma: how to be a courteous killer? , 2014 .

[58]  A. Vainikka,et al.  Dose-dependent mortality of the noble crayfish (Astacus astacus) to different strains of the crayfish plague (Aphanomyces astaci). , 2014, Journal of invertebrate pathology.

[59]  A. Schrimpf,et al.  First evidence of crayfish plague agent in populations of the marbled crayfish (Procambarus fallax forma virginalis) , 2014 .

[60]  A. Petrusek,et al.  Continental-wide distribution of crayfish species in Europe: update and maps , 2014 .

[61]  R. Kortet,et al.  Aphanomyces astaci PsI-genotype isolates from different Finnish signal crayfish stocks show variation in their virulence but still kill fast , 2013 .

[62]  C. Chucholl Invaders for sale: trade and determinants of introduction of ornamental freshwater crayfish , 2012, Biological Invasions.

[63]  P. Vennerström,et al.  Persistent infection by crayfish plague Aphanomyces astaci in a noble crayfish population – a case report , 2011 .

[64]  G. Rasool,et al.  ANALYSIS OF GENETIC DIVERSITY IN , 2008 .

[65]  Michael Marten,et al.  Der Marmorkrebs (Cambaridae, Decapoda) in Deutsch­ land - ein weiteres Neozoon im Einzugsgebiet des Rheins The Marbled Crayfish (Cambaridae, Decapoda) in Germany - another Neozoon in the River Rhine encatchment , 2004 .

[66]  G. Vogt,et al.  Ecology: Parthenogenesis in an outsider crayfish , 2003, Nature.

[67]  Y. Benjamini,et al.  Controlling the false discovery rate: a practical and powerful approach to multiple testing , 1995 .

[68]  T. Unestam,et al.  Blood reactions in vitro in crayfish against a fungal parasite , 1972 .

[69]  D. W. Weiss,et al.  The host-parasite relationship between freshwater crayfish and the crayfish disease fungus Aphanomyces astaci: responses to infection by a susceptible and a resistant species. , 1970, Journal of general microbiology.