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Silkworm model for Francisella novicida infection

Publication Date - 2017-10-21 00:00:00

Publication Title : Silkworm model for Francisella novicida infection

Publicationed By : Shib Shankar Saha

Publication Publication Date : 2017-10-21 00:00:00

Publication Online Link : http://dx.doi.org/10.1016/j.micpath.2017.10.036

Publication Description :

Understanding the virulence and pathogenesis of human pathogens using insect models is an increasingly  popular method. Francisella novicida, which is virulent in mice but non-pathogenic to immunocompetent humans,  is widely used as an ideal candidate for Francisella research. In this study, we developed a silkworm  (Bombyx mori) infection model for F. novicida by inoculating the hemocoels of silkworms with F. novicida. We  found that silkworms died within 3–7 days of F. novicida infection. However, the deletion mutant of DotU, the  core part of type VI secretion systems, failed to kill silkworm. In whole silkworm bodies, the bacterial load of the  DotU deletion mutant was significantly less than that of the wild-type strain. Approximately 10-fold increase in  bacterial load was recorded in hemolymph and subcutaneous tissues compared with that in the silk gland,  Malpighian tubule, and reproductive organs. The CFU count of the DotU deletion mutant in all organs was  similar results to the whole body CFU count. Confocal microscopy further confirmed the arrested growth of the  mutant strain within hemocytes. The intracellular growth of F. novicida strains was also analyzed using the  silkworm ovary-derived cell line BmN4. In BmN4, both CFU count assay and confocal microscopy revealed  extensive growth of the wild-type strain compared with that of the mutant strain. Francisella DotU has already  been proven as a virulence factor in mammals, and it was also found to be an essential virulence factor in our  silkworm infection model. Therefore, this silkworm infection model is suitable for identifying new virulence  factors of Francisella.



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