Microsatellite markers for direct genotyping of the crayfish plague pathogen Aphanomyces astaci (Oomycetes) from infected host tissues

Frédéric Grandjean, Trude Vrålstad, Javier Diéguez-Uribeondo, Mišel Jelić, Joa Mangombi, Carine Delaunay, Lenka Filipová, Svetlana Rezinciuc, Eva Kozubíková-Balcarová, Daniel Guyonnet, Satu Viljamaa-Dirks, Adam Petrusek

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Aphanomyces astaci is an invasive pathogenic oomycete responsible for the crayfish plague, a disease that has devastated European freshwater crayfish. So far, five genotype groups of this pathogen have been identified by applying random amplified polymorphic DNA analysis on axenic cultures. To allow genotyping of A. astaci in host tissue samples, we have developed co-dominant microsatellite markers for this pathogen, tested them on pure cultures of all genotype groups, and subsequently evaluated their use on tissues of (1) natural A. astaci carriers, i.e., North American crayfish species, and (2) A. astaci-infected indigenous European species from crayfish plague outbreaks. Out of over 200 potential loci containing simple sequence repeat (SSR) motifs identified by 454 pyrosequencing of SSR-enriched library, we tested 25 loci with highest number of repeats, and finally selected nine that allow unambiguous separation of all known RAPD-defined genotype groups of A. astaci from axenic cultures. Using these markers, we were able to characterize A. astaci strains from DNA isolates from infected crayfish tissues when crayfish had a moderate to high agent level according to quantitative PCR analyses. The results support the hypothesis that different North American crayfish hosts carry different genotype groups of the pathogen, and confirm that multiple genotype groups, including the one originally introduced to Europe in the 19th century, cause crayfish plague outbreaks in Central Europe. So far undocumented A. astaci genotype seems to have caused one of the analysed outbreaks from the Czech Republic. The newly developed culture-independent approach allowing direct genotyping of this pathogen in both axenic cultures and mixed genome samples opens new possibilities in studies of crayfish plague pathogen distribution, diversity and epidemiology.

Original languageEnglish (US)
Pages (from-to)317-324
Number of pages8
JournalVeterinary Microbiology
Volume170
Issue number3-4
DOIs
StatePublished - Jan 1 2014

Fingerprint

Aphanomyces
Aphanomyces astaci
Oomycetes
Astacoidea
Plague
plague
crayfish
Microsatellite Repeats
genotyping
microsatellite repeats
pathogens
Genotype
Axenic Culture
axenic culture
genotype
Disease Outbreaks
tissues
random amplified polymorphic DNA technique
loci
Czech Republic

All Science Journal Classification (ASJC) codes

  • Microbiology
  • veterinary(all)

Cite this

Grandjean, F., Vrålstad, T., Diéguez-Uribeondo, J., Jelić, M., Mangombi, J., Delaunay, C., ... Petrusek, A. (2014). Microsatellite markers for direct genotyping of the crayfish plague pathogen Aphanomyces astaci (Oomycetes) from infected host tissues. Veterinary Microbiology, 170(3-4), 317-324. https://doi.org/10.1016/j.vetmic.2014.02.020

Microsatellite markers for direct genotyping of the crayfish plague pathogen Aphanomyces astaci (Oomycetes) from infected host tissues. / Grandjean, Frédéric; Vrålstad, Trude; Diéguez-Uribeondo, Javier; Jelić, Mišel; Mangombi, Joa; Delaunay, Carine; Filipová, Lenka; Rezinciuc, Svetlana; Kozubíková-Balcarová, Eva; Guyonnet, Daniel; Viljamaa-Dirks, Satu; Petrusek, Adam.

In: Veterinary Microbiology, Vol. 170, No. 3-4, 01.01.2014, p. 317-324.

Research output: Contribution to journalArticle

Grandjean, F, Vrålstad, T, Diéguez-Uribeondo, J, Jelić, M, Mangombi, J, Delaunay, C, Filipová, L, Rezinciuc, S, Kozubíková-Balcarová, E, Guyonnet, D, Viljamaa-Dirks, S & Petrusek, A 2014, 'Microsatellite markers for direct genotyping of the crayfish plague pathogen Aphanomyces astaci (Oomycetes) from infected host tissues', Veterinary Microbiology, vol. 170, no. 3-4, pp. 317-324. https://doi.org/10.1016/j.vetmic.2014.02.020
Grandjean, Frédéric ; Vrålstad, Trude ; Diéguez-Uribeondo, Javier ; Jelić, Mišel ; Mangombi, Joa ; Delaunay, Carine ; Filipová, Lenka ; Rezinciuc, Svetlana ; Kozubíková-Balcarová, Eva ; Guyonnet, Daniel ; Viljamaa-Dirks, Satu ; Petrusek, Adam. / Microsatellite markers for direct genotyping of the crayfish plague pathogen Aphanomyces astaci (Oomycetes) from infected host tissues. In: Veterinary Microbiology. 2014 ; Vol. 170, No. 3-4. pp. 317-324.
@article{5a77cd9988d0431eaa83621fa77f01b1,
title = "Microsatellite markers for direct genotyping of the crayfish plague pathogen Aphanomyces astaci (Oomycetes) from infected host tissues",
abstract = "Aphanomyces astaci is an invasive pathogenic oomycete responsible for the crayfish plague, a disease that has devastated European freshwater crayfish. So far, five genotype groups of this pathogen have been identified by applying random amplified polymorphic DNA analysis on axenic cultures. To allow genotyping of A. astaci in host tissue samples, we have developed co-dominant microsatellite markers for this pathogen, tested them on pure cultures of all genotype groups, and subsequently evaluated their use on tissues of (1) natural A. astaci carriers, i.e., North American crayfish species, and (2) A. astaci-infected indigenous European species from crayfish plague outbreaks. Out of over 200 potential loci containing simple sequence repeat (SSR) motifs identified by 454 pyrosequencing of SSR-enriched library, we tested 25 loci with highest number of repeats, and finally selected nine that allow unambiguous separation of all known RAPD-defined genotype groups of A. astaci from axenic cultures. Using these markers, we were able to characterize A. astaci strains from DNA isolates from infected crayfish tissues when crayfish had a moderate to high agent level according to quantitative PCR analyses. The results support the hypothesis that different North American crayfish hosts carry different genotype groups of the pathogen, and confirm that multiple genotype groups, including the one originally introduced to Europe in the 19th century, cause crayfish plague outbreaks in Central Europe. So far undocumented A. astaci genotype seems to have caused one of the analysed outbreaks from the Czech Republic. The newly developed culture-independent approach allowing direct genotyping of this pathogen in both axenic cultures and mixed genome samples opens new possibilities in studies of crayfish plague pathogen distribution, diversity and epidemiology.",
author = "Fr{\'e}d{\'e}ric Grandjean and Trude Vr{\aa}lstad and Javier Di{\'e}guez-Uribeondo and Mišel Jelić and Joa Mangombi and Carine Delaunay and Lenka Filipov{\'a} and Svetlana Rezinciuc and Eva Kozub{\'i}kov{\'a}-Balcarov{\'a} and Daniel Guyonnet and Satu Viljamaa-Dirks and Adam Petrusek",
year = "2014",
month = "1",
day = "1",
doi = "10.1016/j.vetmic.2014.02.020",
language = "English (US)",
volume = "170",
pages = "317--324",
journal = "Veterinary Microbiology",
issn = "0378-1135",
publisher = "Elsevier",
number = "3-4",

}

TY - JOUR

T1 - Microsatellite markers for direct genotyping of the crayfish plague pathogen Aphanomyces astaci (Oomycetes) from infected host tissues

AU - Grandjean, Frédéric

AU - Vrålstad, Trude

AU - Diéguez-Uribeondo, Javier

AU - Jelić, Mišel

AU - Mangombi, Joa

AU - Delaunay, Carine

AU - Filipová, Lenka

AU - Rezinciuc, Svetlana

AU - Kozubíková-Balcarová, Eva

AU - Guyonnet, Daniel

AU - Viljamaa-Dirks, Satu

AU - Petrusek, Adam

PY - 2014/1/1

Y1 - 2014/1/1

N2 - Aphanomyces astaci is an invasive pathogenic oomycete responsible for the crayfish plague, a disease that has devastated European freshwater crayfish. So far, five genotype groups of this pathogen have been identified by applying random amplified polymorphic DNA analysis on axenic cultures. To allow genotyping of A. astaci in host tissue samples, we have developed co-dominant microsatellite markers for this pathogen, tested them on pure cultures of all genotype groups, and subsequently evaluated their use on tissues of (1) natural A. astaci carriers, i.e., North American crayfish species, and (2) A. astaci-infected indigenous European species from crayfish plague outbreaks. Out of over 200 potential loci containing simple sequence repeat (SSR) motifs identified by 454 pyrosequencing of SSR-enriched library, we tested 25 loci with highest number of repeats, and finally selected nine that allow unambiguous separation of all known RAPD-defined genotype groups of A. astaci from axenic cultures. Using these markers, we were able to characterize A. astaci strains from DNA isolates from infected crayfish tissues when crayfish had a moderate to high agent level according to quantitative PCR analyses. The results support the hypothesis that different North American crayfish hosts carry different genotype groups of the pathogen, and confirm that multiple genotype groups, including the one originally introduced to Europe in the 19th century, cause crayfish plague outbreaks in Central Europe. So far undocumented A. astaci genotype seems to have caused one of the analysed outbreaks from the Czech Republic. The newly developed culture-independent approach allowing direct genotyping of this pathogen in both axenic cultures and mixed genome samples opens new possibilities in studies of crayfish plague pathogen distribution, diversity and epidemiology.

AB - Aphanomyces astaci is an invasive pathogenic oomycete responsible for the crayfish plague, a disease that has devastated European freshwater crayfish. So far, five genotype groups of this pathogen have been identified by applying random amplified polymorphic DNA analysis on axenic cultures. To allow genotyping of A. astaci in host tissue samples, we have developed co-dominant microsatellite markers for this pathogen, tested them on pure cultures of all genotype groups, and subsequently evaluated their use on tissues of (1) natural A. astaci carriers, i.e., North American crayfish species, and (2) A. astaci-infected indigenous European species from crayfish plague outbreaks. Out of over 200 potential loci containing simple sequence repeat (SSR) motifs identified by 454 pyrosequencing of SSR-enriched library, we tested 25 loci with highest number of repeats, and finally selected nine that allow unambiguous separation of all known RAPD-defined genotype groups of A. astaci from axenic cultures. Using these markers, we were able to characterize A. astaci strains from DNA isolates from infected crayfish tissues when crayfish had a moderate to high agent level according to quantitative PCR analyses. The results support the hypothesis that different North American crayfish hosts carry different genotype groups of the pathogen, and confirm that multiple genotype groups, including the one originally introduced to Europe in the 19th century, cause crayfish plague outbreaks in Central Europe. So far undocumented A. astaci genotype seems to have caused one of the analysed outbreaks from the Czech Republic. The newly developed culture-independent approach allowing direct genotyping of this pathogen in both axenic cultures and mixed genome samples opens new possibilities in studies of crayfish plague pathogen distribution, diversity and epidemiology.

UR - http://www.scopus.com/inward/record.url?scp=84898470749&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84898470749&partnerID=8YFLogxK

U2 - 10.1016/j.vetmic.2014.02.020

DO - 10.1016/j.vetmic.2014.02.020

M3 - Article

VL - 170

SP - 317

EP - 324

JO - Veterinary Microbiology

JF - Veterinary Microbiology

SN - 0378-1135

IS - 3-4

ER -