Version: 7.2014
Appendix 6 – Pre-surgical Preparation Procedures
Species
Mouse and Rat
Pre-surgical preparation description
Aseptic technique is used for Survival Surgery.
Surgery Area: The laboratory area is disinfected before use. A dedicated space for survival surgery
is recommended. Sterile drapes are to be used to drape off the incision area if the abdomen or
thorax are open.
Surgeon Prep: The surgeon wears a mask, cap,sterile surgical gloves, and clean lab coat or
gown. The surgeon performs a thorough hand scrubbing before donning gloves.
Patient Prep: If hair is present over the incision site, it is removed with clippers, razor, or depilatory
to include an area approximately twice as large as the intended surgery site. Skin is disinfected
using three alternating rounds of surgical disinfectant scrub/solution (betadine, hibiclens,
chlorhexadine, etc.) with 70% isopropyl alcohol or sterile saline rinses. Care is taken to avoid overwetting fur outside of the surgical area as this will increase hypothermia. Consider application of
disinfectant/alcohol using cotton-tip applicators rather than larger gauze squares as it may be harder
to control application in small areas. A sterile surgery field is best maintained through the placement
of a sterile drape over non-sterile parts of the animal and surrounding area. Sterile drapes must be
used if the abdominal or thoracic cavities are opened. Ophthalmic ointment should be placed in
both eyes of anesthetized rodents.
Instrument Prep: Instruments, sutures, wound clips, and implanted devices are sterilized in an
autoclave or ethylene oxide sterilizer prior to surgery and a sterile field is maintained during
surgery. Instruments are re-sterilized between rodents using an autoclave or bead sterilizer.
Instruments will not be used on more than 4-5 rodents prior to re-autoclaving.
USDA-covered
species (including
Hamster, Guinea
Pig, Dog, Pig,
Rabbit Sheep,
Cats, Ferrets and
other covered
rodents)
Aseptic technique is used for Survival Surgery.
Surgery Area: Surgery is done in a dedicated operating room.
Surgeon Prep: The surgeon wears a mask, cap, sterile surgical gloves, and sterile gown. The
surgeon performs a thorough surgical hand scrubbing before donning gloves.
Patient Prep: The hair over incision areas is removed in an area approximately twice as large as the
intended surgical site. A sterile skin prep is done using at least three alternating rounds of surgical
disinfectant scrub (betadine, hibiclens, chlorhexadine, etc.) with 70% isopropyl alcohol or sterile
saline rinses. Sterile drapes must be used to protect the surgical field. Ophthalmic ointment should
be placed in both eyes of anesthetized animals.
Instrument Prep: Instruments, sutures, wound clips, biopsy devices, and implanted devices are
sterile and a sterile field is maintained during the procedure. Instruments are autoclaved between
animals.
Definitions:
Disinfection: cleaning procedures that reduce the number of pathogenic organisms to a harmless level.
Aseptic technique: Aseptic technique is required for all survival surgeries regardless of species. This includes hair removal
and disinfection of the surgical site; preparation of the surgeon, including proper surgical attire and sterile gloves;
sterilization of all instruments, devices, and materials; and the use of operative techniques to reduce the likelihood of
infection
Sterilization: cleaning procedure that kills all living organisms on an object.
(Based on guidelines from NIH)
Table 1. RECOMMENDED HARD SURFACE DISINFECTANTS
(e.g., table tops, equipment)
Always follow manufacturer's instructions for dilution and expiration periods.
1
Version: 7.2014
AGENT
EXAMPLES *
COMMENTS
Alcohols
70% ethyl alcohol
85% isopropyl alcohol
Quaternary Ammonium
Roccal®, Quatricide®
Chlorine
Phenolics
Sodium hypochlorite
(Clorox ® 10% solution)
Chlorine dioxide
(Clidox®, Alcide®, MB-10®)
Glutaraldehydes
(Cidex®, Cetylcide®, Cide Wipes®)
Lysol®, TBQ®
Contact time required is 15 minutes. Contaminated
surfaces take longer to disinfect. Remove gross
contamination before using. Inexpensive.
Rapidly inactivated by organic matter. Compounds
may support growth of gram negative bacteria.
Corrosive. Presence of organic matter reduces
activity. Chlorine dioxide must be fresh; kills
vegetative organisms within 3 minutes of contact.
Chlorhexidine
Nolvasan® , Hibiclens®
Glutaraldehydes
Rapidly disinfects surfaces.
Less affected by organic material than other
disinfectants.
Presence of blood does not interfere with activity.
Rapidly bactericidal and persistent. Effective against
many viruses.
The use of common brand names as examples does not indicate a product endorsement.
Table 2. SKIN DISINFECTANTS
Alternating disinfectants is more effective than using a single agent. For example, an iodophor scrub can be alternated
three times with 70% alcohol, followed by a final soaking with a disinfectant solution. Alcohol, by itself, is not an adequate
skin disinfectant. The evaporation of alcohol can induce hypothermia in small animals.
AGENT
EXAMPLES *
Iodophors
Betadine®, Prepodyne®,
Wescodyne®
Nolvasan®, Hibiclens®
COMMENTS
Reduced activity in presence of organic matter. Wide
range of microbicidal action. Works best in pH 6-7.
Cholorhexidine
Presence of blood does not interfere with activity.
Rapidly bactericidal and persistent. Effective against
many viruses. Excellent for use on skin.
The use of common brand names as examples does not indicate a product endorsement.
Table 3. RECOMMENDED INSTRUMENT STERILANTS
Always follow manufacturer's instructions for dilution, exposure times and expiration periods.
AGENT
EXAMPLES *
COMMENTS
Steam sterilization (moist
heat)
Autoclave
Dry Heat
Hot Bead Sterilizer
Dry Chamber
Gas sterilization
Ethylene Oxide
Chlorine
Chlorine Dioxide
Glutaraldehydes
Glutaraldehyde
(Cidex®, Cetylcide®,
Metricide®)
Effectiveness dependent upon temperature, pressure
and time (e.g., 121oC for 15 min. vs. 131oC for 3
min).
Fast. Instruments must be cooled before contacting
tissue. Only acceptable for instruments between
surgeries, NOT for initial sterilization of tools. Only
tips of instruments are sterilized with hot beads.
Requires 30% or greater relative humidity for
effectiveness against spores. Gas is irritating to
tissue; all materials require safe airing time.
Corrosive to instruments. Instruments must be rinsed
with sterile saline or sterile water before use.
Several hours required for sterilization. Corrosive and
irritating. Instruments must be rinsed with sterile saline
or sterile water before use.
2
Version: 7.2014
Hydrogen peroxide-acetic
acid
Actril®, Spor-Klenz®
Several hours required for sterilization. Corrosive and
irritating. Instruments must be rinsed with sterile saline
or sterile water before use.
The use of common brand names as examples does not indicate a product endorsement.
3