# Cover Page

##### Protocol Classification

Check one of the following boxes:

+--------------------+-------------------------------------------------+
| ##                 | First request for AWRC approval for the         |
| ### *New Protocol* | research project.                               |
+--------------------+-------------------------------------------------+
| *Triennial         | Protocol Update, full report required every     |
| Renewal*           | three years.                                    |
+--------------------+-------------------------------------------------+
| *Modification*     | Minor change in the protocol, such as,          |
|                    | anesthesia method, euthanasia procedure, drug   |
|                    | dose, or small increase in number of animals    |
|                    | used.                                           |
+--------------------+-------------------------------------------------+
| *Discontinued      | A summary and progress report of the research.  |
| Protocol*          |                                                 |
+--------------------+-------------------------------------------------+

##### 

## Animal Use Protocol Number

New Protocols - The AUP number will be assigned by the AWRC office.

Existing Protocols - Provide the AUP number cited in the most recent
approval letter.

##### Lead Investigator(s)

The lead investigator is responsible for the following:

-   content of the protocol

-   conduct of individuals in research carried out under the protocol

-   training certification for individuals performing research under the
    protocol

-   filing of yearly reports to the AWRC Committee

Protocol renewal notices and correspondence are sent to the lead
investigator whose name is listed first on the protocol.

##### Title

The protocol title should be unique and brief (35 characters or less).
The title should differ from the grant/proposal titles.

##### Grant /Proposals/Contract Listings

Funding agency information is necessary for certification. Provide name
of sponsoring agency, grant or project number, and grant title. Copies
of the certification ARE NOT sent to the funding agency unless
requested. If you want a copy of the certification sent to the funding
agency, contact Chris Byrne via email at <CEByrne@lbl.gov> or by phoning
510-4586-5507.

##### Biosafety Committee and Radiation Protection Groups

***Biosafety Committee:*** Contact Paul Blodgett via email
<PMCBlodgett@lbl.gov> or phone 510-486-6218.\
***Radiation Protection:*** Contact Chris Donahue via email,
<CADonahue@lbl.gov> or phone 510-486-7736.

# Scientific Merit Review

If a proposal has not been submitted to an external granting agency, but
has undergone internal departmental review at LBNL, attach a letter or
form signed by the department head indicating approval.

# 2. Animal Use Summary

###### a. Specify the type and origin of animals to be used.

*Species and Strain(s)* - The common name is sufficient for standard
laboratory strains (CD-1 mouse; Sprague-Dawley rat; New Zealand White
rabbit; mongrel dog). Provide common and scientific name for exotic
species (clawed frog, *Xenopus laevis*).

*Source(s) of Animals* - Check all sources that apply; fill in the
source name for each category.

###### b. Animal housing

Federal regulation requires that the AWRC inspect long-term housing
sites and locations. List long-term housing sites and locations where
animals are held for eight hours or more.

###### c. Summary, total number of animals

The AWRC acknowledges the difficulty of projecting animal use over a
three-year period for some protocols; however, please make an estimate.
A modification can be filed if the animal use number changes plus or
minus 10%. Provide species/strain and number of animals per each group.
Question 6(b) asks for details about how the number of animals was
derived.

# 3. Summary of Proposed Research

###### a. Abstract

Provide a non-technical description of the research that can be
understood by the general public. The abstract should include the
overview, working hypothesis, and expected outcome of the research. The
abstract should also describe how the research relates to human or
animal health issues.

###### b. Alternatives to Animal Use

Federal law requires that investigators consider alternatives to live
animal use in research. Alternatives to animal use can be divided into
four broad categories.

-   Modification of existing animal use

-   Use of animal-derived material in place of whole animals

-   Replacement of living systems with non-living ones

-   Use of mathematical or computer models

Describe why live animals are needed to meet the goals stated in the
abstract. "None" or "there are no alternatives" will not be accepted as
answers.

Describe alternatives to animal use that are incorporated in the
protocol. Alternatives can include use of surplus animals from other
protocols, use of internal controls such as experiments in which the
glands on one side of is treated and the other side receives a sham
treatment.

###### c. Justify Animal Model Selection

# 4. Summary of Proposed Research

Explain why each species, breed, strain, or mutant animal was chosen.

###### i. Tests a new hypothesis

Federal law requires that researchers avoid, when possible, duplicating
previous work. Conduct a literature search to verify that your research
does not unnecessarily duplicate previous studies.

###### ii. Confirms, or follow-ups, previous work

Provide a brief rationale and a list of pertinent literature for the
study.

###### iii. Production protocol

Provide a brief description of the material produced from animal use
research. Conduct a literature search to verify the material.

*Federal law requires proof that the research does not unnecessarily
duplicate previous work.*

# 5. Summary of Research Results 

###### 5a. Summary of research results ( triennial renewals and discontinued protocols only).

Provide the following information:

-   Number of animals allowed in the previous protocol year.

-   Number of animals used in the previous protocol year.

-   Explanation for any discrepancy in animal use numbers that differs
    by plus or minus 10% from the last approved protocol.

-   List research problems encountered and how they were solved.

###### 

###### 5b. Summary of research progress(triennial renewals, discontinued protocols, and modifications to protocols).

Provide a non-technical summary of your research. Refer to the working
hypothesis and the expected outcome described in the original protocol.
Demonstrate how animal use aided your research. Focus on how the
progress of the research advances understanding in biology or medicine.
If applicable, include copies of the references that support your
summary such as publications, supporting documents, reprints, or grant
progress reports

# 6. Procedures Used

Before answering Question 5, review all applicable Animal Colony
Guidelines.

> *Survival Surgery in Rodents*
>
> *Production of Monoclonal Antibodies/Bleeding*

###### 6a. List of Procedures to Include in Protocol

AWRC must insure that appropriate procedures are used in animal
research. For each species, provide a separate description of every
procedure used, identify the procedure with a short title and designate
a number or letter for each procedure. The designation will facilitate
completion of the table in section 6(b)\]. Include the following
information for each procedure:

> aseptic precautions
>
> anesthesia methods
>
> list of all drugs, include the dosage and administration route
>
> post-op care
>
> details of follow-up care for surgical or other manipulative
> procedures.
>
> how animal care will be monitored and how long the monitoring will
> last
>
> length of time animals survive following procedure(s)
>
> how endpoints are determined
>
> criteria for determining early sacrifice of animals when complications
> or untoward events occur.

If the proposed experiments include animal models that are created by
surgical alteration or by introduction of a transgene, include a
statement describing the animal\'s condition. A description of the
animal\'s condition is required to determine if the animal is
experiencing distress or pain because of conditions induced by creation
of the animal model.

##### 6b. Number of Animals Used for Each Procedure

Provide the number of animals used for each procedure detailed in 6a.

A sample table is provided, to assist you in calculating the figures.

<table>
<colgroup>
<col style="width: 7%" />
<col style="width: 26%" />
<col style="width: 6%" />
<col style="width: 17%" />
<col style="width: 4%" />
<col style="width: 4%" />
<col style="width: 5%" />
<col style="width: 3%" />
<col style="width: 0%" />
<col style="width: 4%" />
<col style="width: 4%" />
<col style="width: 9%" />
</colgroup>
<tbody>
<tr class="odd">
<td colspan="2" rowspan="3"></td>
<td colspan="7"><h5 id="number-of-compounds-tested">Number of compounds
tested</h5></td>
<td colspan="3"></td>
</tr>
<tr class="even">
<td colspan="4"><p id="number-of-time-points" class="heading">Number of
time points</p></td>
<td colspan="2"></td>
<td colspan="2"></td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td colspan="3"><p id="number-ratstime-point" class="heading">Number
rats/time point</p></td>
<td></td>
<td></td>
<td colspan="3"></td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td colspan="5"><strong>No. Rats/time point x no. of time points x no.
of compounds tested = Total</strong></td>
<td colspan="2"></td>
<td colspan="3"></td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td><strong><br />
Year</strong></td>
<td colspan="2"><strong><br />
Procedure</strong></td>
<td><strong>Species/<br />
Strain</strong></td>
<td colspan="2"><strong>No. per<br />
Group</strong></td>
<td colspan="3"><strong>No. of<br />
Groups</strong></td>
<td colspan="2"><strong>No. of<br />
Reps</strong></td>
<td><h6 id="section-2"></h6>
<h6 id="total">Total</h6></td>
</tr>
<tr class="even">
<td>1</td>
<td colspan="2">Whole body distribution studies</td>
<td>Rat<br />
Sprague Dawley</td>
<td colspan="2">4</td>
<td colspan="3">5</td>
<td colspan="2">4</td>
<td>80</td>
</tr>
<tr class="odd">
<td>1</td>
<td colspan="2">Competitive inhibition studies</td>
<td>Rat<br />
Sprague Dawley</td>
<td colspan="2">4</td>
<td colspan="3">4</td>
<td colspan="2">2</td>
<td>32</td>
</tr>
<tr class="even">
<td>1</td>
<td colspan="2">Metabolic Studies</td>
<td>Rat<br />
Sprague Dawley</td>
<td colspan="2">2</td>
<td colspan="3">3</td>
<td colspan="2">2</td>
<td>12</td>
</tr>
<tr class="odd">
<td>2</td>
<td colspan="2">Same as above</td>
<td>Same as above</td>
<td colspan="2"></td>
<td colspan="3"></td>
<td colspan="2"></td>
<td>124</td>
</tr>
<tr class="even">
<td>3</td>
<td colspan="2">Same as above</td>
<td>Same as above</td>
<td colspan="2"></td>
<td colspan="3"></td>
<td colspan="2"></td>
<td>124</td>
</tr>
<tr class="odd">
<td colspan="9"></td>
<td colspan="2"><strong><br />
Sum</strong></td>
<td><strong><br />
372</strong></td>
</tr>
</tbody>
</table>

# 7. Euthanasia Techniques

Review AWRC Guidelines for recommended Euthanasia methods. The
Guidelines are available by contacting Chris Byrne via email at
<CEByrne@lbl.gov> or by phoning 5507.

Indicate euthanasia method used for each species. If drugs are used,
provide name, dosage, and route of administration.

# 8. Pain and/or Distress

AWRC must categorize each project as A, B, or C on the basis of the use
of analgesic, anesthetic, and tranquilizing drugs. In some protocols
more than one category may be involved; if so, indicate number of
animals (or percentage) in each category and explain.

#### A - No pain or distress; no drugs used.

Chose category A when procedures involve momentary pain or distress to
animal subjects (excluding that caused by venipuncture). Justification
must be given when this choice is made.

#### B. Pain and/or distress; drugs used.

Indicate species, dosage, and route of administration for each drug
used. This is a summary listing; provide details in Section 6,
Procedures.

#### C. Pain and/or distress, drugs not used because they would interfere with the protocol goal

If category C is selected, provide a justification for pain and/or
distress

Describe the methods and sources (i.e., AWIC, CATT) used to determine
why there is no alternative to the use of vertebrate animals in the
procedure.

Indicate why alleviating drugs or euthanasia agents will interfere with
experimental results and when animals subjected to such conditions will
be terminated.

# 9. Sharing Animal Resources

*The Committee must evaluate appropriate animal use for each protocol.*

Provide information on the following:

-   surplus tissues/animals from this protocol used in other studies

-   surplus tissues/animals from other protocol used in this study

-   animals used in the protocol that will be used in other studies

# 10. Claim of Confidentiality

Contents of the protocol are available for public disclosure. A
confidential claim can be made when the protocol discloses unpublished
data or contains research procedures for which copyright or patent is
being sought.

# Signature Page

LBNL regulation (NIH Assurance of Compliance) requires signatures of the
Principal Investigator and Division Director. Unsigned documents will
not be reviewed by the AWRC.

##### General Information

Principal Investigator signs and dates protocol

**Required:** List PIs office, laboratory, and emergency phone numbers
where they can be contacted in case of animal emergency.

**Required:** List alternate responsible individual's office,
laboratory, and emergency phone numbers where they can be contacted in
case of animal emergency.

PI's Division Director signs and dates protocol. Unsigned protocols will
not be reviewed.

**AWRC Approval**

Signed by AWRC Chairman, after the protocol is approved. The PI is sent
the AWRC approval letter with a copy of the approved protocol. A copy of
the certification is kept on file in the AWRC office.