October 3, 2013 Christina Nascimento, MS, CPIA

1. IACUC Educational Series kickoff session:IACUC Updates: Discuss regulatory updates, new policies, and protocol writing with the IACUC staff October 3, 2013 Christina Nascimento, MS, CPIA

2. Agenda • New IACUC Policies • Disaster Planning • Occupational Health • Regulatory Updates • The AVMA Guidelines for the Euthanasia of Animals: 2013 Edition • Protocol Writing & Review • Animal Usage, Justification for Use of Animals, & Breeding Colony numbers • Administrative Updates • Other Questions? • Topics for future sessions?

3. Disaster Planning

4. 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.

5. Disaster Planning Emergency 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.

6. Disaster Planning Document training on Disaster Plan

7. Occupational Health • Requirements • All personnel who work in laboratory animal facilities or have frequent contact with animal must enroll in the occupational health and safety program • Procedure • Initial questionnaire • Follow-up • Clearance • Annual updates

8. Occupational Health • Changes • Clearance required for inclusion on animal protocols • No changes for BWH personnel • Will be included in annual renewals • HMS Only • Surveys distributed and returned to OEHN • Contact person at HMS to answer questions

9. Occupational Health Initial Health Questionnaire

10. Occupational Health Annual Update Questionnaire

11. The AVMA Guidelines for the Euthanasia of Animals: 2013 Edition • Relevant changes: • CO2 displacement rate for rodents • Cervical dislocation for rodents • Chilling of zebrafish • OLAW Webinar: http://www.youtube.com/watch?v=c8ivjAysVBI • CO2: acceptable “with conditions”: • CO2: Gradual displacement rate of 10-30% • Flowmeters attach to CO2 source • Suggest post SOP at euthanasia station • Dwyer instruments : VFA-24 (2" scale) or VFB-66 (4" scale) • Airgas: Western® CGA320 12 LPM Flowmeter Regulator WESM1-320-12FM

12. The AVMA Guidelines for the Euthanasia of Animals: 2013 Edition • Cervical Dislocation without anesthesia acceptable “with conditions” • 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” • Amendments approved with the method will be sent to the HCCM Manager of Training. • Lab must set up a training session with the HCCM Manger of Training for cervical dislocation • Training in the procedure must be documented in the lab’s animal procedure training records. • Only after all the steps above are completed, can the lab proceed with performing the procedure

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

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

15. Protocol WritingSection A (Animal Usage, justifications, and breeding) • A.2.c: Justify the number of animals to be used: • 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).

16. Protocol WritingSection A (Animal Usage, justifications, and breeding) • Describe the logic behind the proposed number of animals per group: • 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).

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

18. Protocol WritingSection A (Animal Usage, justifications, and breeding) • Estimate number of breeding adults, offspring produced, and number of offspring euthanized without research procedures performed: • 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.

19. Tracking Breeding Animals in the HCCM Facility

20. Administrative Updates • Section S and Annual Renewal: • No signatures required by lab members other than PI • Research Officers assigned to specific PIs • Rolling Protocol submissions • Questions? • Suggestions for future topics?