Animal Resource Facility
Conducting Research with Animals
Animal Use Guidelines
Animal Research Qualifications and Responsibilities
Anesthesia and Analgesia For Laboratory Animals
Surgery
Minor Procedures
Handling and Restraint
Housing, Animal Husbandry and Care
Euthanasia
Animal Research Qualifications and Responsibilities
In order to use animals for research, teaching or testing, the animal user must fulfill all obligations associated with this privilege. These obligations include:
- Completion of adequate training to provide competency in handling the species to be used and competency in the procedures to be used. This training may be accomplished through on-line training sites, viewing training videos/CDs, etc., and/or "wet labs and workshops" conducted by the OACC and ARF.
- Institutional Animal Care and Use Committee approval to use animals as proposed in the official Animal Care And Use Protocol Form.
Anesthesia and Analgesia For Laboratory Animals
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INTRODUCTION, OVERVIEW, PRINCIPLES
Federal law requires that animal researchers consult with the attending Veterinarian during the planning stages of animal procedures to ensure, among other things, the proper use of anesthetics, analgesics and tranquilizers. When completing the Animal Use Protocol form you will be required to complete a section addressing the use of anesthetics and analgesics, including the dosage and route of administration. The most commonly employed anesthetics and analgesics for laboratory animals; their dose, route of administration, and approximate duration of action are listed below by species. This information should only be used as guidelines, since there may be substantial variation among strains or breeds within a species. Anesthesia dose-response variability has become increasingly common with the advent of genetically modified mice. Consulting with the institutional veterinarian can assist you in choosing the appropriate agent for your particular needs. Additional considerations related to proper use, mechanisms of action, accepted routes of administration, monitoring anesthesia depth, and other technical aspects of using anesthetic agents may be provided by the veterinarian. It is especially important that animal researchers inform the veterinarian if anesthesia-related mortalities are experienced, if the subject's response to anesthetic protocols is inconsistent, if recommended doses of anesthetic or analgesic appear inadequate, etc. Any problems or inconsistencies with analgesia should be considered serious matters, and discussed with the veterinary staff. -
CONTROLLED SUBSTANCE USE
Some anesthetics and analgesics commonly used in veterinary medicine are controlled substances. Most analgesics and many injectable anesthetics are controlled substances. Their use requires the investigator to obtain a DEA (Drug Enforcement Administration) number or obtain the drug on an individually dispensed basis from the ARF. The ARF maintains DEA registration for the purpose of providing animal researchers with controlled substances that are essential for the conduct of IACUC - approved research. An ARF - provided controlled substance usage log must be maintained, and these drugs must be stored in a restricted access, locked drawer, cabinet, etc. To request controlled substances from the ARF you must submit a controlled substance request form to the ARF office. - INHALATION AGENTS
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INJECTABLE ANESTHETIC AGENTS
Injectable anesthetic agents have long been the favorite of laboratory animal researchers, not because of superior anesthetic qualities, but rather because of the relative ease of use. Unfortunately, even when used according to established dosing regimens, anesthesia-related mortalities are much higher for injectable agents, compared to properly administered gas anesthetics. It is much more difficult to control the depth of anesthesia with injectable anesthetics, unless an intravenous catheter is in place. Recovery periods are also generally prolonged, and hypothermia is a common sequella to the use of injectable anesthetics. Whenever possible, inhalant anesthetics should be chosen over injectables for use in laboratory animals. The table below provides some guidelines for the choice of anesthetic. The attending veterinarian must be consulted for specific recommendations. -
AGENTS
Analgesic agents for laboratory animals are generally most effective given by injection. Orally available compounds are generally not recommended because it is difficult to provide accurate, consistent dosing, animals tend to drink less when in pain, analgesics are generally not palatable, and most are insoluble in water. The best regimen for pain management involving painful surgical procedures is to use analgesics preemptively as part of the anesthesia induction. Analgesics should be used for all major surgical procedures, for post-operative pain management. Although, a single dose administered pre-operatively may be sufficient, all animals must be monitored for signs of pain post-operatively to determine whether additional doses are indicated. The use and dose of analgesics is very species and procedure dependent. The tables below provide some species-specific guidelines for the choice of available analgesic agents. The attending veterinarian must be consulted for appropriate, specific analgesic regimens. - DOSAGE RECOMMENDATIONS (BY SPECIES)
Recommendations for the choice of anesthetic are provided in the following sections; divided by species. Note that these are recommended dosages, that have worked with relative consistency in each of the species listed. However, many factors affect the success, or failure, of anesthetics and analgesics including; health of the animal, stress, pain, distress, time of day, metabolic state, etc. All researchers should consult with the veterinary staff for the proper anesthetic/analgesic regimen.
AMPHIBIANS (Frogs)
BIRDS (Chickens, Ducks)
FERRETS
MICE
RABBITS
RATS(Rattus norwegicus)
Surgery
ARF Surgery Facilities
The ARF maintains surgery facilities for non-rodent (large) animals, equipped
with anesthesia machines, monitoring and support equipment. These facilities
must be used for all survival procedures on animals larger than the laboratory
rat. These facilities are also available for non-survival procedures on large
animals.
Survival Surgery Guidelines
Survival Surgery, regardless of species, must be conducted aseptically. Surgical
procedures on non-rodent species, larger than the laboratory rat, must be conducted
in dedicated surgery facilities (ARF surgery facility). General requirements for
surgery, survival and non-survival, for all species is described in a detailed
summary,
"General Surgical Guidelines". A second summary entitled
“Rodent Surgical
Guidelines” describes specific information and requirements for surgery involving
laboratory rodents. In general, aseptic technique, appropriate anesthesia and
analgesia, adequate intra-operative patient monitoring, and provisions for post-operative
care are minimal requirements for conducting survival surgery in all species. Aseptic
surgery principles address the following: sterilization of instruments, surgeon's prep,
surgical site prep, and all surgical procedures from skin incision to closure.
Anesthesia monitoring is imperative and must be documented in species larger than
mice and rats.
Non-Survival (Terminal, Acute) Surgery Guidelines
Non-Survival Surgery is defined as surgery conducted under general anesthesia,
and the patient is not recovered from the anesthesia. Euthanasia is performed at
the end of the experimental surgery procedures,, usually with an overdose of
anesthetic, administered intravenously. Non-survival procedures require the use
of clean instruments, and provisions for adequate monitoring of depth of
anesthesia.
Minor Procedures
Minor procedures such as injections, oral dosing, observations, weighing, etc. may be conducted in the animal rooms. The ARF provides procedure rooms with inhalant anesthesia, induction chambers, nose cones, waste gas scavenging devices, and recovery units for those procedures wherein anesthesia may be desirable for restraint or immobilization, such as blood collection, biopsies, iv injections, etc.
Handling and Restraint
Handling and Restraint of all animals, but especially rodents is a skill developed from practice and proper training. Animals should be restrained in a gentle, firm, and quick manner in order to minimize the potential for stress and harm to both the restrained and the restrainer. The ARF offers "hands-on" workshops for individuals requiring an introduction to proper restraint methods.
TopHousing, Animal Husbandry and Care
All research animal housing and husbandry is provide by the ARF staff in accord with the Guide for the Care and Use of Laboratory Animals. If an IACUC approved protocol describes non-standard housing conditions, the ARF staff must be informed of the exemption. A Standard Operating Procedure (SOP) must be developed by the researcher, approved by the ARF veterinarian, and posted in the animal room. If the PI assumes responsibility for the husbandry of the animals, a log/record sheet must be posted in the room that records husbandry duties performed by the research staff.
Euthanasia
Euthanasia of Laboratory Animals
Euthanasia is the act of inducing a humane death. In laboratory animals, euthanasia
is most commonly the endpoint of any experiment. It is imperative that researchers
conduct euthanasia with respect for the animal's life and with the understanding
that pain, anxiety, and distress must be minimized during the procedure. The technique
employed for euthanasia should ensure rapid unconsciousness, followed by cessation of
cardiac and respiratory functions, and finally, loss of brain activity. Whatever
method is method is chosen, consideration must be given to reducing the stress
and anxiety associated with the procedure. The American Veterinary Medical Association
Guidelines on Euthanasia compiled a report in 2007. This report contains specific
guidelines for each species, and acceptable methods of euthanasia. View this report at:
http://www.avma.org/issues/animal_welfare/euthanasia.pdf. In order to perform
euthanasia on animals, the individual must be adequately trained and competent
in the procedure. Euthanasia methods must be detailed in the IACUC-approved animal
use protocol form. Methods used must include measures to ensure that death has occurred;
that there is no chance for the animal to recover, and regain consciousness.
Training is available through the ARF or the Office of Animal Care and Compliance.
Euthanasia of Mouse and Rat Fetuses and Neonates
Late-term fetuses and neonatal rodents are capable of experiencing pain and
distress. Consideration must be given to additional requirements to perform
humane euthanasia of late-term fetuses and neonates. The NIH Animal Research
Advisory Council has developed
guidelines for the euthanasia of rodent fetuses and neonates. Review these
recommendations for determining acceptable methods of euthanasia for mouse and
rat fetuses and neonates.
Euthanasia of Rodents Using Carbon Dioxide
Carbon Dioxide Asphyxiation is the most common method employed to euthanize
small rodents (e.g., mice and rats). Proper methodology must be employed to
reduce the potential for anxiety, distress and pain associated with high
concentrations of CO2. The NIH Animal Research Advisory Council has
published
Guidelines for Euthanasia of Rodents Using Carbon Dioxide. The ARF has
posted these recommendations as an SOP (Standard Operating Procedure) in the ARF
necropsy suite. If CO2 euthanasia is used in research laboratories,
the PI must request the proper equipment from the ARF, and post the SOP provided
by the ARF.