, Volume 142, Issue 5, pp 381–395 | Cite as

Population structure and genetic analysis of narrow-clawed crayfish (Astacus leptodactylus) populations in Turkey

  • Suleyman Akhan
  • Yusuf Bektas
  • Selcuk Berber
  • Gokhan Kalayci


The genetic differentiation among Turkish populations of the narrow-clawed crayfish was investigated using a partial sequence of cytochrome oxidase subunit I gene (585 bp) of 183 specimens from 17 different crayfish populations. Median joining network and all phylogenetic analyses disclosed a strong haplotype structure with three prominent clades diverged by a range between 20 and 50 mutations and substantial inter-group pairwise sequence divergence (5.19–6.95 %), suggesting the presence of three distinct clades within the Anatolian populations of Astacus leptodactylus. The divergence times among the three clades of Turkish A. leptodactylus are estimated to be 4.96–3.70 Mya using a molecular clock of 1.4 % sequence divergence per million years, pointing to a lower Pliocene separation. The high level of genetic variability (H d  = 95.8 %, π = 4.17 %) and numerous private haplotypes suggest the presence of refugial populations in Anatolia unaffected by Pleistocene habitat restrictions. The pattern of genetic variation among Turkish A. leptodactylus populations, therefore, suggests that the unrevealed intraspecific genetic structure is independent of geographic tendency and congruent with the previously reported geographic distribution and number of subspecies (A. l. leptodactylus and A. l. salinus) of A. leptodactylus.


Astacus leptodactylus Cytochrome oxidase subunit I Genetic structure Turkey 



We would like to thank the editor and the reviewer for their time and valuable remarks. Funding for this research was provided by Scientific Project Unit of Recep Tayyip Erdogan University (Project 2009.103.02.1).

Conflict of interest

The authors declare that there is no conflict of interest between authors and no financial relationship with the organizations that sponsored research.


  1. Ahn DH, Kawai T, Kim SJ, Rho HS, Jung JW, Kim W, Lim BJ, Kim MS, Min GS (2006) Phylogeny of Northern Hemisphere freshwater crayfishes based on 16S rRNA gene analysis. Kor J Genet 28:185–192Google Scholar
  2. Attard J, Vianet R (1985) Variabilité génétique et morphologique de cinq populations de l‘écrevisse européenne Austropotamobius pallipes (Lereboullet 1858) (Crustacea, Decapoda). Can J Zoolog 63:2933–2939CrossRefGoogle Scholar
  3. Balık S, Ustaoğlu MR, Sarı HM, Berber S (2005) Determination of traits some growth and morphometric of crayfish (Astacus leptodactylus Eschscholtz, 1823) at Demirköprü Dam Lake (Manisa). Ege J Fish Aquat Sci 22:83–89Google Scholar
  4. Bandelt HJ, Forster P, Röhl A (1999) Median-joining networks for inferring intraspecific phylogenies. Mol Biol Evol 16:37–48PubMedCrossRefGoogle Scholar
  5. Baran I, Timur M, Oray IK, Timur G, Rahe R, Soylu E (1987) Investigation on a disease causing serious mortality on crayfish (Astacus leptodactylus) populations in Turkey. Eur Aquac Soc Sweden 87:6–7Google Scholar
  6. Bardakcı F, Tatar N, Hrbek T (2004) Genetic Relationships between Anatolian Species and Subspecies of Aphanius (Pisces, Cyprinodontiformes) based on RAPD Markers. Biologia 59:559–566Google Scholar
  7. Bardakcı F, Değerli N, Özdemir Ö, Başıbüyük HH (2006) Molecular phylogeny of the Turkish brown trout (Salmo trutta L.): mitochondrial DNA PCR-RFLP variation. J Fish Biol 68:36–55CrossRefGoogle Scholar
  8. Baric S, Höllrigl A, Füreder L, Dalla Via J (2005) Mitochondrial and microsatellite DNA analyses of Austropotamobius pallipes populations in South Tyrol (Italy) and Tyrol (Austria). B Fr Peche Piscic 376–377:599–612CrossRefGoogle Scholar
  9. Bernatchez L (2001) The evolutionary history of brown trout (Salmo trutta L.) inferred from phylogeographic, nested clade, and mismatch analyses of mitochondrial DNA variation. Evolution 55:351–379PubMedCrossRefGoogle Scholar
  10. Bilgin R (2011) Back to the suture: the distribution of intraspecific genetic diversity in and around anatolia. Int J Mol Sci 12(6):4080–4103PubMedCrossRefPubMedCentralGoogle Scholar
  11. Bott R (1950) Freshwater crayfish of Europe (Decapoda, Astacidae). Abh Senckenberg Naturf Ges 483:1–36Google Scholar
  12. Braband A, Kawai T, Scholtz G (2007) The phylogenetic position of the East Asian freshwater crayfish Cambaroides within the Northern Hemisphere Astacoidea (Crustacea, Decapoda, Astacida) based on molecular data. J Zool Syst Evol Res 44:17–24CrossRefGoogle Scholar
  13. Buhay JE, Crandall KA (2005) Subterranean phylogeography of freshwater crayfishes shows extensive gene flow and surprisingly large population sizes. Mol Ecol 14:4259–4273PubMedCrossRefGoogle Scholar
  14. Cataudella R, Paolucci M, Delaunay C, Ropiquet A, Hassanin A, Balsamo M, Grandjeand F (2010) Genetic variability of Austropotamobius italicus in the Marches region: implications for conservation. Aquat Conserv 20:261–268CrossRefGoogle Scholar
  15. Chiesa S, Scalici M, Negrini R, Gibertini G, Nonnis Marzano F (2011) Fine-scale genetic structure, phylogeny and systematics of threatened crayfish species complex. Mol Phylogenet Evol 61:1–11PubMedCrossRefGoogle Scholar
  16. Crandall KA, Fitzpatric JF Jr (1996) Crayfish molecular systematic: using a combination of procedures to estimate phylogeny. Syst Biol 45:1–26CrossRefGoogle Scholar
  17. Crandall KA, Harris DJ, Fetzner JW Jr (2000) The monophyletic origin of freshwater crayfish estimated from nuclear and mitochondrial DNA sequenceses. P Roy Soc B Biol Sci 267:1679–1686CrossRefGoogle Scholar
  18. Duman E, Pala M (1998) Investigation on growth characteristics of cryfish population (Astacus leptodactylus salinus Nordmann, 1842) which live in Ağın region of Keban Dam Lake. Ege J Fish Aquat Sci 15(1–2):9–17Google Scholar
  19. Durand JD, Ünlü E, Doadrio I, Pipoyan S, Templeton AR (2000) Origin, radiation, dispersion and allopatric hybridization in the chub Leuciscus cephalus. Proc R Soc Biol Sci Ser B 267:1687–1697CrossRefGoogle Scholar
  20. Durna S, Bardakci F, Degerli N (2010) Genetic diversity of Garra rufa Heckel, 1843 (Teleostei: Cyprinidae) in Anatolia. Biochem Syst Ecol 38(1):83–92CrossRefGoogle Scholar
  21. Erençin Z, Köksal G (1977) Studies on the freshwater crayfish (Astacus leptodactylus Eschscholtz, 1823) in Anatolia. Freshwater Crayfish 3:187–192Google Scholar
  22. Erinç S (1978) Changes in the physical environment in Turkey since the end of the last glacials. In: Brice WC (ed) The environmental history of the near and middle east since the last ice age. Academic Press, London, pp 87–100Google Scholar
  23. Excoffier L, Smouse PE, Quattro JM (1992) Analysis of molecular variance inferred from metric distances among DNA haplotypes: application to human mitochondrial DNA restriction data. Genetics 131:479–491PubMedPubMedCentralGoogle Scholar
  24. Excoffier L, Laval G, Schneider S (2005) Arlequin ver. 3.11: an integrated software package for population genetics data analysis. Evol Bioinform 1:47–50Google Scholar
  25. Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791CrossRefGoogle Scholar
  26. Fevolden SE, Taugbøl T, Skurdal J (1994) Allozymic variation among populations of noble crayfish, Astacus astacus L., in southern Norway: implications for management. Aquac Res 25:927–935CrossRefGoogle Scholar
  27. Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Biotech 3:294–299Google Scholar
  28. Fratini S, Zaccara S, Barbaresi S, Grandjean F, Souty-Grosset C, Crosa G, Gherardi F (2005) Phylogeography of the threatened crayfish (genus Austropotamobius) in Italy: implication for its conservation. Heredity 94:108–118PubMedCrossRefGoogle Scholar
  29. Geldiay R (1949) Çubuk Barajı ve Emir Gölünün makro ve mikro faunasının mukayeseli incelenmesi. Ankara Üniversitesi Fen Fakültesi Mecmuası 2:146–252Google Scholar
  30. Geldiay R, Kocataş A (1970) The preliminary report about the taxonomy and distribution of Astacus (Decapoda) of Turkey. Scientific Report of the Faculty Science, Ege Universitiy vol 94, pp 1–7Google Scholar
  31. Gouin N, Souty-Grosset C, Ropiquet A, Grandjean F (2002) High dispersal ability of Austropotamobius pallipes revealed by microsatellite markers in a French brook. B Fr Peche Piscic 367:681–689CrossRefGoogle Scholar
  32. Gouin N, Grandjean F, Pain S, Souty-Grosset C, Reynolds R (2003) Origin and colonization history of the white-clawed crayfish, Austropotamobius pallipes, in Ireland. Heredity 91:70–77PubMedCrossRefGoogle Scholar
  33. Gouin N, Grandjean F, Souty-Grosset C (2006) Population genetic structure of the endangered crayfish Austropotamobius pallipes in France based on microsatellite variation: biogeographical inferences and conservation implications. Freshw Biol 51:1369–1387CrossRefGoogle Scholar
  34. Gouin N, Souty-Grosset C, Borquez J, Bertin A, Grandjean F (2011) Disentangling the impact of demographic factors on population differentiation of an endangered freshwater crayfish (Austropotamobius pallipes) using population density and microsatellite data. Freshw Biol 56:2105–2118CrossRefGoogle Scholar
  35. Grandjean F, Souty-Grosset C, Holdich DM (1997) Mitochondrial DNA variation in four British populations of the white-clawed crayfish, Austropotamobius pallipes: implications for management. Aquat Living Resour 10:121–126CrossRefGoogle Scholar
  36. Grandjean F, Harris DJ, Souty-Grosset C, Crandall KA (2000) Systematics of the European endangered crayfish species Austropotamobius pallipes (Decapoda: Astacidae). J Crustacean Biol 20:522–529CrossRefGoogle Scholar
  37. Harlıoğlu MM (2004) The present situation of freshwater crayfish, Astacus leptodactylus (Eschscholtz, 1823) in Turkey. Aquaculture 230:181–187CrossRefGoogle Scholar
  38. Harlıoğlu MM (2008) The harvest of the freshwater crayfish Astacus leptodactylus Eschscholtz in Turkey: harvest history, impact of crayfish plague, and present distribution of harvested populations. Aquacult Int 16:351–360CrossRefGoogle Scholar
  39. Harlıoğlu MM, Güner U (2007) A new record of recently discovered crayfish, Austropotamobius torrentium (Shrank, 1803), in Turkey. B Fr Peche Piscic 387:1–5Google Scholar
  40. Harlıoğlu MM, Harlıoğlu AG (2004) The harvest of freshwater crayfish, Astacus leptodactylus (Eschscholtz, 1823) in Turkey. Rev Fish Biol Fish 14:415–419CrossRefGoogle Scholar
  41. Harlıoğlu MM, Holdich DM (2001) Meat yields in the introduced crayfish, Pacifastacus leniusculus and Astacus leptodactylus, from British waters. Aquac Res 32:411–417CrossRefGoogle Scholar
  42. Hasegawa M, Kishino H, Yano TA (1985) Dating of the human–ape splitting by a molecular clock of mitochondrial DNA. J Mol Evol 22:160–174PubMedCrossRefGoogle Scholar
  43. Hewitt GM (1996) Some genetic consequences of ice ages, and their role in divergence and speciation. Biol J Linn Soc 58:247–276CrossRefGoogle Scholar
  44. Hobbs HH Jr (1989) An illustrated checklist of the American Crayfishes (Decapoda: Astacidae, Cambaridae, and Parastacidae). Smithsonian Contrib Zool 480:1–236CrossRefGoogle Scholar
  45. Holdich DM (1993) A review of astaciculture: freshwater crayfish farming. Aquat Living Resour 6:307–317CrossRefGoogle Scholar
  46. Holdich DM (2002) Present distribution of crayfish in Europe and some adjoining countries. B Fr Peche Piscic 367:611–650CrossRefGoogle Scholar
  47. Holdich DM, Haffner P, Noël P (2006) Species files. In: Souty-Grosset C, Holdich DM, Noël PY, Reynolds JD, Haffner P (eds) Atlas of crayfish in Europe. Muséum national d’Histoire naturelle, Paris, (Patrimoines naturels, 64), pp 76–81Google Scholar
  48. Holthuis LB (1961) Report on a collection of Crustacea Decapoda and Stomatopoda from Turkey and the Balkans. Zool Verh 47:1–67Google Scholar
  49. Hrbek T, Meyer A (2003) Closing of the Tethys Sea and the phylogeny of Eurasian killifishes (Cyprinodontiformes: Cyprinodontidae). J Evol Biol 16:7–26CrossRefGoogle Scholar
  50. Hrbek T, Küçük F, Frickey T, Stölting KN, Wildekamp RH, Meyer A (2002) Molecular phylogeny and historical biogeography of the Aphanius (Pisces, Cyprinodontiformes) species complex of central Anatolia, Turkey. Mol Phylogenet Evol 25:125–137PubMedCrossRefGoogle Scholar
  51. Hrbek T, Stölting KN, Bardakcı F, Küçük F, Wildekamp RH, Meyer M (2004) Plate tectonics and biogeographical patterns of the Pseudophoxinus (Pisces: Cypriniformes) species complex of central Anatolia, Turkey. Mol Phylogenet Evol 32:297–308PubMedCrossRefGoogle Scholar
  52. Huelsenbeck JP, Ronquist F (2001) MrBayes: Bayesian inference of phylogeny. Bioinformatics 17:754–755PubMedCrossRefGoogle Scholar
  53. Iorgu EI, Popa OP, Petrescu AM, Popa LO (2011) Cross-amplification of microsatellite loci in the endangered stone-crayfish Austropotamobius torrentium (Crustacea: Decapoda). Knowl Manag Aquat Ec 401:08CrossRefGoogle Scholar
  54. Karaman MS (1962) Ein Beitrag zur Systematik des Astacidae (Decapoda). Crustaceana 3:173–191CrossRefGoogle Scholar
  55. Karaman MS (1963) Studie der Astacidae (Crustacea, Decapoda). Hydrobiologia (The Hague) 22:111–132Google Scholar
  56. Kimura M (1980) A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16(2):111–120PubMedCrossRefGoogle Scholar
  57. Klobučar GIV, Podnar M, Jelić M, Franjević D, Faller M, Štambuk A, Gottstein S, Simić V, Maguire I (2013) Role of the Dinaric Karst (western Balkans) in shaping the phylogeographic structure of the threatened crayfish Austropotamobius torrentium. Freshw Biol 58:1089–1105CrossRefGoogle Scholar
  58. Knowlton N, Weigt LA (1998) New dates and new rates for divergence across the Isthmus of Panama. Proc R Soc Lond 265:2257–2263CrossRefGoogle Scholar
  59. Kõiv K, Gross R, Paaver T, Kuehn R (2008) Isolation and characterization of first microsatellite markers for the noble crayfish, Astacus astacus. Conserv Genet 9:1703–1706CrossRefGoogle Scholar
  60. Kõiv K, Gross R, Paver T, Kuehn R (2009) Isolation and characterization of 11 novel microsatellite DNA markers in the noble crayfish, Astacus astacus. Anim Genet 40(1):124PubMedCrossRefGoogle Scholar
  61. Köksal G (1988) Astacus leptodactylus in Europe. In: Holdich DM, Lowery RS (eds) Freshwater crayfish: biology. Management and Exploitation, Croom Helm, London, pp 365–400Google Scholar
  62. Matallanas B, Ochando MD, Vivero A, Beroiz B, Alonso FC, Callejas C (2011) Mitochondrial DNA variability in Spanish populations of A. italicus inferred from the analysis of a COI region. Knowl Manag Aquat Ec 401:30Google Scholar
  63. Matallanas B, Callejas C, Ochando MD (2012) A genetic approach to Spanish populations of the threatened Austropotamobius italicus located at three different scenarios. Sci World J 2012:975930CrossRefGoogle Scholar
  64. Mathews LM, Adams L, Anderson E, Basile M, Gottardi E, Buckholt MA (2008) Genetic and morphological evidence for substantial hidden biodiversity in a freshwater crayfish species complex. Mol Phylogenet Evol 48(1):126–135PubMedCrossRefGoogle Scholar
  65. Mesquita N, Carvalho G, Shaw P, Crespo E, Coelho MM (2001) River basin-related genetic structuring in an endangered fish species, Chondrostoma lusitanicum, based on mtDNA sequencing and RFLP analysis. Heredity 86:253–264PubMedCrossRefGoogle Scholar
  66. Oray I (1990) The crayfish situation in Turkey. In: De Pauw N, Billard R (eds) Aquaculture Europe’89 business joins science. Aquaculture Society Special Publication, vol 12. Bredene, Belgium, pp 250–251Google Scholar
  67. Por FD, Dimentman C (1989) The legacy of Tethys. An aquatic biogeography of the Levant (Monographiae Biologicae 63). Kluwer Academic, DordrechtGoogle Scholar
  68. Posada D, Buckley TR (2004) Model selection and model averaging in phylogenetics: advantages of the AIC and Bayesian approaches over likelihood ratio tests. Syst Biol 53:793–808PubMedCrossRefGoogle Scholar
  69. Posada D, Crandall KA (1998) Modeltest: testing the model of DNA substitution. Bioinformatics 14(9):817–818PubMedCrossRefGoogle Scholar
  70. Posada D, Crandall KA (2001) Selecting the best-fit model of nucleotide substitution. Syst Biol 50:580–601PubMedCrossRefGoogle Scholar
  71. Rahe R, Soylu E (1989) Identification of the pathogenic fungus causing destruction to Turkish crayfish stocks (Astacus leptodactylus). J Invertebr Pathol 54:10–15CrossRefGoogle Scholar
  72. Rozas J, Sanches-DelBarrio JC, Messeguer X, Rozas R (2003) DnaSP, DNA polymorphism by the coalescent and other methods. Bioinformatics 19:2496–3497PubMedCrossRefGoogle Scholar
  73. Schrimpf A, Schulz HK, Theissinger K, Pârvulescu L, Schulz R (2011) The first large-scale genetic analysis of the vulnerable noble crayfish Astacus astacus reveals low haplotype diversity in central European populations. Knowl Manag Aquat Ec 401:35CrossRefGoogle Scholar
  74. Şengör AMC, Altıner D, Cin A, Ustaömer T, Hsu KJ (1988) Origin and assembly of the Tethyside orogenic collage at the expence of Gondwana Land. In: Audley-Charles MG, Hallam A (eds) Gondwana and Tethys. Oxford University Press, New York, pp 119–181Google Scholar
  75. Shull HC, Perez-Losada M, Blair D, Sewell K, Sinclair EA, Lawler S, Ponniah M, Crandall KA (2005) Phylogeny and biogeography of the freshwater crayfish Euastacus (Decapoda: Parastacidae) based on nuclear and mitochondrial DNA. Mol Phylogenet Evol 37:249–263PubMedCrossRefGoogle Scholar
  76. Sinclair EA, Fetzner JW Jr, Buhay J, Crandall KA (2004) Proposal to complete a phylogenetic taxonomy and systematic revision for freshwater crayfish (Astacidea). Freshw Crayfish 14:21–29Google Scholar
  77. Song H, Buhay JE, Whiting MF, Crandall KA (2008) Many species in one: DNA barcoding overestimates the number of species when nuclear mitochondrial pseudogenes are coamplified. P Nat Acad Sci Biol 105:13486–13491CrossRefGoogle Scholar
  78. Souty-Grosset C, Grandjean F, Raimond R, Frelon M, Debenest C, Bramard M (1997) Conservation genetics of the white-clawed crayfish Austropotamobius pallipes: the usefulness of the mitochondrial DNA marker. B Fr Peche Piscic 347:677–692CrossRefGoogle Scholar
  79. Stefani F, Zaccara S, Delmastro GB, Buscarino M (2011) The endangered white-clawed crayfish Austropotamobius pallipes (Decapoda, Astacidae) east and west of the Maritime Alps: a result of human translocation? Conserv Genet 12(1):51CrossRefGoogle Scholar
  80. Swofford DL (2003) PAUP*. Phylogenetic analysis using parsimony (*and other methods). Version 4 beta 10. Sinauer Associates, Sunderland, MassachusettsGoogle Scholar
  81. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739PubMedCrossRefPubMedCentralGoogle Scholar
  82. Thompson J, Higgins D, Gibson T (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4690PubMedCrossRefPubMedCentralGoogle Scholar
  83. Tortonese E (1952) Relazione preliminare di un viaggio a scopo zoologico attraverso l’Asia Minore. Bollettino dell’Istituto e Museo di Zoologia della Università di Torino 3:81–97Google Scholar
  84. Triantafyllidis A, Leonardos I, Bista I, Kyriazis ID, Stoumboudi MT, Kappas I, Amat F, Abatzopoulos TJ (2007) Phylogeography and genetic structure of the Mediterranean killifish Aphanius fasciatus (Cyprinodontidae). Mar Biol 152:1159–1167CrossRefGoogle Scholar
  85. Trontelj P, Yoichi M, Boris S (2005) Phylogenetic and phylogeographic relationships in the crayfish genus Austropotamobius inferred from mitochondrial COI gene sequences. Mol Phylogenet Evol 34:212–226PubMedCrossRefGoogle Scholar
  86. Yue GH, Li J, Bai Z, Wang CM, Feng F (2010) Genetic diversity and population structure of the invasive alien red swamp crayfish. Biol Invasions 12:2697–2706CrossRefGoogle Scholar
  87. Yüksel F (2007) The investigation of the crayfish (Astacus leptodactylus Eschscholtz, 1823) population amplitude in Keban dam lake, PhD Thesis, Instutue of Science, Fırat University, Elazığ, TurkeyGoogle Scholar
  88. Zaccara S, Stefani F, Galli P, Nardi PA, Crosa G (2004) Taxonomic implications in conservation management of white-clawed crayfish (Austropotamobius pallipes) (Decapoda, Astacidae) in Northern Italy. Biol Conserv 120:1–10CrossRefGoogle Scholar
  89. Zaccara S, Stefani F, Crosa G (2005) Diversity of mitochondrial DNA of the endangered white-clawed crayfish (Austropotamobius pallipes) in the Po River catchment. Freshw Biol 50:1262–1272CrossRefGoogle Scholar
  90. Zengin M, ve Kurtoğlu IZ (2006) A Preliminary assessment on the status of crayfish populations in Trabzon-Sera Lake. YUNUS Res Bull 6(2):6–7Google Scholar

Copyright information

© Springer International Publishing Switzerland 2014

Authors and Affiliations

  • Suleyman Akhan
    • 1
  • Yusuf Bektas
    • 2
  • Selcuk Berber
    • 3
  • Gokhan Kalayci
    • 2
  1. 1.Department of Aquaculture, Faculty of FisheriesAkdeniz UniversityAntalyaTurkey
  2. 2.Department of Basic Sciences, Faculty of FisheriesRecep Tayyip Erdogan UniversityRizeTurkey
  3. 3.Faculty of FisheriesÇanakkale 18 Mart UniversityÇanakkaleTurkey

Personalised recommendations