Molecular and morphological characterization of the metacercariae of two species of diplostomid trematodes (Platyhelminthes, Digenea) in freshwater fishes of the Batalha River, Brazil

  • Larissa Sbeghen Pelegrini
  • Thayana Gião
  • Diego Henrique Mirandola Dias Vieira
  • Maria Isabel Müller
  • Reinaldo José da Silva
  • Gerardo Pérez-Ponce de León
  • Rodney Kozlowiski de Azevedo
  • Vanessa Doro AbdallahEmail author
Fish Parasitology - Original Paper


The Diplostomidae include a large group of flatworms with complex life cycles and are frequently found parasitizing the eyes and central nervous system of freshwater fishes. The morphological identification of the metacercariae at species level is not always possible. Thus, molecular tools have become essential to assist in the parasite species determination. This study was aimed at describing two diplostomid metacercariae found in freshwater fish in São Paulo, Brazil, based on morphological characters and in the genetic characterization of COI sequences. Our results showed that the two recognized taxa (Tylodelphys sp. and Diplostomidae gen. sp.) appear to be different from the species already described in South America. Tylodelphys sp. differs morphologically from Tylodelphys xenopi, T. mashonense, T. jenynsiae, and T. scheuringi. The metacercariae of T. clavata and T. conifera are smaller than Tylodelphys sp., while T. podicipina is larger than the metacercariae described here. The phylogenetic analysis of COI sequences yielded Tylodelphys sp. as the sister species of Tylodelphys sp. 4, a species reported from the brain of the eleotrid Gobiomorus maculatus in Oaxaca, Mexico. The metacercariae identified as Diplostomidae gen. sp. are morphologically different from the known diplostomid metacercariae and did not match with other diplostomid sequences available. Diplostomidae gen. sp. is recovered as the sister species of Diplostomum ardeae. Although the morphological evidence and the COI sequences differentiate the metacercariae found, the absence of adult specimens of both species precludes the specific designation. This is one of the first papers that use an integrative taxonomy approach to describe the species diversity of diplostomid trematodes in Brazil.


Fish parasites Flatworms Trematoda Cytochrome c oxidase Morphological analysis Bayesian analysis 



The authors would like to thank Tiago dos Santos Tardivo and Carolina Massucci M. da Silva from the Center for Electronic Microscopy in the Department of Biosciences of UNESP Botucatu for their assistance with the scanning electron microscope (SEM) preparations.

Funding information

The present work was supported by a research fellowship from the São Paulo Research Foundation, or FAPESP (V. D. A., 2012/23655-0), (R. K. A., 2014/12862-0), (R. J. S., 2016/50377-1); M.I.M. (grant number CAPES AUX-PE-PNPD 3005/2010 and the Young Researcher Program PROPE-UNESP 02/2016, FAPESP 2017/16546-3) by the Conselho Nacional de Desenvolvimento Científico e Tecnológico, or CNPq (R. J. S., 309125/2017-0) and by CNPq-PROTAX (R. J. S., 440496/2015-2). G.P.P.L. received support from the program PAPIIT-UNAM IN202617.

Compliance with ethical standards

The euthanasia methodologies of the host fish were carried out following the guidelines of the National Council of the Animal Experimentation Control (CONCEA), and the research project was submitted to the Ethical Committee on Animal Use (CEUA) of the Universidade do Sagrado Coração (USC) (authorization no. 3353050417).

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

436_2019_6362_MOESM1_ESM.doc (89 kb)
ESM 1 (DOC 89 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Larissa Sbeghen Pelegrini
    • 1
  • Thayana Gião
    • 2
  • Diego Henrique Mirandola Dias Vieira
    • 1
  • Maria Isabel Müller
    • 1
    • 3
  • Reinaldo José da Silva
    • 1
  • Gerardo Pérez-Ponce de León
    • 4
  • Rodney Kozlowiski de Azevedo
    • 5
  • Vanessa Doro Abdallah
    • 5
    Email author
  1. 1.Instituto de Biociências de BotucatuUniversidade Estadual Paulista (UNESP)BotucatuBrazil
  2. 2.Pró-reitoria de Pesquisa e Pós-graduação, Laboratório de Ictioparasitologia, Rua Irmã ArmindaUniversidade do Sagrado Coração (USC)BauruBrazil
  3. 3.Departamento de Ciências BiológicasUniversidade Federal de São Paulo (UNIFESP)DiademaBrazil
  4. 4.Instituto de BiologiaUniversidad Nacional Autónoma de México (UNAM)Mexico CityMexico
  5. 5.Programa de Pós-Graduação em Análise de Sistemas AmbientaisCentro Universitário CESMACMaceióBrazil

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