Preparation and Inactivation of C. burnetii and S. aureus

Bacteria (Nine Mile and Guyana strains of C. burnetii) were prepared as previously reported (12 (link)). Avirulent variants of Nine Mile bacteria were obtained after repeated passages in L929 cells. Bacteria were stored at −80°C, their concentration was determined by Gimenez staining, and bacterial viability was assessed using the live/dead BacLight bacterial viability kit (Molecular Probes, Life Technologies) (51 (link)). Bacteria were inactivated at 95°C for 30 min. S. aureus (ATCC 25923) bacteria were grown on blood agar plates (bioMérieux) and quantified by flow cytometry (FACS BD Fortessa).

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Mezouar S., Vitte J., Gorvel L., Ben Amara A., Desnues B, & Mege J.L. (2019). Mast Cell Cytonemes as a Defense Mechanism against Coxiella burnetii. mBio, 10(2), e02669-18.

Publication 2019

BacLight bacterial viability kit blood agar plates FACS BD Fortessa

Agar Bacteria Bacterial viability Blood Flow cytometry L929 cells Molecular probes S aureus Strains

Corresponding Organization :

Other organizations : Méditerranée Infection Foundation, Aix-Marseille Université, Institut de Recherche pour le Développement, Inserm, Centre National de la Recherche Scientifique

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Variable analysis

independent variables

Strains of Coxiella burnetii bacteria (Nine Mile and Guyana strains)
Passage of Nine Mile bacteria in L929 cells to obtain avirulent variants

dependent variables

Bacterial concentration determined by Gimenez staining
Bacterial viability assessed using the live/dead BacLight bacterial viability kit
Quantification of Staphylococcus aureus (ATCC 25923) bacteria by flow cytometry

control variables

Storage of bacteria at -80°C
Inactivation of bacteria at 95°C for 30 minutes

positive controls

Staphylococcus aureus (ATCC 25923) bacteria as a control sample

negative controls

Inactivation of bacteria at 95°C for 30 minutes to serve as a control for subsequent experiments

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