Position an anesthetized rat laterally on a heating pad to maintain normal body temperature.
Disinfect the shaved skin, then make an incision below the rib cage to expose the abdominal cavity.
Gently displace the liver to visualize the right kidney and externalize the kidney.
Take a syringe containing a suspension of pathogenic E. coli.
Inject the suspension into the renal pelvis, which is a funnel-shaped structure in the kidney, to induce a kidney infection or pyelonephritis.
Cover the injection site with sterile gauze to prevent bacterial spread beyond the injection site, thereby reducing the risk of off-target infection.
Slowly withdraw the needle and rinse the kidney with sterile saline. Then, return the kidney to its anatomical position and close the wound.
The rat is now ready for assessment of host–bacterial interactions.
Prior to starting this procedure, prepare the bacterial injection bolus containing between 1 and 10 million cells. After anesthetizing the rat and preparing its skin for surgery, apply vet eye ointment to the eyes to prevent dryness. Position the rat on a warmed surgical platform in a left decubitus position with its right flank facing up.
Scrub the skin with a disinfectant. Repeat three times and then scrub the surgical site with 70% alcohol swab until the skin is clear. To begin the surgery, using a Number 10 scalpel, make a small, two to three-centimeter right dorsal retroperitoneal incision beginning at the bottom of the rib cage.
Continue by dissecting away the subcutaneous tissue, fat, and muscles to ultimately visualize and access the abdominal cavity.
Once the liver is clearly visible and accessible, use blunted forceps to retract it upwards. Then, using a second pair of blunted forceps, expose the right kidney so it sits just outside of the abdominal cavity.
Now, position the kidney for injection in the nondominant hand. Using the dominant hand, precisely target the injection needle on the renal pelvis. Then, slowly and steadily, inject 0.1 milliliters of bacterial or saline solution. Here, saline is being injected. The bolus will look like a white bubble in the tissue.
Before retracting the needle, place a strip of absorbable hemostat over the injection site to prevent an outflow of the inoculum into the peritoneum. Then, slowly pull the needle out. Now, using a large syringe, thoroughly rinse the kidney with normal saline. Then, return the kidney to the abdominal cavity.
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