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Prevalence, molecular identification and genotyping of the crayfish plague pathogen, Aphanomyces astaci in major narrow-clawed crayfish (Pontastacus leptodactylus Eschscholtz, 1823) populations from Türkiye

Süleyman Akhan

Süleyman Akhan

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Akhan, Süleyman
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İfakat Tülay Çağatay

İfakat Tülay Çağatay

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Çağatay, İfakat Tülay
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Selçuk Berber

Selçuk Berber

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Berber, Selçuk
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Büşra Taştan

Büşra Taştan

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Taştan, Büşra
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Yiğit Taştan

Yiğit Taştan

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Taştan, Yiğit
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Tuba Dalar

Tuba Dalar

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Dalar, Tuba
  
Nov 06, 2024
Prevalence, molecular identification and genotyping of the crayfish plague pathogen, Aphanomyces astaci in major narrow-clawed crayfish (Pontastacus leptodactylus Eschscholtz, 1823) populations from Türkiye's Cover Image
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Published Online: Nov 06, 2024
Page range: 515 - 524
Received: May 07, 2024
Accepted: Oct 08, 2024
DOI: https://doi.org/10.2478/jvetres-2024-0061
Keywords
© 2024 Süleyman Akhan et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1.

Map of sampling stations where Turkish crayfish (Pontastacus leptodactylus) were caught to determine the prevalence of crayfish plague
Map of sampling stations where Turkish crayfish (Pontastacus leptodactylus) were caught to determine the prevalence of crayfish plague

Fig. 2.

PCR sensitivity test results for detection of Aphanomyces astaci M – ladder; N – negative control; Lanes: 1 – 50 ng; 2 – 5 ng; 3 – 500 pg; 4 – 50 pg; 5 – 5 pg; 6 – 500 fg; 7 – 50 fg; a – 30-cycle PCR; b – 40-cycle PCR
PCR sensitivity test results for detection of Aphanomyces astaci M – ladder; N – negative control; Lanes: 1 – 50 ng; 2 – 5 ng; 3 – 500 pg; 4 – 50 pg; 5 – 5 pg; 6 – 500 fg; 7 – 50 fg; a – 30-cycle PCR; b – 40-cycle PCR

Fig. 3.

PCR amplification results for Aphanomyces astaci from the tissue of Turkish crayfish symptomatic for crayfish plague a – abdominal cuticle tissue (n = 20); b – telson tissue (n = 20); c – melanised tissue (n = 20)
PCR amplification results for Aphanomyces astaci from the tissue of Turkish crayfish symptomatic for crayfish plague a – abdominal cuticle tissue (n = 20); b – telson tissue (n = 20); c – melanised tissue (n = 20)

Fig. 4.

PCR amplification results for Aphanomyces astaci from the tissue of Turkish crayfish asymptomatic for crayfish plague a – abdominal cuticle tissue (n = 20); b – telson tissue (n = 20)
PCR amplification results for Aphanomyces astaci from the tissue of Turkish crayfish asymptomatic for crayfish plague a – abdominal cuticle tissue (n = 20); b – telson tissue (n = 20)

Fig. 5.

National Center for Biotechnology Information basic local alignment search tool results of amplified partial internal transcribed spacer region of Aphanomyces astaci from GenBank P – positive control; S-A – symptomatic abdomen tissue; S-T – symptomatic telson tissue, S-M – symptomatic melanised tissue, AS-A – asymptomatic abdomen tissues, AS-T – asymptomatic telson tissues
National Center for Biotechnology Information basic local alignment search tool results of amplified partial internal transcribed spacer region of Aphanomyces astaci from GenBank P – positive control; S-A – symptomatic abdomen tissue; S-T – symptomatic telson tissue, S-M – symptomatic melanised tissue, AS-A – asymptomatic abdomen tissues, AS-T – asymptomatic telson tissues

The prevalence of the crayfish plague pathogen Aphanomyces astaci in samples of Turkish crayfish populations from 41 locations

Sampling station Latitude (N) Longitude (E) Sampling date n Symptomatic crayfish PCR-positive crayfish Symptomatic crayfish (%) PCR-positive crayfish (%)
1 Altınyazı reservoir 41°04′45.48″ 26°35′16.08″ 27/09/2016 20 5 0 25 0
2 Apa reservoir 37°21′34.92″ 32°32′43.44″ 18/08/2016 20 7 1 35 5
3 Lake Apolyont 40°08′55.98″ 28°36′53.32″ 14/08/2015 20 5 2 25 10
4 Arpaçay reservoir 40°37′09.38″ 43°41′37.67″ 26/08/2016 20 4 3 20 15
5 Lakes Bafra and Balık 41°34′26.34″ 36°04′49.91″ 7/06/2016 20 5 6 25 30
6 Lake Bayramşah 41°07′31.66″ 27°11′57.34″ 18/04/2016 5 0 0 0 0
7 Lake Beyşehi 37°46′20.64″ 31°31′16.36″ 11/11/2016 20 4 8 20 40
8 Bıyıkali reservoir 41°00′26.23″ 27°23′30.33″ 18/04/2016 20 5 1 25 5
9 Lake Çavuşçu 38°21′02.00″ 31°52′37.00″ 18/08/2016 20 3 2 15 10
10 Lake Çıldır 41°03′28.77″ 43°12′41.70″ 26/08/2016 20 4 1 20 5
11 Lake Eğirdir 38°01′25.52″ 30°52′22.30″ 26/10/2015 21 8 9 38 42.9
12 Gelingüllü reservoir 39°36′33.84″ 35°02′37.32″ 18/12/2015 20 15 1 75 5
13 Lake Gölmarmara 38°37′17.76″ 28°01′49.15″ 12/01/2016 20 1 0 5 0
14 Hirfanlı reservoir 39°11′58.37″ 33°33′01.47″ 18/08/2016 20 8 1 40 5
15 Lake Işıklı 38°14′07.70″ 29°53′35.13″ 28/08/2015 20 5 12 25 55
16 Lake İznik 40°26′59.71″ 29°32′02.30″ 31/08/2016 23 6 8 26.1 34.8
17 Kadıköy reservoir 40°47′39.87″ 26°46′28.57″ 27/09/2016 20 6 4 30 20
18 Karababa reservoir 41°12′41.35″ 27°02′56.03″ 18/04/2016 20 3 3 15 15
19 Karacakılavuz reservoir 41°06′52.31″ 27°21′39.29″ 8/08/2015 20 3 0 15 0
20 Karaidemir reservoir 40°57′21.47″ 27°00′38.23″ 8/08/2015 20 2 1 10 5
21 Karamanlı reservoir 37°23′59.50″ 29°50′12.04″ 30/09/2016 20 4 1 20 5
22 Lake Karataş 37°23′11.35″ 29°58′05.78″ 3/09/2015 20 3 3 15 15
23 Lake Karpuzlu 40°49′11.88″ 26°18′28.67″ 2/09/2015 20 4 2 20 10
24 Keban reservoir 38°48′08.03″ 38°43′45.74″ 10/11/2015 20 10 1 50 5
25 Mamasın reservoir 38°24′06.07″ 34°07′55.84″ 13/07/2016 20 3 1 15 5
26 Lake Manyas 40°12′10.56″ 27°56′53.99″ 14/08/2015 24 1 0 4 0
27 Porsuk reservoir 39°38′07.80″ 30°16′45.84″ 17.06.2015 20 14 0 70 0
28 Lake Poyrazlar 40°50′18.01″ 30°28′01.87″ 14.08.2015 15 1 4 6.7 26.7
29 Lake Sera 40°59′10.85″ 39°36′55.41″ 10.11.2015 20 3 6 15 30
30 Seyhan reservoir 37°02′24.00″ 35°19′55.00″ 1.11.2016 20 5 1 25 5
31 Lake Gölhisar 37°06′51.19″ 29°36′02.34″ 16.03.2016 20 4 1 20 5
32 Yenice reservoir 39°56′21.01″ 27°12′50.26″ 22.04.2016 15 2 0 13.3 0
33 Lake Yeniçağa 40°46′51.95″ 32°01′22.14″ 10.10.2016 20 8 3 40 15
34 Durusu reservoir 41°20′00.00″ 28°34′00.00″ 15.09.2015 20 3 4 15 20
35 Lake Suğla 37°20′15.00″ 32°01′56.00″ 22.10.2015 20 9 7 45 35
36 Lake Taşkısığı 40°52′07.00″ 30°24′12.00″ 10.10.2016 22 3 15 13.6 68.2
37 Lake Gölcük 38°18′45.25″ 28°01′43.88″ 28.10.2015 20 2 7 10 35
38 Hanoğlu reservoir 41°11′24.19″ 27°22′08.37″ 2.11.2016 20 3 8 15 40
39 Atikhisar reservoir 40°07′26.04″ 26°31′24.24″ 30.10.2016 15 3 3 20 20
40 Lake Bostancılar 41°27′04.56″ 32°56′59.07″ 11.10.2016 20 3 3 15 15
41 Lake Aktaş 41°13′00.06″ 43°12′56.87″ 12.11.2016 20 6 4 30 20
∑ = 800 ∑ = 193 ∑ = 137 x = 23.6 x = 16.5

Crayfish samples from which Aphanomyces astaci was isolated and genotyped by multi-locus microsatellite analysis and compared to reference strains

Locations of examined crayfish Allele sizes of amplified microsatellite loci
Aast2 Aast4 Aast6 Aast7 Aast9 Aast10 Aast12 Aast13 Genotypes
Genotype A1 (reference strain) 160 103 157 205 179 142 226/240 194 A
Genotype B2 (reference strain) 142 87 148 214 164/182 132/134 226/240 202 B
Eğirdir Lake 143 87 148 211 164/182 134 226/241 194 B
Karababa reservoir 159 87 158 214 164/182 132 226/241 194 B
Karaidemir reservoir 142 87 148 211 163/185 134 226/240 194 B
Mamasın reservoir 142 87 148 211 164/183 134 226/240 194 B
Lake Suğla 158 101 158 211 180 142 226/240 194 A
Hanoğlu reservoir 142 87 148 214 164/181 132 226/240 194/202 B
Atikhisar reservoir 142 87 149/158 211 164/183 134 226/240 194 B

PCR detection of Aphanomyces astaci from various crayfish tissue

Sample type Tissue type n Number of PCR-positive samples Detection rate (%)
Symptomatic specimens Telson 20 10 50
Abdominal cuticle 20 12 60
Melanised cuticle 20 8 40
Asymptomatic specimens Telson 20 1 5
Abdominal cuticle 20 2 10
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