Phylogenetic relationships, expanded diversity and distribution of Crassiphiala spp. (Digenea, Diplostomidae), agents of black spot disease in fish

  • Tyler J. Achatz
  • Eric E. Pulis
  • Alan Fecchio
  • Isaac J. Schlosser
  • Vasyl V. TkachEmail author
Fish Parasitology - Original Paper


Crassiphiala is a monotypic genus of diplostomid digeneans and is the type genus of the subfamily Crassiphialinae. The type species Crassiphiala bulboglossa parasitizes kingfishers in the Nearctic and has a Neascus-type metacercaria that encysts on fish intermediate hosts, often causing black spot disease. While recent molecular phylogenetic studies included some members of the Crassiphialinae, no DNA sequence data of Crassiphiala is currently available. Our molecular and morphological study of adult and larval crassiphialines from the Americas revealed the presence of at least three lineages of Crassiphiala from the Nearctic and two lineages from the Neotropics. This is the first record of Crassiphiala from the Neotropics. Herein, we provide the first molecular phylogeny of the Diplostomoidea that includes Crassiphiala. Our data revealed 0.2–2.4% divergence among 28S sequences and 11–19.8% among CO1 sequences of lineages of Crassiphiala. The results of our analyses did not support the monophyly of Crassiphialinae. Our results clearly demonstrated that the diversity of Crassiphiala has been underestimated.


Diplostomidae Crassiphiala Molecular phylogeny Diversity Black spot disease 



We are grateful to Dr. João B. Pinho (Universidade Federal de Mato Grosso, Cuiabá, Mato Grosso, Brazil), Dr. Francisco Tiago de Melo (Federal University of Pará, Belém, Pará, Brazil), and Dr. Jeffrey A. Bell (University of North Dakota) for their invaluable help with obtaining permits and field collecting. We are grateful to Mary Jaros-Gourneau (University of North Dakota) for her assistance with processing of some of the samples.

Conflict of interest

The authors declare that they have no conflict of interests.

Funding information

Collecting and processing of the specimens were supported by the grant DEB-1120734 from the National Science Foundation and grant R15AI092622 from the National Institutes of Health, USA to VVT, and the Joe K. Neel Memorial Award from the University of North Dakota and Willis A. Reid, Jr. Student Research Grant from the American Society of Parasitologists to TJA. AF was supported by a postdoctoral fellowship (PNPD scholarship) from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).

Compliance with ethical standards

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards (University of North Dakota IACUC protocol 0610-1). This article does not contain any studies with human participants performed by any of the authors.


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Copyright information

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

Authors and Affiliations

  1. 1.Department of BiologyUniversity of North Dakota, Starcher HallGrand ForksUSA
  2. 2.Department of Science and MathematicsNorthern State UniversityAberdeenUSA
  3. 3.Programa de Pós-Graduação em Ecologia e Conservação da BiodiversidadeUniversidade Federal de Mato GrossoCuiabáBrazil

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