Development of acute Pseudomonas aeruginosa and Acinetobacter baumannii lung mono-challenge models in mice using oropharyngeal instillation.

Antimicrobial resistant pathogens such as Pseudomonas aeruginosa and Acinetobacter baumannii can cause potentially fatal infections in susceptible individuals, with respiratory tract infections among the most common clinical presentation. The development of novel treatments or prophylactic interventions to combat these infections is urgently needed, and requires robust, reliable animal models for their preclinical evaluation. In particular, the bacterial burden needs to be accurately determined before and after administration of the potential therapy under evaluation to quantitate the effectiveness of the treatment. We provide two reliable, non-invasive murine acute lung challenge models with either P. aeruginosa or A. baumannii using an oropharyngeal aspiration technique, which has been widely overlooked in studies testing vaccines or treatments for these pathogens. Here, we show that this non-surgical technique to deliver suspensions into mouse lungs does not significantly impact animal welfare (based on welfare monitoring and weight) and allows uniform bilateral distribution of the bacterial dose resulting in even bioburden in both lungs. The optimal timepoint for humane killing and organ harvest was 24 h after challenge for both pathogens, and at least 4x106 and 107 CFU/mouse were needed to obtain a reproducible P. aeruginosa or A. baumannii bioburden, respectively. These mouse challenge models offer a valuable tool to assess therapeutic interventions against P. aeruginosa or A. baumannii infections.