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Identification “by eye”: integrative character assessment informs regional field identification of greater fritillary butterflies (Nymphalidae: Speyeria)


The pronounced morphological variability exhibited by Speyeria butterflies can hinder the identification of these taxa, and thus presents a challenge to their management and conservation. While several studies have documented declines in American populations of Speyeria due to habitat change, the response of Canadian populations of Speyeria to widespread anthropogenic pressures remains poorly understood due to their difficult identification. Here, we assessed the utility of morphological characters recommended in regional field guides for the identification of Speyeria butterflies in Canada, focusing on three morphologically variable subspecies that inhabit boreal forests subject to widespread oil sands extraction: S. aphrodite manitoba, S. atlantis hollandi, and S. hesperis beani. We scored 159 specimens for 11 morphological characters, and complimented this data by sequencing the barcode region of the COI gene for 15 of these specimens. Our results indicate a high level of intraspecific variability in several characters, and the COI gene revealed that initial morphological identifications were incorrect. A further assessment of character reliability identified sets of morphological characters that, in combination with specimen sex and species natural history, improved subspecies identification. Notably, we found that a relatively novel field marker—in vivo eye color—consistently distinguished S. aphrodite manitoba from S. hesperis beani and S. atlantis hollandi. Our results emphasize the importance of using an integrative approach for the accurate identification of morphologically variable species, particularly in situations where molecular methods are not readily available, such as citizen science programs.

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  1. Acorn JH (2017) Entomological citizen science in Canada. Can Entomol.

    Article  Google Scholar 

  2. Acorn JH, Sheldon I (2006) Butterflies of British Columbia. Lone Pine, Edmonton, AL

    Google Scholar 

  3. Bird CD, Hilchie GJ, Kondla NG et al (1995) Alberta butterflies. Alberta Public Affairs Bureau/Queens Printer, The Provincial Museum of Alberta, Edmonton, AL

    Google Scholar 

  4. Breed GA, Stichter S, Crone EE (2013) Climate-driven changes in northeastern US butterfly communities. Nat Clim Chang.

    Article  Google Scholar 

  5. Brock JP, Kaufman K, Bowers R et al (2003) Kaufman field guide to butterflies of North America. Houghton Mifflin, Oak Harbor, OH

    Google Scholar 

  6. Campbell EO, Davis CS, Dupuis JR, Muirhead K, Sperling FAH (2017) Cross-platform compatibility of de novo-aligned SNPs in a non-model butterfly genus. Mol Ecol Res.

    Article  Google Scholar 

  7. Caven AJ, King KC, Wiese JD, Brinley Buckley EM (2017) A descriptive analysis of Regal Fritillary (Speyeria idalia) habitat utilizing biological monitoring data along the big bend of the Platte River. J Insect Conserv, NE.

    Book  Google Scholar 

  8. Chermock FH, Chermock RL (1940) Some new diurnal lepidoptera from the riding mountains and the sand ridge. Can Entomol, Manitoba.

    Book  Google Scholar 

  9. dos Passos CF, Grey LP (1947) Systemic catalogue of Speyeria (Lepidoptera, Nymphalidae) with designations of type and fixations of type localities. Am Museum Novit 404–421

  10. Dunford JC (2009) Taxonomic overview of the greater fritillary genus Speyeria Scudder and the atlantis-hesperis species complexes, with species accounts, type images, and relevant literature (Lepidoptera: Nymphalidae). Insecta Mundi

  11. Giesy JP, Anderson JC, Wiseman SB (2010) Alberta oil sands development. Proc Natl Acad Sci USA 107:951–952.

    CAS  Article  PubMed  Google Scholar 

  12. Glassberg J (2001) Butterflies through binoculars: the west. Oxford University Press, Oxford

    Google Scholar 

  13. Glassberg J (2017) A swift guide to butterflies of North America. Princeton University Press, Princeton

    Book  Google Scholar 

  14. Guppy CS, Shepard J, Royal British Columbia Museum (2001) Butterflies of British Columbia : including western Alberta, southern Yukon, the Alaska Panhandle, Washington, northern Oregon, northern Idaho, northwestern Montana. UBC Press, Vancouver

    Google Scholar 

  15. Hammond PC, Mccorkle DV, Bergman W (2013) Hybridization studies of genomic compatibility and phenotypic expression in the greater fritillary butterflies (Nymphalidae: Argynnini). J Lepid Soc 67:263–273.

    Article  Google Scholar 

  16. Hebert PDN, Cywinska A, Ball SL, DeWaard JR (2003) Biological identifications through DNA barcodes. Proc R Soc B Biol Sci.

    Article  Google Scholar 

  17. Hill RI, Ganeshan M, Wourms L et al (2018) Effectiveness of DNA barcoding in Speyeria butterflies at small geographic scales. Diversity.

    Article  Google Scholar 

  18. Katoh K, Rozewicki J, Yamada KD (2017) MAFFT online service: multiple sequence alignment, interactive sequence choice and visualization. Brief Bioinform.

    Article  PubMed Central  Google Scholar 

  19. Layberry RA, Hall PW, Lafontaine JD, Canada Institute for Scientific and Technical Information (1998) The butterflies of Canada. University of Toronto Press, Toronto

    Book  Google Scholar 

  20. Maddison WP, Maddison DR (2018) Mesquite: a modular system for evolutionary analysis

  21. McHugh A, Bierzychudek P, Greever C et al (2013) A molecular phylogenetic analysis of Speyeria and its implications for the management of the threatened Speyeria zerene hippolyta. J Insect Conserv.

    Article  Google Scholar 

  22. Moeck AH (1975) Geographic variability in Speyeria: comments, records, and description of a new subspecies. Entomological Reprint Specialists, Los Angeles

    Google Scholar 

  23. Nielsen SE, Boyce MS, Stenhouse GB (2004) Grizzly bears and forestry: I. Selection of clearcuts by grizzly bears in west-central Alberta, Canada. For Ecol Manage.

  24. Pelham JP (2012) A catalogue of the butterflies of the United States and Canada, with a complete bibliography of the descriptive and systematic literature. J Res Lepid 40:1–658

    Google Scholar 

  25. R Core Team (2017) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria.

  26. Ríos-Saldaña CA, Delibes-Mateos M, Ferreira CC (2018) Are fieldwork studies being relegated to second place in conservation science? Glob Ecol Conserv.

    Article  Google Scholar 

  27. Riva F, Acorn JH, Nielsen SE (2018a) Localized disturbances from oil sands developments increase butterfly diversity and abundance in Alberta’s boreal forests. Biol Conserv 217:173–180.

    Article  Google Scholar 

  28. Riva F, Acorn JH, Nielsen SE (2018b) Narrow anthropogenic corridors direct the movement of a generalist boreal butterfly. Biol Lett.

    Article  PubMed  PubMed Central  Google Scholar 

  29. Riva F, Pinzon J, Acorn JH, Nielsen SE (2019) Composite effects of cutlines and wildfire result in fire refuges for plants and butterflies in boreal treed peatlands. Ecosystems.

    Article  Google Scholar 

  30. Shuey J, Jacquart E, Orr S et al (2016) Landscape-scale response to local habitat restoration in the regal fritillary butterfly (Speyeria idalia) (Lepidoptera: Nymphalidae). J Insect Conserv.

    Article  Google Scholar 

  31. Soroye P, Ahmed N, Kerr JT (2018) Opportunistic citizen science data transform understanding of species distributions, phenology, and diversity gradients for global change research. Glob Chang Biol.

    Article  PubMed  Google Scholar 

  32. Soulé ME (1985) What is conservation biology? Bioscience 35:727–734.

    Article  Google Scholar 

  33. Swengel SR, Schlicht D, Olsen F, Swengel AB (2011) Declines of prairie butterflies in the midwestern USA. J Insect Conserv 15:327–339.

    Article  Google Scholar 

  34. Williams BL (2002) Conservation genetics, extinction, and taxonomic status: a case history of the regal fritillary. Conserv Biol.

    Article  Google Scholar 

  35. Williams BL, Brawn JD, Paige KN (2003) Landscape scale genetic effects of habitat fragmentation on a high gene flow species: Speyeria idalia (Nymphalidae). Mol Ecol 12:11–20.

    CAS  Article  PubMed  Google Scholar 

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We thank eButterfly and all of their affiliated citizen scientists for collecting, reporting, and vetting butterfly observation data used in this publication. We additionally thank F. and T. Sperling, B. Acorn, and S. Ferguson for specimen collection, and the Molecular Biology Services Unit at the University of Alberta for sequencing support. Funding for the research was provided by two Alberta Conservation Association Grant in Biodiversity to EC and FR, an NSERC Discovery Grant to Felix Sperling, and a DeWind Award from the Xerces Society to FR.

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FR and EOC contributed equally as first authors to the manuscript; All authors designed the study and contributed to draft revisions; FR and FC collected Speyeria butterflies in the Alberta boreal forest; FC scored all specimens and prepared the data for analysis; FR and EOC conducted the analyses and prepared the R script.

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Correspondence to Federico Riva.

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The authors have no conflicts of interest to declare.

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This research did not involve human participants, and thus an informed consent form was not necessary. The study complies with the University of Alberta Animal Ethics Policy.

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Riva, F., Campbell, E.O., Carroll, F. et al. Identification “by eye”: integrative character assessment informs regional field identification of greater fritillary butterflies (Nymphalidae: Speyeria). J Insect Conserv 24, 259–267 (2020).

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  • COI gene
  • Speyeria
  • Morphology
  • Species identification
  • Citizen science