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The study of Cytochrome B (CYTB): species-specific detection and phylogenetic relationship of Echinostoma revolutum, (Froelich, 1802)

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Abstract

Echinostoma revolutum is known as a significant intestinal trematode in various species of animals and humans. It presents complexities in terms of both the morphological and molecular biological data. This is the first study of the application of Cytochrome B gene (CYTB) as a target for studying the phylogeny and designing species-specific primer of E. revolutum. Adult trematodes were harvested from experimentally infected hamsters at 18 days of post-infection. Each worm was identified based on their morphological appearance. The novel CYTB primers were designed from other Echinostoma species to initially amplify CYTB region in E. revolutum. All sequence data of E. revolutum in five provinces of Central Thailand were used as the target for designing the species-specific primer for E. revolutum. The results revealed that CYTB gene can separate E. revolutum into two sister groups by geographical distribution, comprising the eastern and western area groups. Moreover, it also separates E. revolutum from other Echinostoma species, including two sibling species; E. caproni and E. paraensei. In addition, we developed the high performance species-specific primer of E. revolutum. It can detect DNA from a single egg, as well as cercaria, metacercaria and adult stages of this trematode with no cross-reactions to other trematodes and their hosts. Therefore, this research is a positive initial step for the future study of E. revolutum CYTB. The future studies based on this gene should be continued with all species in revolutum complex to overcome the problems of systemic classification that arise in this complex group.

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References

  • Blasco-Costa I, Cutmore SC, Miller TL, Nolan MJ (2016) Molecular approaches to trematode systematics: ‘best practice’ and implications for future study. Syst Parasitol 93(3):295–306

    Article  PubMed  Google Scholar 

  • Chai JY (2009) Echinostomes in humans. In: Fried B, Toledo R (eds) The biology of echinostomes. Springer, Berlin, pp 147–183

    Chapter  Google Scholar 

  • Chai JY, Sohn WM, Na BK, Van De N (2011) Echinostoma revolutum: metacercariae in Filopaludina snails from Nam Dinh province, Vietnam, and adults from experimental hamsters. Korean J Parasitol 49(4):449–455

    Article  PubMed  PubMed Central  Google Scholar 

  • Chantima K, Chai JY, Wongsawad C (2013) Echinostoma revolutum: freshwater snails as the second intermediate hosts in Chiang Mai, Thailand. Korean J Parasitol 51(2):183–189

    Article  PubMed  PubMed Central  Google Scholar 

  • Detwiler JT, Bos DH, Minchella DJ (2010) Revealing the secret lives of cryptic species: examining the phylogenetic relationships of echinostome parasites in North America. Mol Phylogenet Evol 55(2):611–620

    Article  PubMed  Google Scholar 

  • Freedman D, Pisani R, Purves R (2007) Statistics (International Student Edition), 4th edn. WW Norton & Company, New York

    Google Scholar 

  • Fried B, Graczyk TK (2004) Recent advances in the biology of Echinostoma species in the “revolutum” group. Adv Parasitol 58:139–195

    Article  PubMed  Google Scholar 

  • Fried B, Graczyk TK, Tamang L (2004) Food-borne intestinal trematodiases in humans. Parasitol Res 93(2):159–170

    Article  PubMed  Google Scholar 

  • Georgieva S, Selbach C, Faltýnková A, Soldánová M, Sures B, Skírnisson K, Kostadinova A (2013) New cryptic species of the ‘revolutum’ group of Echinostoma (Digenea: Echinostomatidae) revealed by molecular and morphological data. Parasit Vectors 6(1):64

    Article  PubMed  PubMed Central  Google Scholar 

  • Georgieva S, Faltýnková A, Brown R, Blasco-Costa I, Soldánová M, Sitko J, Scholz T, Kostadinova A (2014) Echinostoma ‘revolutum’ (Digenea: Echinostomatidae) species complex revisited: species delimitation based on novel molecular and morphological data gathered in Europe. Parasit Vectors 7(1):520

    PubMed  PubMed Central  Google Scholar 

  • Hanelt B, Schmidt-Rhaesa A, Bolek MG (2015) Cryptic species of hairworm parasites revealed by molecular data and crowdsourcing of specimen collections. Mol Phylogenet Evol 82:211–218

    Article  PubMed  Google Scholar 

  • Hsieh HM, Chiang HL, Tsai LC, Lai SY, Huang NE, Linacre A, Lee JCI (2001) Cytochrome b gene for species identification of the conservation animals. Forensic Sci Int 122(1):7–18

    Article  CAS  Google Scholar 

  • Huffman JE, Fried B (1990) Echinostoma and echinostomiasis. Adv Parasitol 29:215–269

    Article  CAS  PubMed  Google Scholar 

  • Hung NM, Anh NTL, Van PT, Thanh BN, Van HN, Van HH (2015) Current status of fish-borne zoonotic trematode infections in Gia Vien district, Ninh Binh province, Vietnam. Parasit Vectors 8(1):21

    Article  PubMed  PubMed Central  Google Scholar 

  • Hüttner M, Nakao M, Wassermann T, Siefert L, Boomker JD, Dinkel A, Sako Y, Mackenstedt U, Romig T, Ito A (2008) Genetic characterization and phylogenetic position of Echinococcus felidis Ortlepp, 1937 (Cestoda: Taeniidae) from the African lion. Int J Parasitol 38(7):861–868

    Article  CAS  PubMed  Google Scholar 

  • Huyse T, Oeyen M, Larmuseau MH, Volckaert FA (2017) Co-phylogeographic study of the flatworm Gyrodactylus gondae and its goby host Pomatoschistus minutus. Parasitol Int 66(2):119–125

    Article  PubMed  Google Scholar 

  • Kato H, Gomez EA, Martini-Robles L, Muzzio J, Velez L, Calvopiña M, Daniel RA, Mimori T, Uezato H, Hashiguchi Y (2016) Geographic distribution of Leishmania species in Ecuador based on the cytochrome b gene sequence analysis. PLoS Negl Trop Dis 10(7):e0004844

    Article  PubMed  PubMed Central  Google Scholar 

  • Kechemir N, Jourdane J, Mas-Coma S (2002) Life cycle of a new African echinostome species reproducing by parthenogenesis. J Nat Hist 36(15):1777–1784

    Article  Google Scholar 

  • Kostadinova A, Herniou E, Barrett J, Littlewood DTJ (2003) Phylogenetic relationships of Echinostoma Rudolphi, 1809 (Digenea: Echinostomatidae) and related genera re-assessed via DNA and morphological analyses. Syst Parasitol 54(3):159–176

    Article  CAS  PubMed  Google Scholar 

  • Leary SL, Underwood W, Anthony R, Gwaltney-Brant S, Poison A, Meyer R (2013) AVMA guidelines for the euthanasia of animals, 2013th edn. American Veterinary Medical Association Schaumburg, Schaumburg

    Google Scholar 

  • Morgan J, Blair D (1998) Mitochondrial ND1 gene sequences used to identify echinostome isolates from Australia and New Zealand. Int J Parasitol 28(3):493–502

    Article  CAS  PubMed  Google Scholar 

  • Nagataki M, Tantrawatpan C, Agatsuma T, Sugiura T, Duenngai K, Sithithaworn P, Andrews RH, Petney TN, Saijuntha W (2015) Mitochondrial DNA sequences of 37 collar-spined echinostomes (Digenea: Echinostomatidae) in Thailand and Lao PDR reveals presence of two species: Echinostoma revolutum and E. miyagawai. Infect Genet Evol 35:56–62

    Article  CAS  PubMed  Google Scholar 

  • Noikong W, Wongsawad C (2014) Epidemiology and molecular genotyping of echinostome metacercariae in Filopaludina snails in Lamphun province, Thailand. Asian Pac J Trop Med 7(1):26–29

    Article  PubMed  Google Scholar 

  • Noikong W, Wongsawad C, Chai JY, Saenphet S, Trudgett A (2014) Molecular analysis of echinostome metacercariae from their second intermediate host found in a localised geographic region reveals genetic heterogeneity and possible cryptic speciation. PLoS Negl Trop Dis 8:e2778

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Parson W, Pegoraro K, Niederstätter H, Föger M, Steinlechner M (2000) Species identification by means of the cytochrome b gene. Int J Legal Med 114(1–2):23–28

    Article  CAS  PubMed  Google Scholar 

  • Qureshi AW, Tanveer A, Mas-Coma S (2016) Epidemiological analysis of human fascioliasis in northeastern Punjab, Pakistan. Acta Trop 156:157–164

    Article  PubMed  Google Scholar 

  • Sah R, Khadka S, Hamal R, Poudyal S (2018) Human echinostomiasis: a case report. BMC Res Notes 11(1):17

    Article  PubMed  PubMed Central  Google Scholar 

  • Saijuntha W, Sithithaworn P, Duenngai K, Kiatsopit N, Andrews RH, Petney TN (2011) Genetic variation and relationships of four species of medically important echinostomes (Trematoda: Echinostomatidae) in South-East Asia. Infect Genet Evol 11(2):375–381

    Article  PubMed  Google Scholar 

  • Saijuntha W, Duenngai K, Tantrawatpan C (2013) Zoonotic echinostome infections in free-grazing ducks in Thailand. Korean J Parasitol 51(6):663–667

    Article  PubMed  PubMed Central  Google Scholar 

  • Simon C, Frati F, Beckenbach A, Crespi B, Liu H, Flook P (1994) Evolution, weighting, and phylogenetic utility of mitochondrial gene sequences and a compilation of conserved polymerase chain reaction primers. Ann Entomol Soc Am 87(6):651–701

    Article  CAS  Google Scholar 

  • Sohn WM, Chai JY, Yong TS, Eom KS, Yoon CH, Sinuon M, Socheat D, Lee SH (2011) Echinostoma revolutum infection in children, Pursat province, Cambodia. Emerg Infect Dis 17(1):117–119

    Article  PubMed  PubMed Central  Google Scholar 

  • Toledo R, Esteban J (2016) An update on human echinostomiasis. Trans R Soc Trop Med Hyg 110(1):37–45

    Article  CAS  PubMed  Google Scholar 

  • Toledo R, Esteban JG, Fried B (2009) Recent advances in the biology of echinostomes. Adv Parasitol 69:147–204

    Article  PubMed  Google Scholar 

  • Toledo R, Muñoz Antoli C, Esteban JG (2014) Intestinal trematode infections. In: Toledo R, Muñoz-Antoli C, Esteban JG (eds) Digenetic trematodes. Springer, Berlin, pp 201–240

    Chapter  Google Scholar 

  • Valadas SY, da Silva JI, Lopes EG, Keid LB, Zwarg T, de Oliveira AS, Sanches TC, Joppert AM, Pena HF, Oliveira TM (2016) Diversity of Sarcocystis spp. shed by opossums in Brazil inferred with phylogenetic analysis of DNA coding ITS1, cytochrome B, and surface antigens. Exp Parasitol 164:71–78

    Article  CAS  PubMed  Google Scholar 

  • Xu W, Morris U, Aydin-Schmidt B, Msellem MI, Shakely D, Petzold M, Björkman A, Mårtensson A (2015) SYBR green real-time PCR-RFLP assay targeting the plasmodium cytochrome b gene: a highly sensitive molecular tool for malaria parasite detection and species determination. PLoS ONE 10(3):e0120210

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yamaguti S (1958) Systema helminthum. The digenetic trematodes of vertebrates, vol 1. Interscience, New York

    Google Scholar 

Download references

Acknowledgements

We greatly acknowledge Srinakharinwirot University, Thailand for providing funding and relevant facilities (Grant No. 239/2560). Finally, we would like to thank Mr. Simon McIver and Mr. Russell Kirk Hollis for their assistance in editing this manuscript.

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Authors and Affiliations

  1. Applied Parasitology Research Laboratory, Department of Biology, Faculty of Science, Srinakharinwirot University, Bangkok, Thailand

    Sothorn Anucherngchai, Thapana Chontananarth & Thanawan Tejangkura

  2. Center of Excellence in Animal, Plant and Parasitic Biotechnology, Srinakharinwirot University, Bangkok, Thailand

    Thapana Chontananarth & Thanawan Tejangkura

  3. Korean Association of Health Promotion, Seoul, South Korea

    Jong-Yil Chai

Authors
  1. Sothorn Anucherngchai
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  2. Thapana Chontananarth
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  3. Thanawan Tejangkura
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  4. Jong-Yil Chai
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Contributions

AS and CT: Conception and design of study. CJY and AS: Support specimens. AS: Acquisition of data. AS, TT and CT: Analysis and/or interpretation of data. AS and CT: Drafting the manuscript. AS, CT and CJY: Revising the manuscript critically for important intellectual content. AS, TT, CT and CJY: Approval of the version of the manuscript to be published.

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Correspondence to Thapana Chontananarth.

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Anucherngchai, S., Chontananarth, T., Tejangkura, T. et al. The study of Cytochrome B (CYTB): species-specific detection and phylogenetic relationship of Echinostoma revolutum, (Froelich, 1802). J Parasit Dis 43, 66–74 (2019). https://doi.org/10.1007/s12639-018-1057-0

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  • DOI: https://doi.org/10.1007/s12639-018-1057-0

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