15. Using tissue forceps (two tooth dissecting/Adson-Brown),
pinch and hold up the abdominal muscle along the linea alba
and make a small incision.
16. Using scissors, extend the opening in abdominal muscle to
~4 cm (Fig. 3d).
17. Using the same tissue forceps, raise one side of the abdominal
incision and place a DMC inside rat peritoneal cavity using the
blunt-end forceps (Fig. 3e). Gently push the DMC toward
either side of the peritoneal cavity to prevent it from being
nicked/ruptured during suturing. Be sure to position it so that
it does not become entangled in the intestines or interfere with
bladder expansion.
18. Close the abdominal incision site by suturing (Fig. 3f). Begin
by suturing each end of the incision with double knots. Working upward, close the remaining incision by placing sutures
~2–3 mm apart (Fig. 3g). Trim excess suture material using
scissors.
19. Bring both sides of skin incision together using the two forceps, aligning them (Fig. 3h), and close the skin incision site
using a contiguous line of wound clips (Fig. 3i) (see Note 15).
20. Place the rat on top of a clean surgical drape in a clean cage
containing fresh bedding. Place the cage on top of the circulating water heating pad to maintain the appropriate core body
temperature. Monitor the animal continuously until alert and
responsive and then turn off heating pad.
21. Analgesia should be administered for at least 2 days
postoperatively.
22. Animals should be monitored at least once daily for the first
week postoperatively and then every other day thereafter. If
animals show any sign of distress or discomfort, consult institutional veterinary staff immediately (see Note 16).
3.4 Recovery
of Mammalian
Host-Adapted
Organisms from DMCs
1. At 7–12 days after implantation, euthanize animals by CO 2
asphyxiation or other approved method.
2. Place animal in a BSC in a supine position (Fig. 4a).
3. Using sterile surgical scissors, expose the abdominal wall.
4. Using sterile tissue forceps, lift one side of the sutured abdominal incision site and reopen using sterile scissors.
5. Locate and remove the DMC using blunt-end forceps.
6. Using sterile forceps to hold the DMC by one knot, remove
contents using an 18-G, 1.5 in. needle attached to a sterile
10 ml syringe. Insert needle at the top of DMC to avoid
leakage (Fig. 4b).
7. Transfer contents to a sterile 15 ml tube (see Note 17).
238
Andre Alex Grassmann and Melissa J. Caimano
pinch and hold up the abdominal muscle along the linea alba
and make a small incision.
16. Using scissors, extend the opening in abdominal muscle to
~4 cm (Fig. 3d).
17. Using the same tissue forceps, raise one side of the abdominal
incision and place a DMC inside rat peritoneal cavity using the
blunt-end forceps (Fig. 3e). Gently push the DMC toward
either side of the peritoneal cavity to prevent it from being
nicked/ruptured during suturing. Be sure to position it so that
it does not become entangled in the intestines or interfere with
bladder expansion.
18. Close the abdominal incision site by suturing (Fig. 3f). Begin
by suturing each end of the incision with double knots. Working upward, close the remaining incision by placing sutures
~2–3 mm apart (Fig. 3g). Trim excess suture material using
scissors.
19. Bring both sides of skin incision together using the two forceps, aligning them (Fig. 3h), and close the skin incision site
using a contiguous line of wound clips (Fig. 3i) (see Note 15).
20. Place the rat on top of a clean surgical drape in a clean cage
containing fresh bedding. Place the cage on top of the circulating water heating pad to maintain the appropriate core body
temperature. Monitor the animal continuously until alert and
responsive and then turn off heating pad.
21. Analgesia should be administered for at least 2 days
postoperatively.
22. Animals should be monitored at least once daily for the first
week postoperatively and then every other day thereafter. If
animals show any sign of distress or discomfort, consult institutional veterinary staff immediately (see Note 16).
3.4 Recovery
of Mammalian
Host-Adapted
Organisms from DMCs
1. At 7–12 days after implantation, euthanize animals by CO 2
asphyxiation or other approved method.
2. Place animal in a BSC in a supine position (Fig. 4a).
3. Using sterile surgical scissors, expose the abdominal wall.
4. Using sterile tissue forceps, lift one side of the sutured abdominal incision site and reopen using sterile scissors.
5. Locate and remove the DMC using blunt-end forceps.
6. Using sterile forceps to hold the DMC by one knot, remove
contents using an 18-G, 1.5 in. needle attached to a sterile
10 ml syringe. Insert needle at the top of DMC to avoid
leakage (Fig. 4b).
7. Transfer contents to a sterile 15 ml tube (see Note 17).
238
Andre Alex Grassmann and Melissa J. Caimano