Coxiella burnetii, a small, gram negative bacterium, is the causative agent of Q fever, a disease with flu-like symptoms that has both acute and chronic stages of infection. Q fever is zoonotic with reservoirs in many domesticated animals such as sheep and cattle. As with many other obligate intracellular parasites, a Coxiella infection can be difficult to treat and, even with treatment, chronic infection may still occur. In order to identify small molecules that inhibit bacterial growth, the nonpathogenic C. burnetii nine mile phase II Clone 4 (CBNMIIC4) was prepared in axenic media under hypobaric conditions and was characterized by proteomic analysis and mammalian cell infection followed by antibody staining. Methods to enumerate bacteria using OD600 density, colony forming units and genomic equivalents were developed to standardize the multiplicity of infection (MOI). RAW cell, or mouse macrophage cell, monolayers infected with CBNMIIC4 were used to test the effect of the antibiotic doxycycline and the mycotoxin cytochalasin B on CBNMIIC4 infection for the purpose of establishing positive drug controls for the high throughput assay. Characterized CBMNIIC4 stocks (stored at -70°C) are now ready to be transferred for use in a high throughput assay.

Development of a Coxiella burnetii culturing method for high throughput assay to identify growth inhibitory small molecules

255th National Meeting (2018)

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