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VILLANOVA UNIVERSITY
OFFICE OF RESEARCH AND SPONSORED PROJECTS
ANIMAL USE PROTOCOL
(VERTEBRATES ONLY)
(REVISED January 8, 2014)
INSTRUCTIONS: Please read carefully
This form is to be completed for all projects proposing the use of vertebrate animals in research. In
addition, any significant changes in previously approved research procedures must be submitted (as
addendum) for additional IACUC review and approval. The completed protocol must describe
research procedures and adhere to standards and requirements as directed by federal oversight
agencies including the NIH and USDA. Competed forms are to be submitted to the Villanova
IACUC chairperson for full committee review PRIOR to the undertaking of any lab or field activities
to which this protocol applies.
The names of all researchers who will be using animals under this protocol, including research
assistants and associates, undergraduate and graduate students, visiting scientists and postdoctoral
fellows, must be kept on file by the principal investigator. In addition, ALL research students must
complete appropriate supervised training in animal handling procedures and provide the IACUC with
health documentation and training certification record as verified by the principal investigator
PRIOR to participation in any lab or field activity involving live animal manipulations.
Protocols are to be submitted by campus mail to the IACUC chair at the address listed below or by
E-mail as an MSWord file attachment. In the case of E-mail submission, the investigator is asked to
send a print copy of the approval page with their signature by campus mail to the IACUC chair.
Faculty will receive a hard copy of the protocol for their files upon approval. If the proposed
research is undertaken, the protocol will become a public document. Investigators will be required to
submit annual protocol for IACUC review.
Dr. Louise Russo
Department of Biology
E-mail: louise.russo@villanova.edu
If you have any questions about completion of the protocol, please contact the IACUC chair.
Supportive research materials may also be accessed directly via the IACUC website
(http://www1.villanova.edu/villanova/vpaa/orgp/research/ora/compliance/institutional_animalcareandusecommitt
ee.html).
THIS FORM MUST BE TYPED
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1. PRINCIPAL INVESTIGATOR (s) and CAMPUS ADDRESS:
NAME
TELEPHONE
E-mail
2. PROJECT TITLE (required):
3. PROPOSED SPONSOR
GRANT/CONTRACT NO. (If known)
4. ESTIMATED DATES OF PROTOCOL: From
to
(Not to exceed 3 years)
5. ACADEMIC DEPARTMENT
6. SUPERVISOR IN CHARGE OF ANIMAL COLONY
7. PROVIDE A NON-TECHNICAL STATEMENT/DESCRIPTION of the GOALS and IMPORTANCE OF
THIS STUDY (should be written for a lay person’s/ non-scientist’s understanding):
8. PERSON(s) WHO WILL PROVIDE PRIMARY ANIMAL CARE (to check boxes in Word, double click
on the appropriate box and select “checked” from the option list):
Departmental Animal Caretaker: please identify
Faculty member/student submitting this protocol
Student worker: please identify
Animals will not be housed in Villanova facilities
9. EMERGENCY CONTACT IF PRINCIPAL INVESTIGATOR IS ABSENT:
Name (s)
daytime telephone
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10. INVENTORY AND FACILITIES
Indicate the species and animal numbers likely to be used in described procedures as well as building
and specific room location(s) where they will be housed if applicable. NOTE: Prior approval must be
obtained if LIVE animals are to be kept for more than 12 hours at sites other than those listed below.
Field investigation only, live animals will not be housed in Villanova facilities
Live animals will be housed in Villanova facilities
Preserved or museum specimens will be used
ANIMAL SPECIES
(Genus species)
NUMBER
to be
used
LOCATION WHERE
STORED/HOUSED
Include building AND room numbers
11. PROPOSED SOURCE(s) OF ACQUIRED ANIMALS / SPECIMENS:
12. PROCEDURES TO BE USED: Describe all experimental procedures to be performed utilizing animals.
Differentiate between species. For field investigations that involve capture and manipulation of animals, describe
methods of capture, type of invasive or noninvasive manipulation that is planned, and final disposition of captured
specimens. For surgical procedures, describe methods of anesthesia including drug compounds and routes of
administration, and post-operative care when appropriate.
NOTE: federal guidelines now specifically stipulate that if animals are to be administered drugs/chemical agents,
formulations must include pharmaceutical-grade compounds (PGCs) for both veterinary and research applications.
However, investigators may request an exception to this mandate based upon acceptable scientific justifications for
the use of non-PGCs. Please refer to Appendix B to this protocol which includes a summary of the Guidelines for
the Use of Non-Pharmaceutical Grade Compounds in Laboratory Animals. Within your experimental procedure
narrative, you MUST include specific information for each treatment compound as to whether it is a PGC or nonPGC compound in addition to source information (supplier/place of purchase). If you expect to use a non-PGC in
your protocol, you MUST include specific scientific reasoning for that option in your experimental
procedures for IACUC review. Use information outlined in the Guidelines for the Use of Non-Pharmaceutical
Grade Compounds in Laboratory Animals (Appendix B) for support of your request making clear why you cannot
use a PGC compound for the protocol.
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13. SELECT APPLICABLE STATEMENT REGARDING PAIN and DISTRESS. Refer to Appendix A for
categories of animals based upon pain and distress level.
Pain and/or distress will not be induced by the described procedures
Some level of distress will be induced ; pain will not be induced
Pain and/or distress will occur; pain will be relieved by drugs listed below as appropriate:
ANIMAL SPECIES
DRUG
DOSE
ROUTE
If drug intervention would interfere with the protocol, clearly describe the reasons why:
If pain and/or distress are induced during the laboratory procedure an alternative procedure(s)
must be considered.
No alternative is acceptable. Reference source:
An alternative is available but will not be used, explain:
14. If this protocol is categorized as inducing pain and/or distress to the animal (see page#7 for Categories B,
C, D), by law you must consider an alternative. These alternatives include less stressful procedures, replacing
the species selected with one believed to be less likely to suffer from the procedure, and reducing the number of
animals you are requesting. Please indicate the source(s) you examined for the alternatives and answer the
following questions. Guidance on completion of literature review is available from ORSP or the IACUC
chairperson upon request.
Animal Welfare Information Center (AWIC)
Current Research Information Center (CRIC)
Biological Abstracts
PsychInfo
Literature Search
Other (Please specify)
The USDA and NIH require the following information: date of Literature Search
Years covered in the literature search: from
Key words used in literature search:
to
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15. IF SURVIVAL SURGERY IS REQUIRED, IN WHICH ROOM(s) WILL SURGERY AND POSTOPERATIVE CARE BE PERFORMED?
16. DOES THE USE OF ANIMALS AS DESCRIBED IN THE PROTOCOL DUPLICATE PREVIOUS
EXPERIMENTS?
No, based upon my present knowledge; Reference sources used (AWIC, Biol.
Abstracts etc.):
Yes, explain why duplication is necessary:
17. PROPOSED EUTHANASIA TECHNIQUES: List by species and technique, including drug type, dose, and
route of administration where applicable.
ANIMAL SPECIES
MODE of EUTHANASIA
What criteria will the investigator use to assure that death has occurred (be very specific: i.e.
cessation of respiration and heart beat, exsanguination, etc.)?
18. DISPOSITION OF ANIMAL REMAINS
Provide specific information on where (room locations) and under what conditions animal remains are to be
stored and how those remains will be processed for disposal.
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19. IS THERE AN ALTERNATIVE to the USE of LIVE ANIMALS for the PURPOSE of this RESEARCH?
No, explain :
Yes, explain why alternative is not being used
20. JUSTIFY the NUMBER of ANIMALS and SPECIES THAT WILL BE USED in the PROPOSED
STUDY. Include important information related to study design, data analysis including statistical
measures, etc. that are important in defining animal numbers for the project.
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21. APPROVAL
I have determined that students, staff, and/or faculty who carry out the procedures described herein are qualified
to complete these tasks in a humane and scientifically acceptable manner.
Date
Signature of Principal Investigator (PI)
Name and Title of PI (Type or Print Clearly)
COMMENTS BY CAMPUS VETERINARIAN
APPROVED BY CAMPUS VETERINARIAN
Date
Signature of Veterinarian
APPROVED BY ANIMAL CARE COMMITTEE
Date
Signature of IACUC Chair
CATEGORY OF ANIMALS APPROVED FOR USE: (to be completed by the IACUC)
A
B
C
D
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Appendix A
CATEGORIES of ANIMAL USE ACCORDING to the LEVEL of INDUCED
PAIN & DISTRESS
Category A. Studies on live, vertebrate animals causing little or no pain or distress. These include:
housing and restraining animals for observation or examination, blood sampling, injection of nontoxic materials.
Category B. Studies involving some pain or distress of short duration. These include cannulation
of vessels or body cavities performed under anesthesia, minor surgical procedures under
anesthesia such as biopsies, overnight food or water deprivation, behavioral studies on awake
animals that involve short-term restraint. Studies on anesthetized animals that do not regain
consciousness are included in this category.
Category C. Studies using animals involving moderate to severe pain or stress or the potential for
moderate to severe pain/stress. These include major surgical procedures under anesthesia,
permitting recovery with adherence to acceptable veterinary practices, i.e. post-op analgesia, fluid
therapy and required veterinary nursing care. Animals should not show signs of prolonged clinical
distress such as behavioral abnormalities, dehydration, abnormal vocalization, or infections.
Should these clinical abnormalities develop and cannot be alleviated, the animals must be
euthanized with minimal delay.
Category D. Projects that involve significant stress or pain without the benefit of pain-relieving
drugs or appropriate therapy. Such studies include: application of noxious stimuli from which
escape is impossible, exposure to noxious stimuli or agents whose effects are unknown, footpad or
intradermal injections of Freund=s complete adjuvant, the use of euthanasia methods not
approved by AVMA. These projects are considered by some to be highly questionable or
unacceptable irrespective of the significance of the results. It is the explicit responsibility of the
investigator to explore alternative methods before proceeding with such studies. Veterinary
involvement in these studies is essential.
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Appendix B
Guidelines for the Use of Non-Pharmaceutical Grade Compounds in
Laboratory Animals
Reference: http://oacu.od.nih.gov/ARAC/documents/Pharmaceutical_Compounds.pdf
Investigators are expected to use pharmaceutical grade compounds whenever possible. The Guidelines
presented here are a synthesis of the requirements and expectations for the use of non-pharmaceutical
grade compounds when they are necessary. This document references the regulations, policies, and
guidelines of the major animal research oversight organizations.1-4
Definitions:
Pharmaceutical-grade compound: A pharmaceutical-grade compound (PGC) is defined as any active or
inactive drug, biologic or reagent, for which a chemical purity standard has been established by a
recognized national or regional pharmacopeia (e.g., the U.S. Pharmacopeia (USP), British Pharmacopeia
(BP), National Formulary (NF), European Pharmacopoeia (EP), Japanese Pharmacopeia (JP), etc.). These
standards are used by manufacturers to help ensure the products are of the appropriate chemical purity
and quality, in the appropriate solution or compound, to ensure stability, safety, and efficacy. 2 The
Food and Drug Administration (FDA) maintains a database listing of FDA approved commercial
formulations for both FDA approved human drugs (the Orange Book) and veterinary drugs (the Green
Book).
Availability: Refers to compounds that are commercially available from an active U.S. vendor. Note:
OLAW has stated that the exorbitant cost of Nembutal (pentobarbital 50 mg/ml injection) has placed it
logistically into the unavailable category. 4
New investigational compound: A compound supplied by its manufacturer for testing in an
experimental setting only and for this reason would not have chemical purity standards established and
by default is considered a non-pharmaceutical grade compound (Non-PGC).
Requirements:
When compounds are used for the clinical treatment of animals or to prevent or reduce/eliminate
animal pain or distress PGCs must be used whenever possible.2
When compounds are used to accomplish the scientific aims of the study PGCs are preferred if available
and suitable. 2
The use of Non-PGCs in laboratory animals must be described and justified in the animal use protocol
and/or covered by an IACUC policy developed for their use and approved by the IACUC.
The AAALAC states that the investigator and the IACUC should consider the following factors when using
Non-PGCs2 :
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
The provided scientific justification such as:
 A PGC is not available; this includes new investigational compounds.
 A PGC is not available in the appropriate concentration or formulation or the
appropriate vehicle control is unavailable
 The Non-PGC is required to generate data that are part of an ongoing study or to
generate data that are comparable to previous work
• Whether the chemical properties of the compound are appropriate for the study (e.g., the purity,
grade, stability in and out of solution, solution vehicle properties, pH, osmolality, and
compatibility of the solvent and other components of final preparation) and the route of
administration.
• The method of preparation, labeling (preparation and use-by dates), administration and storage of
formulations should be appropriately considered with the aim of maintaining their stability and
quality;
• Use must be compliant with applicable national or regional regulatory guidelines and
requirements and the requirements of relevant funding agencies.
The last three factors are not specific to Non-PGCs and therefore should be considered by the ACUC in
general terms as is done for all pharmaceuticals and reagents and will only be considered further in this
guideline briefly.
Although the potential animal welfare consequences of complications are less evident in non-survival
studies, the scientific issues remain the same as in survival studies and therefore apply to non-survival
studies. The use of a non-pharmaceutical-grade euthanasia agent must meet the same standards as for
use in any other application.3
The guidelines pertain to all components, both active and inactive, contained in the preparation to be
administered. Therefore, the vehicle used to facilitate administration of a compound is as important of a
consideration as the active compound in the preparation. Veterinary and human drugs that are
reconstituted in a manner not in accord with the product insert are considered Non-PGCs.
Recommendations for use of non-pharmaceutical-grade compounds:
Where the use of Non-PGCs may be essential for the conduct of science, the goal of the ACUC should be
to consider the health and well-being of the animals while aiding the researcher in minimizing
potentially confounding experimental variables and maximizing reproducibility of the research.2
As stated by OLAW, this Guideline suggests that the IACUC, in making its evaluation may consider factors
including, for example grade, purity, sterility, acid-base balance, pyrogenicity, osmolality, stability, site
and route of administration, compatibility of components, side effects and adverse reactions, storage,
and pharmacokinetics.3 As these considerations should be made for pharmaceuticals and reagents of all
kinds, this guidance from OLAW is generally applicable.
When selecting compounds the following order of choice should be applied:
1. FDA approved veterinary or human pharmaceutical compounds;
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2. FDA approved veterinary or human pharmaceutical compounds used to compound a needed
dosage form;
3. USP/NF, BP, or other pharmacopeia recognized PGCs used in a needed dosage form;
4. Analytical grade bulk chemical used to compound a needed dosage form (requires justification);
5. Other grades and sources of compounds (requires justification).
The NIH Veterinary Pharmacist can provide assistance in availability, procurement, and formulation of
various PGCs.
The IACUC may use a variety of administrative methods to review and approve the use of such agents.
For example, the IACUC may establish acceptable scientific criteria within the institution, rather than on
a case-by-case basis. Investigators and IACUCs should consider relevant animal welfare and scientific
issues including safety, efficacy, availability of PGCs, and the inadvertent introduction of new variables.3
With the exception of pentobarbital4, and presumably any other comparable situations, cost savings
alone is not an adequate justification for the use of Non-PGCs.
Acceptable scientific justifications for the use of non-pharmaceutical-grade compounds:4
1. No equivalent veterinary or human drug is available for experimental use. The highest-grade
equivalent chemical reagent should be used and formulated aseptically, with a non-toxic vehicle,
as appropriate for the route of administration.
2. Although an equivalent veterinary or human drug is available for experimental use, the chemical
grade reagent might be required to replicate methods from previous studies if it is the only
option to produce results that are directly comparable.
3. Although an equivalent veterinary or human drug is available, dilution or change in formulation is
required.
• If the formulation as provided must be diluted, altered by addition, or otherwise changed,
there may be no additional advantage to be gained by using the USP formulation.
• In this situation, use of the highest grade reagent may have the advantage of single-stage
formulation and also result in purity that is equal to or higher than the human or veterinary
drug.
• Professional judgment should be used to determine the appropriate test material and to
ensure use of an agent with the least likelihood for causing adverse effects.
4. The available human or veterinary drug is not concentrated enough to meet experimental
requirements.
5. The available human or veterinary drug does not meet the non-toxic vehicle requirements for the
specified route of administration.
References:
1
U.S. Department of Agriculture, Animal and Plant Health Inspection Service, Animal Care, Policy 3Veterinary Care, March 25, 2011.
2
AAALAC Frequently asked questions about Non-Pharmaceutical Grade Compounds
3 OLAW Position Statement- Non-Pharmaceutical Grade Substances
4 Transcript of OLAW On-line Seminar broadcast on March 1, 2012 - Use of Non-PharmaceuticalGrade Chemicals and Other Substances in Research with Animals
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Approved – 12/03/2008
Revised – 12/08/2010; 04/10/13
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