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Intron sequence variation of the echinostomes (Trematoda; Echinostomatidae): implications for genetic investigations of the 37 collar-spined, Echinostoma miyagawai Ischii, 1932 and E. revolutum (Fröelich, 1802)

  • Genetics, Evolution, and Phylogeny - Original Paper
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Abstract

Echinostomes are a diverse group of digenetic trematodes that are difficult to classify by predominantly traditional techniques and contain many cryptic species. Application of contemporary genetic/molecular markers can provide an alternative choice for comprehensive classification or systematic analysis. In this study, we successfully characterized the intron 5 of domain 1 of the taurocyamine kinase gene (TkD1Int5) of Artyfechinostomum malayanum and the other two species of the 37 collar-spined group, Echinostoma revolutum and Echinostoma miyagawai, whereas TkD1Int5 of Hypoderaeum conoideum cannot be amplified. High levels of nucleotide polymorphism were detected in TkD1Int5 within E. revolutum and E. miyagawai, but not in A. malayanum. Thus, TkD1Int5 can be potentially used as genetic marker for genetic investigation of E. miyagawai and E. revolutum. We therefore used TkD1Int5 to explore genetic variation within and genetic differentiation between 58 samples of E. miyagawai and five samples of E. revolutum. Heterozygosity was observed in 17 and two samples with 16 and three insertion/deletion (indel) patterns in E. miyagawai and E. revolutum, respectively. Heterozygous samples were then cloned and nucleotide sequence was performed revealing the combined haplotypes in a particular sample. Based on nucleotide variable sites (excluding indels), the 72 E. miyagawai and seven E. revolutum haplotypes were subsequently classified. The haplotype network revealed clear genetic differentiation between E. miyagawai and E. revolutum haplogroups, but no genetic structure correlated with geographical localities was detected. High polymorphism and heterogeneity of the TkD1Int5 sequence found in our study suggest that it can be used in subsequent studies as an alternate independent potential genetic marker to investigate the population genetics, genetic structure, and possible hybridization of the other echinostomes, especially the 37 collar-spined group distributed worldwide.

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Funding

This research was supported by Faculty of Medicine, Thammasat University, Thailand, to C. Tantrawatpan (grant no. 034/2560).

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Authors and Affiliations

  1. Walai Rukhavej Botanical Research Institute, Mahasarakham University, Maha Sarakham, 44150, Thailand

    Weerachai Saijuntha

  2. Division of Cell Biology, Department of Preclinical Sciences, Faculty of Medicine, Thammasat University, Rangsit Campus, Pathumthani, 12120, Thailand

    Chairat Tantrawatpan

  3. Department of Environmental Medicine, Kochi Medical School, Kochi University, Oko, Nankoku, Kochi, 783-8505, Japan

    Takeshi Agatsuma

  4. Department of Public Health, Faculty of Science and Technology, Phetchabun Rajabhat University, Phetchabun, 67000, Thailand

    Kunyarat Duenngai

  5. Cholangiocarcinoma Research Institute, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand

    Paiboon Sithithaworn & Ross H. Andrews

  6. Department of Zoology and Paleontology and Evolution, State Museum of Natural History Karlsruhe, Erbprinzenstrasse 13, 76133, Karlsruhe, Germany

    Trevor N. Petney

  7. Faculty of Medicine, Imperial College London, St Mary’s Campus, South Wharf Street, London, W2 1NY, UK

    Ross H. Andrews

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  1. Weerachai Saijuntha
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  2. Chairat Tantrawatpan
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  3. Takeshi Agatsuma
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  4. Kunyarat Duenngai
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  5. Paiboon Sithithaworn
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  6. Trevor N. Petney
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  7. Ross H. Andrews
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Correspondence to Chairat Tantrawatpan.

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Saijuntha, W., Tantrawatpan, C., Agatsuma, T. et al. Intron sequence variation of the echinostomes (Trematoda; Echinostomatidae): implications for genetic investigations of the 37 collar-spined, Echinostoma miyagawai Ischii, 1932 and E. revolutum (Fröelich, 1802). Parasitol Res 119, 2485–2494 (2020). https://doi.org/10.1007/s00436-020-06734-z

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