Protocol #:

Amendment
Approval Date:

TUFTS UNIVERSITY & TUFTS MEDICAL CENTER

INSTITUTIONAL ANIMAL CARE AND USE COMMITTEE

http://www.tufts.edu/central/research/AnimalCare.htm

Email: /Phone: 617-636-4109 /Fax: 617-636-8354

Supplement A

Animal Use Protocol Amendment

Amendments to protocols require Institutional Animal Care and Use Committee (IACUC) review and approval prior to initiation. The IACUC reserves the right to determine whether proposed changes require more information, full committee review, or submission of a new protocol. When submitting an amendment, the Principal Investigator is required to review all of the details of the original protocol to assure the IACUC that all unamended details remain identical to the original protocol. Please note that certain changes to protocols may affect other aspects of the protocol. Those changes need to be reflected in this amendment also.

I. General Information

PRINCIPAL INVESTIGATOR: / DEGREE(S):
ACADEMIC POSITION/TITLE: / DEPARTMENT:
MAILING ADDRESS:
DIRECT PHONE #: / EMERGENCY PH #:
EMAIL:
LABORATORY MANAGER: / DEGREE(S):
DIRECT PHONE #: / EMERGENCY PH #:
EMAIL:
PROTOCOL TITLE:

II. Proposed Modifications

Not all Sections in the amendment are relevant for the type of amendment requested. Please complete all areas that will require changes or additions to your original protocol. For all checkboxes, double click on the box to mark.

additional animals needed

species of animal used

procedure(s) used

none of the above

III. Verification of Regulatory Approvals

Institutional Biosafety Committee (IBC) for =

Infectious Agent Registration Number(s) =

Recombinant DNA Registration Number(s) =

Environmental Health and Safety Chemical Hazard

Chemical Hazard name(s) =

Health Physics for Radioisotopes or Irradiation

Hazard name(s) =

Federal or State Wildlife Permit for =

Species and Permit Agency name(s) =

IV. Justification for Additional Animals or Change in Species

A. Change Requested:

additional animals (complete E and D, if applicable)

change in species (complete B, C, and D, if applicable)

B. Type of Species Requested (IF NEW):

Species: / Species:

C. Justify Use of the New Species: Explain why the particular animal model was selected. Describe the unique characteristics that the new species has that are necessary for your investigations. The description needs to be understandable to a lay person.

D. Use of Genetically Modified Animals: Required for each genetically modified strain used, if not already approved in the original protocol. “Use” includes creation of the strain OR breeding of already created mutants. For strains created at this institution, IBC approval is required PRIOR to initiation. For strains that are created elsewhere and used for further breeding here, IBC notification is required. For strains that are created elsewhere and will not be involved in breeding at this institution, no IBC involvement is necessary.

Confirm here that neither parental transgenic rodent contains the following genetic modifications: (i) incorporation of more than one-half of the genome of an exogenous eukaryotic virus from a single family of viruses; or (ii) incorporation of a transgene that is under the control of a gammaretroviral long terminal repeat (LTR); and the transgenic rodent that results from this breeding is not expected to contain more than one-half of an exogenous viral genome from a single family of viruses.

Please list the gene(s) (or family) that will be (or has been) introduced into the germ line and provide a brief description of its encoded gene products and known function. Also, provide the: 1) transgene source, 2) vector used, and 3) if a toxin or other hazardous agent is encoded

4) and describe the expected phenotype. [If unknown, please provide an educated guess based on the known function of the gene(s)]

E. Justification for Additional Animals:

V. Justification for Number of Animals

A.  Assessment of pain and distress: For each species, total the number requested and then identify the number of animals that will be utilized per USDA category. Remember to include all animals used for breeding/maintenance in the total.

USDA Category C: procedures with minimal, momentary, or no distress

USDA Category D: use of appropriate anesthetics, tranquilizers, or analgesics to alleviate pain and/or distress

USDA Category E: animals may experience unrelieved pain and/or distress without intervention

Species 1 / Species 2 / Species 3 / Species 4
Name of species
Total number
requested
Category C
Category D
Category E

B.  Justification for the Number of Animals Requested BY EXPERIMENT: Explain how the number of animals requested was determined. Include justification for the group sizes, the number of groups per experiment, the number of repetitions, etc. The number of animals should be the minimum number required to obtain statistically valid results. Please include a brief description of the statistical analyses, including tests, power and probability levels utilized, if applicable. Include which animals belong in which pain/distress category (C, D, or E). You are encouraged to include a table or flowchart. Assistance in determining appropriate sample sizes can be found from The Center for Statistical Consultation and Research (CSCAR).

C.  Number of Animals Obtained from Breeding: Provide a table/chart that organizes the number expected from breeding.

Include all parents and offspring not directly used in experimental procedures. Include how many litters per female, how many animals born that are culled, and the litter size. The IACUC suggests estimating high for 10 pups per pregnancy, if unknown. Please use this guideline or give justification for a different estimate. All animals born have to be accounted for in the protocol even if not used in the experiments.

VI. Change in Procedure

A. Procedure to be Added/Changed:

B. Describe and Justify: (For survival surgeries, please save the details for Supplement S)

C. Justification for Category E Procedures (if applicable):

Please provide a scientific justification to explain why the use of anesthetics, analgesics, sedatives or tranquilizers during and/or following painful or distressing procedures is contraindicated:

VII. PROCEDURAL DETAILS

A. EXPERIMENTAL ADMINISTRATIONS

For each substance injected, provide: 1) Name, 2) Volume, 3) Site of injection and how it is performed (i.e. needle or gavage gauge, 4) How sterility of the injection(s) is ensured, and 5) Dosage (if appropriate).

B. BLOOD AND/OR BODY FLUID COLLECTION

Maximum survival blood collection volume is 7.7 ml/kg for mice and 64 ml/kg for rats. You can contact DLAM/DTRR for other species maximum collection volumes, if necessary.

1) Blood draw procedure(s) and anatomical area used
2) Total amount for each blood draw
3) Maximum number of draws/animal
4) Frequency of draws/animal
5) Provide maximum volume drawn and timeframe

C. IMPLANTATION OF NON-BIOLOGICS (only for in-dwelling devices over 24 hours)

1) Type and material of implant
2) Site(s) of implantation
3) Size of implant
4) Method of sterilization
5) Length of time of implantation
6) Removal procedure

7) If prolonged implantation is necessary, what methods will be used to avoid tissue infection, inflammation, erosion, or accidental removal of the implant?

VIII. REGULATORY EXCEPTIONS

If yes, please provide justification for exceptions listed below.

1. Are multiple major survival surgeries performed on the same animal? Also include the timeframe between each surgery, including the terminal procedure.

2. Will there be any non-standard housing conditions? Justify the conditions necessary and explain how husbandry will be performed and who will be responsible.

3. Are unanesthetized animals restrained OVER 30 minutes? Indicate the method, duration, and frequency.

4. Will water or food be restricted during any portion of the project? If yes, please describe (substance restricted, duration, location, monitoring, etc.). Do not include overnight fasting for surgery.

5. Describe and justify any other exceptions to the Guide that are not addressed above:

IX. Post-Procedural Care and Monitoring

NOTE: Provide the potential adverse effects and analgesic details of survival surgeries in Supplement S

A. What adverse effects may occur as a result of the experiments to the animals? (Describe distress, pain, significant discomfort, morbidity, etc.) If adverse effects occur, how will they be alleviated?

B.  Humane endpoint criteria (e.g., tumor size, % body wt gain/loss, body condition, inability to eat or drink, behavioral abnormalities, clinical symptoms, signs of toxicity, etc.) must be specified when the experimental manipulations could cause significant adverse effects or are potentially lethal. Clearly list the criteria used to determine when euthanasia will be performed even if prior to the experimental endpoint.

C. What is the frequency and the length of time that ALL animals will be observed in order to evaluate pain/distress during the lifespan of the protocol? Be clear about increasing observations during the time course of an experiment, if appropriate. Also, provide information about general observations for animals during lapses in experimental procedures.

X. Therapeutic Restrictions

This section is only applicable for a new procedure. In an emergency, animals will be treated to relieve suffering and preserve life, or if necessary, euthanized. Investigators will be contacted prior to diagnostic testing, therapy, or euthanasia whenever possible. In the event that contact is not possible, please respond below, and list restrictions on therapy where applicable:

A. No therapeutic restrictions exist.

B. Do not use the following medications (e.g., corticosteroids, antihistamines, antibiotics):

C. If emergency euthanasia is necessary, please save the following specimens: (NOTE: Special arrangements must be made by the responsible investigator with DLAM before onset of the project. Unless otherwise noted, specimen will be placed in DLAM refrigerator for a maximum of 3 days.)

XI. Disposition of Animals Following Study

This section needs to be completed for each new procedure and/or species. Provide details of euthanasia for each species. If animals are not euthanized please describe their final disposition. Even if the experimental plan does not include euthanasia, protocols must include an emergency plan in case euthanasia becomes necessary. (If a physical method of sacrifice is to be used, such as cervical dislocation or decapitation, without prior anesthesia or sedation, please provide scientific justification. This is considered a conditional method based on preserving experimental data).

·  For the selection boxes, double-click on box, click on your selection, and then by the using the arrows move your selection the top of the list, and click OK. If your choice isn’t an option, select NONE and provide further information in text box

·  Only euthanasia methods for smaller species are provided in the text boxes. If a larger species is used, please provide the information in each relevant area beside the box.

·  Copies can be made for each line if euthanasia for more than one species needs to be described.

No change from current protocol.

The euthanasia method below is requested:

Primary euthanasia method: OR ______

Secondary euthanasia method: OR ______

All/Some animals described in this protocol will NOT be euthanized upon completion.

However, emergency euthanasia needs to be included for all animals. Please note no animal may be given away w/o permission from DTRR/DLAM.

Primary euthanasia method: OR ______

Secondary euthanasia method: OR ______

XII. Location of Animals

1. Are live animals ever removed from the central housing facilities? yes no N/A (Field study)

NOTE: Animals can only be out of the centralized facility for less than 24 hours.

Purpose for which room is used / Room number / Is room IACUC approved? / Longest period of time the
animals would be present
sacrifice/tissue harvest / yes no
surgery (non-survival) / yes no
non-surgical procedures (survival) / yes no
surgery (survival) / yes no
other (specify): / yes no

If yes, please respond to the following:

2. Is the location listed above a change from what is currently approved in this protocol? yes no

NOTE: New areas must be inspected by the IACUC prior to use.

3. Justification for removing animals from central housing facility:

XIII. Duplication and Consideration of Alternative Methods

This section is required for a new procedure.

A. Duplication:

These experiments have not been conducted previously

Previously performed experiments were inconclusive

Although similar experiments have been performed, these studies extend our knowledge (explain below)

None of the above apply (provide specific justification below)

B. Alternative Methods Verification for USDA Category D & E Procedures: The search needs to be specific for the procedure and the species. For example: [(anesthesia or anaesthesia or anesthesiology) and (mice or mouse) and (alternative or alternatives)] or [(skin incision) and (rat or rats) and (alternative or alternatives)]

Database Search:

Painful/Distressful Procedures Added in this Amendment: (describe)
Date Search was Conducted: (month/year)
Years Covered in Search: / Database(s) Used:
Provide the key word combinations searched with the # of references for each

C. Narrative statement that alternatives to potentially painful/distressful procedures were evaluated:

XIV. Principal Investigator Assurance of Compliance

As the individual responsible for this project, I confirm that:

The information contained in this amendment is true and accurate, and that, to the best of my knowledge, it conforms with Tufts University/Tufts Medical Center IACUC, NIH, USDA, and MDPH policies on the use of animals in research and teaching.

I have considered alternatives to the biological models used in this project, and have found these other methods unacceptable on scientific or educational grounds.

I certify that I have determined that the research proposed herein is not unnecessarily duplicative of previously reported research.

All personnel listed on this protocol have agreed to participate in this study and are aware of procedures that are Approved. All individuals who will be involved with the animals used in the project have been instructed in the humane care, handling, and use of animals, and I have reviewed their qualifications.