PROTOCOL #
BROWN UNIVERSITY
Institutional Animal Care and Use Committee
Animal Use Form
Please complete all applicable sections of the Animal Use Form within this document. Once complete, save as
a PDF document and upload into the Coeus Protocol record.
1. Detailed description of the proposed use of laboratory animals.
a. Provide a clear and concise sequential description of the procedures involving the use of animals that is
easily understood by all members of the committee.
b. Describe nonstandard housing and husbandry requirements.
c. Provide a description and rationale for humane endpoints.
d. Describe the use of hazardous materials and provision of a safe working environment.
e. Describe the use of restraint.
f.
Justify the omission of pain-relieving agents.
g. If animals are to be moved through non-research areas, including between buildings, describe the
mode of transportation and measures taken to protect the animals and minimize public exposure.
h. If a member of the Animal Care staff finds that one of your animals has died, describe how the carcass
should be handled and whether a member of your staff be contacted immediately.
2. Surgical Procedures
a. Name and describe in detail each surgical procedure as listed in the Coeus surgical procedures.
Identify on which animal each procedure is being performed.
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b. Multiple Survival Surgeries
i.
Provide a complete scientific justification for performing more than one survival surgery on an
individual animal.
ii.
Provide the interval(s) between the multiple surgeries and the rationale for choosing the interval(s).
c. Pre-Operative Procedures
i.
Identify and describe all pre-operative procedures, including all preparations of the animal(s) for
surgery.
ii.
Describe how the surgical site(s) will be prepared prior to surgery.
iii.
Aseptic Procedures
1. Indicate each method that will be used to ensure the sterility of materials, excluding materials
that have been sterilized by the manufacturer.
High-pressure/temperature (autoclave)
Gas sterilization (ethylene oxide at BioMedical Center)
Dry heat (hot bead sterilizer, between multiple rodent surgeries after initial
sterilization)
Plasma sterilization (Lifespan, by special arrangement)
Chemical sterilant, (e.g., glutaraldehyde, peracetic acid):
Other (Explain):
2. Indicate which of the following procedures will be used to maintain a sterile field during surgery.
Sterile instruments
Sterile gloves
Sterile drapes
Face mask
Surgeon cap
Surgeon scrub
Sterile gown
Other (Explain):
d. Intraoperative Procedures
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i.
Identify and describe all intraoperative procedures
ii.
If you plan on using implanted materials (sutures, staples, wound clips, etc.) or experimental
devices (catheters, electrodes, pins, pumps, etc.), complete the following.
Material
Describe size and
composition
What is the tissue
closure?
Duration
(e.g. days)
iii.
Physical Support - Indicate any physical methods used to support the animal(s) during surgery (e.g.
heating pads, blankets, etc.)
iv.
Monitoring - Describe methods used to monitor the state of anesthesia and general well-being of
the animal(s) during surgery.
e. Post-Operative Care (for survival surgery only)
i.
How long will the animal(s) survive after surgery? (If multiple surgeries are planned, answer for the
last surgery before euthanasia.)
ii.
Give the frequency of post-operative monitoring and how long the monitoring will continue.
iii.
Who will be responsible for post-operative care until the animal can ambulate without danger to
itself and thereafter (including after-hours, weekends, and holidays)?
iv.
List any physical methods used to support the animals in the immediate post-operative period (e.g.,
heating pads, blankets, fluids, etc.)
v.
If you do not plan to use analgesics to provide post-operative pain relief to the animals following
surgery, please justify.
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vi.
f.
Please describe wound management measures that will be used to prevent chronic infections
around any implanted medical device(s).
Post-Operative Complications
i.
Describe any possible or expected post-operative complications and what will be done if these
complications arise.
ii.
Provide criteria by which a decision to euthanize a surgical patient post-operatively will be made.
iii.
In case there is an emergency medical situation and you or your staff cannot be reached, identify
drugs or classes of drugs that should not be used as part of the treatment plan.
g. Maintenance of Post-surgical Medical Records
i.
Identify the individuals who will be responsible for maintaining accurate, daily, post-surgical written
medical records.
ii.
Describe where post-surgical records will be kept.
3. Euthanasia
Unless a deviation is justified for scientific or medical reasons, methods should be consistent with the
current AVMA Guidelines on Euthanasia. Guidelines can be found at:
https://www.avma.org/KB/Policies/Pages/Euthanasia-Guidelines.aspx
a. If physical method, please describe.
b. If chemical method, please describe.
c. Describe endpoint criteria.
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d. Describe disposal methods.
e. Describe contingency plan.
f.
Justify any method that is not consistent with current AVMA guidelines.
4. Emergency Contact Information
Provide a contact name and telephone number for the individual to be contacted in case of an emergency.
Name:
Telephone Number:
5. Drugs
a. List all drugs identified in the Coeus procedures section.
Dose
Drug
mg/kg
mL
Route
Frequency
(e.g. times/day)
Duration
(e.g. days)
b. Describe the record system and security measures to be employed for maintenance of each drug.
6. Non-Hazardous Agents/Materials
List all non-hazardous agents/materials used in the protocol.
Agent/material
Dose and/or Volume
Route of admin.
7. Biological Materials
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a. List all biological materials identified in the Coeus procedures section to be used in animals.
Material (e.g. fluid, cells, tissues)
Route of admin.
b. Describe how these materials were screened for infectious agents.
8. Infectious Agents
a. List all infectious agents, identified in the Coeus procedures section, to be used in animals.
Agent and strain or construct
CDC Biosafety Level of agent
Route of admin.
b. Describe whether an antibiogram, anti-viral drug sensitivity screen, or other appropriate drug sensitivity
panel has been determined for the agent(s) listed to assist physicians in selecting proper therapy if an
inadvertent human infection occurs.
c. If any of these agents are on the CDC list of "Select Agents", describe whether they pose a bioterrorism
risk.
d. Describe whether any infectious agent contains recombinant nucleic acid.
9. Toxic Agents/Materials
a. List all toxic agents/materials, identified in the Coeus procedures section, to be used in animals.
Agent/material
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Nature of Hazard*
Route of admin.
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b. If any of these agents are on the CDC list of "Select Agents", describe whether they pose a bioterrorism
risk.
10. Radioactive Agents
a. List all radioactive agents, identified in the Coeus procedures section, to be used in animals.
Radioactive Agent (include
isotope)
Agent dose
mg/kg
mL
Activity (e.g. mCi/kg)
Route of admin.
b. List which investigator has been given permission by the Radiation Safety Committee to utilize the
isotope(s) indicated.
11. Specimen Collection
a. List any fluids or tissues that will be collected from animals postmortem.
b. If blood is to be collected ante mortem, complete the table below.
*
Site of blood
collection
Amount of blood collected
expressed as volume (mL)/
and % of body weight
(assume 1 ml = 1 gram)
mL
%
mL
%
mL
%
Number of blood
collections
Interval between
collections
weeks
weeks
weeks
c. List all tissue and fluid to be collected ante mortem.
Tissue or fluid
collected
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Site & method of
collection
Amount
(G)
Volume
(mL)
Number of
collections
Interval between
collections
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12. Breeding
a. Justify the need for breeding.
b. Describe breeding scheme/plan.
c. Anticipated total number of postembryonic animals that will be produced (For all but amphibian and
fish):
Species
Strain
# of Male
breeders
needed
# of
Female
breeders
needed
Expected
# of
Offspring
Estimated #
of Offspring
used for this
protocol
Estimated # Estimated #
of offspring
of
transferred
Offspring
to another Euthanized
protocol
without use
d. Describe what will be done with surplus animals.
e. If purchasing pregnant females, explain whether the dams will be allowed to give birth to live pups and
will the pups be used for this project. If pups will not be used for this project, describe what will be done
with the surplus.
f.
Transgenic/Knockout Breeding Programs
i.
DNA/transgene or gene to be disrupted.
ii.
Anticipated consequences to the animal of genetic manipulation.
iii.
Method of monitoring presence of transgene in the animals.
iv.
Describe any special care or monitoring that may be required.
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v.
Describe the disposition of the founders and for how long will they be maintained.
13. Antibody Production
a. Monoclonal Antibody Production:
i.
If monoclonal antibodies will be produced in animals or harvested from hybridoma cell lines as part
of this project, is antibody harvest limited to existing hybridoma cell lines with no further
immunizations or lymphocyte fusions planned?
Yes
No
If not, complete the following table regarding the immunization protocol for the animals prior to
lymphocyte harvest for hybridoma creation. For each antigen for which multiple immunization days
will be used, use a separate row in the table for each immunization day.
Injection
day (e.g. day
0, 7, 30, etc.)
ii.
Antigen
Adjuvant
Total
injection
volume
Amount
antigen
mg
mL
mg
mL
mg
mL
Number
of divided
injections
Injection route
and anatomical
location
(e.g., SC, dorsum)
If feeder cells for supporting hybridoma colony growth will be collected from animals, describe the
exact procedures that will be used to collect the feeder cells and the number of animals needed for
this purpose.
b. Alternatives: Consider alternate research methods that can replace the use of animals. lf any animals
will be used to expand hybridoma cell lines so that antibodies can be harvested from ascites fluid,
complete items i and ii below; then proceed to item iii.
i.
Explain why in vitro cell culture systems for harvesting monoclonal antibodies are not adequate to
meet the research objectives.
ii.
Complete the following table.
Hybridoma
cell line
designation
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Number of
animals used
for ascites
production
Priming
agent and
volume(mL)
Number and
timing of
priming
injections
Volume of
injected
hybridoma
cells (mL)
Number of
abdominal
taps before
euthanasia
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iii.
mL
mL
mL
mL
mL
mL
Euthanasia - Describe the criteria that will be used to determine the terminal abdominal tap:
c. Polyclonal Antibody Production: If polyclonal antibodies will be produced in this species of animal as a
part of this project, complete items i. and ii. then go to item d. If not, go to item d below.
i.
Complete the following table. For each antigen for which multiple immunization days will be used.
Use a separate row in the table for each day.
Injection
day (e.g.
day 0, 7, 30,
etc.)
ii.
Antigen
Adjuvant
Amount
antigen
Total
injection
volume
mg
mL
mg
mL
mg
mL
mg
mL
mg
mL
mg
mL
Number of
divided
injections
Injection route and
anatomical
location
(e.g., SC, dorsum)
Possible adverse effects in animals that might be seen from the proposed antigen or adjuvant
injections and what measures will be taken should these adverse effects occur.
d. Terminal blood collection: If animals used for antibody production will be exsanguinated as a method of
euthanasia, complete items i. through iii. below, then go to item e. If not, go to item e.
i.
Describe the method of exsanguination.
ii.
If anesthetics, tranquilizers, or analgesics will be used prior to exsanguination, describe the
administration of pain-relieving agents including dose (mg/kg), volume (ml), route, and
frequency/duration here; then proceed to item 5. If not, justify the omission of pain-relieving agents
below; then proceed to item e.
iii.
Describe method for ensuring animals are dead following blood withdrawal.
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e. Describe how the antigens or cell lines listed above will be screened to ensure they do not harbor
infectious agents that could infect other laboratory animals or people after injection.
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