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[Staining of microsporidian spores with diamidine phenylindole]

Iu S Tokarev, Iu M Malysh, Iu A Zakharova

    Parazitologiia
    |July 28, 2012
    PubMed

    Insights

    DNA-specific fluorochrome DAPI aids in diagnosing microsporidian infections in insects. It stains key spore structures, improving identification and viability assessment of these pathogens.

    Area of Science:

    • Microbiology
    • Parasitology
    • Cell Biology

    Context:

    • Microsporidian infections pose challenges in diagnosing invertebrate and vertebrate hosts.
    • Microscopy and staining techniques are crucial for pathogen identification.
    • DNA-specific fluorochrome 4',6-diamidino-2-phenylindole (DAPI) is a common cellular stain.

    Purpose:

    • To evaluate the utility of DAPI staining for diagnosing microsporidian infections in insect hosts.
    • To investigate DAPI's binding characteristics beyond nuclear DNA within microsporidian spores.
    • To assess DAPI's effectiveness in differentiating microsporidian spores and evaluating spore viability.

    Summary:

    • DAPI staining was applied to microsporidian spores from various insect orders (Orthoptera, Coleoptera, Diptera, Lepidoptera).
    • While DAPI primarily stains diplokarya (nuclei), it also binds to other spore components like the exospore, cytoplasm, polar filament, and polaroplast.
    • These non-nuclear staining patterns, though weaker than DNA staining, aid in distinguishing true microsporidian spores and identifying unstained endospore regions, facilitating diagnosis of mono- and diplokaryotic spores.
    • Staining of the extruded polar filament provides insights into spore viability and the extrusion process.

    Impact:

    • DAPI staining offers a valuable, accessible method for diagnosing microsporidian infections in insects.
    • The technique enhances the differentiation of microsporidian spores from non-pathogenic structures.
    • Observing polar filament extrusion via DAPI staining can serve as an indicator of spore viability and infection stage.

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