Bioluminescent Imaging of Bacterial Biofilm Infections In Vivo - Bacterial Pathogenesis: Methods and Protocols - page 220

3.8. Experimental Model of Infection
The experimental catheter-associated UTI in a rat model developed by
Kurosaka et al. (19) was established in mice with modifications for BPI.
1. Cut polyethylene catheter material PE 50 into 6-mm long segments and fit onto
a flexible metal wire.
2. Wrap this around a similar metal wire in a spiral and place in boiling water for
1 min.
3. Remove the segment and sterilize each piece with 70% ethanol and air dry
overnight.
4. Bacterial biofilm is grown on the catheter material as described above in
Subheading 3.4 .
5. Cut the metal stylet of a 24-gauge Teflon catheter up to 5 cm and fit either
sterile PE tubing or tubing that has been colonized with bacteria.
6. Anesthetize CF-1 female mice weighing 26-30 g with 2.5% isoflurane gas and
place in the supine position.
7. Clean the periurethral area with 70% ethanol.
8. Grasp the papilla with light pressure using forceps and lift away from the
abdomen.
9. While maintaining the papilla in this position, place the end of the stylet bearing
the 6-mm-long tubing into the urethral opening and advance until the leading
end is in the bladder.
10. Push the 5-cm Teflon catheter until the shorter segment is pushed off the stylet,
thereby leaving the 6-mm-long PE tubing free in the bladder lumen (the PE
tubing will revert spontaneously to its spiral form thus remain in place until the
end of the monitoring period).
11. Remove the Teflon catheter with the stylet.
12. Image animals using a highly sensitive low-light optical imaging system (see
imaging procedure below).
13. Remove Teflon catheter post-inoculation.
To induce post-catheterize infection, place a sterile tubing in the bladder as
described above and inoculate the bladder with defined quantity of pathogen
in 50 μL phosphate-buffered saline over 30 s by insertion of Teflon catheter
into the bladder through the urethra ( see Note 5 for challenge doses required
to establish chronic UTI for each pathogen).
3.9. Central Venous Catheter-Associated Infection in Mice
Long-term implantation of intravascular devices is one of the major advances
in modern medical and surgical practice. A common complication with these
implanted foreign bodies is their susceptibility to pyogenic infections, especially
due to Staphylococci (20) . Establishment of such infections is due to either
perioperative bacterial contamination or postoperative colonization of the