Abstract
Bacterial loads can be determined as colony forming units (CFU) at any point of the infection by culturing spleen homogenates on agar plates. This is a reliable method for comparing the kinetics of infection in various mouse strains, estimating the virulence of different bacterial mutants or isolates and for vaccine testing and vacine estandarization. Although this method has been designed to recover Brucella or Salmonella organisms from spleen, the procedure may be applicable for other bacteria such as Listeria and Mycobacterium as well as to count bacterial loads in other organs such as liver or lymph nodes.
Keywords: Spleen, CFU, Bacteria, Mouse
Materials and Reagents
Equipment
Procedure
Notes
Recipes
Acknowledgments
We thank María-Jesús Grilló for helping in the standardization of this technique. This protocol was adapted from the following original published papers Barquero-Calvo et al. (2007) and Nagy et al. (2007), and from Covelli (2013). This work was funded by grants from FIDA-2006, FS-CONARE (UNA/UCR), MICIT/CONICIT (FI-487-09), 8-N-2005 and B/3456-1 (NeTropica), B/3456-2 (IFS), AGL2004-01162/GAN (Spain), CNRS and INSERM (France) and MASTERSWITCH projects (European Communities).
References
If you have any questions/comments about this protocol, you are highly recommended to post here. We will invite the authors of this protocol as well as some of its users to address your questions/comments. To make it easier for them to help you, you are encouraged to post your data including images for the troubleshooting.