IACUC Educational Series kickoff session:

IACUC Updates: Discuss regulatory updates, new policies, and protocol writing with the

IACUC staffOctober 3, 2013

Christina Nascimento, MS, CPIA

Agenda New IACUC Policies

o Disaster Planningo Occupational Health

Regulatory Updateso The AVMA Guidelines for the Euthanasia of Animals: 2013

Edition Protocol Writing & Review

o Animal Usage, Justification for Use of Animals, & Breeding Colony numbers

Administrative Updates Other Questions? Topics for future sessions?

Disaster Planning

Disaster PlanningLaboratory-Level Emergency Response Resource Guide : Approved May 2013

Applies to: All users of research animals in laboratories.

Pre-Emergency Planning Steps: Know your building, department and local emergency plans. Register for Harvard’s Message Me notification Take weather forecasts into account when planning experiments. Maintain sufficient quantities of euthanasia agent(s) to euthanize all

animals in the lab. Maintain paper copies of emergency contact lists and call trees in both

home and work areas.. Verify that critical animal-related equipment is plugged into functioning

emergency outlets, if needed. Cryopreserve mouse strains (embryos or sperm) that are irreplaceable, If you have specific animals which cannot be replaced, contact facility

supervisor to identify them.

Disaster PlanningEmergency Action Steps: Obey all alarms and emergency notifications. Human safety is the top priority. Evacuate first, then return animals to animal housing facility if possible If animals cannot be returned, secure the animals in cages and leave

immediately. Euthanize animals that are in imminent danger As soon as building re-entry is authorized, assess health and safety status of

the animals and make efforts to optimize their care Return animals to their designated housing facility, if possible; if not possible,

maintain contact with the animal facility manager/supervisor and: Check animals more frequently in affected areas and report status changes to

animal facility supervisors or veterinary staff. Monitor temperature and humidity.

Animals that are found to be affected by extreme heat/cold/other environmental conditions as a result of the emergency should be removed from the environment and treated or euthanized

Call the Facilities Emergency Number if animal lives may be at risk from a facilities-related emergency.

Disaster Planning Document training on

Disaster Plan

Occupational Health Requirements

o All personnel who work in laboratory animal facilities or have frequent contact with animal must enroll in the occupational health and safety program

Procedureo Initial questionnaireo Follow-upo Clearanceo Annual updates

Occupational Health Changes

o Clearance required for inclusion on animal protocolso No changes for BWH personnel o Will be included in annual renewals

HMS Onlyo Surveys distributed and returned to OEHNo Contact person at HMS to answer questions

Occupational HealthInitial Health Questionnaire

Occupational HealthAnnual Update Questionnaire

The AVMA Guidelines for the Euthanasia of Animals: 2013 Edition

Relevant changes:o CO2 displacement rate for rodentso Cervical dislocation for rodentso Chilling of zebrafisho OLAW Webinar: http://

www.youtube.com/watch?v=c8ivjAysVBI

CO2: acceptable “with conditions”:o CO2: Gradual displacement rate of 10-30%o Flowmeters attach to CO2 sourceo Suggest post SOP at euthanasia stationo Dwyer instruments : VFA-24 (2" scale) or VFB-66 (4" scale)o Airgas: Western® CGA320 12 LPM Flowmeter Regulator WESM1-320-12FM

The AVMA Guidelines for the Euthanasia of Animals: 2013 Edition

Cervical Dislocation without anesthesia acceptable “with conditions” o Animal Protocol must be amended and approved by the IACUC

before performing cervical dislocation• Scientific justification no longer needed• Performing the euthanasia method without IACUC approval would

be considered “off protocol”o Amendments approved with the method will be sent to the

HCCM Manager of Training. o Lab must set up a training session with the HCCM Manger of

Training for cervical dislocationo Training in the procedure must be documented in the lab’s

animal procedure training records. o Only after all the steps above are completed, can the lab

proceed with performing the procedure

Protocol WritingSection A (Animal Usage, justifications, and breeding)

Species # Used per year

CATEGORY B -Teaching, research, or tests that involve breeding or housing of animals only Breeding colony Housing ONLY (no other procedures) Total number of animals to be used in Category B only

CATEGORY C -Teaching and/or research experiments that do not involve more than momentary pain and/or distress or require the use of pain relieving drugs. Alert animals (behavioral observation or brief restraint) Anesthetize and release for non-invasive procedures (e.g. imaging, bandage change) Change in environmental parameters (diet, light cycle, room temperature, etc.) Chemical carcinogens (e.g. BrdU) Euthanasia with or without tissue/organ harvest Forced exercise Gavage Irradiation Microbiological agents Non-surgical collection of body fluids (blood, urine, etc.) Radioisotopes Simple injections (IP, IV, etc.) Tail biopsy for genotyping (mice under 21 days) Use as a parasitic host Total number of animals to be used in Category C

Note: For all procedures in Categories D & E (below) a search must be completed in Section P

CATEGORY D – Teaching and/or research experiments or tests that will involve pain and/or distress and for which appropriate anesthetic, analgesic or tranquilizing drugs will be used. Anesthetize and release for non-surgical painful procedures (e.g. retro-orbital bleeds) Antibody production: ascites Antibody production: polyclonal (non-ascites, no footpad) Burns Controlled drugs or substances (as test material, e.g. cocaine) Electric shock Food or water deprivation other than pre-surgical fasting Footpad injections (antibody production or microorganism) Introduction of illness (including the administration of toxins, microbiological agents,

clinical transgenic phenotypes)

Introduction of illness (including the administration of toxins, microbiological agents, clinical transgenic phenotypes)

Lavage Multiple MAJOR survival surgery Multiple MINOR survival surgery Non-survival surgery Physical trauma Survival surgery: MAJOR Survival surgery: MINOR Tail biopsy for genotyping (mice over 21 days) Toxic substances (e.g. streptozotocin, MPTP, cisplatin, doxycycline) Transgenic mouse production (embryo manipulations to produce novel strains, etc. Tumor induction or implantation Unusual or prolonged restraint (e.g. pie chamber, chair) Use of paralytic agents Total number of animals to be used in Category D

CATEGORY E – Teaching and/or research experiments, surgery or tests that involve PAIN and/or DISTRESS, and for which appropriate anesthetic, analgesics or tranquilizing drugs are NOT used because they would adversely affect the results or interpretation of data. Death-as-an-endpoint* studies Lethal dose studies* Noxious stimuli from which there is no escape Pain study Withholding of post-operative analgesia for any surgical procedure Total number of animals to be used in Category E

*You must follow the Guidelines for ‘Death as an Endpoint’ or ‘Lethal Dose’ studies at: https://www.hms.harvard.edu/ORSP/Secure/animal_guidelines.html

EXCEPTIONS to STANDARDS – Select any policy deviations that will be required in this protocol. Scientific justification must be included in the indicated section of the protocol and/or separate exemption form. Single housing of animals Section D.4 Withholding of enrichment from singly housed animals Section D.4.c Euthanasia methods that are not consistent with the AVMA Guidelines Section E & Exemption Ascites collection Section G & Exemption Non-pharmaceutical grade drugs and other compound suspensions Section I.7 Withholding postoperative analgesia** Section J.2.k & Exemption Food/water withholding or scheduling (not ad-lib) Section N Smaller caging than the recommended space allotment (e.g.: CLAMS) Exemption form Deviations from housing density standards Exemption form Toe-clipping for identification purposes Exemption form Deviations from the 12/12 hour light/dark cycle Section N Any other exceptions to the standards of the PHS Policy, the Guide, or the

AWA. - Please specify: Exemption form

Protocol WritingSection A (Animal Usage, justifications, and

breeding)2. JUSTIFICATION FOR THE USE OF ANIMALS

Federal Assurances: You are required by federal law to provide the following justifications.

a. Why are animals needed for this study? (i.e.: Could the same information be obtained by experiments using tissue culture or computer models? If you are generating antibodies, could you use in vitro methods?)

b. Justify choice of each species:

c. Justify the number of animals to be used: (You must convince the Committee that the number of animals is appropriate to the work being proposed. Provide groups and statistical analysis to be used if relevant.)

d. Discuss your strategy to reduce the number of animals and to minimize pain and distress to your animals: (examples include: earlier euthanasia, decreasing the number of time points, changing the methods to those that are less invasive):

Protocol WritingSection A (Animal Usage, justifications, and

breeding) A.2.c: Justify the number of animals to be used:

o Example: We estimate we will need a maximum number of 3175 animals over the 3-year approval period. This number is based on an estimated number of 60 novel agents that will be screened for anti-inflammatory properties. This total number is based on 60 studies/3 years with 50 animals/study; 6 groups per study (2 control groups with 5 animals/group and 4 test groups with10 animals/group). Control group 1 will receive the inflammatory agent followed by treatment with vehicle only. Control Group 2 will receive inflammatory agent followed a well-established anti-inflammatory agent. Test groups will receive the inflammatory agent followed by treatment with a potential therapeutic (novel) agent at 4 different concentrations, respectively. Since we anticipate that as many as 5% of the animals will not exhibit a sufficient minimum initial inflammatory response to be entered in the study, we are requesting an additional 150 animals. In addition, we are asking for an additional 25 animals for the purpose of training research staff in the proposed manipulations (e.g. foot pad injections, iv administration of test compounds, measurements of inflammatory response).

Protocol WritingSection A (Animal Usage, justifications, and

breeding) Describe the logic behind the proposed number of animals per

group:o Example: We will be testing one novel agent at 4 different

concentrations per study (i.e. 4 test groups per study). The selected dose concentrations will be based on what we estimate to be the smallest concentration necessary to confirm anti-inflammatory properties, up to the highest concentration necessary to maximize anti-inflammatory properties without causing toxicity, and two additional intermediate doses to see if a maximum anti-inflammatory threshold can be achieved at lower doses. We will also treat one control group with vehicle only, and a second control with a well-established anti-inflammatory compound at its most efficacious dose. The number of animals per group is based in a student’s t-test or a one-way ANOVA. In addition, the necessary number of animals per test and control groups in this screening approach is well established in the literature: (reference relevant publications).

Protocol WritingSection A (Animal Usage, justifications, and

breeding)

e. Breeding Colony users only: The estimated numbers of animals in breeding colonies must be provided and justified. These numbers must be also be tracked on a log sheet in the animal housing room.

i. Estimated number of breeding adults:

ii. Estimated number of offspring produced:

iii. Estimated total number of offspring euthanized without research use:

iv. Criteria for offspring euthanized without research use:

Protocol WritingSection A (Animal Usage, justifications, and

breeding) Estimate number of breeding adults, offspring produced,

and number of offspring euthanized without research procedures performed:o Example: Assuming each breeder pair will produce an average of 1

litter of 5 pups per month and will breed for 3 months, 15 pairs should produce 15x5x3=225 mice in 3 months, i.e. 75 mice per month or 900 mice per year X 3 years of the approval period is 2700 mice that can be used in experiments or to refresh the breeders. After this initial phase, 360 of the offspring will be used to refresh breeders, leaving an estimated 2340 mice (1170 males, 1170 females) for experiments. However, since we are only interested in female mice for our study; of the 2340 mice only 1170 will be used in experiments and the other 1170 males will be euthanized without use. We also anticipate that approximately 50 offspring will be produced at a time when we cannot use them and therefore will be euthanized without use. We will try to keep the number of animals produced in excess to a minimum by separating breeding pairs as needed to reduce production.

Tracking Breeding Animals in the HCCM Facility

Administrative Updates Section S and Annual Renewal:

o No signatures required by lab members other than PI Research Officers assigned to specific PIs Rolling Protocol submissions

Questions? Suggestions for future topics?