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In vitro culture of Parascaris equorum larvae and initial investigation of parasite excretory-secretory products

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Abstract

Currently, diagnosis of Parascaris equorum infection in equids is limited to patent infections. The goals of this study were to culture P. equorum larvae in vitro and identify excretory-secretory (ES) products for prepatent diagnostic testing. Parascaris equorum L2/L3 larvae were hatched and cultured for up to 3 weeks for ES product collection. Fifth stage (L5) P. equorum were also cultured for ES product collection. Examination of ES fractions by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and silver stain revealed L2/L3 products ranging from 12–94 kDa and L5 products ranging from 12–189 kDa. Western blot analyses were conducted using polyclonal antibodies produced against P. equorum or Baylisascaris procyonis L2/L3 ES products, sera from rabbits inoculated with B. procyonis or Toxocara canis eggs, and sera from animals naturally infected with P. equorum or T. canis. Western blot results indicated parasite antigens migrating at 19 and 34 kDa may be useful for specifically detecting P. equorum infections.

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Acknowledgments

We thank the University of Kentucky’s College of Agriculture for funding this research. The authors are grateful to Frank Berry, Bryan Cassill, Dr. Chris Christensen, Kristen Fox, Dr. Eugene Lyons, Dr. Martin Nielsen, Dr. William Silvia, Laura Strasinger, Dr. Elizabeth Ubelhor, and Stacy White for their assistance with specimen collection. We also thank Dr. Noel Inocencio, Jessica Gould, Lynn Ennis, and Sara Tanner for sharing their laboratory expertise.

This investigation (Manuscript No. 14-07-056) was approved by the director of the University of Kentucky Agricultural Experiment Station. All research conducted was in compliance with the current laws of the United States.

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Authors and Affiliations

  1. Department of Animal and Food Sciences, University of Kentucky, W.P. Garrigus Building, Lexington, KY, 40546-0215, USA

    Steffanie V. Burk, Tammy Brewster-Barnes, Eric S. Vanzant, Robert J. Harmon & Mary G. Rossano

  2. Department of Veterinary Science, Gluck Equine Research Center, University of Kentucky, Lexington, KY, 40546-0099, USA

    Sriveny Dangoudoubiyam & Daniel K. Howe

  3. Veterinary Diagnostic Laboratory, Department of Veterinary Science, University of Kentucky, Lexington, KY, 40511, USA

    Uneeda K. Bryant & Craig N. Carter

  4. Department of Comparative Pathobiology, College of Veterinary Medicine, Purdue University, 625 Harrison Street, West Lafayette, IN, 47907, USA

    Kevin R. Kazacos

  5. Equine Science Department, Otterbein University, 1 South Grove Street, Westerville, OH, 43081, USA

    Steffanie V. Burk

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  1. Steffanie V. Burk
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  2. Sriveny Dangoudoubiyam
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Correspondence to Steffanie V. Burk.

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Burk, S.V., Dangoudoubiyam, S., Brewster-Barnes, T. et al. In vitro culture of Parascaris equorum larvae and initial investigation of parasite excretory-secretory products. Parasitol Res 113, 4217–4224 (2014). https://doi.org/10.1007/s00436-014-4097-0

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  • DOI: https://doi.org/10.1007/s00436-014-4097-0

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