Source: Hoover, J. L., et al., A Robust Pneumonia Model in Immunocompetent Rodents to Evaluate Antibacterial Efficacy against S. pneumoniae, H. influenzae, K. pneumoniae, P. aeruginosa or A. baumannii. J. Vis. Exp. (2017)
This video demonstrates the establishment of lung infection in an immunocompetent rodent model via intubation and installation of bacterial suspensions into lung lobes. This model enables antibacterial efficacy studies in immunocompetent animals.
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Anesthetize, Intubate, and Inoculate Animals
NOTE: Specific pathogen-free (SPF) immunocompetent male Sprague-Dawley rats weighing 100 – 120 g are recommended. Provide all animals with food and water ad libitum and house them socially on wood chips or other absorbent bedding with standard 12-hour light-dark cycles.
Figure 1: Nonsurgical Intratracheal Intubation. Tools are shown for intubating the rat (A) and the mouse (B). Once anesthetized, the animal should be positioned as shown in C, with the head to the right and tail to the left if the individual performing the technique is right-handed. Place the left forefinger gently on the throat to help palpate the tracheal rings for confirmation of correct cannula placement. The inoculum should be placed deep into the left lung, as shown in the ventral view of a rat lung (D). Colonies growing on agar after serial dilution and plating of a sample should appear similar to that shown in E. Figure 1D, Copyright © Gill Smith [Laboratory Animals, Volume 25 (1991), G. Smith, A simple non-surgical method of intrabronchial instillation for the establishment of respiratory infections in the rat, pages 46 – 49]. Reprinted with permission.
The authors have nothing to disclose.
Guide Tool | Intek Services Ltd | GT01 | |
1 mL TB syringe slip tip | Becton Dickinson and Company | 309659 | |
PrecisionGlide Needle 25 G x 5/8 | Becton Dickinson and Company | 305122 | |
Intramedic polyethylene tubing | Clay Adams brand – Becton Dickinson and Company | 427401 | Plastic cannula for mouse |
Animal feeding needle – straight 20 x 3" 2-1/4 | Popper and Sons, Inc | 7903 | Used to create metal cannula for mice |
75 TN 5.0 µL syringe 26s/2"/2 | Hamilton | 87930 | HPLC glass injection syringe |
Difco Nutrient Agar | Becton Dickinson and Company | 213000 | |
Pyrex tube, culture 25×150 screwcap with rubber line | Corning | 9825-25 | To autoclave/store metal cannulae |
PrecisionGlide Needle 25 G x 5/8 | Becton Dickinson and Company | 305122 | |
70% isopropyl alcohol | Vi-Jon (Swan) | NDC 0869-0810-43 | |
Isoflurane, USP | Piramal Healthcare | NDC 66794-013-10 | |
50 mL Polypropylene Conical Tube 30 x 115 mm | Falcon, a Corning Brand | 352070 | |
75 TN 5.0 µL syringe 26s/2"/2 | Hamilton | 87930 | HPLC glass injection syringe |