Appendix A
Environmental Enhancement / Enrichment
STEVENS INSTITUTE OF TECHNOLOGY (SIT) has an active plan of environmental enrichment that
may include animal socializing directly or indirectly through the cage with other animals as well as
with technician. Unless otherwise, specified, the SIT will provide all available forms of enrichment.
1. Enrichment techniques Are there any additional enrichment forms/methods/techniques that are
included in this protocol?
Yes
No
If YES, please describe.
2. No enrichment? Are there any forms/methods/techniques of enrichment/enhancement that
should not be used in this study?
Yes
No
If YES, provide complete justification for this exemption?
Page 1 of 24
Appendix B
Breeding Programs
Complete Appendix B for all proposals planning on establishing a breeding colony or for those
studies utilizing time-pregnant animals. Studies incorporating breeding programs or offspring from
time-pregnant animals will be required to report annual production (number of offspring used) at the
time of IACUC protocol annual renewal.
1. Description: Provide a specific description of the type of breeding program to be utilized (harem,
monogamous pair, etc).
2. Personnel responsible: Identify and provide contact information for personnel responsible for
the breeding program.
3. Records: Identify and provide contact information for personnel responsible for maintaining
breeding program records.
4. Adults: How many adults (annually) will be utilized on average for this study?
5. Final Disposition of adults: What is the final disposition of these adults at the conclusion of
their breeding program?
6. Offspring: What is the approximate litter size anticipated from each breeding or time-pregnancy?
7. Final Disposition of offspring: What is the final disposition of any offspring not utilized in the
experimental program?
Page 2 of 24
Appendix C
Management of Surgical Pain and Distress
NOTE: Surgical procedures can be categorized as major or minor.
Major survival surgery penetrates and exposes a body cavity, produces substantial impairment of
physical or physiologic functions, or involves extensive tissue dissection or transection action (e.g.;
laparotomy, thoracotomy, joint replacement, and limb amputation).
Minor survival surgery does not expose a body cavity and causes little or no physical impairment (e.g.;
wound suturing, injections, ear notching, peripheral vessel cannulation, percutaneous biopsy, and most
procedures routinely done on an outpatient basis in veterinary clinical practice). Animals undergoing a
minor survival procedure typically do not show significant signs of postoperative pain, have minimal
complications, and quickly return to normal function.
1. This is:
Non-survival surgery (respond to 2-7 only)
Survival surgery (respond to 2-12)
2. Provide a complete formulary of medications related to surgical procedure(s):
Note: Please consult with Attending Veterinarian for appropriate medication and dosage(s)
for each surgery and or procedure.
Species
Agent
Dose
Route
Frequency/Duration
Pre-anesthetic
Anesthetics
Analgesics
Fluid/blood
replacement
Antibiotics
a. Who will conduct the anesthesia procedure(s)?
b. What criteria will be used to assess anesthetic depth and how will this be monitored?
c. Describe the training/experience, including species of animals, of personnel conducting the
anesthetic procedure?
3. Check items to be used:
Item
Yes
No
Item
Yes
No
Item
Endotracheal Tube
Fluids
Sterile drapes
Heat Supplement
Anesthetic vaporizer
Sterile gloves
Hair removal
Mask and Cap
Sterile gown
Disinfection of skin
Stereotaxic device
Sterile instruments
Yes
No
Page 3 of 24
4.
Describe ALL the surgical procedure(s):
a. Surgical site preparation
b. Surgical approach
c. Any unique or special techniques
d. Wound closure method, suture materials and suture removal plan
e. Who will conduct the surgical procedure(s)?
f. Describe the training/experience, including species of animals, of personnel
conducting the surgical procedure?
5.
Where will surgery take place (building and room numbers)?
(Note: Rodents from barrier rooms’ surgeries are to take place in the Laminar Flow
Hood)
6.
What is the anticipated duration of surgery and/or anesthesia?
7.
Will you be using neuromuscular blocking agents?
If Yes, Complete table below.
Species
NM Agent
Dose
No
Yes
Method/s to Assure
Maintenance of
Anesthesia
Justification
Page 4 of 24
COMPLETE ONLY IF SURVIVAL SURGERY
8. What is the length of time animals will survive following recovery from
anesthesia.
9. Will more than one survival surgical procedure be performed on the same animal?
 No  Yes — Please complete Appendix J
Multiple major survival surgical procedures on a single animal are discouraged but may be
permitted if they are related components of a research project, are scientifically justified by
the investigator, or if they are needed for clinical reasons. Cost savings is not an adequate
reason for performing multiple major survival surgical procedures.
10.
Will postoperative analgesics used?
Species
Analgesics
No - Justify:
Route
Yes — complete table below
Dose
Monitoring Method
11. Describe potential post-operative complications.
12. Specify all post-surgical monitoring and care procedures, including suture removal
(mandatory) and any measures in addition to analgesics to alleviate post-operative pain
or discomfort.
Page 5 of 24
Appendix D
Management of Non-Surgical Pain and Distress
(Note: Some examples of these procedures would be re-bandaging, measurements, pictures,
observations, equipment monitoring (telemetry), etc.)
1. This is:
Non-survival procedure (respond to 2-7 only).
Survival procedure (respond to 2-10):
2. Provide a complete formulary of medications related to procedure:
Note: Please consult with Attending Veterinarian for appropriate medication and dosage(s).
Species
Agent
Dose
Route
Frequency/Duration
Pre-anesthetic
Anesthetics
Analgesics
Fluid/blood
replacement
Antibiotics
a. Who will conduct the anesthesia procedure(s)?
b. What criteria will be used to assess anesthetic depth and how will this be monitored?
c. Describe the training/experience, including species of animals, of personnel
conducting the anesthetic procedure?
3. Check items to be used:
Item
Yes
No
Item
Yes
No
Item
Endotracheal Tube
Fluids
Sterile drapes
Heat Supplement
Anesthetic vaporizer
Sterile gloves
Hair removal
Mask and Cap
Sterile gown
Disinfection of skin
Stereotaxic device
Sterile instruments
Yes
No
4. Describe the procedure/s:
a. Site preparation
b. Procedure to be used
c. Any unique or special techniques
d. Post procedure events
Page 6 of 24
e. Who will conduct the procedure/s?
f. Describe the training/experience, including species of animals, of personnel conducting the
procedure?
5. Where will procedure take place (building and room numbers)?
6. What is the anticipated duration of procedure and/or anesthesia?
7. Will you be using neuromuscular blocking agents?
Species
NM Agent
Dose
No
Yes Complete table
Method/s to Assure
Maintenance of
Anesthesia
Justification
Page 7 of 24
COMPLETE ONLY IF SURVIVAL PROCEDURE
8. What is the length of time animals will survive following recovery from anesthesia and/or
procedure.
9. Will post-procedure analgesics be used?
table below
Species
Analgesics
Route
No - Justify:
Dose
Yes — complete
Monitoring Method
10. Describe potential post-procedure complications.
11. Specify all post-procedure monitoring and care procedures, and any measures in
addition to analgesics to alleviate post-procedure pain or discomfort.
Page 8 of 24
Appendix E __________________ Collection of Biological Samples
Biological samples include blood collection, urine collection, ascites, cerebrospinal fluid, biopsy,
organ tissue etc.
1.
Indicate when the samples are being collect
2.
Are any tissues being donated, transferred, used and/or requested to be used by another
researcher, institution, principal investigator, vendor etc.
No
Ante-mortem
Post-mortem.
Yes — complete Application for Animal Tissue(s) Use
3.
Indicate the body fluid or material to be collected.
4.
Indicate the method and site of collection.
5.
Indicate the volume of fluid of material to be collected.
6.
Indicate the frequency of collection
7.
Will the animal(s) be anesthetized or sedated during this procedure?
Yes — complete table
No
Agent
Dose
Route
Frequency Duration
Page 9 of 24
Appendix F
Antibody Formation / Hybridoma & Ascites/ Tumor Induction
Antibody Formation  N/A
1. Indicate what antigen will be used.
2. Indicate what vehicle/adjuvant will be used:
a . Initial immunization:
b . Subsequent immunizations:
c . Anticipated complications/side effects:
3. Indicate sites for immunization:
4. Indicate route of administration:
5. What is the total and per site injection volume?
6. What is the frequency/duration of immunization (e.g., 1 injection every 2 weeks for 3 months)?
Ascites Production
 N/A
Fluid accumulation associated with ascites/hybridomas should not become greater than 10% of body weight.
Animals should be euthanized if they become moribund.
7. Indicate the maximum volume of ascites fluid to be collected per sampling (ml/mouse) and
the method of collection (skin prep, gauge needed, gravity vs. suction, etc)
8. Indicate the number of fluid collections and anticipated frequency of collection.
9. Describe procedures used to care for and monitor the health of animals
with ascites and the point of euthanasia.
Page 10 of 24
Tumor Induction
 N/A
10. Are cells lines free of:
Adventitious viruses?
Mycoplasmas?
Yes provide documentation
Yes provide documentation
No
No
Unknown
Unknown
11. Complete as applicable
12. Describe the pathogenesis of tumor development with specific description of the pain
Species
Carcinogen
Route
Tumor Type
Site of Development
and distress expected:
13. What is the expected endpoint (e.g., tumor size, ulcerations)?
14. Does the agent represent a hazard to humans?
No
Yes, Respond to 15 & 16 below and provide Institutional Biosafety Committee Approval
15. List methods for protection of personnel and containment of hazard.
16. List decontamination procedures and means of disposal of contaminated animals and tissues.
Page 11 of 24
(Agent, cells, compound, drug, etc that comes from outside that produces an effect or condition.)
Appendix G
Exogenous Substances
All agents including experimental (Test) compounds listed in Application to Use Animals for Research or
Teaching (questions 17 and 18) must be listed in this section. This section is not for the listing of veterinary
pharmaceuticals (antibiotics, anesthetics, analgesics).
1. Provide the following information:
Agent/Cmpd
Test/Novel Cmpd
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Yes
No
Dose
Volume
Vehicle
Route
Frequency
For all Test/Novel Compounds please answer questions 2- 4.
2. Which of the following classes applies to test/novel compound(s)?
In vitro or in vivo data exist and indicate no known toxicity.
In vitro or in vivo data exist and indicate probable known toxicity — submit information with
protocol application. [This information can come from testing the compound in question or compounds with
similar structure. It may include information such as chemical class (stimulant, depressant, etc.) mechanism/site
of action, or cytotoxicity]
Toxicity testing part of the protocol.
3. Provide the literature or structure search strategy that was used to determine class:
4. For each box checked in 2 above, describe and indicate your procedures for determining the
following:
Initial Dose
Route
Intervals for
Increasing the dose
(e.g. half log)
Reasons for deviating
from the plan
Rationale for target dose
(e. g., adverse effects and/or
dose needed to treat the
disorder)
Page 12 of 24
Questions 5 - 12 pertain to all agents, compounds and/or hazards mentioned above, please
answer and provide information below.
5. Describe any potential adverse side effects that may result from the administration of any of
the agents/materials listed above. If agents are unknown or their potential side effects are not
documented, provide a reasonable estimate of the effects of the general class of chemicals
(e.g., compound may have sedative properties, compound will likely produce diarrhea, etc).
6. What special procedures, containment or protection does SIT and Research staffs need to
assume when working with these animals or their bedding/cages?
7. List decontamination procedures and means of disposal of contaminated animals and wastes:
8. What is the length of time animals or caging are considered hazardous?
9. Have the protocols for containment and disposal of hazards and protection of personnel been
approved by the appropriate authority?
No
Yes (Approval(s) must be submitted prior to application submission)
10. Establish a data collection procedure for monitoring animals on the treatment, and attach it to
this protocol. This procedure should include an observational checklist of signs of pain and
distress in the species in this study, and the frequency of observations to be made during the
treatment period.
11. List personnel who will monitor the animals. Indicate the training/background that provides
them with the expertise to do this.
12. Ideally, humane endpoints (predictive signs, indicators of an irreversible deteriorating
condition) should be established prior to beginning animal work. Realizing the difficulty in
predicting these for studies which involve compounds with unknown adverse effects, Pls
should try to develop humane endpoints if severe pain, severe distress, or death is observed.
Guidelines should be in place to euthanize animals 1) when humane endpoints are reached, 2)
when the study objectives have been realized, 3) if it becomes clear that they cannot be
realized, or 4) whenever the degree of suffering is not required or justified by the protocol.
A. Explain how animals with severe responses to treatment will be handled.
B. List the signs of pain or distress that may be severe enough to indicate the need for
euthanization.
C. State the method of euthanasia.
All animal monitoring forms should be kept in a location accessible to the Attending Veterinarian
Page 13 of 24
and animal care staff. The PI will notify the Attending Veterinarian in a timely manner when
Unanticipated Significant Adverse events are observed. The PI will complete the relevant
monitoring sheet and submit a copy of it to the IACUC Office.
Include the following information in the Protocol Annual Review Form:
(1) number of compounds used;
(2) number of animals used;
(3) number of animals categorized as something other than the original pain
category;
(4) number of animals euthanized after reaching the humane endpoint.
NOTE: Approval from the appropriate authorities must occur prior to commencing animal work.
(Safety Department, Institutional Biosafety Committee and/or Radiation Safety)
Page 14 of 24
Appendix H
Experimental Disease Induction
1. Infectious Induction information
{What is causing the disease induction? (Includes chemical or physical methods, cells or
tissue, exogenous agents, etc)}
Species
Induction given
(i.e. cells, virus,
agent)
No
Vehicle
Expected
Endpoint
Expected
Mortality (%)
2.
Can
the disease
induction
be
transmitted
to other
animals?
Yes: Explain
3. Describe the pathogenesis of illness with specific description of the pain and distress
expected.
4. Does the disease induction represent a hazard to personnel?
No
Yes
If Yes, please respond to 5 & 6 below & provide Institutional Biosafety Committee Approval.
5.
List methods for protection of personnel and containment of hazards:
6.
List decontamination procedures and means of disposal of contaminated animals and wastes:
Page 15 of 24
Appendix I
Prolonged Physical Restraint
The Guide for the Care and Use of Laboratory Animals provides the following definition of physical restraint:
“Physical restraint is the use of manual or mechanical means to limit some or all of an animal’s normal
movement for the purpose of examination, collection of samples, drug administration, therapy or experimental
manipulation.” (1). Brief physical restraint of agricultural animals for examination, collection of samples, and a
variety of other experimental and clinical manipulations can be accomplished manually or with devices such as
stocks, head gates, stanchions, or squeeze chutes (2). If routine restraint does not cause distress or discomfort
to the animal, a detailed description is NOT required in the IACUC protocol. Prolonged Restraint is defined as
physical restraint of a conscious animal lasting longer than 30 minutes.
1.
Justify the need for prolonged physical restraint.
2. Describe the restraint device.
3. Describe how the animal(s) will be adapted to the restraint device.
4. What is the duration of a restraint period?
5. Are animals monitored during the restraint period?
How often?
Yes
No
6. Are there any anticipated problems as a result of the restraint device (e.g., skin lesion from
harness, moist dermatitis, etc)
Page 16 of 24
Appendix J
Multiple Major Survival Surgical Procedures
Consult with the attending Veterinarian
A major surgical procedure is defined as a surgical intervention that penetrates or exposes a body cavity
(peritoneal, thoracic, cranium) or produces substantial impairment of physical or physiologic function. Multiple
major procedures are those whereby an animal will regain consciousness after each procedure. Procedures
must be described in Appendix C. A major surgery followed by a second procedure where the animal is
sacrificed is not considered multiple major surgical procedures.
1.
Justify the need for multiple major surgical events in a single animal.
2.
What is the time interval between the surgical events?
Page 17 of 24
Appendix K
Food and/or Fluid Restriction
Note: NPO (nil per os / nothing by mouth) procedures for pre-surgical fasting are not included in this
consideration. NPO procedures shall not extend for greater than 24 hours; if surgical delays are
encountered, the animals should be fed and re-fasted prior to the next scheduled procedure.
1. Will
FOOD or
FLUIDS be restricted?
Justify the need to restrict food and/or fluid.
2. Check all methods that will be used to ensure adequate nutritional intake and hydration
METHOD
Body Weight
Urine Output
Fecal Output
Blood Urea Nitrogen
Hematocrit
Food Intake
FREQUENCY OF CHECKS
3. Restriction protocols typically base the restriction amount relative to a baseline, (free-choice
consumption) parameter (body weight, intake amount). What will this restriction amount use
as the baseline?
What is the maximum % restriction to be done for any animal?
4. Growing animals must be frequently re-assessed to ensure normal growth patterns. What
provisions will be made for these animals to assure that their nutritional needs are
maintained?
5. The Animal Welfare Act states that if "water is not continually available, it must be offered to
them as often as necessary to ensure their health and well-being, but no less than twice daily
for at least 1 hour each time, unless otherwise required by the attending veterinarian."
Will animals be provided unrestricted access to water/fluids twice daily for at
least 1 hour each time?
Yes
No
If NO, explain and justify
Page 18 of 24
Appendix L
Animal Pain and/or Distress
Appendix L should be completed if there are any procedures that are proposed that may cause more than
momentary, slight pain or distress during which the appropriate sedatives, analgesics, or anesthetics will
be withheld or in which chronic pain or distress is induced. The management of post-procedural pain or
distress is typically addressed with the use of appropriate pharmacological and non-pharmacologic agents (see
Appendix C). Proposals which incorporate animal manipulations or procedures which may create more than
momentary pain and distress (noxious injections, tumor growth, sequel to compound administration, etc)
should also be addressed.
1.
Justify the scientific need to withhold appropriate drugs or induce the pain/ distress. (Include
references)
2.
What is the duration of time that an animal may experience this pain/distress?
3.
Describe non-pharmaceutical means to alleviate pain/distress (soft bedding, social housing,
supplemental heat, etc.) that will be provided.
4.
Describe situations where an animal may be removed prematurely from a study.
NON-SURGICAL PAIN/DISTRESS
5.
Describe those proposals whereby animals are likely to experience more than momentary
pain or distress as a result of manipulations or procedures (noxious injections, tumor growth,
sequelae to compound administration, etc).
6.
Will any anesthetics, analgesics, or tranquilizing drugs be used to reduce this pain or
distress?
7.
What non-pharmaceutical measures will be used to minimize discomfort, distress, pain, or
suffering (e.g., fluids, supplemental heat, soft bedding, etc)?
Page 19 of 24
Appendix M
Behavioral Training and Testing
Useful Resources:
NIH Publication: Methods and Welfare Considerations in Behavioral Research with Animals NIH
Publication No. 02-5083, March 2002 http://www.nimh.nih.gov/researchfunding/grants/animals.pdf
American Physiological Society Publication: Resource Book for the Design of Animal Exercise
Protocols, Feb. 2006
http://www.the-aps.org/mm/SciencePolicy/AnimalResearch/Publications/Animal-ExerciseProtocols/book14824.pdf
1. What form(s) of behavioral training/testing will be used?
2. Describe how the behavioral training/test is conducted [include descriptions of the devices,
preliminary animal training, fluid/food restriction, reward/ positive reinforcement, duration of
trial, frequency of behavioral testing, etc]
3. If an unexpected problem or event occurs in the performance of the above described
behavioral training/testing procedure(s) that directly impacts the live animal, what steps will
be taken to ensure appropriate treatment is provided?
4. Will animal be observed/attended throughout the duration of the trial/test?
Yes
No
If NO, provide rationale
5. Describe any unique post-trial animal husbandry that may be required. (e.g. dry/warm
environment for animals in the Morris Water Maze, soft padding for animals on the Rod Test,
etc)
6. List Personnel involved with the actual training and indicate her/his level of knowledge as it
relates to the training/testing used in the lab.
Page 20 of 24
Appendix N
Genetically Altered or Manipulated Animal Welfare Profile
See guidance notes for help with completion.
1.
Exact nomenclature
2.
Background strain
3.
Number of backcross generations
4.
Details of modification
Include: type of modification (if microinjection
include copy number where known), gene
affected, inheritance pattern.
5.
References and websites
(where applicable)
6.
Contact Name
7.
Address of contact
8.
Email of contact
9.
Where did the strain originate?
10.
Genotyping method
(at exporting facility)
(please attach protocol)
Send control DNA if possible.
11.
Immune status
Summary of husbandry and welfare issues
12.
General appearance (include photograph if appropriate)
 Coat color
 Any physical abnormalities?
 Is any remedial action necessary?
13.
Behavioral traits
E.G. aggression, tremor, over-grooming etc.
Is any remedial action necessary?
14.
Other abnormalities:
Include typical known
post mortem findings.
15.
16.
Was a structured welfare assessment
carried out for the period that adults
are normally maintained? If ‘yes’ how
long was this?
Husbandry recommendations:
 What diet are the mice fed on?
 Housing system e.g. Ventilated Racks
 Environmental enrichments
17.
What is the current breeding strategy?
Average litter size
Pre/post-weaning mortality
18.
Growth rate
19.
Have specific phenotypic tests been performed?
Breeding life-span
(attach/include data if available)
(please attach results)
Yes
No
Profile Guidance Notes
Page 21 of 24
The animal profile is intended to provide husbandry and welfare information for establishments receiving
genetically altered animal. The profile does not replace any requirements of STEVENS INSTITUTE OF
TECHNOLOGY or regulatory agencies.
The profile is provided for information and guidance and the originating establishment accepts no liability
arising from the use of information provided in this document.
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
Details of correct strain nomenclature can be found at www.jax.org/jaxmice or originating source for
the particular animal.
Details of the original genetic background should be given.
Details of any subsequent breeding should be given.
Please give details of the transgene (and, where known, copy number if generated by pronuclear
microinjection or random integration) or targeting construct and ES cell details. If animal are genetic
mutants, give details of the nature of the mutation and whether original animal were spontaneous or
induced mutants.
Please give details of any publications or websites that deal specifically with the strain. If substantial
information is available please give the most comprehensive or recent reference.
Name of primary contact (e.g. facility manager, director)
Address of originating establishment
Email address of primary contact
This should refer to the originator of the GA animal and the contact details in order to obtain
permission to use the animal. Details should include where and when the animal was generated.
Please give details of the genotyping procedure. Is screening by Southern Blotting, PCR, ELISA,
physical appearance or phenotypic expression? Please attach a protocol for the screening method.
Any known microbiological contamination? Include any information on what microorganisms were
included in the screen, the length of time between each screen and the method used (e.g. sentinels
or littermates).
General appearance should include details such as the range of coat colors to be expected, coat
condition and texture (e.g. downy or fine), any problems with hair growth, any physical abnormalities
(e.g. shortened tails, extra digits, shortened face, obesity, overgrown or under developed teeth). Body
weight at different ages and growth rate information should be given, where available.
Any abnormal behavioral patterns should be noted (e.g. posture, movement, circling, tremors,
aggression, submissive, over-grooming).
Any other abnormalities should be included.
Welfare assessments should be carried out for the period that adults are normally maintained for that
particular line, and the limit to this should be noted.
Details of husbandry and advice should be provided including any special precautions that should be
taken. Points to consider include:
 What containment category does the animal fall into (1-4)?
 Are the animal immunosuppressed?
 What is the most appropriate housing system? For example, conventional housing, Individual
Ventilated Caging, isolators, containment or quarantine.
 Type of consumables required i.e. fiber-free diet, atherogenic diet, powdered diet, irradiated,
autoclaved (please state consumable supplier).
 Are any dietary supplements required e.g. Lactated rehydration therapy? Please provide a
reason as to why this may be necessary.
 The type of bedding and / or nesting materials required.
 Any environmental enrichment devices found to be useful or inappropriate.
 Provide current environmental parameters including temperature, humidity and photoperiod.
This is especially important if conditions differ from Department of Biological Resources or
regulatory agencies guidelines.
Breeding recommendations: It is important to state which animals (male, female, all) carry the relevant
transgene for breeding and welfare purposes. The transgenic status of the breeding colony should be
stated i.e. homozygous or heterozygous, as well as details of the system used to breed animal and
any comments relating to breeding management (e.g. age for first mating, pairs or harems, inbred or
outbred, etc.). Also, include average litter size, average number born live / dead, pre and post-
Page 22 of 24
weaning losses.
18. Where available, body weight at different ages and growth rate information should be given.
19. Specific phenotypic tests include post mortem data, histology and any other analyses performed on
the animal.
Page 23 of 24
Appendix O
Three Year Summary Report
(For application resubmission)
Provide a brief update on the progress made in achieving the specific aims of the protocol. Please also include
the number/amount of test compound used (if any).
PROBLEMS/ADVERSE EVENTS. Describe any unanticipated adverse events, morbidity or mortality, the
cause(s), if known, and how these problems were resolved. If NONE, this should be indicated.
Page 24 of 24