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

Parasitology research (2014-09-12)
Steffanie V Burk, Sriveny Dangoudoubiyam, Tammy Brewster-Barnes, Uneeda K Bryant, Daniel K Howe, Craig N Carter, Eric S Vanzant, Robert J Harmon, Kevin R Kazacos, Mary G Rossano
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.

MATERIALS
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