The Experts below are selected from a list of 720 Experts worldwide ranked by ideXlab platform
Adam Petrusek - One of the best experts on this subject based on the ideXlab platform.
-
real time pcr assays for rapid identification of common Aphanomyces Astaci genotypes
Frontiers in Ecology and Evolution, 2021Co-Authors: Marco Di Domenico, Agata Mrugala, Cesare Camma, Riccardo Caprioli, Valentina Curini, Carla Giansante, Michaela Mojžisova, Adam PetrusekAbstract:Crayfish plague is an acute disease caused by the oomycete Aphanomyces Astaci that seriously impacts the populations of European native crayfish species. The introduction of non-indigenous crayfish of North American origin and their wide distribution across Europe has largely contributed to further spread of crayfish plague in areas populated by indigenous species. Tracking A. Astaci genotypes may thus represent a useful tool to investigate the crayfish plague natural history in its European range, as well as the sources and introduction pathways of the pathogen. In this study we describe the development of genotype-specific TaqMan real-time PCR assays to distinguish the five genotype groups of A. Astaci previously defined by their distinct RAPD patterns. The method was evaluated using DNA extracts from pure A. Astaci cultures representing the known genotype groups, and from A. Astaci-positive crayfish clinical samples collected during crayfish plague outbreaks recently occurred in Central Italy and Czechia. Group A, B, D and E assays are extremely specific, while group C assay (not yet documented from crayfish plague outbreaks) showed cross-reactivity with A. fennicus. The unusual A. Astaci genotype Up documented from crayfish plague outbreaks in Czechia is detected by the group B real-time PCR, while the genotype rust1 described in USA from Faxonius rusticus is detected by the group C assay. Analysis of additional markers (such as sequencing of the nuclear internal transcribed spacer or mitochondrial ribosomal subunits) may complement these cases when the real-time PCR-based genotyping is not conclusive. Overall, the method represents a robust and fast tool to genotype A. Astaci during outbreaks or latent infections.
-
Aphanomyces Astaci genotypes involved in recent crayfish plague outbreaks in central italy
Diseases of Aquatic Organisms, 2018Co-Authors: Riccardo Caprioli, Agata Mrugala, Cesare Camma, Marco Di Domenico, Valentina Curini, Carla Giansante, Adam PetrusekAbstract:The oomycete Aphanomyces Astaci is the causative agent of crayfish plague in native European freshwater crayfish. Molecular analyses showed that several distinct genotype groups of this pathogen, apparently associated with different original host taxa, are present in Europe. Tracking their distribution may contribute to understanding the introduction pathways of A. Astaci. We used microsatellite markers to genotype the pathogen strains involved in 7 mass mortalities of the endangered indigenous crayfish Austropotamobius pallipes that occurred between 2009 and 2016 in the Abruzzi and Molise regions, central Italy. Three A. Astaci genotype groups (A, B, and D, with the latter represented by 2 distinct multilocus genotypes) were identified, suggesting the existence of multiple infection sources even in a relatively small area. Most crayfish plague episodes were due to genotype groups associated with the North American host species Pacifastacus leniusculus and Procambarus clarkii, although these crayfish are not widespread in the study area. A. Astaci genotype group A was detected not only in crayfish plague outbreaks but also in apparently healthy Astacus leptodactylus imported for human consumption from Armenia and kept alive in an aquaculture facility. Imports of chronically infected A. leptodactylus from Armenia, Turkey, and possibly Eastern Europe are an underestimated introduction pathway for A. Astaci. Although we cannot exclude the presence of latently infected native populations of A. pallipes in the region, A. Astaci infections in legally imported crayfish species considered vulnerable to crayfish plague may represent further reservoirs of A. Astaci; this should be reflected in the policies regulating the trade of live crayfish.
-
recent acute crayfish mortality reveals Aphanomyces Astaci presence in bosnia and herzegovina
Journal of Invertebrate Pathology, 2017Co-Authors: Agata Mrugala, Adam Petrusek, Radek Sanda, Dario Maric, Jasna VukicAbstract:Although the introduction of the crayfish plague pathogen Aphanomyces Astaci to Europe is responsible for substantial declines in native crayfish populations throughout the whole continent, its presence has never been officially confirmed in many European regions, including most of the Balkan Peninsula. We demonstrate that the recent crayfish mortality observed in Bosnia and Herzegovina (Mostarsko blato karst field, Neretva river drainage) was caused by A. Astaci. The causative strain is known only from European crayfish, indicating that A. Astaci poses a threat to native species in this region, even in the absence of its main vectors, the North American crayfish.
-
apparent interspecific transmission of Aphanomyces Astaci from invasive signal to virile crayfish in a sympatric wild population
Journal of Invertebrate Pathology, 2017Co-Authors: Joanna James, Agata Mrugala, Birgit Oidtmann, Adam Petrusek, Joanne CableAbstract:The crayfish plague pathogen (Aphanomyces Astaci) causes mass mortalities of European crayfish when transmitted from its original North American crayfish hosts. Little is known, however, about interspecific transmission of the pathogen between different American crayfish species, although evidence from trade of ornamental crayfish suggests this may happen in captivity. We screened signal and virile crayfish for A. Astaci at allopatric and sympatric sites in a UK river. Whilst the pathogen was detected in signal crayfish from both sites, infected virile crayfish were only found in sympatry. Genotyping of A. Astaci from virile crayfish suggested the presence of a strain related to one infecting British signal crayfish. We conclude that virile crayfish likely contracted A. Astaci interspecifically from infected signal crayfish. Interspecific transmission of A. Astaci strains differing in virulence between American carrier species may influence the spread of this pathogen in open waters with potential exacerbated effects on native European crayfish.
-
the prevalence of Aphanomyces Astaci in invasive signal crayfish from the uk and implications for native crayfish conservation
Parasitology, 2017Co-Authors: Joanna James, Agata Mrugala, S Nutbeamtuffs, N Vinuelarodriguez, Adam Petrusek, Joanne Cable, Birgit OidtmannAbstract:The crayfish plague agent, Aphanomyces Astaci, has spread throughout Europe, causing a significant decline in native European crayfish. The introduction and dissemination of this pathogen is attributed to the spread of invasive North American crayfish, which can act as carriers for A. Astaci. As native European crayfish often succumb to infection with A. Astaci, determining the prevalence of this pathogen in non-native crayfish is vital to prioritize native crayfish populations for managed translocation. In the current study, 23 populations of invasive signal crayfish (Pacifastacus leniusculus) from the UK were tested for A. Astaci presence using quantitative PCR. Altogether, 13 out of 23 (56·5%) populations were found to be infected, and pathogen prevalence within infected sites varied from 3 to 80%. Microsatellite pathogen genotyping revealed that at least one UK signal crayfish population was infected with the A. Astaci genotype group B, known to include virulent strains. Based on recent crayfish distribution records and the average rate of signal crayfish population dispersal, we identified one native white-clawed crayfish (Austropotamobius pallipes) population predicted to come into contact with infected signal crayfish within 5 years. This population should be considered as a priority for translocation.
Birgit Oidtmann - One of the best experts on this subject based on the ideXlab platform.
-
improved method for genotyping the causative agent of crayfish plague Aphanomyces Astaci based on mitochondrial dna
Parasitology, 2019Co-Authors: Diana Minardi, Birgit Oidtmann, David J Studholme, Tobia Pretto, Mark Van Der GiezenAbstract: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.
-
new genotyping method for the causative agent of crayfish plague Aphanomyces Astaci based on whole genome data
Journal of Invertebrate Pathology, 2018Co-Authors: David J Studholme, Diana Minardi, Tobia Pretto, Mark Van Der Giezen, Birgit OidtmannAbstract:Abstract The oomycete Aphanomyces Astaci causes crayfish plague, the most important disease of European freshwater crayfish species. Presumably introduced into Europe 150 years ago with the import of North American crayfish, A. Astaci is highly pathogenic to European crayfish species. Five genotypes (A, B, C, D, and E) have been defined based on random amplified polymorphic DNA analysis (RAPD-PCR) from A. Astaci pure cultures. The distinction of genotypes is an essential tool to conduct molecular epidemiological studies on crayfish plague and it has been used to clarify and better understand the history and spread of this disease in Europe. Whereas RAPD-PCR requires DNA from pure culture isolates, the development of genotyping tools that can be applied to DNA extracted from clinical samples allows a much wider application of genotyping studies, including revisiting historic samples. In this study, we present a new approach that adds to currently available methods for genotyping A. Astaci strains directly from clinical crayfish samples. Whole-genome sequencing of A. Astaci strains representing all currently known genotypes was employed, genomic regions unique to the respective genotype identified, and a PCR-based genotyping assay designed, which focuses on the presence/absence of PCR product after amplification with the genotype-specific primers. Our diagnostic methodology was tested using DNA extracts from pure A. Astaci cultures, other Aphanomyces species and additional oomycetes, samples from a recent Italian crayfish plague outbreak and additional historical samples available in the Centre for Environment, Fisheries and Aquaculture Science laboratory. The new markers were reliable for pure culture and clinical samples from a recent outbreak and successfully discriminated genotype A, B, D, and E. The marker for genotype C required an additional sequencing step of the generated PCR product to confirm genotype.
-
apparent interspecific transmission of Aphanomyces Astaci from invasive signal to virile crayfish in a sympatric wild population
Journal of Invertebrate Pathology, 2017Co-Authors: Joanna James, Agata Mrugala, Birgit Oidtmann, Adam Petrusek, Joanne CableAbstract:The crayfish plague pathogen (Aphanomyces Astaci) causes mass mortalities of European crayfish when transmitted from its original North American crayfish hosts. Little is known, however, about interspecific transmission of the pathogen between different American crayfish species, although evidence from trade of ornamental crayfish suggests this may happen in captivity. We screened signal and virile crayfish for A. Astaci at allopatric and sympatric sites in a UK river. Whilst the pathogen was detected in signal crayfish from both sites, infected virile crayfish were only found in sympatry. Genotyping of A. Astaci from virile crayfish suggested the presence of a strain related to one infecting British signal crayfish. We conclude that virile crayfish likely contracted A. Astaci interspecifically from infected signal crayfish. Interspecific transmission of A. Astaci strains differing in virulence between American carrier species may influence the spread of this pathogen in open waters with potential exacerbated effects on native European crayfish.
-
the prevalence of Aphanomyces Astaci in invasive signal crayfish from the uk and implications for native crayfish conservation
Parasitology, 2017Co-Authors: Joanna James, Agata Mrugala, S Nutbeamtuffs, N Vinuelarodriguez, Adam Petrusek, Joanne Cable, Birgit OidtmannAbstract:The crayfish plague agent, Aphanomyces Astaci, has spread throughout Europe, causing a significant decline in native European crayfish. The introduction and dissemination of this pathogen is attributed to the spread of invasive North American crayfish, which can act as carriers for A. Astaci. As native European crayfish often succumb to infection with A. Astaci, determining the prevalence of this pathogen in non-native crayfish is vital to prioritize native crayfish populations for managed translocation. In the current study, 23 populations of invasive signal crayfish (Pacifastacus leniusculus) from the UK were tested for A. Astaci presence using quantitative PCR. Altogether, 13 out of 23 (56·5%) populations were found to be infected, and pathogen prevalence within infected sites varied from 3 to 80%. Microsatellite pathogen genotyping revealed that at least one UK signal crayfish population was infected with the A. Astaci genotype group B, known to include virulent strains. Based on recent crayfish distribution records and the average rate of signal crayfish population dispersal, we identified one native white-clawed crayfish (Austropotamobius pallipes) population predicted to come into contact with infected signal crayfish within 5 years. This population should be considered as a priority for translocation.
-
a comparative study of molecular diagnostic methods designed to detect the crayfish plague pathogen Aphanomyces Astaci
Veterinary Microbiology, 2011Co-Authors: Stephen W Tuffs, Birgit OidtmannAbstract:Abstract Crayfish plague is the most important disease of freshwater crayfish with a significant impact on European species. We compared the analytical test sensitivity and specificity of three published PCR assays for the detection of Aphanomyces Astaci , the causative agent of crayfish plague: a conventional PCR assay targeting the ITS region and two TaqMan ® real time assays, targeting either the ITS region or the chitinase gene. We also tested a variation of the conventional assay, by changing one of the primers. Test specificity was assessed using DNA from a range of A. Astaci strains and an array of closely related Oomycetes, host tissue and DNA from other organisms that may be present in a diagnostic sample. Sensitivity was assessed using genomic A. Astaci DNA from mycelium and zoospores. All assays were found to be of good to excellent sensitivity with levels of detection ranging from 1 (real time assay targeting the ITS region), over 10 (conventional PCR) to 100 zoospores (real time assay targeting the chitinase gene). All three published assays were also specific for A. Astaci and did not cross-react with any other test organisms included in this study. The tested variation of the conventional PCR assay with a changed forward primer led to amplification of some non A. Astaci DNA. Advantages and disadvantages, including suitable application are discussed for each assay.
Jenny Makkonen - One of the best experts on this subject based on the ideXlab platform.
-
controlled infection experiment with Aphanomyces Astaci provides additional evidence for latent infections and resistance in freshwater crayfish
Frontiers in Ecology and Evolution, 2021Co-Authors: Caterina Francesconi, Harri Kokko, Japo Jussila, Jenny Makkonen, Anne Schrimpf, Kathrin TheissingerAbstract: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 towards 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.
-
comparative transcriptome analysis of noble crayfish and marbled crayfish immune response to Aphanomyces Astaci challenges
bioRxiv, 2021Co-Authors: L L Bostjancic, Japo Jussila, Jenny Makkonen, C Francesconi, C Rutz, L Hoffbeck, Laetitia Poidevin, Arnaud Kress, B Feldmeyer, Miklos BalintAbstract:Introduction of invasive North American crayfish species and their pathogen Aphanomyces Astaci has significantly contributed to the decline of European freshwater crayfish populations. In this study, noble crayfish, a susceptible native European species, and marbled crayfish, an invasive disease-resistant species, were challenged with haplogroup A (low virulence) and haplogroup B (high virulence) strain of A. Astaci. Hepatopancreatic tissue was isolated 3 and 21 days post-challenge. Our results revealed strong up-regulation in expression levels of the prophenoloxidase cascade immune-related genes in the haplogroup B challenged noble crayfish 3 days post-challenge. In the marbled crayfish, we observed an up-regulation of immune system relevant genes (DSCAM, AP, ALFs, CTLs and hemocyanin) 3 days post-challenge. This response highlights the marbled crayfish capability of building the immune tolerance. Furthermore, we successfully characterised several novel immune related gene groups in both crayfish species, contributing to our current understanding of crayfish immune related genes landscape. Graphical abstracta) Study species noble crayfish (Astacus astacus) in purple and marbled crayfish (Procambarus virginalis) in green challenged with the pathogen Aphanomyces Astaci haplogroup A (Hap A) strain of low virulence and haplogroup B (Hap B) strain of high virulence. b) Sampling scheme of the infection experiment: 5 individuals were taken from the experiment three- and 21-days post-challenge. From each individual, a hepatopancreas sample was taken, followed by RNA isolation and sequencing. c) De novo transcriptome assembly and annotation were conducted for each species. d) Differential gene expression analysis revealed the distinct immune response in the noble crayfish 3 days post-challenge with the Hap B strain of A. Astaci and marbled crayfish 3 days post-challenge with the Hap A strain of A. Astaci. Immune related DEGs were not present in either species 21 days post-challenge with A. Astaci. e) Noble crayfish challenged with the Hap B strain of A. Astaci were acutely infected and ultimately moribund, while the A. Astaci Hap A challenged marbled crayfish showed high resistance to the pathogen, resulting infected without any mortality. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=103 SRC="FIGDIR/small/445163v1_ufig1.gif" ALT="Figure 1"> View larger version (53K): org.highwire.dtl.DTLVardef@d1830aorg.highwire.dtl.DTLVardef@f2196forg.highwire.dtl.DTLVardef@63c8f2org.highwire.dtl.DTLVardef@11da1b6_HPS_FORMAT_FIGEXP M_FIG C_FIG
-
narrow clawed crayfish in finland Aphanomyces Astaci resistance and genetic relationship to other selected european and asian populations
Knowledge and Management of Aquatic Ecosystems, 2020Co-Authors: Japo Jussila, Harri Kokko, Ivana Maguire, Vesa Tiitinen, Jenny MakkonenAbstract:The narrow-clawed crayfish (Pontastacus leptodactylus ) is an alien species in Finland with only a few populations reported from the southeastern region during the last century. We discovered a productive population in the lake Jangynjarvi, which is upstream from the previously reported wild narrow-clawed crayfish population in that region. Preliminary studies indicated that this population is not infected with Aphanomyces Astaci . We collected narrow-clawed crayfish samples from the lake Jangynjarvi population for both infection challenge and genetic studies, in order to investigate possible A. Astaci resistance among this Finnish population and to evaluate their phylogenetic position that would enable us to speculate different scenarios of distribution pathways or origin of the population. The infection studies indicated that the narrow-clawed crayfish in this population were more resistant against A. Astaci infection (B haplogroup A. Astaci ) compared to the noble crayfish (Astacus astacus ) from the lake Rytky in North Savo, while all crayfish of both species in the B haplogroup A. Astaci challenged groups died within 58 days post-infection. Results of the phylogenetic reconstruction indicate that the lake Jangynjarvi narrow-clawed crayfish are closely related to narrow-clawed crayfish from the lake Bolshoye near Krasnoye, located on the White Sea island of Solovestky and also populations from Tyumen region, both in Russia. This could confirm previous speculations about introduction of the narrow-clawed crayfish from Russia into Finland or could indicate previous hydrological connection.
-
non destructive method for detecting Aphanomyces Astaci the causative agent of crayfish plague on the individual level
Journal of Invertebrate Pathology, 2020Co-Authors: Dora Pavic, Jenny Makkonen, Milan Cankovic, Ines Petric, Sandra Hudina, Ivana Maguire, Tomislav Vladusic, Lidija Sver, Reno Hrascan, Karla OrlicAbstract:Abstract The pathogenic oomycete Aphanomyces Astaci, transmitted mainly by invasive North American crayfish, causes the crayfish plague, a disease mostly lethal for native European crayfish. Due to its decimating effects on native crayfish populations in the last century, A. Astaci has been listed among the 100 worst invasive species. Importantly, detecting the pathogen in endangered native crayfish populations before a disease outbreak would provide a starting point in the development of effective control measures. However, current A. Astaci-detection protocols either rely on degradation-prone eDNA isolated from large volumes of water or, if focused on individual animals, include killing the crayfish. We developed a non-destructive method that detects A. Astaci DNA in the microbial biofilm associated with the cuticle of individual crayfish, without the need for destructive sampling. Efficiency of the new method was confirmed by PCR and qPCR and the obtained results were congruent with the traditional destructive sampling method. Additionally, we demonstrated the applicability of the method for A. Astaci monitoring in natural populations. We propose that the new method should be used in future monitoring of A. Astaci presence in endangered European native crayfish individuals as an alternative to eDNA-based monitoring.
-
mtdna allows the sensitive detection and haplotyping of the crayfish plague disease agent Aphanomyces Astaci showing clues about its origin and migration
Parasitology, 2018Co-Authors: Jenny Makkonen, Japo Jussila, Raine Kortet, Laura Martintorrijos, Jorn Panteleit, Nina Sophie Keller, Kathrin Theissinger, Jose Vladimir Sandovalsierra, Javier DieguezuribeondoAbstract:The oomycete Aphanomyces Astaci, the causative agent of crayfish plague, is listed as one of the 100 worst invasive species in the world, destroying the native crayfish populations throughout Eurasia. The aim of this study was to examine the potential of selected mitochondrial (mt) genes to track the diversity of the crayfish plague pathogen A. Astaci. Two sets of primers were developed to amplify the mtDNA of ribosomal rnnS and rnnL subunits. We confirmed two main lineages, with four different haplogroups and five haplotypes among 27 studied A. Astaci strains. The haplogroups detected were (1) the A-haplogroup with the a-haplotype strains originating from Orconectes sp., Pacifastacus leniusculus and Astacus astacus; (2) the B-haplogroup with the b-haplotype strains originating from the P. leniusculus; (3) the D-haplogroup with the d1 and d2-haplotypes strains originating from Procambarus clarkii; and (4) the E-haplogroup with the e-haplotype strains originating from the Orconectes limosus. The described markers are stable and reliable and the results are easily repeatable in different laboratories. The present method has high applicability as it allows the detection and characterization of the A. Astaci haplotype in acute disease outbreaks in the wild, directly from the infected crayfish tissue samples.
Japo Jussila - One of the best experts on this subject based on the ideXlab platform.
-
controlled infection experiment with Aphanomyces Astaci provides additional evidence for latent infections and resistance in freshwater crayfish
Frontiers in Ecology and Evolution, 2021Co-Authors: Caterina Francesconi, Harri Kokko, Japo Jussila, Jenny Makkonen, Anne Schrimpf, Kathrin TheissingerAbstract: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 towards 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.
-
comparative transcriptome analysis of noble crayfish and marbled crayfish immune response to Aphanomyces Astaci challenges
bioRxiv, 2021Co-Authors: L L Bostjancic, Japo Jussila, Jenny Makkonen, C Francesconi, C Rutz, L Hoffbeck, Laetitia Poidevin, Arnaud Kress, B Feldmeyer, Miklos BalintAbstract:Introduction of invasive North American crayfish species and their pathogen Aphanomyces Astaci has significantly contributed to the decline of European freshwater crayfish populations. In this study, noble crayfish, a susceptible native European species, and marbled crayfish, an invasive disease-resistant species, were challenged with haplogroup A (low virulence) and haplogroup B (high virulence) strain of A. Astaci. Hepatopancreatic tissue was isolated 3 and 21 days post-challenge. Our results revealed strong up-regulation in expression levels of the prophenoloxidase cascade immune-related genes in the haplogroup B challenged noble crayfish 3 days post-challenge. In the marbled crayfish, we observed an up-regulation of immune system relevant genes (DSCAM, AP, ALFs, CTLs and hemocyanin) 3 days post-challenge. This response highlights the marbled crayfish capability of building the immune tolerance. Furthermore, we successfully characterised several novel immune related gene groups in both crayfish species, contributing to our current understanding of crayfish immune related genes landscape. Graphical abstracta) Study species noble crayfish (Astacus astacus) in purple and marbled crayfish (Procambarus virginalis) in green challenged with the pathogen Aphanomyces Astaci haplogroup A (Hap A) strain of low virulence and haplogroup B (Hap B) strain of high virulence. b) Sampling scheme of the infection experiment: 5 individuals were taken from the experiment three- and 21-days post-challenge. From each individual, a hepatopancreas sample was taken, followed by RNA isolation and sequencing. c) De novo transcriptome assembly and annotation were conducted for each species. d) Differential gene expression analysis revealed the distinct immune response in the noble crayfish 3 days post-challenge with the Hap B strain of A. Astaci and marbled crayfish 3 days post-challenge with the Hap A strain of A. Astaci. Immune related DEGs were not present in either species 21 days post-challenge with A. Astaci. e) Noble crayfish challenged with the Hap B strain of A. Astaci were acutely infected and ultimately moribund, while the A. Astaci Hap A challenged marbled crayfish showed high resistance to the pathogen, resulting infected without any mortality. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=103 SRC="FIGDIR/small/445163v1_ufig1.gif" ALT="Figure 1"> View larger version (53K): org.highwire.dtl.DTLVardef@d1830aorg.highwire.dtl.DTLVardef@f2196forg.highwire.dtl.DTLVardef@63c8f2org.highwire.dtl.DTLVardef@11da1b6_HPS_FORMAT_FIGEXP M_FIG C_FIG
-
narrow clawed crayfish in finland Aphanomyces Astaci resistance and genetic relationship to other selected european and asian populations
Knowledge and Management of Aquatic Ecosystems, 2020Co-Authors: Japo Jussila, Harri Kokko, Ivana Maguire, Vesa Tiitinen, Jenny MakkonenAbstract:The narrow-clawed crayfish (Pontastacus leptodactylus ) is an alien species in Finland with only a few populations reported from the southeastern region during the last century. We discovered a productive population in the lake Jangynjarvi, which is upstream from the previously reported wild narrow-clawed crayfish population in that region. Preliminary studies indicated that this population is not infected with Aphanomyces Astaci . We collected narrow-clawed crayfish samples from the lake Jangynjarvi population for both infection challenge and genetic studies, in order to investigate possible A. Astaci resistance among this Finnish population and to evaluate their phylogenetic position that would enable us to speculate different scenarios of distribution pathways or origin of the population. The infection studies indicated that the narrow-clawed crayfish in this population were more resistant against A. Astaci infection (B haplogroup A. Astaci ) compared to the noble crayfish (Astacus astacus ) from the lake Rytky in North Savo, while all crayfish of both species in the B haplogroup A. Astaci challenged groups died within 58 days post-infection. Results of the phylogenetic reconstruction indicate that the lake Jangynjarvi narrow-clawed crayfish are closely related to narrow-clawed crayfish from the lake Bolshoye near Krasnoye, located on the White Sea island of Solovestky and also populations from Tyumen region, both in Russia. This could confirm previous speculations about introduction of the narrow-clawed crayfish from Russia into Finland or could indicate previous hydrological connection.
-
mtdna allows the sensitive detection and haplotyping of the crayfish plague disease agent Aphanomyces Astaci showing clues about its origin and migration
Parasitology, 2018Co-Authors: Jenny Makkonen, Japo Jussila, Raine Kortet, Laura Martintorrijos, Jorn Panteleit, Nina Sophie Keller, Kathrin Theissinger, Jose Vladimir Sandovalsierra, Javier DieguezuribeondoAbstract:The oomycete Aphanomyces Astaci, the causative agent of crayfish plague, is listed as one of the 100 worst invasive species in the world, destroying the native crayfish populations throughout Eurasia. The aim of this study was to examine the potential of selected mitochondrial (mt) genes to track the diversity of the crayfish plague pathogen A. Astaci. Two sets of primers were developed to amplify the mtDNA of ribosomal rnnS and rnnL subunits. We confirmed two main lineages, with four different haplogroups and five haplotypes among 27 studied A. Astaci strains. The haplogroups detected were (1) the A-haplogroup with the a-haplotype strains originating from Orconectes sp., Pacifastacus leniusculus and Astacus astacus; (2) the B-haplogroup with the b-haplotype strains originating from the P. leniusculus; (3) the D-haplogroup with the d1 and d2-haplotypes strains originating from Procambarus clarkii; and (4) the E-haplogroup with the e-haplotype strains originating from the Orconectes limosus. The described markers are stable and reliable and the results are easily repeatable in different laboratories. The present method has high applicability as it allows the detection and characterization of the A. Astaci haplotype in acute disease outbreaks in the wild, directly from the infected crayfish tissue samples.
-
growth survival and spore formation of the pathogenic aquatic oomycete Aphanomyces Astaci and fungus fusarium avenaceum are inhibited by zanthoxylum rhoifolium bark extracts in vitro
Fishes, 2018Co-Authors: Caterina Pagliarulo, Japo Jussila, Jenny Makkonen, Daniela Sateriale, Elisa Scioscia, Nunziatina De Tommasi, Roberta Colicchio, Chiara Pagliuca, Elena Scaglione, Paola SalvatoreAbstract:This study aimed to evaluate the in vitro activity of Zanthoxylum rhoifolium bark (Zr-b) extracts against pathogenic aquatic oomycete/fungal isolates that cause different diseases in native European crayfish resulting in an elevated mortality rate and severe economic repercussions. n-hexane, chloroform, chloroform–methanol (9:1) and methanol extracts of Zr-b were used to evaluate the antifungal activity against the strain UEF88662 of Aphanomyces Astaci (oomycete) and the strain SMM2 of Fusarium avenaceum (fungus). The anti-oomycete and antifungal activity was quantitatively evaluated by growth, survival and sporulation microbiological assays. The extracts tested demonstrated a dose-dependent inhibitory effect on oomycete and fungal growth and survival, as well as on the production of oomycete and fungal spores. This work presents alternatives for the treatment and prevention of the spreading of Aphanomyces Astaci and Fusarium avenaceum, the etiological agents of the diseases crayfish plague and brown spot disease, respectively. The antifungal properties of Zanthoxylum rhoifolium bark extracts warrant further research on their use in the prevention and treatment of both oomycete and fungal diseases. The antifungal properties of Zanthoxylum rhoifolium bark extracts, shown in vitro, indicate the possibility of their use in new therapeutic and prophylactic strategies, providing perspectives for the design of in vivo studies.
Harri Kokko - One of the best experts on this subject based on the ideXlab platform.
-
controlled infection experiment with Aphanomyces Astaci provides additional evidence for latent infections and resistance in freshwater crayfish
Frontiers in Ecology and Evolution, 2021Co-Authors: Caterina Francesconi, Harri Kokko, Japo Jussila, Jenny Makkonen, Anne Schrimpf, Kathrin TheissingerAbstract: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 towards 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.
-
narrow clawed crayfish in finland Aphanomyces Astaci resistance and genetic relationship to other selected european and asian populations
Knowledge and Management of Aquatic Ecosystems, 2020Co-Authors: Japo Jussila, Harri Kokko, Ivana Maguire, Vesa Tiitinen, Jenny MakkonenAbstract:The narrow-clawed crayfish (Pontastacus leptodactylus ) is an alien species in Finland with only a few populations reported from the southeastern region during the last century. We discovered a productive population in the lake Jangynjarvi, which is upstream from the previously reported wild narrow-clawed crayfish population in that region. Preliminary studies indicated that this population is not infected with Aphanomyces Astaci . We collected narrow-clawed crayfish samples from the lake Jangynjarvi population for both infection challenge and genetic studies, in order to investigate possible A. Astaci resistance among this Finnish population and to evaluate their phylogenetic position that would enable us to speculate different scenarios of distribution pathways or origin of the population. The infection studies indicated that the narrow-clawed crayfish in this population were more resistant against A. Astaci infection (B haplogroup A. Astaci ) compared to the noble crayfish (Astacus astacus ) from the lake Rytky in North Savo, while all crayfish of both species in the B haplogroup A. Astaci challenged groups died within 58 days post-infection. Results of the phylogenetic reconstruction indicate that the lake Jangynjarvi narrow-clawed crayfish are closely related to narrow-clawed crayfish from the lake Bolshoye near Krasnoye, located on the White Sea island of Solovestky and also populations from Tyumen region, both in Russia. This could confirm previous speculations about introduction of the narrow-clawed crayfish from Russia into Finland or could indicate previous hydrological connection.
-
Aphanomyces Astaci colonization and immune reaction in North American crayfish.
2018Co-Authors: Laura Martín-torrijos, Harri Kokko, Japo Jussila, Jenny Makkonen, Tadashi Kawai, Javier Diéguez-uribeondoAbstract:North American crayfish species showing immune responses to Aphanomyces Astaci infection. Photographs of (A, B) P. clarkii and (C, D) P. leniusculus specimens. Melanin formation, visualized as melanized patches (arrows), characterizes a strong immune response against A. Astaci infections on the (A) joints of a chela and (B) subabdominal cuticle of a P. clarkii specimen, and on the (C) ventral and (D) dorsal chela surfaces of a P. leniusculus specimen.
-
Aphanomyces Astaci isolate from latently infected stone crayfish austropotamobius torrentium population is virulent
Journal of Invertebrate Pathology, 2017Co-Authors: Japo Jussila, Jenny Makkonen, Al Vrezec, Tina Jaklic, Hobo Kukkonen, Harri KokkoAbstract:Aphanomyces Astaci infection is the cause of crayfish plague in European crayfish. Here the virulence of an A. Astaci As strain isolated from apparently healthy stone crayfish (Austropotamobius torrentium) from Slovenia was compared to that of the Psl-Puujarvi A. Astaci isolate in 3 crayfish species: noble crayfish (Astacus astacus), signal crayfish (Pacifastacus leniusculus) from Finland and stone crayfish from Slovenia. All 3 crayfish species were challenged with PsI-Puujarvi A. Astaci and succumbed to crayfish plague, with both noble crayfish and stone crayfish showing 100% mortality, while 25% of the signal crayfish died during the challenge. In comparison, the As-Slovenia A. Astaci isolate was pathogenic for noble crayfish but not for signal crayfish or stone crayfish. This finding suggests that A. Astaci virulence could be species specific and a strain from latent A. Astaci infection in one native European crayfish species could be detrimental to other native European crayfish species.
-
eroded swimmeret syndrome in female crayfish pacifastacus leniusculus associated with Aphanomyces Astaci and fusarium spp infections
Diseases of Aquatic Organisms, 2015Co-Authors: Lennart Edsman, Harri Kokko, Jenny Makkonen, Per Nystrom, Alfred Sandstrom, Marika Stenberg, Vesa Tiitinen, Japo JussilaAbstract:We describe a novel syndrome in crayfish, eroded swimmeret syndrome (ESS), affecting wild female signal crayfish Pacifastacus leniusculus. ESS causes partial or total swim- meret erosion. We observed ESS only in female signal crayfish larger than 40 mm carapace length, i.e. sexually mature and probably having carried eggs at least once. The eroded swimmerets were melanised, indicating a crayfish immune system response. We isolated Fusarium tricinctum spe- cies complex (SC), F. sambucinum SC, Saprolegnia parasitica and S. australis from the melanised tissue of the eroded swimmerets. ESS includes chronic Aphanomyces Astaci infection and a sec- ondary infection by Fusarium sp. In Sweden, we found female signal crayfish with ESS in 6 out of 11 populations with a prevalence below 1% in lakes with commercially productive signal crayfish populations and higher than 29% in lakes with documented signal crayfish population crashes. In Finland, the ESS prevalence was from 3.4 to 6.2% in a commercially productive population. None of the sampled male signal crayfish showed signs of ESS. A caging experiment indicated that females with at least 1 lost swimmeret carried on average 25% fewer fertilized eggs compared to females with intact swimmerets. ESS could significantly reduce individual female fecundity and thus could also affect fecundity at the population level. The decline in reproductive success due to ESS could be among the factors contributing to fluctuations in wild signal crayfish populations.