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

Akhan, Süleyman; Çağatay, İfakat Tülay; Berber, Selçuk; Taştan, Büşra; Taştan, Yiğit; Dalar, Tuba

doi:10.2478/jvetres-2024-0061

Figures & Tables

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

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

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)

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)

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
Locations of examined crayfish	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
Asymptomatic specimens	Abdominal cuticle	20	2	10

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

Figures & Tables

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Fig. 5.

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

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

PCR detection of Aphanomyces astaci from various crayfish tissue

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