Molecular detection of microsporidia Vairimorpha ceranae and Nosema bombycis growth in the lepidopteran Sf9 cell line

Construction of scFv Antibodies against the Outer Loops of the Microsporidium Nosema bombycis ATP/ADP-Transporters and Selection of the Fragment Efficiently Inhibiting Parasite Growth

“…Previously, we produced polyclonal Abs against N. bombycis β-tubulin overexpressed in E. coli and demonstrated the promise of Western blotting for detecting intracellular growth of the parasite in infected Sf9 cell cultures [ 22 ]. In this study, we attempted to use immunoblotting with these Abs to further confirm the N. bombycis -inhibitory efficacy of scFv5 compared to other selected recombinant Abs.…”

“…When cell cultures were infected with 10 or 20 spores per cell, Abs against N. bombycis β-tubulin stained two protein bands. The lower band corresponded to the parasite β-tubulin, and the upper one, to the homologous S. frugiperda protein [ 22 ]. When cell cultures were infected with 50 spores per cell, the parasite protein band was stained more brightly.…”

“…Previously, it was shown that the analysis of microsporidia transcripts involved in sporogenesis but not expressed at the initial stages of infection may be successfully used to assay the parasite growth in cell cultures, regardless of a multiplicity of their infection [ 25 ]. In the case of infection of Sf9 cells with N. bombycis spores, the level of expression of the gene encoding the parasite PTP2 may serve as a such marker [ 22 ]. To match the number of parasite and insect host transcripts in the same cDNA samples, we used the expression level of the S. frugiperda COXI gene as a reference.…”

“…Although the formation of N. bombycis spores in B. mori ovarian tissue cultures was observed as early as 2 days after infection [ 28 ], qPCR analysis of the parasite β-tubulin gene copies [ 13 ] and immunoblotting with Abs against the same protein [ 22 ] demonstrated the parasite growth in Sf9 cells between days 4 and 7 after inoculation. This is most likely due to the immediate germination of first-generation spores to infect new insect cells.…”

The Use of Parthenogenetic Clones to Create Highly Heterogeneous Hybrids of the Silkworm (Bombyx mori L.)

“…For example, the use of clones to study the nature of motor activity [2]. It is proved that traditional methods of breeding in combination with methods of cloning, obtaining polyploid forms and elimination of lethal are suitable for creating an open parthenozygotic population of silkworms optimal for solving problems of sericulture [3][4][5][6], it was also possible to prove the possibility of using parthenoclones in breeding and for hybridization [7][8][9]. As the mother breed for the preparation of hybrids, a female parthenoclone is used, consisting of females with strictly the same heredity.…”