, Volume 785, Issue 1, pp 249–260 | Cite as

Phylogeography of stream-dwelling trout in the Republic of Macedonia and a molecular genetic basis for revision of the taxonomy proposed by S. Karaman

  • Saša Marić
  • Simona Sušnik Bajec
  • Johannes Schöffmann
  • Vasil Kostov
  • Aleš Snoj
Primary Research Paper


To evaluate whether molecular data support the distinctiveness of Salmo macedonicus (Karaman, 1924) and Salmo pelagonicus Karaman, 1938, and to examine a possible impact of non-native trout translocated from the Drim drainage on the indigenous trout of the Vardar drainage, 187 individuals from 15 populations sampled across both drainages were studied by analysing the complete mitochondrial DNA control region and 12 microsatellite DNA loci. On the basis of both marker systems, the analysed populations were divided into two main genetic groups: I, native populations of the Drim drainage, along with some introduced populations of the Vardar drainage, and II, native Vardar populations, along with some populations exhibiting introgressed genotypes. The populations assigned to group I correspond taxonomically to Salmo farioides Karaman, 1938, among which those from the Drim were indigenous and those detected in the Vardar drainage were introduced. Four native Vardar populations from group II indicated two distinct clusters whose distribution matched the proposed range of S. macedonicus (two populations from the upper Vardar system) and S. pelagonicus (two populations from the Crna Reka system). Based upon the results of the study, some conservation genetic guidelines are proposed to help propagate and sustain the non-introgressed native trout populations.


Vardar drainage Genetic diversity mtDNA Microsatellites Translocations Conservation 



This study was supported by the Slovenian Research Agency, and the Ministry of Education, Science and Technological Development of the Republic of Serbia (Grant No. 173045). Many thanks go to Simo Georgiev who provided very useful information about trout translocations in Republic of Macedonia, and Iain F. Wilson for English revision of the manuscript.

Supplementary material

10750_2016_2930_MOESM1_ESM.docx (26 kb)
Supplementary material 1 (DOCX 26 kb)


  1. Apostolidis, A. P., D. Loukovitis & C. S. Tsigenopoulos, 2008a. Genetic characterization of brown trout (Salmo trutta) populations from the Southern Balkans using mtDNA sequencing and RFLP analysis. Hydrobiologia 600: 169–176.CrossRefGoogle Scholar
  2. Apostolidis, A. P., M.-J. Madeira, M. M. Hansen & A. Machodrom, 2008b. Genetic structure and demographic history of brown trout (Salmo trutta) populations from the southern Balkans. Freshwater Biology 53: 1555–1566.CrossRefGoogle Scholar
  3. Apostolidis, A. P., M. Th. Stoumboudi, E. Kalogianni, G. Cote & L. Bernatchez, 2011. Genetic divergence among native trout Salmo trutta populations from southern Balkans based on mitochondrial DNA and microsatellite variation. Journal of Fish Biology 79: 1950–1960.CrossRefPubMedGoogle Scholar
  4. Bandelt, H. J., P. Forster & A. Röhl, 1999. Median-joining networks for inferring intraspecific phylogenies. Molecular Biology and Evolution 16(1): 37–48.CrossRefPubMedGoogle Scholar
  5. Belkhir, K., P. Borsa, L. Chikhi, N. Raufaste & F. Bonhomme, 1996–2004. GENETIX v. 4.04, Logiciel sous WindowsTM pour la Génétique des Populations. Université Montpellier 2, Laboratoire Génome et Population, Montpellier.Google Scholar
  6. Berrebi, P., C. Tougard, S. Dubois, Zh Shao, I. Koutseri, S. Petkovski & A. J. Crivelli, 2013. Genetic diversity and conservation of the Prespa trout in the Balkans. International Journal of Molecular Sciences 14: 23454–23470.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 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: 351–379.CrossRefPubMedGoogle Scholar
  8. Buroker, N. E., J. R. Brown, T. A. Gilbert, P. J. O’Hara, A. T. Beckenbach, W. K. Thomas & M. J. Smith, 1990. Length heteroplasmy of sturgeon mitochondrial DNA: an illegitimate elongation model. Genetics 124: 157–163.PubMedPubMedCentralGoogle Scholar
  9. Cortey, M., C. Pla & J. L. García-Marín, 2004. Historical biogeography of Mediterranean trout. Molecular Phylogenetics and Evolution 33: 831–844.CrossRefPubMedGoogle Scholar
  10. Delpino, I., 1935. La diffusione e la distribuzione in Italia della Trutta genivittata (Heckel-Kner 1858). Bollettino di Pesca Piscicoltura e Idrobiologia Roma 11: 196–210.Google Scholar
  11. Delling, B., 2003. Species diversity and phylogeny of Salmo with emphasis on southern trouts (Teleostei, Salmonidae). Unpublished D. Phil. Thesis, Stockholm University.Google Scholar
  12. Dimovski, A. & R. Grupče, 1971. L’Ichtyofaune de la riviere Brégalnitsa. Izdanija, Institut de Pisciculture de la RS de Macedonie. Skopje IV(7): 1–37.Google Scholar
  13. Evanno, G., S. Regnaut & J. Goudet, 2005. Detecting the number of clusters of individuals using the software STRUCTURE: a simulation study. Molecular Ecology 14: 2611–2620.CrossRefPubMedGoogle Scholar
  14. Frankham, R., J. D. Ballou & D. A. Briscoe, 2004. A primer of conservation genetics, 1st ed. University Press, Cambridge.CrossRefGoogle Scholar
  15. Fumagalli, L., A. Snoj, D. Jesenšek, F. Balloux, T. Jug, O. Duron, F. Brossier, A. Crivelli & P. Berrebi, 2002. Extreme genetic differentiation among the remnant populations of marble trout (Salmo marmoratus) in Slovenia. Molecular Ecology 11(12): 2711–2716.CrossRefPubMedGoogle Scholar
  16. Georgiev, S., 2011. Study of Salmo genus (Pisces, Salmonidae) on the Balkan Peninsula. Ribarstvo 69: 11–20.Google Scholar
  17. Goudet, J., 1995. FSTAT (Version 1.2): a computer program to calculate F-statistics. Journal of Heredity 86: 485–486.Google Scholar
  18. Hewitt, G. M., 1999. Post-glacial re-colonization of European biota. Biological Journal of the Linnean Society 68: 87–112.CrossRefGoogle Scholar
  19. Karaman, S., 1924. Pisces Macedoniae. Hrvatska Stamparija, Split: 90.Google Scholar
  20. Karaman, S., 1927. Salmonidi Balkana. Glasnik Skopskog Naučnog Društva 2: 253–268.Google Scholar
  21. Karaman, S., 1938. Beitrag zur Kenntnis der Süsswasserfische Jugoslawiens. Bulletein de la Société Scientifique de Skoplje 18: 131–139.Google Scholar
  22. Karaman, S., 1957. The Radika river trouts. Folia Balcanica 10: 57–70.Google Scholar
  23. Kohout, J., A. Šedivá, A. Apostolou, T. Stefanov, S. Marić, M. Gaffaroğlu & V. Šlechta, 2013. Genetic diversity and phylogenetic origin of brown trout Salmo trutta populations in eastern Balkans. Biologia 68(6): 1229–1237.CrossRefGoogle Scholar
  24. Kostov, V., K. Rebok, V. Slavevska-Stamenković & M. Ristovska, 2010: Fish fauna of River Bregalnica (R. Macedonia). In BALWOIS, Proceedings of the Fourth Conference on Water Observation and Information System for Decision Support. Ohrid, 1–8.Google Scholar
  25. Kottelat, M. & J. Freyhof, 2007. Handbook of European Freshwater Fishes. Kottelat, Cornol, Switzerland & Freyhof, Berlin: 646 pp.Google Scholar
  26. Lerceteau-Kohler, E. & S. Weiss, 2006. Development of multiplex PCR microsatellite assay in brown trout Salmo trutta, and its potential application for the genus. Aquaculture 258: 641–645.CrossRefGoogle Scholar
  27. Lo Brutto, S., N. Hristovski & M. Arculeo, 2010. Genetic divergence between morphological forms of brown trout Salmo trutta L. in the Balkan region of Macedonia. Journal of Fish Biology 76: 1220–1227.CrossRefPubMedGoogle Scholar
  28. Marić, S., S. Sušnik, P. Simonović & A. Snoj, 2006. Phylogeographic study of brown trout from Serbia, based on mitochondrial DNA control region analysis. Genetics Selection Evolution 38: 411–430.CrossRefGoogle Scholar
  29. Marić, S., B. Kalamujić, A. Snoj, A. Razpet, L. Lukić-Bilela, N. Pojskić & S. Sušnik Bajec, 2012. Genetic variation of European grayling (Thymallus thymallus) populations in the Western Balkans. Hydrobiologia 691: 225–237.CrossRefGoogle Scholar
  30. Phillips, R. B., M. P. Matsuoka, I. Konon & K. M. Reed, 2000. Phylogenetic analysis of mitochondrial and nuclear sequences supports inclusion of Acantholingua ohridana in the genus Salmo. Copeia 2: 546–550.CrossRefGoogle Scholar
  31. Pritchard, J. K., M. Stephens & P. Donnelly, 2000. Inference of population structure using multilocus genotype data. Genetics 155: 945–959.PubMedPubMedCentralGoogle Scholar
  32. Pustovrh, G., S. Sušnik Bajec & A. Snoj, 2011. Evolutionary relationship between marble trout of the northern and southern Adriatic basin. Molecular Phylogenetics and Evolution 59: 761–766.CrossRefPubMedGoogle Scholar
  33. Pustovrh, G., A. Snoj & S. Sušnik Bajec, 2014. Molecular phylogeny of Salmo of the western Balkans, based upon multiple nuclear loci. Genetics Selection Evolution 46: 7.CrossRefGoogle Scholar
  34. Razpet, A., S. Sušnik, T. Jug & A. Snoj, 2007. Genetic variation among trout in the River Neretva basin, Bosnia and Herzegovina. Journal of Fish Biology 70: 94–110.CrossRefGoogle Scholar
  35. Ristovska, M., V. Kostov, D. Prelic, V. Slavevska-Stamenković & J. Arsovska, 2011. Fish community structure and water quality assessment of Babuna River. Journal of International Environmental Application & Science 6(4): 508–517.Google Scholar
  36. Sambrook, J., E. F. Fritsch & T. Maniatis, 1989. Molecular cloning. A laboratory manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.Google Scholar
  37. Sell, J. & Z. Spirkovski, 2004. Mitochondrial DNA differentiation between two forms of trout Salmo letnica, endemic to the Balkan Lake Ohrid, reflects their reproductive isolation. Molecular Ecology 13: 3633–3644.CrossRefPubMedGoogle Scholar
  38. Sidorovski, M., 1955. Nekoi Morfološki karakteri na pastrmkite od Mavrovskoto ezero i reka Radika. Izdanija, Institut de Pisciculture de la RP Macedoine 1(5): 135–147. (in Macedonian).Google Scholar
  39. Sidorovski, M., 1960. Plodnost pastrmki iz Mavrovskog Jezera i reke Radike. Publikacije stručnog udruženja za unapređenje slatkovodnog ribarstva Jugoslavije 1: 60. (in Serbian).Google Scholar
  40. Snoj, A., E. Melkič, S. Sušnik, S. Muhamedagić & P. Dovč, 2002. DNA phylogeny supports revised classification of Salmothymus obtusirostris. Biological Journal of the Linnean Society 77(3): 399–411.CrossRefGoogle Scholar
  41. Snoj, A., S. Marić, P. Berrebi, A. Crivelli, S. Shumka & S. Sušnik, 2009. Genetic architecture of trout from Albania as revealed by mtDNA control region variation. Genetics Selection Evolution 41: 22.CrossRefGoogle Scholar
  42. Snoj, A., B. Glamuzina, A. Razpet, J. Zablocki, I. Bogut, E. Lerceteau-Köhler, N. Pojskić & S. Sušnik, 2010. Resolving taxonomic uncertainties using molecular systematics: Salmo dentex and the Balkan trout community. Hydrobiologia 651(1): 199–212.CrossRefGoogle Scholar
  43. Stanković, S., 1960. The Balkan Lake Ohrid and its living world. Monographiae Biologicae, Vol. IX. Uitgeverij Dr. W. Junk, Den Haag.Google Scholar
  44. Sušnik, S., I. Knizhin, A. Snoj & S. Weiss, 2006. Genetic and morphological characterization of a Lake Ohrid endemic, Salmo (Acantholina) ohridanus with a comparison to sympatric Salmo trutta. Journal of Fish Biology 68(A): 2–23.CrossRefGoogle Scholar
  45. Sušnik, S., A. Snoj, I. F. Wilson, D. Mrdak & S. Weiss, 2007a. Historical demography of brown trout (Salmo trutta) in the Adriatic drainage including the putative S. letnica endemic to Lake Ohrid. Molecular Phylogenetics and Evolution 44: 63–76.CrossRefPubMedGoogle Scholar
  46. Sušnik, S., S. Weiss, T. Odak, B. Delling, T. Treer & A. Snoj, 2007b. Reticulate evolution: ancient introgression of the Adriatic brown trout mtDNA in softmouth trout Salmo obtusirostris (Teleostei: Salmonide). Biological Journal of the Linnean Society 90: 139–152.CrossRefGoogle Scholar
  47. Šorić, V., 1990. Salmonids in the Ohrid–Drim–Skadar system. Acta Societatis Zoologicae Bohemoslovacaei 54: 305–319.Google Scholar
  48. Tamura, K., G. Stecher, D. Peterson, A. Filipski & S. Kumar, 2013. MEGA6: molecular Evolutionary Genetics Analysis Version 6.0. Molecular Biology and Evolution 30: 2725–2729.CrossRefPubMedPubMedCentralGoogle Scholar
  49. Thompson, J. D., T. J. Gibson, F. Plewniak, F. Jeanmougin & D. G. Higgins, 1997. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 25: 4876–4882.CrossRefPubMedPubMedCentralGoogle Scholar
  50. Uiblein, F., A. Jagsch, W. Honsig-Erlenburg & S. Weiss, 2001. Status, habitat use, and vulnerability of the European grayling in Austrian waters. Journal of Fish Biology 59: 223–247.Google Scholar
  51. Vähä, J. P., J. Erkinaro, E. Niemelä & C. R. Primmer, 2007. Life-history and habitat features influence the within-river genetic structure of Atlantic salmon. Molecular Ecology 16: 2638–2654.CrossRefPubMedGoogle Scholar
  52. Van Oosterhout, C., W. F. Hutchinson, D. P. M. Wills & P. Shipley, 2004. MICRO-CHECKER: software for identifying and correcting genotyping errors in microsatellite data. Molecular Ecology Notes 4: 535–538.CrossRefGoogle Scholar
  53. Wilson, I., 2004. Low genetic variability in the summer Koran (Salmo letnica aestivalis Stefanovic) of Lake Ohrid. The Albanian Journal of Natural & Technical Sciences 16: 3–13.Google Scholar

Copyright information

© Springer International Publishing Switzerland 2016

Authors and Affiliations

  • Saša Marić
    • 1
  • Simona Sušnik Bajec
    • 2
  • Johannes Schöffmann
    • 3
  • Vasil Kostov
    • 4
  • Aleš Snoj
    • 2
  1. 1.Faculty of Biology, Institute of ZoologyUniversity of BelgradeBelgradeSerbia
  2. 2.Department of Animal Science, Biotechnical FacultyUniversity of LjubljanaDomžaleSlovenia
  3. 3.St. Veit/GlanAustria
  4. 4.Institute of Animal ScienceUniversity in Skopje “Ss. Cyril and Methodius”SkopjeRepublic of Macedonia

Personalised recommendations