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Research and Innovation: Laboratory Animal Medicine

Aseptic Survival Surgery – Rodents

Personnel Training and Survival Surgery Certification

  • All personnel must complete the online Survival Surgery training in Workday and have a DLAM veterinarian observe them performing survival surgery before they can be approved and certified to perform survival surgeries on rodents at UAMS or ACRI.
  • Additional training may be required for specific techniques/procedures on a case-by-case basis as determined by the IACUC.

Surgery Location

  1. As recommended by The Guide, rodent survival surgery should be performed in a dedicated space. The surgical area must be easily sanitized. The immediate surgical area should not be used for other purposes during the time of surgery, including animal preparation (e.g., fur removal), and traffic in this area should be minimized.
  2. Preparation of Surgery Surface: Prior to surgeries, clean and disinfect the surface upon which surgery will be performed.
  3. Note that DLAM maintains procedural areas acceptable for rodent surgery in the Barton (1RB21), Cancer Institute (438 and 442), and Biomedical Research Center (B128) animal facilities. These rooms are available to all investigators on a first-come-first serve basis. Reservations are recommended.

Surgical Instruments and Implants

All instruments must be sterilized before use. Proper sterilization methods include:

  • Autoclaving
  • Gas sterilization (e.g. ethylene oxide)
  • Liquid sterilant (e.g. glutaraldehyde)

Between animals, instruments should be cleaned of debris and tips re-sterilized by being placed in a glass bead sterilizer for at least 10-20 seconds, or in 70% isopropyl alcohol for at least 2 minutes.

If using a glass bead sterilizer, allow tips to cool completely before use (cold, sterile water or saline may be used to cool instrument tips). If using alcohol, allow alcohol to evaporate completely or rinse tips in sterile water before use.

While alcohols have limitations as disinfectants, Keen et al. reported that, with sufficient soaking in isopropyl alcohol between surgeries, the same batch of sterilized instruments could be used for up to 5 serial laparotomies in mice without developing bacterial contamination. They note that more complex/invasive IACUC Policy: Rodent Aseptic Survival Surgery Page 3 of 10 surgeries may require more frequent re-sterilization, so the investigator should carefully consider the specifics of their surgical procedure when selecting the re-sterilization frequency. Implants should be sterilized according to manufacturer recommendations

Surgical Supplies

Start each day using sterilized instruments, sterile surgical supplies and sterile wound closure materials. Expired materials cannot be used for survival procedures. They may be used, in some circumstances, for non-survival procedures. If both survival and non-survival procedures are performed in the same area, any expired materials must be marked “For non-survival surgery only” and kept separate from those supplies used for survival surgery.
Sterile surgical glove options include:

  • Single use sterile surgical gloves (pre-packaged)
  • Nitrile exam gloves wrapped (or in pouches) and autoclaved
  • Nitrile exam gloves with fingertips soaked in 0.08% peracetic acid for 90 seconds. Ensure gloves are completely dry prior to use.

Surgeon Preparation

  • Long hair should be tied back.
  • Hands should be washed well with antiseptic surgical scrub or other skin disinfectant soap.
  • Surgeons must wear a mask, clean lab coat or gown, and sterile gloves. For surgical procedures that do not involve the surgeon’s hands coming in contact with sterile skin, instrument surfaces, or supplies (i.e. “tips only” procedure), non-sterile gloves may be worn.
  • Non-sterile exam gloves can be worn during animal prep and changed to sterile gloves prior to beginning the surgical procedure or handling sterile items.
  • Gloves should be changed/re-sterilized between animals or when contamination occurs.
  • Fingertips of clean gloves can be soaked in 70% isopropyl for at least 30 seconds in between animals. Allow gloves to dry before contact with animals.
  • This was sufficient for up to 5 serial laparotomies in mice. However, the investigator should carefully consider the specifics of their surgical procedure when selecting the frequency of glove changing.

Animal Preparation

  • Animals should be anesthetized as described in the approved AUP (Appropriate options for anesthesia can be found in the Rodent Aesthetics Formulary).
  • If applicable, administer pre-operative analgesics as described in the AUP.
  • Apply ophthalmic ointment to both eyes to prevent corneal desiccation/ulceration.
  • Remove hair from the surgical site via shaving or depilatory cream as approved in the AUP.
  • Clean the surgical site moving in a circular motion from inside to out with either of the following combinations 1) betadine scrub solution and 70% isopropyl alcohol or 2) chlorhexidine scrub and sterile saline. Repeat this step 3 times using a clean gauze/swab each time.
  • Move animal to surgery location and apply a sterile drape as described in AUP. Drapes are recommended for relatively invasive procedures that place gloves or instruments near unsanitized animal skin/fur. Drapes are also recommended for procedures involving closure with sutures, since the suture material can easily pass over unsanitized animal fur during application. Draping options include sterile gauze, autoclaved paper drape material, sterile glove paper, or ‘Press ‘N’Seal’ film.
  • The animal must be monitored regularly (e.g. toe pinch or tail pinch) to ensure that an adequate plane of anesthesia is reached before the surgery is started and is maintained throughout the procedure.
  • Animals should be placed on a surface that can maintain their body temperature during the procedure. Circulating water pads or re-usable warming discs are good options.

Post-Operative Care

  • Incision closure should be performed using the method and material approved in the AUP. Appropriate options can be found in Appendix C. All incision closure material should be sterile, including metal clips/staples.
  • After completion of the surgery, as much blood as possible should be removed from the site and adjacent fur.
  • Animals should be placed in a clean cage for recovery.
  • Cage should be placed partially on a warming pad/disc such that the animal can move away from the heat source
  • Animals must be frequently observed (every 10-15 minutes) until ambulatory and clearly awake.
  • Once awake and ambulatory, animals can be returned to the housing area in a clean cage.
  • A Postoperative Care Card (Appendix D) must be completed and placed behind the cage card for each cage containing animals that have been returned to the facility following survival surgery.
  • Post-operative analgesics and frequency of monitoring must be followed as described in the AUP.
  • Non-resorbable sutures or wound clips must be removed 7-14 days after surgery unless otherwise described in the approved AUP.

Documentation

A sample surgical record form can be found in Appendix E. Records must be kept of all survival surgical procedures in research animals, and those records must be available for review by the IACUC on their semi-annual site visits as IACUC Policy: Rodent Aseptic Survival Surgery Page 5 of 10 well as by inspectors/visitors from the various regulatory and accrediting agencies. The records should include at a minimum:

  • Species/Animal ID (individual or at cage level)
  • The procedure performed & date
  • The name, dose, and route of the anesthetic used, and an indication of complications, if any.
  • The name, dose, and route of the analgesic used, if applicable
  • Post-recovery monitoring dates, observations, and treatments.