Production of polyclonal anti-β-tubulin antibodies and immunodetection of Vairimorpha (Nosema) ceranae (Opisthosporidia: Microsporidia) proliferative stages in the midguts of Apis mellifera and in the Sf9 cell culture

Abstract

Summary A number of methods to detect microsporidia infections are based on recovery of spores due to their thick refractive spore walls and presence of chitin that easily binds to fluorogenic and chromogenic stains. Visualization of intracellular stages in infected cells and organs has been always a challenge. In this report a new immunodetection method based on the use of β -tubulin antibodies, has been developed to mark the intracellular life cycle stages of Vairimorpha ( Nosema ) ceranae , an emergent pathogen of Apis mellifera . For this purpose, we carried out the heterologous expression of the V. ceranae β -tubulin gene in E. coli cells and obtained polyclonal antibodies to the recombinant protein. The antibodies were successfully applied for immunolabelling of microsporidian cells in insect midguts in vivo as well as in Sf9 cell culture in vitro .