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Parasitology research2014; 113(11); 4217-4224; doi: 10.1007/s00436-014-4097-0

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

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.
Publication Date: 2014-09-11 PubMed ID: 25209615DOI: 10.1007/s00436-014-4097-0Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

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The research paper presents a study that investigates the cultivation of Parascaris equorum larvae in the laboratory and the identification of excretory-secretory products from these larvae for potential use in early diagnostic testing of P. equorum infection in horses.

Understanding Parascaris equorum

  • Parascaris equorum is a parasitic roundworm found in the small intestine of horses, donkeys, and other equids. Infections are especially harmful to young horses and can lead to severe digestive problems and mortality.
  • Currently, diagnosis of a P. equorum infection can only be made during the patent phase of the infection, when the parasite is mature and releasing eggs that can be detected in the feces of the infected host.
  • The researchers aimed to identify excretory-secretory (ES) products from P. equorum larvae that could potentially be used to detect an infection before it reaches the patent phase.

Lab Cultivation of P. equorum Larvae

  • The researchers hatched and cultured P. equorum larvae in a laboratory setting. They kept the larvae (at the second and third larval stages, or L2/L3) for up to three weeks to collect their ES products.
  • In addition, they also cultured fifth stage (L5) P. equorum larvae to gather their ES products.
  • The ES products from both L2/L3 and L5 larvae were then examined via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and silver staining.

Analysis of ES Products

  • The SDS-PAGE and silver stain analysis revealed the protein composition of the ES products. The L2/L3 products consisted of proteins ranging from 12-94 kDa (kilodaltons), while the L5 products had proteins ranging from 12-189 kDa.
  • The team then performed a Western blot analysis, a technique used to detect specific proteins in a sample, on the ES products. They used polyclonal antibodies generated against P. equorum and Baylisascaris procyonis ES products, as well as serum from rabbits inoculated with B. procyonis or another type of roundworm called Toxocara canis eggs, and serum from animals naturally infected with P. equorum or T. canis.
  • These analyses revealed specific parasite antigens (proteins that trigger an immune response) migrating at 19 and 34 kDa that could potentially be used for specifically detecting P. equorum infections.

This study’s findings might open up new avenues for generating early diagnostic tools for P. equorum infections in horses, potentially reducing infection rates and increasing overall equine health.

Cite This Article

APA
Burk SV, Dangoudoubiyam S, Brewster-Barnes T, Bryant UK, Howe DK, Carter CN, Vanzant ES, Harmon RJ, Kazacos KR, Rossano MG. (2014). In vitro culture of Parascaris equorum larvae and initial investigation of parasite excretory-secretory products. Parasitol Res, 113(11), 4217-4224. https://doi.org/10.1007/s00436-014-4097-0

Publication

ISSN: 1432-1955
NlmUniqueID: 8703571
Country: Germany
Language: English
Volume: 113
Issue: 11
Pages: 4217-4224

Researcher Affiliations

Burk, Steffanie V
  • Department of Animal and Food Sciences, University of Kentucky, W.P. Garrigus Building, Lexington, KY, 40546-0215, USA, sburk@otterbein.edu.
Dangoudoubiyam, Sriveny
    Brewster-Barnes, Tammy
      Bryant, Uneeda K
        Howe, Daniel K
          Carter, Craig N
            Vanzant, Eric S
              Harmon, Robert J
                Kazacos, Kevin R
                  Rossano, Mary G

                    MeSH Terms

                    • Animals
                    • Antibodies, Helminth / blood
                    • Antigens, Helminth / chemistry
                    • Ascaridida Infections / diagnosis
                    • Ascaridoidea / chemistry
                    • Blotting, Western
                    • Electrophoresis, Polyacrylamide Gel
                    • Horses / parasitology
                    • In Vitro Techniques
                    • Larva / chemistry
                    • Rabbits

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                    Citations

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