Development of microsatellite markers in Caryophyllaeus laticeps (Cestoda: Caryophyllidea), monozoic fish tapeworm, using next-generation sequencing approach

Abstract

Caryophyllaeus laticeps (Pallas 1781) (Cestoda: Caryophyllidea) is a monozoic tapeworm of cyprinid fishes with a distribution area that includes Europe, most of the Palaearctic Asia and northern Africa. Broad geographic distribution, wide range of definitive fish hosts and recently revealed high morphological plasticity of the parasite, which is not in an agreement with molecular findings, make this species to be an interesting model for population biology studies. Microsatellites (short tandem repeat (STR) markers), as predominant markers for population genetics, were designed for C. laticeps using a next-generation sequencing (NGS) approach. Out of 165 marker candidates, 61 yielded PCR products of the expected size and in 25 of the candidates a declared repetitive motif was confirmed by Sanger sequencing. After the fragment analysis, six loci were proved to be polymorphic and tested for heterozygosity, Hardy-Weinberg equilibrium and the presence of null alleles on 59 individuals coming from three geographically widely separated populations (Slovakia, Russia and UK). The number of alleles in particular loci and populations ranged from two to five. Significant deficit of heterozygotes and the presence of null alleles were found in one locus in all three populations. Other loci showed deviations from Hardy-Weinberg equilibrium and the presence of null alleles only in some populations. In spite of relatively low polymorphism and the potential presence of null alleles, newly developed microsatellites may be applied as suitable markers in population genetic studies of C. laticeps.

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Acknowledgments

The authors would like to acknowledge Prof. Tomáš Scholz (Biology Centre ASCR, Institute of Parasitology, České Budějovice, Czech Republic) for the donation of C. laticeps material from his collection. The work was financially supported by the Slovak Research and Development Agency under contract APVV-0653-11 and as implementation of projects Revogene Research Centre for Molecular Genetics (ITMS26240220067) and Centre of Excellence for Parasitology (ITMS26220120022) supported by the Research and Development Operational Programme funded by the European Regional Development Fund.

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Correspondence to Ivica Králová-Hromadová.

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Ivica Králová-Hromadová and Gabriel Minárik equally contributed to this work.

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Králová-Hromadová, I., Minárik, G., Bazsalovicsová, E. et al. Development of microsatellite markers in Caryophyllaeus laticeps (Cestoda: Caryophyllidea), monozoic fish tapeworm, using next-generation sequencing approach. Parasitol Res 114, 721–726 (2015). https://doi.org/10.1007/s00436-014-4239-4

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Keywords

  • Population genetics
  • STR marker design
  • Non-segmented fish tapeworms
  • Caryophyllidea