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Evaluation of the Skin Peptide Defenses of the Oregon Spotted Frog Rana pretiosa Against Infection by the Chytrid Fungus Batrachochytrium dendrobatidis

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Abstract

Population declines due to amphibian chytridiomycosis among selected species of ranid frogs from western North America have been severe, but there is evidence that the Oregon spotted frog, Rana pretiosa Baird and Girard, 1853, displays resistance to the disease. Norepinephrine-stimulated skin secretions were collected from a non-declining population of R. pretiosa that had been exposed to the causative agent Batrachochytrium dendrobatidis. Peptidomic analysis led to identification and isolation, in pure form, of a total of 18 host-defense peptides that were characterized structurally. Brevinin-1PRa, -1PRb, -1PRc, and -1PRd, esculentin-2PRa and -PRb, ranatuerin-2PRa, -2PRb, -2PRc, and -2PRe, temporin-PRb and -PRc were identified in an earlier study of skin secretions of frogs from a different population of R. pretiosa known to be declining. Ranatuerin-2PRf, -2PRg, -2PRh, temporin-PRd, -PRe, and -PRf were not identified in skin secretions from frogs from the declining population, whereas temporin-PRa and ranatuerin-2PRd, present in skin secretions from the declining population, were not detected in the current study. All purified peptides inhibited the growth of B. dendrobatidis zoospores. Peptides of the brevinin-1 and esculentin-2 families displayed the highest potency (minimum inhibitory concentration = 6.25–12.5 μM). The study provides support for the hypothesis that the multiplicity and diversity of the antimicrobial peptide repertoire in R. pretiosa and the high growth-inhibitory potency of certain peptides against B. dendrobatidis are important in conferring a measure of resistance to fatal chytridiomycosis.

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Acknowledgments

This work was supported by a Faculty Support Grant and a University/National Research Foundation Grant from U.A.E. University to J.M.C. and NSF grant 1121758 to L.R-S. The authors thank Marko Anderson of the Washington Department of Corrections and Carri LeRoy and Kelli Bush of the Sustainability in Prisons Project, and Allison Abrahamse and Rich Sartor of Northwest Trek, for help in collecting skin secretions.

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Authors and Affiliations

  1. Department of Biochemistry, College of Medicine and Health Sciences, United Arab Emirates University, 17666, Al Ain, United Arab Emirates

    J. Michael Conlon, Milena Mechkarska & Manju Prajeep

  2. Departments of Pathology, Microbiology and Immunology, and Biological Sciences, Vanderbilt University School of Medicine, MCN A-5301, Nashville, TN, 37232-2363, USA

    Laura K. Reinert & Louise A. Rollins-Smith

  3. Department of Chemistry, Faculty of Science, United Arab Emirates University, 17551, Al Ain, United Arab Emirates

    Mohammed A. Meetani

  4. Institute for Research and Innovation in Biomedicine (IRIB), CNRS UMR 6270, University of Rouen, 76821, Mont-Saint-Aignan, France

    Laurent Coquet & Thierry Jouenne

  5. Washington Department of Fish and Wildlife, Habitat Program, Olympia, WA, 98501-1091, USA

    Marc P. Hayes

  6. Southern Illinois University-Carbondale, Carbondale, IL, 62901-2594, USA

    Gretchen Padgett-Flohr

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  1. J. Michael Conlon
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  2. Laura K. Reinert
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  3. Milena Mechkarska
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  4. Manju Prajeep
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  5. Mohammed A. Meetani
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  6. Laurent Coquet
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  7. Thierry Jouenne
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  8. Marc P. Hayes
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  9. Gretchen Padgett-Flohr
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  10. Louise A. Rollins-Smith
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Correspondence to J. Michael Conlon.

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Conlon, J.M., Reinert, L.K., Mechkarska, M. et al. Evaluation of the Skin Peptide Defenses of the Oregon Spotted Frog Rana pretiosa Against Infection by the Chytrid Fungus Batrachochytrium dendrobatidis . J Chem Ecol 39, 797–805 (2013). https://doi.org/10.1007/s10886-013-0294-z

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  • DOI: https://doi.org/10.1007/s10886-013-0294-z

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