IACUC Rodent Survival Surgery Guidance

Rodent Survival Surgery Guidance

This guidance document includes tables of information and references as a resource for PIs. Please review the Brown University Institutional Animal Care and Use Committee (IACUC) Rodent Survival Surgery Policy for more information.

TABLE 1 - RECOMMENDED HARD SURFACE DISINFECTANTS* (e.g., table tops, equipment)
AGENT	EXAMPLES **	COMMENTS
Alcohols	70% ethyl alcohol 85% isopropyl alcohol	Contact time required is 15 minutes. Contaminated surfaces take longer to disinfect. Remove gross contamination before using.
Quaternary Ammonium	Roccal®, Quatricide® Tec-Surf II®	Rapidly inactivated by organic matter. Compounds may support growth of gram negative bacteria.
Chlorine	Sodium hypochlorite (Clorox® 10% solution) Chlorine dioxide (Clidox®, Alcide®, MB-10®) Dispatch®	Corrosive. Presence of organic matter reduces activity. Chlorine dioxide must be fresh; kills vegetative organisms within 3 minutes of contact. Note: chlorox solutions need to be made up fresh daily to maintain activity, whereas Dispatch, available from Environmental Services, is labeled as stable for 1 year.
Glutaraldehydes	Glutaraldehydes (Cidex®, Cetylcide®, Cide Wipes®)	Rapidly disinfects surfaces.
Phenolics	Lysol®, TBQ®	Less affected by organic material than other disinfectants.
Chlorhexidine	Nolvasan® , Hibiclens®	Presence of blood does not interfere with activity. Rapidly bactericidal and persistent. Effective against many viruses.

*Always follow manufacturer's instructions for dilution and expiration periods.
**The use of common brand names as examples does not indicate a product endorsement.

TABLE 2 - SKIN DISINFECTANTS*
AGENT	EXAMPLES**	COMMENTS
Iodophors	Betadine®, Prepodyne®, Wescodyne®	Reduced activity in presence of organic matter. Wide range of microbicidal action. Works best in pH 6-7.
Cholorhexidine	Nolvasan®, Hibiclens®	Presence of blood does not interfere with activity. Rapidly bactericidal and persistent. Effective against many viruses. Excellent for use on skin.

Alternating disinfectants is more effective than using a single agent. For example, an iodophor scrub (with soap) can be alternated once with 70% alcohol, followed by a final soaking with a disinfectant solution (without soap). Alcohol, by itself, is not an adequate skin disinfectant. Since the evaporation of alcohol can induce hypothermia in small animals, avoid exposing excessively large areas.

*Always follow manufacturer's instructions for dilution and expiration periods.
**The use of common brand names as examples does not indicate a product endorsement.

TABLE 3 - Procedures for Sterilizing Surgical Instruments*
AGENT	EXAMPLES**	COMMENTS
--RECOMMENDED METHODS FOR STERILIZING SURGICAL INSTRUMENTS--
Steam sterilization (moist heat)	Autoclave	Effectiveness dependent upon temperature, pressure and time (e.g., 121oC for 15 min. vs. 131oC for 3 min.).
Dry Heat	Hot Bead Sterilizer Dry Chamber	Fast. Instruments must be cooled before contacting tissue. Only tips of instruments are sterilized with hot beads. Instruments should be cleaned prior to inserting in the hot bead sterilizer. Replace the beads per the manufacture’s recommendations.
--ALTERNATIVE METHODS FOR STERILIZING SURGICAL INSTRUMENTS--
Gas Sterilization	Ethylene Oxide	Requires 30% or greater relative humidity for effectiveness against spores. Gas is irritating to tissue; all materials require safe airing time.
Chlorine	Chlorine Dioxide	Corrosive to instruments. Instruments must be rinsed with a minimum of (3) fresh washes of sterile saline or sterile water before use.
Glutaraldehydes	Glutaraldehyde (Cidex®, Cetylcide®, Metricide®)	Several hours required for sterilization. Corrosive and irritating. Instruments must be rinsed with a minimum of (3) fresh washes of sterile saline or sterile water before use.
Hydrogen Peroxide-Acetic Acid	Actril® Spor-Klenz®	Several hours required for sterilization. Corrosive and irritating. Instruments must be rinsed with a minimum of (3) fresh washes of sterile saline or sterile water before use.

* Always follow manufacturer's instructions for dilution, exposure times and expiration periods.
** The use of common brand names as examples does not indicate a product endorsement.

TABLE 4 - RECOMMENDED INSTRUMENT DISINFECTANTS*
AGENT	EXAMPLES**	COMMENTS
Chlorine	Sodium hypochlorite (Clorox ® 10% solution) Chlorine dioxide (Clidox®, Alcide®)	Corrosive to instruments. Presence of organic matter reduces activity. Chlorine dioxide must be fresh. Kills vegetative organisms within 3 min. Instruments must be rinsed with a minimum of (3) fresh washes of sterile saline or sterile water before use.
Chlorhexidine	Nolvasan® Hibiclens®	Presence of blood does not interfere with activity. Rapidly bactericidal and persistent. Effective against many viruses. Instruments must be rinsed with a minimum of (3) fresh washes of sterile saline or sterile water before use.

*Always follow manufacturer's instructions for dilution, exposure times and expiration periods.
** The use of common brand names as examples does not indicate a product endorsement.

TABLE 5 - WOUND CLOSURE SELECTION
MATERIAL*	CHARACTERISTICS AND FREQUENT USES
Polyglactin 910 (Vicryl®),	Absorbable; 60-90 days. Ligate or suture tissues where an absorbable suture is desirable.
Polydiaxanone (PDS®), or Polyglyconate (Maxon®)	Absorbable; 6 months. Ligate or suture tissues especially where an absorbable suture and extended wound support is desirable
Polypropylene (Prolene®)	Non-absorbable. Inert.
Nylon (Ethilon®)	Non-absorbable. Inert. General skin closure.
Silk	Non-absorbable. Restrict the use of silk to cardiovascular procedures or where silk’s excellent handling properties are critical. Avoid for such purposes as routine skin closure since it may wick microorganisms into the wound, and cause tissue reactivity.
Chromic Gut	Absorbable. Versatile material.
Stainless Steel Wound Clips, Staples	Non-absorbable. Requires instrument for removal.
Cyanoacrylate (Vetbond®, Nexaband®)	Skin glue. For non-tension bearing wounds. Note: use only products labeled for surgical use, super glue is not acceptable for surgery.

Suture gauge selection: Use the smallest gauge suture material that will perform adequately.
Cutting and reverse cutting needles: Provide edges that will cut through dense, difficult to penetrate tissue, such as skin.
Non-cutting, taper point or round needles: Have no edges to cut through tissue; used primarily for suturing easily torn tissues such as peritoneum or intestine.

* The use of common brand names as examples does not indicate a product endorsement.

TABLE 6 – ANESTHESIA
Inhaled anesthetics: Isoflurane delivered by mask or endotracheal tube via a precision vaporizer is recommended for all species. Vaporizers are available for use in Ship St. and Biomed procedure rooms. Please contact ACF for information regarding vaporizer availability and training. For very brief procedures, (e.g., tail biopsies for genotyping), it may be acceptable to use isoflurane or other inhalant anesthetics, without a precision vaporizer, in a “bell-jar” placed in a fume hood while precluding direct contact of animal skin with inhalant anesthetics. In all cases the anesthetic vapors must be adequately vented to prevent inadvertent exposure of personnel. Injectable anesthetics: Injectable anesthetics are appropriate for many procedures. There is, however, a great deal of variation in depth and duration of anesthesia among rodent strains and individual animals.
ANESTHETIC	MOUSE	RAT	GUINEA PIG
Ketamine/ Dexmedetomidine Cocktail1	75 mg kg-1 i/p 0.5 mg kg-1 i/p SQ or IP, may not produce surgical plane of anesthesia in some mice	75 mg kg-1 i/p 0.25 mg mg kg-1 i/p	40 mg kg-1 i/p 0.25 mg mg kg-1 i/p
Ketamine/ Xylazine Cocktail2	** **	40 - 90 mg kg-1 i/p 13 mg kg-1 i/p	35 mg kg-1 0.2 – 0.5 mg kg-1
Pentobarbital3	40 – 85 mg kg-1 i/p	40 – 85 mg kg-1 i/p	28 mg kg-1 IP

1 The 2 components can be mixed with water for injection (WFI) (USP water) – see table for Rodent Cocktail Solutions below.
2 Xylazine is a potent respiratory depressant. Supplemental dosing, if necessary, should be done with ½ the original dose of ketamine alone.
3 Consider replacing with inhalant or another injectable anesthetic.
**** Consult veterinarians for information.

TABLE 7 – RODENT COCKTAIL SOLUTIONS
	MOUSE	RAT	GUINEA PIG		MOUSE	RAT	GUINEA PIG
Ketamine mL (100 mg mL-1)	****	3.75 mL	3.75 mL	Ketamine mL (100 mg mL-1)	0.5 mL	3.75 mL	2.0 mL
Xylazine (10 mg mL-1)	****	2.5 mL	2.5 mL	Dexmedetomidine (0.5 mg mL-1)	0.5 mL	2.5 mL	2.5mL
WFI	****	3.75 mL	3.75 mL	WFI	4.0 mL	3.75 mL	5.5 mL
Total Volume	****		10.0 mL	Total Volume	5.0 mL	10.0 mL	10.0 mL
Dosage	****	0.2 mL per 100gm	0.2 mL per 100gm	Dosage	mL per 10 gm	mL per 100gm	2.0mL per kg

**** Consult veterinarians for information.

TABLE 8 - ANALGESICS/ANTI-INFLAMMATORY AGENTS*
AGENT	MOUSE	RAT	GUINEA PIG
Buprenorphine	0.05 – 0.1 mg kg-1 SQ Every 12 hours	0.05 mg kg-1 SQ Every 12 hours	0.05 mg kg-1 SQ Every 12 hours
Butorphanol	0.2 – 2.0 mg kg-1 SQ Every 4 hours	2 mg kg-1 SQ Every 4 hours	****
Meloxicam	0.3 mg kg-1 PO	1 mg kg-1 PO	0.2 mg kg-1 PO
Carprofen	5 mg kg-1 SQ Every 24 hours	5 mg kg-1 SQ Every 24 hours	****

* The IACUC must approve the exclusion of analgesics/anti-inflammatory agents for experimental surgery. If you feel that these agents are likely to interfere with the objectives of your research, provide a scientific justification for their exclusion in your protocol.
****Consult veterinarians for information.
References:

NRC (National Research Council), Institute of Laboratory Animal Resources, Commission on Life Sciences. 1996. Guide for the Care and Use of Laboratory Animals, Washington D.C., National Academy Press.
The Animal Welfare Act, 7USC Sec 2131 et seq. Title 9, Code of Federal Regulations, parts 1, 2, & 3, January 1998.
Laboratory Animal Anesthesia, A practical introduction for research workers and technicians., 2nd Edition, P.A. Flecknell (1996)

PDF Attached:

Rodent Survival Surgery Guidance.pdf

IACUC Rodent Survival Surgery Guidance

Rodent Survival Surgery Guidance

TABLE 7 – RODENT COCKTAIL SOLUTIONS