Abstract

Microsporidia are eukaryotic, unicellular parasites that have been studied for more than 150 years. These organisms are extraordinary in their ability to invade a wide range of hosts including vertebrates and invertebrates, such as human and commercially important animals. A lack of appropriate labeling methods has limited the research of the cell cycle and protein locations in intracellular stages. In this report, an easy fluorescent labeling method has been developed to mark the proliferative and sporogonic phases of microsporidia Nosema bombycis in host cells. Based on the presence of chitin, Calcofluor White M2R was used to label the sporogonic phase, while β-tubulin antibody coupled with fluorescence secondary antibody were used to label the proliferative phase by immunofluorescence. This method is simple, efficient and can be used on both infected cells and tissue slices, providing a great potential application in microsporidia research.

Highlights

  • Microsporidia are obligate intracellular parasites that infect many animal species, including humans and several commercially important organisms, such as bees, silkworms, chickens and aquatic animal [1,2,3,4]

  • SDS-PAGE analysis indicated that recombinant β-Tubulin protein was expressed at a molecular mass of nearly 53 kDa, which was consistent with the deduced size (Fig 1B)

  • Immunoblot assays indicated that the β-Tubulin antiserum recognized a 50 kDa protein in the extract of N. bombycis mature spores (Fig 1C)

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Summary

Introduction

Microsporidia are obligate intracellular parasites that infect many animal species, including humans and several commercially important organisms, such as bees, silkworms, chickens and aquatic animal [1,2,3,4]. The first identified Microsporidia, is the pathogen of silkworm Bombyx mori which was found by Louis Pasteur [6, 7]. It causes pebrine disease though horizontal transmission and vertical transmission. Vertical transmission of N.bombysis via eggs could bring big losses to sericulture, has been the quarantine pathogen of sericulture in every country that cultivate silkworms. The parasite infect the silkworms and some other insects like Drosophila, locusts, Pieris rapae, etc. [8,9,10]

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