rat
Content
Animal Care and Use Committee
Ver.2, Effective: August 9, 2012
WVU ACUC POLICY
Anesthesia, Surgery and Post-Operative Care of Rats and Mice
Table of Contents
I.
II.
III.
IV.
V.
VI.
Introduction
Monitoring & Documentation
Aseptic Technique in Survival Surgery
Postoperative Monitoring & Care
Surgeries on Multiple Animals
Clean Technique in Non-Survival Surgery
I. INTRODUCTION
A. Purpose
This document describes appropriate procedures for anesthesia, aseptic surgery, and post-operative
care of laboratory rats and mice at WVU. Other rodents and other warm-blooded mammals regulated
by the United States Department of Agriculture (USDA) are covered under the USDA Species Policy
for Anesthesia and Surgery.
According to the Guide for the Care and Use of Laboratory Animals, 8th edition (2011; henceforth the
Guide), the following areas must be given attention when performing surgery on laboratory animals:
pre-surgical planning, aseptic and surgical technique, assessment of animal well-being during and
after the procedure, appropriate use of analgesics, and each animal’s physiologic status during all
phases of a protocol involving surgery and postoperative care. In addition, the Guide states that
survival surgery on rodents must be performed in facilities intended for that purpose, using aseptic
procedures to prevent clinical infections. The OLAR veterinary staff is always available to provide
assistance with anesthetic or surgical issues, as well as with surgical complications, and challenging
or prolonged anesthetic recoveries. For additional information, OLAR can be contacted by email at
[email protected] or [email protected] or by telephone at 304-293-2721 for other
information. OLAR is located in Room 186G in the Health Sciences Center North.
B. Preparing Animal Care and Use Protocols
This document describes the expectations of the WVU Animal Care and Use Committee (ACUC) for
(a) anesthesia; (b) aseptic surgical technique; (c) appropriate pre-, intra-, and post-operative care;
and (d) documentation for rats and mice undergoing anesthesia or anesthesia and surgery
procedures.
Investigators who adhere to the requirements of this document should provide a brief description of
Phone: 304-293-9368
Phone: 304-293-1050
Fax: 304-293-3098
http://oric.research.wvu.edu
Chestnut Ridge Research Building
886 Chestnut Ridge Road
PO Box 6845
Morgantown, WV 26506-6845
Equal Opportunity/Affirmative Action Institution
the surgical approach, intraoperative actions, and closure technique in their Animal Care as Use
protocol (henceforth, protocol), and then go on to state that this policy document will be followed (i.e.,
the details listed here need not be repeated if this policy is referenced in the protocol).
Deviations from the procedures described herein are subject to the approval of the ACUC. Any
deviations must be described and scientifically justified in the protocol.
C. General Issues
Investigators should be familiar with the following issues that may apply to their research.
1. Acclimation period: A minimum 48-hour acclimation period is required for laboratory mice and
rats prior to experimental use; in other words, animals cannot be used until the third calendar day
after their arrival. See the ACUC Policy on Acclimation of New Arrivals for detailed information and
exceptions.
2. Anesthesia without surgery: Animals undergoing anesthesia must be observed and monitored
until fully ambulatory as described for survival surgery below, including those anesthetized for brief
time periods and/or for minor or non-invasive procedures. During the anesthetic recovery period,
animals must have appropriate monitoring (e.g. temperature), and active animal surveillance such as
contact and stimulation by the observer to aid in arousal and anesthetic recovery is preferred.
Animals can never be left unattended unless the animal has fully regained consciousness. Criteria
used to define complete consciousness include: (a) a body temperature within 1-2 degrees of normal
(normal is ~99 degrees F) and (b) ability of the animal to right itself (i.e. stand on all 4 limbs) from
being placed in full dorsal recumbency (i.e. placed on their back).
3. Analgesic considerations: Please consult with a veterinarian from the WVU Office of Laboratory
Animal Resources (OLAR) for guidance on appropriate surgery-specific analgesic options, including
doses, timing and administration routes when preparing surgical animal protocols. In general,
analgesics are given preemptively to avoid pain “wind-up.” However, the potential for an adverse
effect of analgesic selection on the stability of the anesthetic procedure may affect the plan for
analgesic use in some cases.
4. Antibiotic use: Postoperative antibiotics may be indicated in conjunction with aseptic technique.
Antibiotic administration prior to surgery is more effective, because the blood supply is still
uncompromised to the surgical area at this time. When needed, post-operative antibiotic
administration can be used, but requires an extended treatment period. Contact an OLAR
veterinarian to discuss antibiotic use as it applies to your specific needs.
5. Fluid administration: All animals undergoing anesthesia and/or surgery > 20 minutes duration
benefit from warm intraperitoneal and/or subcutaneous fluids to insure adequate hydration postoperatively and to assist in a faster recovery following injectable anesthetics, such as ketamine, that
are cleared by the kidneys. Contact an OLAR veterinarian for information on appropriate fluid
volumes for your application. A safe rule of thumb for routine use is to deliver a balanced electrolyte
solution volume of 10-12 ml to an adult rat, and 1-2 ml to an adult mouse, either subcutaneously or
intraperitoneally, or divided by both routes.
6. Eye lubricant use: For must general anesthetic procedures lasting more than 20 minutes, eye
lubricant (i. e., Lacrilube®, opthalmic antibiotic ointment) must be placed in each eye to avoid corneal
desiccation.
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 2
Both fluids and eye lubricant may be applied immediately after anesthetic induction for ease of
administration.
II. MONITORING & DOCUMENTATION
To comply with applicable laws and regulations, surgical records must be maintained for all surgeries
performed and must include all anesthetics and analgesics (drug, dose, frequency) administered,
and surgical complications encountered. The OLAR-supplied pink surgery and anesthesia cards
(“pink cards” found in vivarium animal rooms, or obtained from OLAR), if completely filled out and
placed on rodent cages, may suffice for this purpose.
All surgical records must be accessible and readily available upon request to OLAR, the
ACUC, the federal Office of Laboratory Animal Welfare (OLAW) or – in the case of certain
rodents – the USDA (see the USDA Species Anesthesia and Surgery Policy). Investigators
may submit copies of surgical records to OLAR, which will retain them for inspectors.
Sample anesthetic monitoring and surgery record forms for use by investigators can be found in the
Appendix below and on the OLAR website (see contact information above). Although use of these
particular forms is not required, the type of information contained on the sample forms is required.
A. Pre-Operative Records
Initially, visual assessment of animal health must be made. For example, is the animal normally
hydrated, or does it require additional fluids? If injectable premedications (e.g. anticholinergics,
antibiotics), or anesthetics or analgesics are used, the animal’s body weight or an equivalent
standard should be used to calculate dosages. It is important to note and address any abnormalities
that might affect the animal’s procedural outcome.
B. Intra-Operative Records
Surgery records must contain the following information: PI name, surgeon name, ACUC protocol
number, animal or cage ID, species/strain, and surgery date. In addition, for surgeries lasting
more than 15 minutes, a meaningful assemblage of signs must be monitored and recorded:
anesthetic depth (required-response to toe pinch); mucous membrane color (eye color in
albinos); body temperature; respiratory rate (and respiratory depth when relevant); in some cases,
monitoring of heart rate is also indicated. For short procedures, and in smaller animals, it may be
acceptable to monitor only certain parameters, or to note only changes in these parameters. More
complicated procedures and those of longer duration require more detailed monitoring. All
anesthetics, medications, fluids, and/or test substances administered during surgery must be
recorded. Completed intraoperative records or cage pink cards may be submitted to OLAR for safekeeping or they may be maintained in the laboratory provided they are readily available for inspection,
(see below; the back of the card can be used to record added details).
C. Post-Operative Records
Following surgery, all cages must be clearly identified (e.g. completely filled out “pink card”) during
the postsurgical monitoring period within the housing room. Analgesic doses must be recorded on
pink cards (i.e. including drug-circled or written in, dose, frequency, and initials of the person
administering the drug). If the sutures are non-absorbable, an anticipated suture removal date
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 3
should be listed. If relevant, additional postoperative follow-up information relevant to the animal’s
recovery must also be recorded and maintained in the animal room throughout the post-operative
period, including all animal observations, and medications and/or experimental substances
(cells, drugs, biologicals, etc.) administered spanning the surgical and postoperative periods (see
the Post-Operative Monitoring and Care section below).
III. ASEPTIC TECHNIQUE IN SURVIVAL SURGERY
The following standards must be applied to all survival surgeries. As noted in the Introduction, any
deviation must be approved, in advance, by the ACUC.
A. General Considerations
Surgery may only be performed at a site dedicated to that activity at that time (time or space
separated), preferably with a separate area for patient preparation, and must take place on a clean,
uncluttered surface (e.g. no air ducts impinging on the site or overhead shelved materials
likely to fall on the sterile field). Those entering the space during surgery must abide by aseptic
practices such as donning masks and/or personal protective equipment. Traffic in the area during
surgery should be minimized by using appropriate signage. In addition, the cleaned surface must be
covered with an appropriately cleaned heated surface, barrier cloth, or drape. All drugs, suture, and
sterile gloves used for survival surgery must be used before their expiration date.
B. Instrument Preparation
The sterilization method used must be attuned to the durability of the materials to be sterilized, and
sterilization indicators must be used to validate sterilization completeness (outside and preferably
inside sterile packs). Appropriate techniques include: gas (H202 or ethylene oxide); steam
(autoclave); and liquid, chemical sterilants with proper contact time (e.g. 2% glutaraldehyde x 10
hr, or 8% formaldehyde with 70% alcohol x 18 hr). When liquid sterilants are used, instruments must
be rinsed in sterile saline before use. When gas sterilization is used, proper degassing times (e.g. 12
hr for ethylene oxide) should be observed.
The technique of “tip sterilization” using bead or hot dry sterilizers may also be used with care, as
this requires careful attention to maintenance of aseptic technique. Also, excessive heating of
instruments can damage tissue upon contact. All in-house sterilized items must be used within 6
months of the sterilization date (sterilization dates are inscribed on each sterilization pack). Seventy
percent (70%) alcohol is NOT a sterilant or a high-level disinfectant and, by itself, is not
acceptable for surgical instrument sterilization. Novasan® (chlorhexidine gluconate) Solution (not
scrub) may be used for this purpose. Contact OLAR for information on sterilization services that are
available for delicate instruments or equipment that cannot be autoclaved or tip sterilized. Other items
used in surgery such as gauze, suture material, irrigation fluid, syringes, needles, intravascular
catheters, etc. must also be sterile. Only rust-free instruments may be used for rodent surgery. To
preserve the life of instruments, “Surgical milk” can be used prior to autoclaving. Contact OLAR for
additional information on surgical instrument maintenance.
C. Animal Preparation
Use of a protocol approved anesthetic approach, appropriate for the species and procedure is
required. Animals must be maintained at a surgical plane of anesthesia (documented as
unresponsive to toe pinch, or without physiologic evidence of pain response) and and the anesthetist
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 4
must assure this state throughout the surgical procedure.
Thermal support via an external heat source (e.g. circulating warm water blanket) is recommended
when the duration of anesthesia or surgical conditions indicate a significant drop in body temperature
of more than 3 degrees is likely, (e.g. any procedure where a body cavity is opened); if in doubt you
should supply external heat. Frequency of temperature monitoring should be sufficient to avoid large
fluctuations in physiological parameters that need rapid correction, (e.g. an animal’s body
temperature should be monitored at least every 10–15 minutes during the procedure). When using
external heat sources, minimize insulation between the heat source and the animal to get the best
heating effect. If electric pads are used, a layer between the animal and the heat source may be
appropriate, and the use of pads that develop hot spots must be avoided. When autoclavable pads or
hot water-filled gloves are used, care must be taken to monitor and rewarm them regularly so as not
to draw heat from the animal as they cool.
Sterile ophthalmic ointment (e.g. Puralube®) must be applied to the eyes when animals are
unconscious for more than 15 minutes, and re-applied at least every 1.5 hour (or more frequently if
needed), until the animal has recovered. This treatment will keep the eyes lubricated and prevent
corneal injury.
Hair removal from the surgical site, including a sufficient margin, can be achieved using electric
clippers, disposable razors, depilatory creams (e.g. Nair®- must be completely rinsed from the skin)
or by manual plucking under anesthesia. When using depilatories, contact times may have to be
modified with the species and conditions of use.
Soiled skin may require cleaning before surgical skin disinfection can start. Skin disinfection
of the operative site must include three alternating wipes of an approved soap disinfectant scrub (e.g.
chlorhexidine [Novasan® Scrub] or iodine [Betadine Scrub®]) with wipes of 70 % alcohol or saline
(i.e. for Betasept® or Hibiclens® Scrub) followed by a final application of an antiseptic solution (e.g.
iodine solution without detergent). Do not expose the eyes to the scrub, alcohol or disinfectant
solutions. Excessive animal wetting will speed cooling. Before or after disinfection, secure the animal
at the surgery site and cover with a sterile drape, leaving only the disinfected skin exposed. In rats
and mice, use only light drapes to prevent breathing difficulty in the anesthetized animal.
D. Surgeon/Support Personnel Preparation
Hands and arms should be washed thoroughly using an acceptable surgical scrub/brush (e.g.
chlorhexidine [Novasan® Scrub, Betasept®, or Hibiclens® Scrub] or iodine [Betadine® Scrub]) in the
correct manner (contact OLAR for assistance if this is unfamiliar to you). Surgeons and assistants
must don appropriate surgical attire including a sterile or clean gown, scrub shirt, or lab coat, a
surgical mask, appropriate hair covering, and sterile surgical gloves. Surgical or anesthetic
assistants may wear a clean covering, mask, cap and clean examination gloves, rather than sterile
gloves, if they will not be touching the sterile field or instruments.
E. Operative Techniques
Sterile suture material must be selected specifically for the intended purpose at hand (absorbable
vs. non-absorbable, braided vs. monofilament, size and needle type). For example, skin closure
usually requires a monofilament non-absorbable suture such as nylon. Because rodents tend to
chew out sutures, another selection such a metal staples or stainless steel suture may be more
appropriate in some cases
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 5
It is critical to maintain a sterile surgical field at all times. If a non-sterile object comes in contact
with the surgeon’s gloves, a new pair of sterile surgical gloves must be donned. A non-sterile
surgical assistant should be present when needed to assist in maintaining the surgeon’s aseptic
technique (e.g. adjust anesthesia machine, manipulate non-sterile instrument surfaces, provide
supplies, etc.). Contaminated instruments (generally those touching anything outside the sterile field)
are discarded from use.
When surgery involves the opening of a body cavity, at least a two-layer closure is required whereby
the body wall is closed separately using absorbable suture, and the skin closed with some
combination of wound clips, staples, surgical adhesive, stainless steel suture or another nonabsorbable monofilament suture in a simple interrupted pattern. A subcuticular pattern may be used
for skin closure in larger rodents, such as rats and guinea pigs. Consult an OLAR veterinarian for
additional information. Never use silk for skin closure. Silk may be used internally, but this is
discouraged.
Wound clips or non-absorbable skin sutures must be removed when the wound is healed
sufficiently, typically by 14 days postoperative. Failure to remove sutures by postoperative day 21 is
considered noncompliance unless the ACUC-approved protocol calls for euthanasia of the animal
before then (in which case “n/a” should be entered for the suture removal date on the pink card.
IV. POST-OPERATIVE MONITORING & CARE
Oversight of anesthetic recovery monitoring, record keeping and postoperative care is the
responsibility of the investigator and their staff doing the procedure. Post-anesthetic recovery services
may be contracted out to the OLAR veterinary technicians for a fee (contact
[email protected] for additional information).
A. Anesthetic Recovery Period
The recovery cage should be free of particulate bedding and lined with paper towels, “isopads”, etc.
to prevent pica (ingestion of nonfood items) and gastrointestinal or tracheal obstructions during
anesthetic recovery. This is especially important when using buprenorphine. To avoid cannibalism
and suture chewing by cage mates in the recovery cage, rodents must be housed individually or with
no more than two other individuals who have had the same procedure and are convalescing. Grouphoused animals must be separated when recovered to avoid suture chewing until the time of suture
removal. Only when animals are fully ambulatory can they be returned to the vivarium.
Supervised heat support must be provided with an external heat source for any rodent with a
depressed body temperature (e.g. 3 or more degrees below normal) or one that is slow to recover,
and should remain in place until the animal is fully ambulatory. NOTE: when not in direct attendance
at the recovery cage, always ensure that approx. 1/3 to 1/2 of the cage is unheated so the animal
can move away from the heat source if necessary to avoid hyperthermia. If heating bulbs or elements
are used, these should never be closer than 18” from the animal.
Animals must be directly observed while in the recovery cage at least every 5-10 minutes until
ambulatory. These observations include body temperature measurement via rectal probe or
thermometer and contact stimulation of the recovering animals so they generate their own heat.
Additional monitoring includes respiratory rate/quality, mucous membrane color,
activity/anesthetic depth, and incisional integrity, including any leakage or bleeding.
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 6
Analgesic agents to be given at the time of surgery are preferably administered prior to anesthetic
recovery, unless scientifically justified in the approved ACUC protocol, or under direct veterinary
instruction.
Only when animals are able to freely move around the cage can they be returned to the animal
housing room and left unattended. Provision of moistened rodent food and/or dry food on the cage
floor is strongly recommended for post-operative animals at least 24 hours following surgery,
especially when any of the following is present: postoperative disorientation: hindleg, caudal body or
abdominal surgery incisions; or overall debility. Typical situations for extended supplementation
include surgeries that limit mobility or activity, such as hind limb or abdominal surgery.
B. Postoperative Observations
Following surgery, the investigative staff must examine postoperative animals at least once daily until
the animal is stabilized. Daily examination includes at least observation of the incision for heat,
swelling, drainage, or dehiscence (opening), assessment of overall animal activity, and any signs of
pain or distress (described in detail in the ACUC policy entitled Recognition of Pain and Distress,
Humane Endpoints). Observations are noted on the pink card on the animal’s cage.
All postsurgical examinations must be recorded, including any complications encountered. If signs
of pain or distress are observed, a staff member from OLAR must be notified so that appropriate and
timely veterinary care can be provided.
Anesthesia and post-operative monitoring records (including analgesic administration and suture
removal information) must be maintained in the animal room throughout the postoperative period as
defined above.
V. SURGERIES ON MULTIPLE ANIMALS
When performing surgeries on more than one animal consecutively, the following additional
considerations apply:
A. Instruments
You must begin with a sterile instrument pack. Between animals, the instruments can be wiped
clean and/or rinsed with sterile saline then placed in a chemical disinfectant such as chlorhexidine
(Novasan®) solution, then preferably re-rinsed with sterile water or saline. When possible, instrument
tips can be re-sterilized in a glass bead sterilizer, after any gross tissue contamination has been
removed. A new sterile pack must be used after each 10 surgeries or three hours (whichever
comes first), or any time aseptic technique is breached. The chemical disinfectant must be replaced
when discolored or contaminated with blood or other body fluids.
B. Surgical Gloves
The surgeon may wipe his/her gloves for at least 30 seconds with sterile gauze pads soaked in
chlorhexidine or iodine solution between animals that have already been scrubbed for surgery;
otherwise, a new sterile (or clean for nonsurvival surgeries) pair must be donned between animals. If
the surgeon is also performing animal surgical preparation (scrub) between survival surgeries, s/he
must don new sterile gloves before the next animal.
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 7
C. Monitoring Multiple Anesthetized Animals
If multiple animals will be anesthetized at one time, attention to anesthetic monitoring and supportive
care (e. g. temperature support) is required for each animal, and extra personnel may be necessary
to provide appropriate oversight.
VI. CLEAN TECHNIQUE IN NON-SURVIVAL SURGERY
The following considerations apply to surgeries in which animals remain under anesthesia until they
are euthanized. Whereas aseptic technique is encouraged for all experimental animal surgeries, the
following minimum standards must be applied to non-survival (acute) procedures.
A. General Considerations
Non-survival rodent surgeries must be performed on a clean, uncluttered surface in a room or
portion of a room that is not used for any other purpose during the time of surgical preparation and
the surgical procedure itself. All anesthetic, sedative and analgesic drugs must be used before
their expiration date. Steps to minimize personnel exposure to rodent allergens should be taken.
Other expired drugs, instruments or suture can be used if they are identified as such (see below) and
their use does not lead to an adverse outcome.
B. Instrument Preparation
Surgical instruments must be clean, or sterilized according to the needs of the study. See Section
III.B, Instrument Preparation, for additional information on care of surgical instruments.
C. Animal Preparation
Use of a protocol approved anesthetic agent is required, and animals must be maintained at a
surgical plane of anesthesia (unresponsive to toe pinch, etc.) at all times. The same surgical
monitoring and pre-operative steps are required as for survival surgery, including the continued
verification of anesthetic depth. Site preparation should include hair removal, but a single cleaning
with a disinfectant scrub is sufficient.
D. Surgeon Preparation
Protective gear is in large measure to protect the animal handler and surgeon rather than the animal,
and can consist of standard personal protective equipment as is typically worn in the vivarium, or
surgical attire as described under survival surgery above.
E. Operative Technique
Clean but not necessarily sterile instruments and supplies can be used, but anesthetics, euthanasia
agents, analgesics or sedative must be used before their expiration date. The use of nonpharmaceutical drugs must follow the ACUC Policy on the Use of Non-pharmaceutical Drugs. Drugs,
gloves, suture, and fluids used for non-survival surgeries may be expired if labeled “for acute use
only” or “for non-survival use only.”
F. Multiple Non-Survival Surgeries
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 8
When surgery is performed on consecutive animals, the procedures are largely the same as for other
non-survival procedures. Surgical areas, instruments, and attire, including the surgeon’s gloves, must
be kept clean of accumulated debris by the prudent use of glove changes or cleaning. If more than
one animal is anesthetized, careful attention to monitoring and support of multiple individuals is
crucial, and may require additional personnel.
Reference
National Research Council. 2011. Guide for the Care and Use of Laboratory Animals, 8th Ed. National
Academies Press, Washington DC. pp. 115-121.
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 9
APPENDIX: Cage card option for surgery record, followed by full-page option
Post-surgical records must be completed and kept in the animal housing room during the post-op period,
then returned to OLAR as part of the animal’s medical record, unless they remain on file for inspectors with
the investigator’s study records.
NOTE: Individual cages must be marked with an appropriately completed “pink anesthesia/surgery card” even
if a full-page record is used (and referenced thereon). This will alert the animal care and veterinary staffs of the
anesthetic history of the animal(s).
Surgery cards may be obtained from the OLAR staff.
FRONT:
WVU RODENT ANESTHESIA/ SURGERY RECORD
Principal Investigator:
Date:
Surgeon:
ACUC Protocol No:
Species/Strain:
Animal/Cage ID:
Surgical Procedure:
Pre-Op animal condition:
Suture/Clip removal date:
Preanesthetic Drug(s) Administered
Anesthetic Drug(s) Administered
Drug
Dose/Route
Drug
Dose/Route/Rate
Analgesic(s)
Drug(s)
Dose / Route
Time given
Other medication(s) Drug(s)
Dose / Route
Time given
All medications and procedures above are as stated in the approved animal use protocol
BACK:
Date/Time
Date/Time
WVU RODENT POSTOPERATIVE RECORD
WVU RODENT POSTOPERATIVE RECORD
Animal
Substance(s) given
Surgical Site
Animal
Substance(s)
Condition*
(drug, dose, given
route)
Surgical Site
Condition*
(drug, dose, route)
Initials
Initials
*Animal condition: 1 = Active 2 = Inactive 3 = Moribund 4 = Found Dead
*Animal condition: 1 = Active 2 = Inactive 3 = Moribund 4 = Found Dead
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 10
MULTI-RODENT SURGICAL RECORD
Please ensure that all medications and procedures listed below are as stated in the approved animal use protocol.
GENERAL PROCEDURE INFORMATION
Principal Investigator:
Date:
Surgeon:
ACUC Protocol No.:
Species:
Pre-Surgical Animal Condition:
Evaluation
Preanesthetic Drug(s) Administered
Drug
Dose/Route
Animal/Cage ID:
Analgesic(s):
Dose (mg)/ Route
Drug
Drug
Other Medications:
Drug
Anesthetic Drug(s) Administered
Dose/Route
Dose (mg)/ Route
INTRAOPERATIVE AND ANESTHETIC RECOVERY INFORMATION
Animal ID:
TIME
Animal Weight:
ANIMAL TEMP
RESP RATE
Eye lubricant applied ❑❑❑❑ Righting time:
Anesthesia start (time):
ANESTHETIC
DEPTH
OTHER
INITIALS
Time returned to home cage:
POSTOPERATIVE INFORMATION
DATE / TIME
SURGICAL SITE
(Norm/ Abnormal)
OTHER OBSERVATIONS
SUBSTANCE(S) GIVEN
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
INITIALS
Page 11
INTRAOPERATIVE AND ANESTHETIC RECOVERY INFORMATION
Animal ID:
TIME
AnimalWt:
ANIMAL TEMP
Anesthesia start (time):
RESP RATE
Eye lubricant applied ❑❑❑❑ Righting time:
ANESTHETIC
DEPTH
OTHER
INITIALS
Time returned to home cage:
POSTOPERATIVE INFORMATION
DATE / TIME
SURGICAL SITE
(Norm/ Abnormal)
OTHER OBSERVATIONS
SUBSTANCE(S) GIVEN
INITIALS
INTRAOPERATIVE AND ANESTHETIC RECOVERY INFORMATION
Animal ID:
TIME
AnimalWt:
ANIMAL TEMP
Anesthesia start (time):
RESP RATE
Eye lubricant applied ❑❑❑❑ Righting time:
ANESTHETIC
DEPTH
OTHER
INITIALS
Time returned to home cage:
POSTOPERATIVE INFORMATION
DATE / TIME
SURGICAL SITE
(Norm/ Abnormal)
OTHER OBSERVATIONS
SUBSTANCE(S) GIVEN
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
INITIALS
Page 12
INTRAOPERATIVE AND ANESTHETIC RECOVERY INFORMATION
Animal ID:
TIME
AnimalWt:
ANIMAL TEMP
Anesthesia start (time):
RESP RATE
Eye lubricant applied ❑❑❑❑ Righting time:
ANESTHETIC
DEPTH
OTHER
INITIALS
Time returned to home cage:
POSTOPERATIVE INFORMATION
DATE / TIME
SURGICAL SITE
(Norm/ Abnormal)
OTHER OBSERVATIONS
SUBSTANCE(S) GIVEN
INITIALS
INTRAOPERATIVE AND ANESTHETIC RECOVERY INFORMATION
Animal ID:
TIME
AnimalWt:
ANIMAL TEMP
Anesthesia start (time):
RESP RATE
Eye lubricant applied ❑❑❑❑ Righting time:
ANESTHETIC
DEPTH
OTHER
INITIALS
Time returned to home cage:
POSTOPERATIVE INFORMATION
DATE / TIME
SURGICAL SITE
(Norm/ Abnormal)
OTHER OBSERVATIONS
SUBSTANCE(S) GIVEN
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
INITIALS
Page 13
Ver.2, Effective: August 9, 2012
WVU ACUC POLICY
Anesthesia, Surgery and Post-Operative Care of Rats and Mice
Table of Contents
I.
II.
III.
IV.
V.
VI.
Introduction
Monitoring & Documentation
Aseptic Technique in Survival Surgery
Postoperative Monitoring & Care
Surgeries on Multiple Animals
Clean Technique in Non-Survival Surgery
I. INTRODUCTION
A. Purpose
This document describes appropriate procedures for anesthesia, aseptic surgery, and post-operative
care of laboratory rats and mice at WVU. Other rodents and other warm-blooded mammals regulated
by the United States Department of Agriculture (USDA) are covered under the USDA Species Policy
for Anesthesia and Surgery.
According to the Guide for the Care and Use of Laboratory Animals, 8th edition (2011; henceforth the
Guide), the following areas must be given attention when performing surgery on laboratory animals:
pre-surgical planning, aseptic and surgical technique, assessment of animal well-being during and
after the procedure, appropriate use of analgesics, and each animal’s physiologic status during all
phases of a protocol involving surgery and postoperative care. In addition, the Guide states that
survival surgery on rodents must be performed in facilities intended for that purpose, using aseptic
procedures to prevent clinical infections. The OLAR veterinary staff is always available to provide
assistance with anesthetic or surgical issues, as well as with surgical complications, and challenging
or prolonged anesthetic recoveries. For additional information, OLAR can be contacted by email at
[email protected] or [email protected] or by telephone at 304-293-2721 for other
information. OLAR is located in Room 186G in the Health Sciences Center North.
B. Preparing Animal Care and Use Protocols
This document describes the expectations of the WVU Animal Care and Use Committee (ACUC) for
(a) anesthesia; (b) aseptic surgical technique; (c) appropriate pre-, intra-, and post-operative care;
and (d) documentation for rats and mice undergoing anesthesia or anesthesia and surgery
procedures.
Investigators who adhere to the requirements of this document should provide a brief description of
Phone: 304-293-9368
Phone: 304-293-1050
Fax: 304-293-3098
http://oric.research.wvu.edu
Chestnut Ridge Research Building
886 Chestnut Ridge Road
PO Box 6845
Morgantown, WV 26506-6845
Equal Opportunity/Affirmative Action Institution
the surgical approach, intraoperative actions, and closure technique in their Animal Care as Use
protocol (henceforth, protocol), and then go on to state that this policy document will be followed (i.e.,
the details listed here need not be repeated if this policy is referenced in the protocol).
Deviations from the procedures described herein are subject to the approval of the ACUC. Any
deviations must be described and scientifically justified in the protocol.
C. General Issues
Investigators should be familiar with the following issues that may apply to their research.
1. Acclimation period: A minimum 48-hour acclimation period is required for laboratory mice and
rats prior to experimental use; in other words, animals cannot be used until the third calendar day
after their arrival. See the ACUC Policy on Acclimation of New Arrivals for detailed information and
exceptions.
2. Anesthesia without surgery: Animals undergoing anesthesia must be observed and monitored
until fully ambulatory as described for survival surgery below, including those anesthetized for brief
time periods and/or for minor or non-invasive procedures. During the anesthetic recovery period,
animals must have appropriate monitoring (e.g. temperature), and active animal surveillance such as
contact and stimulation by the observer to aid in arousal and anesthetic recovery is preferred.
Animals can never be left unattended unless the animal has fully regained consciousness. Criteria
used to define complete consciousness include: (a) a body temperature within 1-2 degrees of normal
(normal is ~99 degrees F) and (b) ability of the animal to right itself (i.e. stand on all 4 limbs) from
being placed in full dorsal recumbency (i.e. placed on their back).
3. Analgesic considerations: Please consult with a veterinarian from the WVU Office of Laboratory
Animal Resources (OLAR) for guidance on appropriate surgery-specific analgesic options, including
doses, timing and administration routes when preparing surgical animal protocols. In general,
analgesics are given preemptively to avoid pain “wind-up.” However, the potential for an adverse
effect of analgesic selection on the stability of the anesthetic procedure may affect the plan for
analgesic use in some cases.
4. Antibiotic use: Postoperative antibiotics may be indicated in conjunction with aseptic technique.
Antibiotic administration prior to surgery is more effective, because the blood supply is still
uncompromised to the surgical area at this time. When needed, post-operative antibiotic
administration can be used, but requires an extended treatment period. Contact an OLAR
veterinarian to discuss antibiotic use as it applies to your specific needs.
5. Fluid administration: All animals undergoing anesthesia and/or surgery > 20 minutes duration
benefit from warm intraperitoneal and/or subcutaneous fluids to insure adequate hydration postoperatively and to assist in a faster recovery following injectable anesthetics, such as ketamine, that
are cleared by the kidneys. Contact an OLAR veterinarian for information on appropriate fluid
volumes for your application. A safe rule of thumb for routine use is to deliver a balanced electrolyte
solution volume of 10-12 ml to an adult rat, and 1-2 ml to an adult mouse, either subcutaneously or
intraperitoneally, or divided by both routes.
6. Eye lubricant use: For must general anesthetic procedures lasting more than 20 minutes, eye
lubricant (i. e., Lacrilube®, opthalmic antibiotic ointment) must be placed in each eye to avoid corneal
desiccation.
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 2
Both fluids and eye lubricant may be applied immediately after anesthetic induction for ease of
administration.
II. MONITORING & DOCUMENTATION
To comply with applicable laws and regulations, surgical records must be maintained for all surgeries
performed and must include all anesthetics and analgesics (drug, dose, frequency) administered,
and surgical complications encountered. The OLAR-supplied pink surgery and anesthesia cards
(“pink cards” found in vivarium animal rooms, or obtained from OLAR), if completely filled out and
placed on rodent cages, may suffice for this purpose.
All surgical records must be accessible and readily available upon request to OLAR, the
ACUC, the federal Office of Laboratory Animal Welfare (OLAW) or – in the case of certain
rodents – the USDA (see the USDA Species Anesthesia and Surgery Policy). Investigators
may submit copies of surgical records to OLAR, which will retain them for inspectors.
Sample anesthetic monitoring and surgery record forms for use by investigators can be found in the
Appendix below and on the OLAR website (see contact information above). Although use of these
particular forms is not required, the type of information contained on the sample forms is required.
A. Pre-Operative Records
Initially, visual assessment of animal health must be made. For example, is the animal normally
hydrated, or does it require additional fluids? If injectable premedications (e.g. anticholinergics,
antibiotics), or anesthetics or analgesics are used, the animal’s body weight or an equivalent
standard should be used to calculate dosages. It is important to note and address any abnormalities
that might affect the animal’s procedural outcome.
B. Intra-Operative Records
Surgery records must contain the following information: PI name, surgeon name, ACUC protocol
number, animal or cage ID, species/strain, and surgery date. In addition, for surgeries lasting
more than 15 minutes, a meaningful assemblage of signs must be monitored and recorded:
anesthetic depth (required-response to toe pinch); mucous membrane color (eye color in
albinos); body temperature; respiratory rate (and respiratory depth when relevant); in some cases,
monitoring of heart rate is also indicated. For short procedures, and in smaller animals, it may be
acceptable to monitor only certain parameters, or to note only changes in these parameters. More
complicated procedures and those of longer duration require more detailed monitoring. All
anesthetics, medications, fluids, and/or test substances administered during surgery must be
recorded. Completed intraoperative records or cage pink cards may be submitted to OLAR for safekeeping or they may be maintained in the laboratory provided they are readily available for inspection,
(see below; the back of the card can be used to record added details).
C. Post-Operative Records
Following surgery, all cages must be clearly identified (e.g. completely filled out “pink card”) during
the postsurgical monitoring period within the housing room. Analgesic doses must be recorded on
pink cards (i.e. including drug-circled or written in, dose, frequency, and initials of the person
administering the drug). If the sutures are non-absorbable, an anticipated suture removal date
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 3
should be listed. If relevant, additional postoperative follow-up information relevant to the animal’s
recovery must also be recorded and maintained in the animal room throughout the post-operative
period, including all animal observations, and medications and/or experimental substances
(cells, drugs, biologicals, etc.) administered spanning the surgical and postoperative periods (see
the Post-Operative Monitoring and Care section below).
III. ASEPTIC TECHNIQUE IN SURVIVAL SURGERY
The following standards must be applied to all survival surgeries. As noted in the Introduction, any
deviation must be approved, in advance, by the ACUC.
A. General Considerations
Surgery may only be performed at a site dedicated to that activity at that time (time or space
separated), preferably with a separate area for patient preparation, and must take place on a clean,
uncluttered surface (e.g. no air ducts impinging on the site or overhead shelved materials
likely to fall on the sterile field). Those entering the space during surgery must abide by aseptic
practices such as donning masks and/or personal protective equipment. Traffic in the area during
surgery should be minimized by using appropriate signage. In addition, the cleaned surface must be
covered with an appropriately cleaned heated surface, barrier cloth, or drape. All drugs, suture, and
sterile gloves used for survival surgery must be used before their expiration date.
B. Instrument Preparation
The sterilization method used must be attuned to the durability of the materials to be sterilized, and
sterilization indicators must be used to validate sterilization completeness (outside and preferably
inside sterile packs). Appropriate techniques include: gas (H202 or ethylene oxide); steam
(autoclave); and liquid, chemical sterilants with proper contact time (e.g. 2% glutaraldehyde x 10
hr, or 8% formaldehyde with 70% alcohol x 18 hr). When liquid sterilants are used, instruments must
be rinsed in sterile saline before use. When gas sterilization is used, proper degassing times (e.g. 12
hr for ethylene oxide) should be observed.
The technique of “tip sterilization” using bead or hot dry sterilizers may also be used with care, as
this requires careful attention to maintenance of aseptic technique. Also, excessive heating of
instruments can damage tissue upon contact. All in-house sterilized items must be used within 6
months of the sterilization date (sterilization dates are inscribed on each sterilization pack). Seventy
percent (70%) alcohol is NOT a sterilant or a high-level disinfectant and, by itself, is not
acceptable for surgical instrument sterilization. Novasan® (chlorhexidine gluconate) Solution (not
scrub) may be used for this purpose. Contact OLAR for information on sterilization services that are
available for delicate instruments or equipment that cannot be autoclaved or tip sterilized. Other items
used in surgery such as gauze, suture material, irrigation fluid, syringes, needles, intravascular
catheters, etc. must also be sterile. Only rust-free instruments may be used for rodent surgery. To
preserve the life of instruments, “Surgical milk” can be used prior to autoclaving. Contact OLAR for
additional information on surgical instrument maintenance.
C. Animal Preparation
Use of a protocol approved anesthetic approach, appropriate for the species and procedure is
required. Animals must be maintained at a surgical plane of anesthesia (documented as
unresponsive to toe pinch, or without physiologic evidence of pain response) and and the anesthetist
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 4
must assure this state throughout the surgical procedure.
Thermal support via an external heat source (e.g. circulating warm water blanket) is recommended
when the duration of anesthesia or surgical conditions indicate a significant drop in body temperature
of more than 3 degrees is likely, (e.g. any procedure where a body cavity is opened); if in doubt you
should supply external heat. Frequency of temperature monitoring should be sufficient to avoid large
fluctuations in physiological parameters that need rapid correction, (e.g. an animal’s body
temperature should be monitored at least every 10–15 minutes during the procedure). When using
external heat sources, minimize insulation between the heat source and the animal to get the best
heating effect. If electric pads are used, a layer between the animal and the heat source may be
appropriate, and the use of pads that develop hot spots must be avoided. When autoclavable pads or
hot water-filled gloves are used, care must be taken to monitor and rewarm them regularly so as not
to draw heat from the animal as they cool.
Sterile ophthalmic ointment (e.g. Puralube®) must be applied to the eyes when animals are
unconscious for more than 15 minutes, and re-applied at least every 1.5 hour (or more frequently if
needed), until the animal has recovered. This treatment will keep the eyes lubricated and prevent
corneal injury.
Hair removal from the surgical site, including a sufficient margin, can be achieved using electric
clippers, disposable razors, depilatory creams (e.g. Nair®- must be completely rinsed from the skin)
or by manual plucking under anesthesia. When using depilatories, contact times may have to be
modified with the species and conditions of use.
Soiled skin may require cleaning before surgical skin disinfection can start. Skin disinfection
of the operative site must include three alternating wipes of an approved soap disinfectant scrub (e.g.
chlorhexidine [Novasan® Scrub] or iodine [Betadine Scrub®]) with wipes of 70 % alcohol or saline
(i.e. for Betasept® or Hibiclens® Scrub) followed by a final application of an antiseptic solution (e.g.
iodine solution without detergent). Do not expose the eyes to the scrub, alcohol or disinfectant
solutions. Excessive animal wetting will speed cooling. Before or after disinfection, secure the animal
at the surgery site and cover with a sterile drape, leaving only the disinfected skin exposed. In rats
and mice, use only light drapes to prevent breathing difficulty in the anesthetized animal.
D. Surgeon/Support Personnel Preparation
Hands and arms should be washed thoroughly using an acceptable surgical scrub/brush (e.g.
chlorhexidine [Novasan® Scrub, Betasept®, or Hibiclens® Scrub] or iodine [Betadine® Scrub]) in the
correct manner (contact OLAR for assistance if this is unfamiliar to you). Surgeons and assistants
must don appropriate surgical attire including a sterile or clean gown, scrub shirt, or lab coat, a
surgical mask, appropriate hair covering, and sterile surgical gloves. Surgical or anesthetic
assistants may wear a clean covering, mask, cap and clean examination gloves, rather than sterile
gloves, if they will not be touching the sterile field or instruments.
E. Operative Techniques
Sterile suture material must be selected specifically for the intended purpose at hand (absorbable
vs. non-absorbable, braided vs. monofilament, size and needle type). For example, skin closure
usually requires a monofilament non-absorbable suture such as nylon. Because rodents tend to
chew out sutures, another selection such a metal staples or stainless steel suture may be more
appropriate in some cases
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 5
It is critical to maintain a sterile surgical field at all times. If a non-sterile object comes in contact
with the surgeon’s gloves, a new pair of sterile surgical gloves must be donned. A non-sterile
surgical assistant should be present when needed to assist in maintaining the surgeon’s aseptic
technique (e.g. adjust anesthesia machine, manipulate non-sterile instrument surfaces, provide
supplies, etc.). Contaminated instruments (generally those touching anything outside the sterile field)
are discarded from use.
When surgery involves the opening of a body cavity, at least a two-layer closure is required whereby
the body wall is closed separately using absorbable suture, and the skin closed with some
combination of wound clips, staples, surgical adhesive, stainless steel suture or another nonabsorbable monofilament suture in a simple interrupted pattern. A subcuticular pattern may be used
for skin closure in larger rodents, such as rats and guinea pigs. Consult an OLAR veterinarian for
additional information. Never use silk for skin closure. Silk may be used internally, but this is
discouraged.
Wound clips or non-absorbable skin sutures must be removed when the wound is healed
sufficiently, typically by 14 days postoperative. Failure to remove sutures by postoperative day 21 is
considered noncompliance unless the ACUC-approved protocol calls for euthanasia of the animal
before then (in which case “n/a” should be entered for the suture removal date on the pink card.
IV. POST-OPERATIVE MONITORING & CARE
Oversight of anesthetic recovery monitoring, record keeping and postoperative care is the
responsibility of the investigator and their staff doing the procedure. Post-anesthetic recovery services
may be contracted out to the OLAR veterinary technicians for a fee (contact
[email protected] for additional information).
A. Anesthetic Recovery Period
The recovery cage should be free of particulate bedding and lined with paper towels, “isopads”, etc.
to prevent pica (ingestion of nonfood items) and gastrointestinal or tracheal obstructions during
anesthetic recovery. This is especially important when using buprenorphine. To avoid cannibalism
and suture chewing by cage mates in the recovery cage, rodents must be housed individually or with
no more than two other individuals who have had the same procedure and are convalescing. Grouphoused animals must be separated when recovered to avoid suture chewing until the time of suture
removal. Only when animals are fully ambulatory can they be returned to the vivarium.
Supervised heat support must be provided with an external heat source for any rodent with a
depressed body temperature (e.g. 3 or more degrees below normal) or one that is slow to recover,
and should remain in place until the animal is fully ambulatory. NOTE: when not in direct attendance
at the recovery cage, always ensure that approx. 1/3 to 1/2 of the cage is unheated so the animal
can move away from the heat source if necessary to avoid hyperthermia. If heating bulbs or elements
are used, these should never be closer than 18” from the animal.
Animals must be directly observed while in the recovery cage at least every 5-10 minutes until
ambulatory. These observations include body temperature measurement via rectal probe or
thermometer and contact stimulation of the recovering animals so they generate their own heat.
Additional monitoring includes respiratory rate/quality, mucous membrane color,
activity/anesthetic depth, and incisional integrity, including any leakage or bleeding.
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 6
Analgesic agents to be given at the time of surgery are preferably administered prior to anesthetic
recovery, unless scientifically justified in the approved ACUC protocol, or under direct veterinary
instruction.
Only when animals are able to freely move around the cage can they be returned to the animal
housing room and left unattended. Provision of moistened rodent food and/or dry food on the cage
floor is strongly recommended for post-operative animals at least 24 hours following surgery,
especially when any of the following is present: postoperative disorientation: hindleg, caudal body or
abdominal surgery incisions; or overall debility. Typical situations for extended supplementation
include surgeries that limit mobility or activity, such as hind limb or abdominal surgery.
B. Postoperative Observations
Following surgery, the investigative staff must examine postoperative animals at least once daily until
the animal is stabilized. Daily examination includes at least observation of the incision for heat,
swelling, drainage, or dehiscence (opening), assessment of overall animal activity, and any signs of
pain or distress (described in detail in the ACUC policy entitled Recognition of Pain and Distress,
Humane Endpoints). Observations are noted on the pink card on the animal’s cage.
All postsurgical examinations must be recorded, including any complications encountered. If signs
of pain or distress are observed, a staff member from OLAR must be notified so that appropriate and
timely veterinary care can be provided.
Anesthesia and post-operative monitoring records (including analgesic administration and suture
removal information) must be maintained in the animal room throughout the postoperative period as
defined above.
V. SURGERIES ON MULTIPLE ANIMALS
When performing surgeries on more than one animal consecutively, the following additional
considerations apply:
A. Instruments
You must begin with a sterile instrument pack. Between animals, the instruments can be wiped
clean and/or rinsed with sterile saline then placed in a chemical disinfectant such as chlorhexidine
(Novasan®) solution, then preferably re-rinsed with sterile water or saline. When possible, instrument
tips can be re-sterilized in a glass bead sterilizer, after any gross tissue contamination has been
removed. A new sterile pack must be used after each 10 surgeries or three hours (whichever
comes first), or any time aseptic technique is breached. The chemical disinfectant must be replaced
when discolored or contaminated with blood or other body fluids.
B. Surgical Gloves
The surgeon may wipe his/her gloves for at least 30 seconds with sterile gauze pads soaked in
chlorhexidine or iodine solution between animals that have already been scrubbed for surgery;
otherwise, a new sterile (or clean for nonsurvival surgeries) pair must be donned between animals. If
the surgeon is also performing animal surgical preparation (scrub) between survival surgeries, s/he
must don new sterile gloves before the next animal.
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 7
C. Monitoring Multiple Anesthetized Animals
If multiple animals will be anesthetized at one time, attention to anesthetic monitoring and supportive
care (e. g. temperature support) is required for each animal, and extra personnel may be necessary
to provide appropriate oversight.
VI. CLEAN TECHNIQUE IN NON-SURVIVAL SURGERY
The following considerations apply to surgeries in which animals remain under anesthesia until they
are euthanized. Whereas aseptic technique is encouraged for all experimental animal surgeries, the
following minimum standards must be applied to non-survival (acute) procedures.
A. General Considerations
Non-survival rodent surgeries must be performed on a clean, uncluttered surface in a room or
portion of a room that is not used for any other purpose during the time of surgical preparation and
the surgical procedure itself. All anesthetic, sedative and analgesic drugs must be used before
their expiration date. Steps to minimize personnel exposure to rodent allergens should be taken.
Other expired drugs, instruments or suture can be used if they are identified as such (see below) and
their use does not lead to an adverse outcome.
B. Instrument Preparation
Surgical instruments must be clean, or sterilized according to the needs of the study. See Section
III.B, Instrument Preparation, for additional information on care of surgical instruments.
C. Animal Preparation
Use of a protocol approved anesthetic agent is required, and animals must be maintained at a
surgical plane of anesthesia (unresponsive to toe pinch, etc.) at all times. The same surgical
monitoring and pre-operative steps are required as for survival surgery, including the continued
verification of anesthetic depth. Site preparation should include hair removal, but a single cleaning
with a disinfectant scrub is sufficient.
D. Surgeon Preparation
Protective gear is in large measure to protect the animal handler and surgeon rather than the animal,
and can consist of standard personal protective equipment as is typically worn in the vivarium, or
surgical attire as described under survival surgery above.
E. Operative Technique
Clean but not necessarily sterile instruments and supplies can be used, but anesthetics, euthanasia
agents, analgesics or sedative must be used before their expiration date. The use of nonpharmaceutical drugs must follow the ACUC Policy on the Use of Non-pharmaceutical Drugs. Drugs,
gloves, suture, and fluids used for non-survival surgeries may be expired if labeled “for acute use
only” or “for non-survival use only.”
F. Multiple Non-Survival Surgeries
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 8
When surgery is performed on consecutive animals, the procedures are largely the same as for other
non-survival procedures. Surgical areas, instruments, and attire, including the surgeon’s gloves, must
be kept clean of accumulated debris by the prudent use of glove changes or cleaning. If more than
one animal is anesthetized, careful attention to monitoring and support of multiple individuals is
crucial, and may require additional personnel.
Reference
National Research Council. 2011. Guide for the Care and Use of Laboratory Animals, 8th Ed. National
Academies Press, Washington DC. pp. 115-121.
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 9
APPENDIX: Cage card option for surgery record, followed by full-page option
Post-surgical records must be completed and kept in the animal housing room during the post-op period,
then returned to OLAR as part of the animal’s medical record, unless they remain on file for inspectors with
the investigator’s study records.
NOTE: Individual cages must be marked with an appropriately completed “pink anesthesia/surgery card” even
if a full-page record is used (and referenced thereon). This will alert the animal care and veterinary staffs of the
anesthetic history of the animal(s).
Surgery cards may be obtained from the OLAR staff.
FRONT:
WVU RODENT ANESTHESIA/ SURGERY RECORD
Principal Investigator:
Date:
Surgeon:
ACUC Protocol No:
Species/Strain:
Animal/Cage ID:
Surgical Procedure:
Pre-Op animal condition:
Suture/Clip removal date:
Preanesthetic Drug(s) Administered
Anesthetic Drug(s) Administered
Drug
Dose/Route
Drug
Dose/Route/Rate
Analgesic(s)
Drug(s)
Dose / Route
Time given
Other medication(s) Drug(s)
Dose / Route
Time given
All medications and procedures above are as stated in the approved animal use protocol
BACK:
Date/Time
Date/Time
WVU RODENT POSTOPERATIVE RECORD
WVU RODENT POSTOPERATIVE RECORD
Animal
Substance(s) given
Surgical Site
Animal
Substance(s)
Condition*
(drug, dose, given
route)
Surgical Site
Condition*
(drug, dose, route)
Initials
Initials
*Animal condition: 1 = Active 2 = Inactive 3 = Moribund 4 = Found Dead
*Animal condition: 1 = Active 2 = Inactive 3 = Moribund 4 = Found Dead
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
Page 10
MULTI-RODENT SURGICAL RECORD
Please ensure that all medications and procedures listed below are as stated in the approved animal use protocol.
GENERAL PROCEDURE INFORMATION
Principal Investigator:
Date:
Surgeon:
ACUC Protocol No.:
Species:
Pre-Surgical Animal Condition:
Evaluation
Preanesthetic Drug(s) Administered
Drug
Dose/Route
Animal/Cage ID:
Analgesic(s):
Dose (mg)/ Route
Drug
Drug
Other Medications:
Drug
Anesthetic Drug(s) Administered
Dose/Route
Dose (mg)/ Route
INTRAOPERATIVE AND ANESTHETIC RECOVERY INFORMATION
Animal ID:
TIME
Animal Weight:
ANIMAL TEMP
RESP RATE
Eye lubricant applied ❑❑❑❑ Righting time:
Anesthesia start (time):
ANESTHETIC
DEPTH
OTHER
INITIALS
Time returned to home cage:
POSTOPERATIVE INFORMATION
DATE / TIME
SURGICAL SITE
(Norm/ Abnormal)
OTHER OBSERVATIONS
SUBSTANCE(S) GIVEN
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
INITIALS
Page 11
INTRAOPERATIVE AND ANESTHETIC RECOVERY INFORMATION
Animal ID:
TIME
AnimalWt:
ANIMAL TEMP
Anesthesia start (time):
RESP RATE
Eye lubricant applied ❑❑❑❑ Righting time:
ANESTHETIC
DEPTH
OTHER
INITIALS
Time returned to home cage:
POSTOPERATIVE INFORMATION
DATE / TIME
SURGICAL SITE
(Norm/ Abnormal)
OTHER OBSERVATIONS
SUBSTANCE(S) GIVEN
INITIALS
INTRAOPERATIVE AND ANESTHETIC RECOVERY INFORMATION
Animal ID:
TIME
AnimalWt:
ANIMAL TEMP
Anesthesia start (time):
RESP RATE
Eye lubricant applied ❑❑❑❑ Righting time:
ANESTHETIC
DEPTH
OTHER
INITIALS
Time returned to home cage:
POSTOPERATIVE INFORMATION
DATE / TIME
SURGICAL SITE
(Norm/ Abnormal)
OTHER OBSERVATIONS
SUBSTANCE(S) GIVEN
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
INITIALS
Page 12
INTRAOPERATIVE AND ANESTHETIC RECOVERY INFORMATION
Animal ID:
TIME
AnimalWt:
ANIMAL TEMP
Anesthesia start (time):
RESP RATE
Eye lubricant applied ❑❑❑❑ Righting time:
ANESTHETIC
DEPTH
OTHER
INITIALS
Time returned to home cage:
POSTOPERATIVE INFORMATION
DATE / TIME
SURGICAL SITE
(Norm/ Abnormal)
OTHER OBSERVATIONS
SUBSTANCE(S) GIVEN
INITIALS
INTRAOPERATIVE AND ANESTHETIC RECOVERY INFORMATION
Animal ID:
TIME
AnimalWt:
ANIMAL TEMP
Anesthesia start (time):
RESP RATE
Eye lubricant applied ❑❑❑❑ Righting time:
ANESTHETIC
DEPTH
OTHER
INITIALS
Time returned to home cage:
POSTOPERATIVE INFORMATION
DATE / TIME
SURGICAL SITE
(Norm/ Abnormal)
OTHER OBSERVATIONS
SUBSTANCE(S) GIVEN
WVU ACUC Policy, Anesthesia, Surgery and Post-Operative Care of Rats and Mice, ver.2, eff. 8/9/2012
INITIALS
Page 13
