Writing and Submitting an
Animal Use Protocol
Understanding the Process
http://tulane.edu/asvpr/iacuc/
The charge of an Institutional Animal Care
and Use Committee (IACUC) as mandated
by
NIH’s Office of Laboratory Animal Welfare
and the USDA is to
ensure the humane care for the use of
animals
used in research and
compliance with guidelines and regulations.
Protocol Breakdown
Each section of the
protocol form will be
explained by
stating what information
is expected and
examples of
acceptable responses.
SECTION I. RENEWAL PROTOCOLS
The committee is asking for a brief summary
of what has been accomplished in the previous
3 years.
Information needed:
•State what aims or experiments have been
completed
•Give a summary of outcomes
•List publications that may have resulted
Renewal summary example
Our studies to date have focused on the molecular mechanisms that underlie dendrite development, with
emphasis on the AMPA glutamate receptor signaling pathway. Since the morphology of neuronal
dendrites is a key determinant of the firing patterns of the neuron, and hence neural network properties,
the ability of AMPA receptor expression to alter dendrite morphology may have critical implications for
firing patterns of brain networks as a whole.
Our studies demonstrated that expression of AMPA glutamate receptor subunits controls dendrite
morphology but in a subunit- and region-specific manner. For example, in motor neurons, expression of
the GluR1 subunit promotes outgrowth of new dendritic branches, whereas expression of GluR2
promotes elongation of existing branches (Prithviraj et al., 2008). In the cortex, expression of either
GluR1 or GluR2 promotes addition of new branch segments (Chen et al., 2009), suggesting that there
are region-specific cellular mechanisms that link glutamate receptors to dendrite branch formation. We
also found that the effects of AMPA receptors on dendrite morphogenesis are distinct from those of other
glutamate receptors (Prithviraj and Inglis, 2008), suggesting distinct roles for specific glutamate receptor
subunits in the developmental regulation of neuronal dendrite outgrowth.
Publications arising from work covered by the previous protocol:
Prithviraj R., Kelly K.M., Clarke A., Hexom T., Espinoza-Lewis R., Inglis, F.M. (2008) Differential
regulation of dendrite morphogenesis by AMPA receptor subunits GluR1 and GluR2 in motor neurons.
Developmental Neurobiology, 68:247-64.
Prithviraj R., Inglis, F.M. (2008) Expression of the motor neuron specific NMDA receptor subunit
NR3B regulates dendrite morphogenesis in spinal motor neurons. Neuroscience, 155:145-53.
SECTION III. SPECIES, NUMBER & CATEGORY OF
ANIMAL USE
1.
2.
3.
4.
•
•
•
•
List the SPECIES
List the STRAIN
List the NUMBER
USDA CATEGORY:
B - animals held, conditioned or bred. No procedures conducted
C – teaching or research involving no pain, distress or use of
analgesia
D – teaching or research involving pain or distress accompanied
with appropriate use of analgesia, anesthetic
E - teaching or research involving pain or distress for which the
appropriate use of analgesia, anesthetic will alter the results
of
the experiment. The justification for not using
appropriate
anesthetic, analgesic or tranquilizing
drugs must be provided.
SECTION IV. REGULATORY INFORMATION
1. Is this an endangered or threatened species?
2. Is short-term (> 30 minutes; < 12 hours) physical restraint of a conscious animal employed?
3. Is long-term (12 hours or more) physical restraint of a conscious animal employed?
4. Will specialized management procedures be required, e.g. specialized caging equipment,
restricted or altered feeding/watering/sanitization schedules, or environmental parameters
such as cage size, temperature, lighting or other fall outside the “Guide for the Care and Use
of Laboratory Animals”?
Information needed: This is the most common one answered. Examples of specialized
management procedures would be using mice that need sterile setups, metabolic cages for
urine collection, drugs given in food or water.
Example Response: Athymic nude mice require sterile housing and autoclaved food and water due
to their immunocompromised status.
5. Will you perform more than one major survival surgery on any animal? PLEASE NOTE: major
survival surgery penetrates and exposes a body cavity or produces substantial impairment of
physical or physiologic functions (such as laparotomy, thoracotomy, craniotomy, joint
replacement or limb amputation)
6. Will animals be housed outside of AAALAC-accredited Tulane facilities for over 12
consecutive hours? If YES, the proposed site must be inspected and approved by the IACUC
prior to use. Please list the location with justification below.
SECTION IV. REGULATORY INFORMATION
7. Will pain or distress without the use of analgesic or anesthetic agents occur during the course of
experimentation? This is Category E and must be scientifically justified with references.
Example Response: The proposed experiments necessitate the full manifestation of bacterial
infection in the selected animals in order to fully characterize the health effects of the disease.
The use of analgesics during the course of the study to alleviate pain might modify the host’s
response to infectious challenge that could compromise the results of the study. Narcotic
analgesics can cause respiratory depression (Butelman et al., 1995) and stimulate the production
of a number of immunomodulatory effects laboratory animals. In-vitro exposure of murine
lymphocytes and macrophages to morphine and its metabolites at a wide range of concentrations
resulted in suppressed B cell proliferation, suppression of IL-2, IL-4, and IL-6, and inhibited
cytotoxic T-lymphocyte induction (Thomas et al., 1995). In addition, opioid analgesics also have
immunomodulatory attributes in mice. Opioids have shown a dose dependent attenuation of the
serum TNF-a response in mice as a result of exposure to LPS and results in a significantly lower
LPS-induced serum TNF-aincrease (Piersma et al., 1999). Opoid analgesics also suppress T- and
B-cells, depress NK activity, and decrease the primary antibody response. Steroidal and
nonsteroidal anti-inflammatories interfere with the inflammatory pathway, which is critical in the
pathogenesis of the infectious disease process.
Butelman ER, Winger G, Zernig G, Woods JH. Butorphanol: characterization of agonist and
antagonist effects in rhesus monkeys. J Pharmacol Exp Ther 1995;272:845-53.
8. Will these procedures be filmed, videotaped or photographed for use outside the institution?
SECTION V. PERSONNEL HAZARDS
A. Is there potential exposure of humans to chemical hazards, physical hazards, radioactive agents, or
biohazardous agents during the course of animal experimentation?
Information needed:
The committee is looking for information on anything used in the study that could be hazardous to lab or
vivarial staff personnel
1. Physical
2. Chemical
3. Radioactive
Describe:
Describe:
Describe:
Please complete Appendix B
Please complete Appendix B
Please complete Appendix B
4. Biohazard
Describe:
Please complete Appendix B
Does this biohazard require IBC approval?.
YES - Proceed to Question B. below
NO
5. Recombinant DNA Describe:
Proceed to Question B. below
B. Have you sought Institutional Biosafety Committee (IBC) review?
Yes, please provide the IBC#
Pending review- date of application to IBC:
SECTION VI. NON-TECHNICAL SUMMARY
Using non technical language (lay) that a layperson with no scientific background
would understand, provide a brief summary that describes the nature and purpose of the
this study and its potential value to human or animal health the advancement of
knowledge or the betterment of society. Do not use scientific jargon, acronyms or
abbreviations and do not cut and paste from grant applications.
Information Needed:
The committee is made up of a diverse group of people and not
all members are scientists. It is important that each member have
a clear understanding of the nature of the proposal and the
benefits of the study.
Example of a Nontechnical summary - scientific terms defined
In general, an artery comprises of three layers: intima (faces lumen, inner most layer, covered by
endothelial cells, and are in contact with blood), media (middle layer, comprises mainly of smooth muscle
cells; SMC) and adventitia (outermost connective tissue). In a normal blood vessel, SMC are surrounded
by basement membranes, and prevent proliferation of SMCs. However, under pathological conditions,
proliferation of SMC leads to a thickened neointima (Neointima is an intima that forms in response to
injury). In about 30% of patients, this tissue remodeling process is so exuberant that it produces stenosis
(narrowing of the lumen) of the vessel and the development of related symptoms. The basement membrane
comprises many components, including elastin, collagens, and proteoglycans, whose expression and
deposition is tightly regulated by a delicate balance between matrix degrading metalloproteinases (MMP)
and tissue-inhibitors of metalloproteinases (TIMP). The purpose of this study is to examine the expression
levels of RECK (reversion-inducing cysteine-rich protein with kazal motifs), an MMP inhibitor and
EMMPRIN (extracellular matrix metalloproteinase inducer), an MMP inducer, in an animal model of
balloon catheter-injured carotid artery. The ultimate goal is to see its expression and regulation during the
course of the disease, and verify whether increasing RECK expression suppresses neointimal hyperplasia.
Example of a nontechnical summary without scientific terms
Exposure to particular types of pathogenic Burkholderia sp. bacteria can result in infection
and disease. Currently, infectious disease caused by this particular class of bacteria is
emerging in parts of the world as a significant source of morbidity and mortality.
Development of new vaccines and therapeutics can greatly benefit in the future prevention
and treatment of disease. One of the essential steps in development and production of new
medical products is the development of animal models of disease. Animal models allow a
better understanding of the disease state and the role of natural and acquired immunity
during infection. Models also provide a test system for evaluation of newly-developed
treatments before clinical use. The objective of this research is to use the mouse as a small
animal model for the study of Burkholderia-related bacterial disease and vaccine testing.
SECTION VII. COMPLETE DESCRIPTION
HYPOTHESES
•This should be brief - one or two sentences that clearly explains the
observation, phenomenon, or scientific problem that you are testing.
•You can use scientific terminology in this section. If however there are terms
that are used solely in your field of expertise it may be best to provide a brief
explanation.
Example 1
If the effect of biological sex on performance of the object-in-place task in rats is
similar to the effect of biological sex on performance of the Silverman-Eals task in
humans, we predict that female rats will outperform male rats. However, because
ovarian steroids can affect performance on cognitive tasks, we predict that
ovariectomized females that receive estradiol and progesterone replacement will
outperform ovariectomized females treated with vehicle, as well as outperform
gonadally-intact males.
Example 2
Long term treatment with sodium nitrite has beneficial effects in attenuating the
progression of experimental PH’.
SECTION VII. COMPLETE DESCRIPTION
Information needed:
• Concise description of the experimental course
of an animal from its ENTRY into the
experiment to the ENDPOINT of the study.
• Chronological order is important
• Provide a table
Example of description utilizing written text
•Animal model: Sprague-Dawley rats, male, age 2 months, weight: ~200 g, will be purchased from Charles River
Laboratories.
•Anesthesia: Isoflurane, 5% for induction and 2% for maintenance in oxygen. We will confirm that a surgical plane of
anesthesia has been reached by testing pedal reflexes (by toe pinching).
•Balloon catheter injury: After one week of stabilization, hair on the thigh will be shaved, scrubbed with alcohol and
sprayed with betadine. Bupivicaine will be used as a local anesthetic. After an incision in the thigh, the femoral artery is
isolated, and a balloon catheter (Maverick® Over-The-Wire catheter (20 × 2.0 mm; Boston Scientific Corporation, Natick,
MA, USA) will be inserted and advanced to the carotid under fluoroscopic guidance. The positioning of the catheter can be
seen on the screen. THESE PROCEDURES HAVE BEEN FOLLOWED FOR SEVERAL YEARS and the co-investigator
has extensive published experience. The balloon will be inflated and maintained at 4.0 atm for exactly 20 sec. The pressure
will then be reduced to 2 atm for denuding the endothelium and the catheter will be dragged to the descending aorta. At this
point, the catheter will be deflated totally and the pressure made negative. The catheter will then be gently withdrawn before
tying the artery. The incision will be closed with Vicryl sutures and the animals will be allowed to recover before being
returned to the vivarium. The animals will be under careful observation and concerns about the well being of the animal
would be brought to the notice of the veterinarian.
•Euthanasia: Animals will be euthanized (SOP 3.5.2) on days 7 and 14 following the procedure. We chose these two time
periods based on the assumption that suppression of RECK gene and protein expression may occur much before ECM
degradation and SMC hyperplasia.
•Tissue collection: Both injured and the contra-lateral carotid arteries (for comparison) will be harvested.
•End points:
•Histology for SMC hyperplasia, lumen size, and localization of RECK and EMMPRIN.
•Expression levels of both RECK and EMMPRIN will be analyzed.
Example of description utilizing written text
1. Subjects. Male and female adult Long-Evans rats (75 days of age) will be obtained from Harlan Sprague-Dawley, Inc., housed
in pairs by sex, and maintained on reverse light-dark cycle (12:12, lights off at 07:00).
2. Ovariectomy. One week after arrival, females will be ovariectomized by a procedure routinely used in our laboratory in the
past and approved under previous protocols. Males will undergo sham surgery. The procedures are conducted under aseptic
conditions using sterile surgical drapes and swabs, and autoclaved instruments, sutures, and wound clips. Experimenters wear
sterile surgical gloves, surgical masks, and laboratory coats. Rats are anesthetized by intraperitoneal injections of ketamine (100
mg/kg) and xylazine (7 mg/kg). Prior to surgery, the incision site on each flank is shaved and cleaned thoroughly with iodinepovidine solution. Following excision of the ovary, ovarian vessels are tied off with sterile suture, the muscle wall is closed with
sterile suture, and the skin is closed with sterile wound clips. Body heat is maintained under heat lamps until consciousness is
regained and animals are returned to vivarium. Ibuprofen (Children’s 100, 25 mg/kg) is added to drinking water for 2-3 days and
rats are monitored regularly over several days. Post-surgical complications from this procedure are rare. Males will be
anesthetized and bilateral flank incisions will be closed with suture and wound clips as described above.
3. Hormone Treatments. Beginning one week after surgery, females will be treated by intramuscular injections of estradiol
benzoate (10 micrograms, 48 hr before cognitive testing) and progesterone (500 micrograms, 4-6 hr before cognitive testing), both
delivered in 0.1 ml of sesame oil vehicle. Control females and males will receive parallel injections of 0.1 ml sesame oil at 48 and
5 hours before testing. Rats will receive these treatments each week prior to administration of cognitive tests.
4. Cognitive Testing. Rats will be evaluated on an object-in-place task described by Barker et al. (J. Neurosci. 27:2948, 2007). For
this task, four different objects (e.g., can, bottles, bowls, cups) will be placed in the corners of a black Plexiglas open field (90 cm x
90 cm x 45cm). Each rat will be introduced into the open field, and the amount of time spent exploring the four objects will be
recorded by DVD during a 5-minute information trial. Each rat then will be transferred to a holding cage for 5 minutes, 1 hour, or
2 hours before being re-introduced to the open field. During the retention trial, two of the original objects will be relocated to new
positions in the open field. Time spent exploring each of the four objects will be recorded during a 5-minute retention trial. The
open field and objects will be wiped cleaned with an ethanol solution and air-dried between each trial. Significantly increased time
spent investigating the objects relocated to novel locations indicates memory for their original location. All rats will be tested at
each of the three delay intervals on three weekly tests.
Group Name
Group Description
Number of Animals
Group 1
Sylvatic DENV serotype 2, strain PM33974
inoculated
3
Group 2
Human DENV serotype 2, strain NGC inoculated
1
Day Post Inoculation
Group 1 (n=3)
Group 2 (n=1)
-3
BLD-Clot (4.9 ml)
BLD-CBC (1.2 ml)
BLD-Chem (1.1 ml)
PE
BLD-Clot (4.9 ml)
BLD-CBC (1.2 ml)
BLD-Chem (1.1 ml)
PE
0
INOC – Subcutaneous (DENV-2 PM33974,
5log10 pfu, 1 ml)
PE
INOC – Subcutaneous (DENV-2 NGC, 5log10
pfu, 1 ml)
PE
1
BLD-Clot (2.2 ml)
PE
BLD-Clot (2.2 ml)
PE
2
BLD-Clot (2.2 ml)
PE
BLD-Clot (2.2 ml)
PE
3
BLD-CBC (1.2 ml)
BLD-Chem (1.1 ml)
BLD-Clot (1.1 ml)
PE
BLD-CBC (1.2 ml)
BLD-Chem (1.1 ml)
BLD-Clot (1.1 ml)
PE
C. STANDARD OPERATING PROCEDURES
D. PROPOSED TREATMENTS
The list below contains some common drugs and doses and must be checked if
they will be administered in this protocol.
E. VETERINARY CARE
• Routine care is the most common choice
• Special care- is only when veterinary
interaction is needed based on the study. This
section is NOT for husbandry or management
issues such as frequent cage changing.
F. ADVERSE EFFECTS /CLINICAL
ENDPOINTS
Example Response:
In the event of post-surgical complications, consultation and treatment by the
attending veterinarian are available. If any rat appears ungroomed or
debilitated during the course of the experiment, the rat will be considered
for euthanasia. If any rat discontinues drinking or eating during the course
of the experiment, this discontinuation must be limited to less than 20%
weight loss, and the rat will be considered for euthanasia. Animals are
weighed one time one week after surgery and then every day during the
food deprivation part of the study. If there is discomfort arising from
surgery that cannot be alleviated (see below), animals will be considered
for euthanasia. In addition, rats that show signs of distress in the water
maze and could be at risk for drowning will not be allowed to continue in
the experiment. If the water maze is the last experiment in the study, we
will consider euthanasia. If the water maze is not the last experiment, rats
will be allowed to continue other non-water learning tasks.
F. ADVERSE EFFECTS /CLINICAL
ENDPOINTS
Example Response:
Based on the current literature radiation of the lungs results in
pneumonitis 6 – 12 weeks post-irradiation that progresses to fibrosis
during weeks 16 – 38 post-irradiation (Fleckenstein et al. Temporal
onset of hypoxia and oxidative stress after pulmonary irradiation. Int J
Radiation Oncology Biol phys, 68:196, 2007). Breathing frequency is
expected to increase approximately 6 weeks post irradiation but the
overall health of the animals is not expected to be impaired during the
course of our experiments.
However the animals will be monitored daily and any that develop
signs of distress, including ruffled fur, obvious lethargy, hunched
posture, cachexia, cyanosis or labored breathing, will be removed from
the study and immediately euthanized by exsanguination.
G. ENDPOINTS
You can choose more than one selection based on the study.
•
The Policy of Humane Experimental Endpoints in Rodent Research is acceptable for
this study.
•
The endpoints for this study are time based and noted on the Experimental Design
Section VII.B. However, if an animal’s physical condition deteriorates the Policy of
Humane Experimental Endpoints in Rodent Research is acceptable for this study.
•
There is NO ENDPOINT to this study other than the approved study period and
euthanasia is not required. However, if an animal’s physical condition deteriorates
the Policy of Humane Experimental Endpoints in Rodent Research is acceptable for
this study.
•
I choose not to use the Policy of Humane Experimental Endpoints in Rodent
Research - Please list the criteria to use to determine when euthanasia will be
performed.
H. EUTHANASIA METHOD
Information needed:
• Make sure to describe the euthanasia method
• You may have more than one method
• Justify any methods not consistent the the
AVMA Guidelines on Euthanasia
www.avma.org/issues/animal_welfare/​euthanasia.pdf
Example Response with more than one method
• CO2 euthanasia per SOP 3.5 by the Vivarium staff.
• Rats will be injected with an anesthetic combination of ketamine (100
mg/kg) and xylazine (5 mg/kg) prior to either intracardiac perfusion.
Intracardiac perfusion is a standard laboratory procedure necessary for
proper fixation of brain tissue that will analyzed by immnocytochemistry.
After the induction of deep anesthesia, the rat will be placed under a fume
hood and a chest incision will be made to expose the pericardial cavity. An
18-gauge needle will be inserted into the left ventricle through which 150
ml of heparinized PBS will be delivered to clear the brain of blood,
followed by 250 ml of a 4% paraformaldehyde solution. A small incision
in the right atrium will allow exit of the perfusate from the body.
Following perfusion, the brain will be removed, cryoprotected, frozen, and
stored at -70 C prior to sectioning for immunocytochemical analysis.
• Rats will be anesthetized with isoflurane inhalation by placing them gently
on a raised platform in a large Bell jar that is saturated with isoflurane
vapors and will be monitored till completely anesthetized. They will be
then rapidly decapitated and brains removed.
SECTION VIII. ANIMAL USE
JUSTIFICATION
A.
RATIONALE FOR ANIMAL USE
Example Response
At the present time, non-animal alternatives are not available
to answer fundamental questions about the neuronal
mechanisms that cause learning-related changes in
behavior. To forge links between neuronal function,
memory, and behavioral output, it is important to
identify not only learning-related changes at the neuronal
level but to determine which of those changes are related
to functional and behavioral consequences. To
accomplish that goal, studies require measurement of
neuronal markers and behavior, both, and thus the use of
intact, behaving animals.
SECTION VIII. ANIMAL USE
JUSTIFICATION
B. APPROPRIATENESS OF THE SPECIES SELECTED- While
question A justified using a live animal, this question should
pertain to the species used.
Example Response:
SD rat has been extensively used to study the mechanisms of
intimal hyperplasia. We also have extensive experience with the
rat model, and have necessary surgical skills and instruments.
Mice have also been used to study intimal hyperplasia, but are
technically challenging and the mortality is much higher.
C. ANIMAL NUMBER JUSTIFICATION
Citation of previous research
Provide enough information to indicate that the previous research is similar enough in concept and
methodology to make it reasonable to use similar sample sizes in the proposed research project.
Example response:
We and others have shown that statistical significance can be best evaluated using a group number of
between 6 and 8 for microdialysis studies (Inglis et al, Verrico et al, Ichikawa et al). Each of these
studies was similar in that probes were implanted into a region of brain such as the prefrontal cortex,
and neurotransmitter release evaluated in perfusate from the dialysis probes. Verrico et al. and
Ichikawa et al. employed similar drug studies, but in different regions of the brain, suggesting that these
studies are feasible with group numbers given. In our extensive experience, numbers required are
greater than the number of animals cited, due to possible misplacement of microdialysis probes (caused
by deflection of the probe tip when it encounters, for example, a blood vessel), blockage or damage to
probe. Thus, we propose to use an n of 10 per microdialysis group. (6 groups = 60 animals).
Refs cited:
•Inglis et al., 1994. Enhanced acetylcholine release in hippocampus and cortex during the anticipation
and consumption of a palatable meal. Neuroscience, 62:1049-1056.
•Verrico et al., 2003. Systemic, but not local, administration of cannabinoid CB1 receptor agonists
modulate prefrontal cortical acetylcholine efflux in the rat. Synapse, 48:178-83.
•Ichikawa et al., 2002. Atypical, but not typical, antipsychotic drugs increase cortical acetylcholine
release without an effect in the nucleus accumbens or striatum. Neuropsychopharmacology
Statistical analysis- if numbers are chosen with the intent of obtaining statistically significant differences, the
most objective tool is usually a power analysis to determine sample size. When groups are compared, the goal
of conducting a power analysis is to determine the appropriate number of animals per group to ensure detection
of a significant difference.
Example of response:
First, we can harvest 4-6 viable segments of the middle or posterior cerebral arteries which do not have
significant side branches. The side branches prevent us from pressurizing the segments and thus they are
not usable for our studies. Although we could also harvest segments of the posterior cerebral arteries, we
prefer to focus on only one cerebral artery because of well documented variations in responsiveness.
Second, we will only use one protocol per arterial segment to avoid “carry over” of effects unless
specifically desired. Third, we will sample arteries for western blotting at different times following
application of drugs to determine the time course of responses. Fourth, we find that an average sample
size of 10 is appropriate for statistical validity for most types of measurements. Fifth, not all possible
experimental manipulations will be done. If some experiments initially appear to be nonproductive, or
less important than other protocols, we will go on to the next set of experiments. Based upon previous
studies, a sample size of 10 for the vascular studies is needed to show a group differences. This number
is based upon:
Vascular studies
1. α=.05
2. power=.80
3. δ=13
4. within group SD=11
5. m=1.0
Pilot study
•Main outcome measure being evaluated (e.g. reduction in tumor size)
•The definition of success (e.g., a 50% reduction in tumor size in 3 animals) that would
indicate that the study should be followed up by a full study.
Example Response:
This is a pilot study and we will use a total of 4 animals. Two
will be used to determine the mechanisms of physiological
high monocyte turnover. The other two will be used to
determine when the monocyte turnover rate returns to that
observed in adults. Outcome will be measured by the turnover
rate of monocytes, macrophages and dendritic cells with BrdU
injection and flow cytometric analysis
SECTION IX. ALTERNATIVES TO PAINFUL AND
DISTRESSFUL PROCEDURES
You must consider alternatives to procedures that may cause
more than momentary or slight pain or distress.
Alternatives are defined as new methods that:
•refine existing usage by minimizing animals’ distress such as
the use of analgesics,
•reduce animal usage such as the use of certain experimental
designs, or
•replace whole animal tests with other procedures such as the
use of in vitro analyses.
• When searching for literature on refinement procedures the
subject matter frequently relates to ANIMAL WELFARE or to
ANIMAL BEHAVIOUR.
• With reduction procedures, the methodology may often relate to
MATHEMATICAL AND STATISTICAL PHENOMENA
concerned with the design and validation of experiments.
• When considering replacement options, the relevant literature
often describes work emphasizing methodology (METHODS,
PROCEDURES and TECHNIQUES) and the use of human
tissue, lower organisms or animal tissue instead of whole animals
(TYPES OF STUDY, ORGANISMS).
Example Literature Search
Database Used
Date of search
Period covered by
search
Keywords used
Number of
relevant
references
Were alternatives
found
Pubmed
06/08/10
1966-2010
animal testing alternatives
AND lung cancer
6
1
Pubmed
06/08/10
1966-2010
animal testing alternatives
AND cigarette smoke
AND asbestos
0
0
PubMed search yielded one item suggesting an alternative. Sci Am. 1997
Feb;276(2):80-2. The authors of the article cite selected instances in the research use of
animals that led to erroneous conclusions. For example, they point out that rodent
inhalation studies in the 1960s indicated that cigarette smoke did not cause lung cancer.
Instead of animal models the authors advocate the use of epidemiological studies.
However, they did not cite more recent studies that demonstrated an association
between lung cancer in rodents and exposure to tobacco smoke.
SECTION X. ENVIRONMENTAL
ENHANCEMENT-Housing
The housing requirements are dependent on the species and we
follow the Guide.
Single housing for any species must be justified
Example of justification for single housing:
Rats may be housed in groups, unless one of the following conditions occurs.
First, aggressive behaviors may be displayed by group-housed males, and this
aggression can lead to injuries. Second, some of the rats will be food-restricted for one
week, a procedure that may promote additional aggression between cage mates. Third,
rats will undergo surgery, and group-housed animals may disturb incision sites on cage
mates. Under these conditions, the animals may need to be housed in single cages for a
brief period of time. The decision to house the animals in group versus single housing
will be made in consultation between the investigators and the Vivarium staff
SECTION X. ENVIRONMENTAL
ENHANCEMENT-Enrichment
3 levels of enrichment
1.Standard Enrichment
2.Specific Enrichment
3.Restricted Enrichment – this one must be justified
Example of Justification for restricted enrichment:
Previous findings from our laboratory indicate that the effects of ovarian
steroids on cognitive performance are affected dramatically when rats are
housed in enriched conditions, masking the effects of hormone treatments on
cognitive performance (Daniel et al., Physiol Behav. 66:11, 1999). If females
in the current study are housed under enriched conditions, it will not be
possible to study the effects of ovarian steroids on cognitive performance.
SECTION XI. PERSONNEL
Information needed:
•The committee only needs personnel that will have direct contact with animals.
•We must know their role i.e. injections, surgery etc. and you must list the training
or experience for that specific role.
Example:
NAME/ TITLE
SPECIFIC ROLE ON PROJECT
TRAINING
Person A - PI
All aspects
12+ Years experience with murine models
Person B- Post doc
Inoculations; anesthesia; tissue collection;
euthanasia
Will be supervised by the PI and
completed rodent wet lab training
Person C- lab staff
Inoculations; anesthesia; tissue collection;
euthanasia
Will be supervised by the PI and
completed rodent wet lab training
SECTION XII. FACILITIES
List any room that is not part of the vivarium where a LIVE
animal is brought for experimentation or euthanasia. You need
to state what procedures are being done in the lab.
Example:
BUILDING
ROOM NUMBER
USE
TMC
7620
Injections, euthanasia
Protocol Submission Process
• Protocols are due on the first of the month ( next business day if first
falls on holiday or weekend)
• If possible an administrative review is done. This review is done to
try to present to the IACUC the most complete protocol
• Protocols missing responses are sent back to the PI and must be
revised one week before the meeting
• Meetings are held 2nd week (UT) and 3rd week (DT)
• After the IACUC meeting, each protocol receives a disposition and a
letter is sent to the PI with the disposition, comments or questions.
This occurs within 1-2 weeks after the meeting.
Disposition Explanation
• Approved as Submitted – The PI has clearly explained the animal
procedures, included all justifications and answered all questions
• Approved with Administrative Notations - These are minor changes that
the IACUC Office will review. This protocol does not need to go back to
the committee or designated reviewer for review
• Modifications to Secure Approval (~90%) – The committee has questions
on the study. These can range from one question to over 10 questions. The
revised protocol is reviewed by one or more designated reviewers.
• Deferred – The committee does not have a clear understanding of what the
PI is proposing and needs to review a revised version as a full committee.
• Withhold approval – The committee rarely ever renders this disposition.
The committee would have to agree that the research should not be
conducted and this could be due to a variety of reasons
Review Outcomes
The IACUC Office
Tidewater
http://tulane.edu/asvpr/iacuc/
504-988-6868 (office)
504-988-1445 (fax)
504-481-9684 (cell)
985-871-6636 (TNPRC Thursday)
iacuc@tulane.edu