Aseptic Technique

University of North Texas Health Science Center

Department of Laboratory Animal Medicine

AsepticTechnique

Purpose: to ensure asepsis during survival surgery

Sterile Tip Technique

•Rodents can develop post-operative infections.

•Infections may confound or alter experimental results.

•Antibiotics may confound and alter experimental results.

Why is aseptic technique important?

Sterile Tip Technique

Infections may not be apparent on casual observation, but may cause changes in physiologic parameters and behavior.

Behavioral effects: •Activity, stress, appetite

Physiological effects:•Sympathetic response = Adrenaline

•Cell dynamics, cytokine expression•Immune axis

In accordance with good scientific practice, ethical principles, and standards set forth in the Public Health service Guide for the Care and Use of Laboratory Animals and the Federal Animal Welfare Act, aseptic surgical procedures must be used.

Sterile Tip Technique

Facilities•Lab approved by IACUC•Separate areas•Clear of clutter•Away from traffic / doors•Air vents•Disinfect surfaces

1. The animal must be maintained in a surgical plane of anesthesia throughout the procedure.

2. Begin surgery with sterile instruments and handle them aseptically.

3. Instruments may be used for a series of similar surgeries provided they are maintained clean and disinfected between animals.

4. Provide analgesics as appropriate.

Points to ensure

Instruments must be sterilized in an autoclave before use. Here are examples of how the instruments can be packed for sterilization.

A glass bead sterilizer can be used to sterilize instruments between rodents. If using a glass bead sterilizer you should remember the following:

1. Only the tips of the instruments are sterilized.2.The tips of delicate instruments may become damaged during immersion in the glass beads.3.The tips of the instruments become extremely hot. The instruments must be allowed to cool before applying them to the skin or other tissues.4.Maximum of ten rodents can be done with one set of instruments sterilized this way.

Place the tips of the sterile instruments on a sterile field (e.g., sterile 4x4 gauze). To avoid contamination of the tips of the instruments during the surgical procedure, always keep the tips on the sterile field and pointed in one direction.

Anesthetics and Analgesics

Injectable anesthesia is given as an IP injection.

The animal is placed in an induction chamber with Isoflurane anesthetic.

While waiting for the animal to become anesthetized,administer pain relief. Administer Buprenorphine bySQ injection at a dose of 0.05 – 0.1 mg/kg.

Apply ophthalmic ointment in the eyes to prevent the corneas from drying during

anesthesia

In the prep area, lay the animal down and place the nose cone over the nose if using Isoflurane.

Shave the entire surgical incision site while the animal is unconscious.

The surgical site is prepped with alternating scrubs of Betadine and 70% alcohol (ending with betadine). Cotton-tipped swabs or 2x2 gauze sponges may be used. It is important to avoid excessively wetting the animal as this will lead to hypothermia and anesthetic complications.

1. Alcohol

2.Betadine scrub

3. Betadine

The animal is now ready to be draped for surgery. A drape is used if any organs are exteriorized during the procedure.

Below is one style of drape. Note that it has been sterilized.

Drape on the animal.

This is another type of drape that can be used. Note that it has also been sterilized.

After donning a clean pair of sterile gloves, an incision through the skin and abdominal musculature is made.

Sterile ophthalmic surgical spears work well to blot blood or fluid from the surgical site. Sterile gauze sponges or cotton-tipped applicators may also be used.

The abdominal musculature is closed with an absorbable suture. Note how only instruments are used to handle the tissue and needle.

The skin is closed with a non-absorbable suture or stapling device. Wound closures must be removed no later than 10 days after surgery by the surgeon.

The animal is placed on a clean surface on a warming tray for recovery from anesthesia, or alternatively in a clean cage on paper towel or similar product, under a heat lamp. Throughout the recovery and post-surgical convalescent period, the animal must be closely monitored until it is awake.

A heating pad or a heat pack can also be used. These can also be used during surgery to keep the animal warm.

If multiple, similar surgeries are performed, the instruments should be rinsed with 70% alcohol and re-sterilized between animals.

Change gloves before performing the next surgery.

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