Animal Care

Animal care and use protocols must address the following: 

The IACUC or research committee should seek to avoid reviewing projects for scientific merit and should instead focus on the welfare of laboratory animals and their caretakers.  In an effort to aid the committee in this respect, the principal investigator should provide the information needed for the committee to make basic judgments about the rationale for the use of animals (including species and numbers), the training and experience of investigators and research personnel, and the ability for the researchers to provide adequate animal care in existing facilities with minimal or reasonable modifications to those facilities.  

Justification for the use of the species 

Alternatives

The US Government Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research, and Training states that “the animals selected for a procedure should be of an appropriate species and quality and the minimum number required to obtain valid results.  Methods such as mathematical models, computer simulations and in vitro biological systems should be considered.”  

The National Research Council uses the term “alternative” to refer to research, testing, or training methods that result in the reduction of number of animals required, that incorporate refinements of procedures which result in the lessening of pain or distress, or that provide for replacement of animals with non-whole animal systems, or the replacement of one animal species with another, particularly if the substitute species in non-mammalian or invertebrate.  

It is important that the researcher provide evidence that he or she has considered whether alternatives are available, and if so, why these alternatives will not be utilized.  The Animal Welfare Act requires that investigators provide an indication of the methods and sources utilized to make this determination, whether it is a bibliography, a relevant database, the investigator’s documented experience in the field, documented consultation with an expert in the field, publications, etc.  Although the Animal Welfare Act only applies to warm-blooded animals (excluding mice, rats, and birds bred for research), following this guideline will provide the committee with a reasonable assurance that the investigator is using the appropriate animal(s) and provide documentation of the determination should such justification be required in the future. 

            Replacements—approaches that might be utilized for replacements in research or teaching

a.      In vitro methods, including cell or tissue culture

b.      Mathematical models or computer simulations

c.      Audiovisual technologies

d.      Replacements of the animal species with “lower” species, particularly non-mammalian or invertebrate species 

Reduction—approaches that might aid in utilizing the minimum number of animals necessary

a.      Increase the ability to determine the effect of interest by increasing the size of the effect, decreasing extraneous variability, optimizing sensitivity of the detection methods, changing the scale of the outcome variable

b.      Minimize the number of treatment groups

c.      Wise use of controls

d.      Repeated measures, replicative sample designs

e.      Tissue sharing with other investigators

             Refinement—approaches that might aid in reducing pain or distress of the animals

a.       Administration of anesthetics or analgesics where pain or distress is unavoidable.

b.      Selection of less stressful endpoints

c.       Careful monitoring to detect pain/stress as early as possible

d.      Careful attention to housing and access to food and water in animals that are impaired

e.       Humane methods of euthanasia

 Duplication of Research

The Animal Welfare Act requires that the committee determine that “The principal investigator has provided written assurance that the activities do not unnecessarily duplicate previous experiments.”  The key word is “unnecessarily.”  The investigator should provide an explanation of how the project differs from previous work, or if it duplicates previous work, provide justification of why this is necessary.  Examples of necessary duplication might include demonstration experiments in a course or repeating experiments as learning tools in student research.

 

Training in care, handling, and use of the species

The investigator should provide an adequate description of the steps that will be taken to adequately train students and other research personnel in the appropriate care, handling, and use of the species.  Training programs must include familiarization with committee guidelines with a focus on the standards for humane care and use.  Training should also include necessary information about diseases that may be passed to personnel by the animals or from personnel to the animals, identification and treatment of those diseases, and general first aid procedures that are commonly required as a result of injuries sustained while caring for the animals.  Evidence of training of students and personnel authorized to handle a given species should be listed in the laboratory records along with the appropriate protocol, and each person on the list should be notified of changes in the care, handling, or use protocol for that species.  

Standards for humane care and use of the species 

Good animal care is essential to good science.  Animals that are sick, stressed, or housed without appropriate attention to the maintenance of a standard, stable environment cannot be expected to provide accurate reproducible data.  Prevention of disease is the primary objective of an animal health maintenance program.  Limited space for animal housing poses special challenges when two or more species are located in the facility, and many of the standards for care are aimed at preventing unintended consequences (disease, death, interference with experiments) of these interactions. 

Quarantine

Newly acquired animals can introduce disease into established colonies.  The first step in the process of disease prevention is therefore procurement of healthy animals.  All incoming animals should be inspected on arrival and housed in quarantine separately from acclimated animals until their health status has been established.  Newly arrived animals should be permitted a stabilization period to allow them to physiologically and behaviorally adapt to their new environment before any experimental use is initiated. 

Separation of Species

Some species of animals carry latent or subclinical infections that can cause serious disease if transmitted to other species.  Because of this consideration, physical separation of species is recommended to reduce the possibility of disease transmission, and intraspecies separation is advisable if the animals are obtained from different vendors, or if the differ in microbiological status. 

Surveillance, Diagnosis, Treatment and Control of Disease

All laboratory animals must be inspected regularly by a trained individual to recognize signs of illness or injury.  Sick or injured animals must receive immediate attention.  Methods of diagnosis, therapy, and disease control should follow appropriate standards for the species.  If a contagious disease is suspected, appropriate measures must be taken to prevent the spread of the infection to healthy animals.  Animals that die unexpectedly should be examined to determine the cause of death.  

Appropriate use of Anesthetics, Analgesics, and Tranquilizers

Animals that are in pain or are severely distressed cannot be expected to demonstrate “normal” physiologic responses.  The appropriate use of anesthetics, analgesics, and tranquilizers provides humane care and assists in collection of valid research data.  Attention should be given to the selection of agents that are appropriate to the species and for effective dosage regimens.  If a painful procedure must be conducted without the use of analgesics, the procedure must be carried out under the direct supervision of the responsible investigator until the students or personnel are adequately trained to recognize and minimize the pain without jeopardizing either the research or the care of the animal.  

Surgery and Post-Surgical Care

Individuals performing survival surgical procedures must provide evidence that the procedures will be conducted with appropriate aseptic techniques, with well-maintained equipment, and with appropriate surgical and post-surgical care strategies outlined in protocol form.  

Field Studies

The investigator has limited control of an animal’s environment in field study situations.  When laboratory animals are to be utilized in field experiments, the investigator should provide evidence that the animals will not be subject to undue stress or harm.  Many professional societies provide guidelines for use of different groups of organisms (i.e. fish, amphibians, etc.) in field studies.  The committee will accept these guidelines (where available) as an attachment to the protocol request in lieu of the standards for humane care and use portion of the application. 

Euthanasia

The following guidelines are based on the 1993 report of the AVMA Panel on Euthanasia.  Students or personnel with job responsibilities requiring animal euthanasia should be well-instructed about theory and technique of the methods used.  Consideration should be given not only to the use of humane methods, but also to the use of methods which produce minimal emotional distress to the personnel carrying out the procedure.  Investigators may request to use a procedure that varies from these guidelines through providing written justification for review and approval by the committee. 

            Acceptable Methods

a.      Anesthetic overdose—barbituates or nonflammable gaseous anesthetics

b.      Carbon dioxide

c.      Commercial euthanasia agents (FP3, T61, etc. administered IV)

d.      Cervical dislocation—poultry, mice, rats, and fish less than 200gm

 

Acceptable Methods under Anesthesia

a.      Exsanguination

b.      Decapitation preceded by loss of consciousness or immediate freezing of head in liquid nitrogen

c.      Air embolism

d.      Potassium chloride

Methods Requiring Specific Justification—cost or convenience are not acceptable justifications.

a.      Rapid freezing in liquid nitrogen—restricted to animals less than 40 g

b.      Stunning immediately by an irreversibly fatal procedure such as decapitation, bilateral pneumothorax, or exsanguinations

c.      Decapitation without prior loss of consciousness

d.      Captive bolt pistol—penetrating type

e.      Electrocution

f.        Microwave irradiation

 

Standards of Animal Husbandry

Laboratory animal husbandry covers (1) facilities and operating procedures in facilities including temperature and humidity, lighting, cage construction and maintenance, cage size, and waste disposal and (2) animal health and husbandry including feeding, water, sanitation, staffing, classification and separation, and veterinary care. 

Housing

Animal housing systems should facilitate animal well-being, meet research requirements and minimize experimental variables.  Minimal space requirements are mandated in the Animal Welfare Act and recommended by various funding agencies.  Housing systems should provide sufficient space to allow freedom of movement and normal postural adjustments, contain a resting area appropriate to the species, confine the animal safely, provide easy access to food and water, be well-ventilated, and meet the biological needs of the species.  All housing should be kept in good repair and allow cage occupants to be inspected with minimal disturbance. 

Environment

Animals should be maintained in an environment appropriate to the species.  Care should be taken to minimize fluctuations in environmental conditions and to ensure that the environment within the containers is measured and not the macroenvironment of the holding facility.  When the environmental conditions for a given species are critical, conditions should be monitored and recorded at regular intervals, and the records should be available in the facility at all times.  

Room Light in Animal Enclosures

Room light should be uniformly diffused and should provide sufficient illumination to allow regular observation and care of the animals and safe working conditions for personnel.  Lighting should meet the biological needs of the animals in terms of intensity and periodicity.  If normal lighting in the facility does not meet the needs of all the species housed there, arrangements must be made to provide appropriate lighting to each species as necessary.  The appropriate light cycle should be noted on the protocol label displayed in the facility. 

Food

Food must be fresh, palatable, uncontaminated with biological or chemical agents, and nutritionally adequate for the intended species.  Feeders must allow easy access to food while minimizing contamination by feces and urine.  Animals should be given enough food to permit normal growth, maintenance of age-appropriate body weight, reproduction, and (where appropriate) lactation.  Food should be stored in designated restricted areas that are cool, clean, dry, and free of vermin.  Food containers must not be stored or prepared in rooms with chemicals or biological agents.  An appropriate feeding schedule should be displayed alongside the protocol label for each species, and feeding records should be maintained in the facility as necessary. 

Water

Clean, fresh water must be available at all times as appropriate for the species.  Empty containers should be replaced, not refilled, to prevent disease. 

Sanitation

Good sanitation is essential to maintaining healthy animal populations.  Cages and tanks must be washed and sanitized before animals are placed in them.  Bedding or water must be replaced and the containers cleaned as often as necessary to keep the animals clean and in an optimal environment.  Cleaning schedules should be posted alongside the protocol label for each species, and cleaning records should be maintained in the facility as necessary. 

Animal facilities and other areas in contact with laboratory animals must be cleaned and disinfected often to keep them free of dirt, debris, and biological or chemical contaminants.  Cleaning utensils must not be used for multiple species, and they must not be moved from room to room.  Deodorizers should not be used to mask animal odors. 

No chemical cleaning agent or pesticide is to be used in the animal facility under any circumstances unless its use has first been approved in writing by all investigators housing animals in the facility.  Failure to follow this guideline may harm the health of the animals or interfere with experiments underway or scheduled in the near future. 

Noise

Unnecessary noise can be stressful to animals and should be minimized.  Whenever possible, noisy activities should be restricted to non-housing areas. 

Animal Identification

Animal identification is important to good animal care and data collection.  Permanent individual identification of animals using tattoos, ear notches, fin clipping, or other methods should be supplemented by identification cards that include stock or strain, vendor, responsible investigator, and protocol number.  Additional information should be included on the protocol labels as needed. 

Care Outside of Normal Working Hours

Emergency, holiday, and weekend care must be available as dictated by the species and care schedule.  Emergency contact and normal care contact information should be posted alongside the protocol care label, in prominent locations in and outside the animal facility, and in the care records. 

Animal Care Records

For each protocol approved by the committee, the investigator must maintain a record of animals received/born, diseases/deaths, care schedules, and any other pertinent information as described above or as necessitated by the species or protocol.  These records must be available for inspection in the animal facility at all times.

          

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