Developing a Mouse Model of Chronic Pseudomonas aeruginosa Skin Wound Infection

Abstract

Source: de Vries, C. R., et.al. A Delayed Inoculation Model of Chronic Pseudomonas aeruginosa Wound Infection. J. Vis. Exp. (2020)

This video demonstrates the delayed inoculation of luminescent Pseudomonas aeruginosa into a mouse bearing two full-thickness dorsal skin wounds covered by a transparent film dressing. This facilitates biofilm formation and the establishment of chronic infection. Regular bioluminescence monitoring enables quantification of bacterial burden and tracking of infection progression over time.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Inoculation with Pseudomonas aeruginosa

1. Dilute overnight PAO1:lux culture to OD₆₀₀ = 0.05 in 75 mL of Lysogeny broth (LB) media containing 100 µg/mL carbenicillin and grow the bacteria until the culture is in early exponential phase (OD₆₀₀≈0.3). This should take approximately 2–3 h.
2. Dilute PAO1:lux in phosphate buffered saline (PBS) to a concentration of (7.5 ± 2.5) x 10² CFU/mL. Be sure to prepare excess inoculum to ensure ....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
0.9% Sodium Chloride injection	Hospira	2484457
C57BL/6J male mice	The Jackson Laboratory	664
Phosphate Buffered Saline (PBS) 1x	Corning	21-040-CV
Press and Seal Cling Wrap	Glad	n/a
Safetyglide Insulin syringe, 1/2 mL, 30 G x 5/16 TW	BD	305934
Scale	Ohaus Scout Pro	SP202
Supplical Nutritional Supplement	Henry Schein Animal Health	29908