labscene: aaalac prep -...
TRANSCRIPT
LabSCENE: AAALAC PREPJanuary 9, 2018
Agenda
Introduction to AAALACKathryn Holthaus
Working in the Animal FacilityAmber Hoggatt, Attending Veterinarian
Working in Use AreasChristina Nascimento, IACUC ManagerPatrick Doyle, IACUC Program CoordinatorJennifer Megyesy, IACUC Protocol Administrator
Working with BL2 AgentsJustin Silva, Laboratory Safety Manager, Environmental Affairs
AAALAC Accreditation
Voluntary Consistent, high standards Commitment to excellence Funding agency recommended
Site Visit (vs. Inspection)
Be excited to share your research! Know protocols, SOPs, monitoring, endpoints, etc.Involve the PI/Lab ManagerYou know your research best, butIt’s okay to say:
“I’m not sure, we should check with …” “That’s the kind of question I usually double check with …” or “I always look that up … before I start”
Institutional Support: Kathryn Holthaus, 617-732-5761
Whistle Blower Policy
Reporting Concerns of Animal Welfare:
Where? Signage posted in animal facilities and lab use areasInfo posted on website
Who?Attending VeterinarianIACUC ManagerIACUC ChairmanBWH Research ComplianceInstitutional Official for the IACUCPartners Compliance Hotline (Anonymous)
Occupational Health Enrollment
All animal users MUST be enrolled in the AESPThose in shared lab spaces should self-identify and enrollUpdate with OHS every 3 years or when health status changes
Riolla RiobeBrigham & Women's Hospital Occupational Health Services
617-732-8502 [email protected]
**PRIOR to the AAALAC site visit**
Amber Hoggatt, Attending Veterinarian
Working in the Animal Facility
Must be labeled with Lab and user contact
Stored in closed, sanitizable container
CLEAN
Compounded drugs/reagents clearly labeled and in date
Expired materials discarded or labeled for acute use only
Compounded drugs and storage
What is the BWH guideline for the shelf-life of a compounded drug?
What is required on compounded drug label?
Experimental compound or generic drug(s)Concentration of active drugStock expiration dateCompounding date or compounding expiration
date
Waste Anesthetic Gas Management
Class II A2 or B2BTM imaging crescentHIM B21Thorn 1633
Waste Anesthetic Gas Management
Waste Anesthetic Gas ManagementBWH Department of Environmental Affairs
Department of Environmental Affairs General Telephone Number: 617-264-3010Department of Environmental Affairs Page ID: 1-5000 (Best Contact Method)
Research Records and retention
Lab notes must be maintained for 7 years after the work is published.
Experimental disease monitoring Rodent surgery reports that demonstrate analgesia
administration Breeding and genotyping records Cell line testing records
Specifically state the experimental model (no abbreviations)
Examples:• Acute DSS colitis
• DSS/AOM induced colonic carcinoma
• Implanted PD melanoma, right flank
• Tumor glioma (Hs 683), IC
• Pulmonary hypertension
• K/BxN serum induced arthritis
Circle ‘Standard CCM Guidelines’ for humane endpoints
OR
Use ‘Other’ field to describe specific exceptions to standard CCM humane endpoint guidelines
Examples:
• Moribund (hypothermia, inactivity, pallor)
• Survival study (death as endpoint)• Tumor ulceration up to 5mm
Describe required care or intervention
Examples:• Humane euthanasia
• Diet gel
• Moist food on floor
• Soft bedding
• Extra enrichment
Indicate when support is provided in notes
Enter minimum monitoring frequency as described in your protocol. Monitoring frequency may change as disease progresses and it is important to update form.
Document each monitoring event and indicate observation or if supportive care or euthanasia is performed
Example Observations:
• No clinical signs• BSC 2-3• Flank tumor <5mm
Example Interventions:
• Humane euthanasia 1/5• Diet gel• Moist food• Soft bedding
Experimental Cages
Rodents must be socially housed Unless protocol is approved for single housing Helpful to write “Social housing exempt” on
cage cards
Rodents must have enrichment Mice = 2 inch nestlet or enviro-dri Rats = rat hide (red lookout, cylinder, or
rectangle)
Have lab level risk assessment and SDS is available
Review BWH Research SOP 502: Administration of hazardous agents to rodentsReview at animal.bwh.harvard.edu
Properly label cages with Hazard Chemical Sticker/Card
Toxic agent use in rodent cages
Biohazard Use in Rodents
Breeding colony
CCM recommended breeding scheme is pair or trio (Allentown cages only)
Proactively manage you breeding colony these following weeks Mark birth dates Separate weanlings at 21-28 days Remove pregnant females in trio or harem cages
Update breeding records Know how many breeders for a specific line Be able to estimate the number of offspring generated on a monthly basis
How to Contact the Vet Staff?
Christina Nascimento, IACUC Manager
Patrick Doyle, IACUC Program Coordinator
Jennifer Megyesy, IACUC Protocol Administrator
Preparing Animal Use Areas
Use Area Preparation
Familiarize all lab animal users with the following documents: Animal protocols they are working on Lab specific training forms Lab-level Disaster Plan SOPs used by the lab for reference All users are listed on protocols they are working on
Use Area Preparation
Clean and tidy lab locations where animals are used Area of lab used for animals should be prepared for animal use at time of site visit
Discard or clean bloody materials
Inventory animal supplies/check for expired materials
Organize documents Archived EIRs and RSRs Breeding colony records Lab member training logs Lab level disaster plan
Training Documentation
BWH Personnel Training Form
Indicate species-specific procedures lab members will perform on IACUC-approved protocols. Include date of training and/or experience. Trainers must initial to confirm each training session, excluding CCM/IACUC training, or courses that provide formal certificates. Personnel cannot perform procedures they are not trained in. Form must be updated as new trainings occur and/or new procedures are added to the protocol. Personnel Training Forms must be maintained in the lab for each person listed on the protocol(s), and availablefor IACUC Inspections, audits, veterinary staff, outside site visitors, etc. Add additional rows as needed.
Name Species
Proc
edur
e
(spe
cify
type
)
Surg
ery
(s
peci
fy ty
pe)
Inje
ctio
ns
(spe
cify
rout
e)
Anes
thes
ia
Euth
anas
ia
Tail
Biop
sy
Bloo
d Co
llect
ion
(spe
cify
rout
e)
Lab-
Leve
l Di
sast
er P
lan
Oth
er (s
peci
fy)
John Doe Mouse
Date Trained and/or experience
9/18/17 Dr. X mouse Colony management
9/20/17 CO2 euthanasia using lab setup
Trained by Dr. X Dr. X
Date Trained and/or experience
Trained by
Always 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 (including storage of research
materials) 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 in the event of an
emergency, please contact your facility supervisor to identify them.
Lab Level Disaster Plan
Immobilize gas cylinders Close door to lab before euthanizing animals Ensure other animals cannot see/smell animals being euthanized Clean area/chamber after each use Properly store carcasses at 7 Celsius and return to facility carcass storage
regularly Keep records for animals after euthanized (7 years after completion of
project) For non-survival surgery, if expired supplies used, they are separated from
survival supplies and labeled “non-survival use only”
Euthanasia Stations
Flowmeter MUST be used with CO2 Gradual displacement rate of 25-30% Calculation determines flow rate per size of chamber (can fill with water
and measure liters) Chamber Height x width x length= liters x 0.25 = flow rate/minute
Post SOP at euthanasia station Clean chamber after each use
CO2 Euthanasia Stations
Guillotines and Decapitation Scissors
Scissors used for decapitation must be kept sharp, clean, and cut easilyScissors that don’t meet these criteria must be removed from service immediately and
replaced or sharpened (replacement is recommended) If you would prefer to sharpen your scissors, we recommend Peter Yao
([email protected]) Cost: $18/pairKeep the receipt! Scissors used for alert decapitation must be sharpened, replaced, or
demonstrated to be appropriately sharp every 12 months All guillotines must be kept sharp, clean, and operate smoothlyGuillotines must be sharpened professionally or demonstrated to be appropriately
sharp at least every 12 monthsWear cut-proof gloves when cleaning a guillotine!
Make sure to brush up on your justification for using alert decapitation if applicable
Survival Surgery Use Areas
Be ready to describe the surgery, from animals/equipment preparation, to recovery Equipment in good working order, no rust
Aseptic technique is followed
Sterilization of instruments/surgical supplies Surgical packs have sterile indicator strip inside, sterile indicator tape outside
Date of sterilization noted on pack (first in, first out- integrity of pack maintained)
Gowning of surgeons (only sterile surgical gloves in use unless USDA covered species)
Maintain sterility during procedure
All surgical supplies in date Supplies used for non-survival procedures should be stored separately and labeled “expired, not for live animal use”
Preparation of site Hair removal
Skin preparation
Draping, fenestration size
Survival Surgery Use Area (cont.)
All drugs are USP grade (unless IACUC exemption approved) Waste gas is scavenged, charcoal canister weighed before each use
Discard canister once weight limit reached
Vaporizers calibrated once yearly Approved heart source is in use (re-circulating water blanket, small animal rated
warming pads) Instruments are sterilized between animals
Bead sterilizer Or, new instruments for each animal
Controlled Substances
Daily usage logs must be up to date Ensure stock in cabinet matches log book
Dilutions MUST be recorded on individual daily usage logs
Complete Inventory form Check correct storage in secondary vialsOnly sterile injectable vials are used
Labeled with contents, concentration, preparation date, and expiration
**Prepare fresh dilutions instead whenever possible
Expired drugs are labeled “expired” and stored in the safe in the expired substances bin Have licenses, packing slips, and DEA form 41 available Only USP grade drugs in use (unless IACUC approved)
What is required before you use surgical instruments on a
second/additional animal for survival surgery?
Quiz Time!
Justin Silva, Laboratory Safety Manager, Environmental Affairs
Working with BL2 Agents
2
BSL-2 is suitable for work involving agents that pose moderate hazards to personnel and the environment.
BSL-2 vs BSL-1 Laboratory personnel have specific training in handling pathogenic agents and are
supervised by scientists competent in handling infectious agents and associated procedures.
Lab access is restricted when work is being conducted.
All procedures in which infectious aerosols or splashes may be created are conducted in BSCs or other physical containment equipment.
BSL-2 Requirements & Best Practices
2
BSL-2 Standard and Special Practices, Safety Equipment, and Facility Requirements
Standard Microbiological Practices: Laboratory supervisor must enforce BWH policies that control access to the lab.
Persons must wash their hands after working with potentially hazardous materials and before leaving the lab.
Eating, drinking, smoking, handling of contact lenses, applying cosmetics, and storing food for human consumption must not be permitted in lab areas.
Policies for the safe handling of sharps must be developed and implemented.
Minimize the creation of splashes and/or aerosols.
2
Standard Microbiological Practices Continued…
Decontaminate work surfaces after completion or work and after any spill or splash of potentially infectious material with appropriate disinfectant.
Anything liquid is decontaminated via bleach before pouring down drain, and anything solid goes into red bag waste and is treated off-site.
The lab supervisor must ensure that lab personnel receive appropriate training regarding their duties, precautions to prevent exposures, and exposure evaluation procedures. Annual retraining, or when policies undergo changes, is required.
BSL-2 Standard and Special Practices, Safety Equipment, and Facility Requirements
Special Practices: All persons entering the laboratory must be advised of the potential hazards and meet
specific entry/exit requirements.
Laboratory personnel must be provided medical surveillance, as appropriate, and offered available immunizations for agents handled or potentially present in the laboratory.
A laboratory-specific biosafety manual must be prepared and adopted as policy. The biosafety manual must be available and accessible.
The laboratory supervisor must ensure that laboratory personnel demonstrate proficiency in standard and special microbiological practices before working with BSL-2 agents.
BSL-2 Standard and Special Practices, Safety Equipment, and Facility Requirements
Special Practices Continued… Potentially infectious materials must be placed in a durable, leak proof container during
collection, handling, processing, storage, or transport within a facility.
Laboratory equipment should be routinely decontaminated, as well as, after spills, splashes, or other potential contamination.
Incidents that may result in exposure to infectious materials must be immediately evaluated and treated according to procedures described in the laboratory biosafety manual. All such incidents must be reported to the laboratory supervisor and Env Affairs. Animals and plants not associated with the work being performed must not be permitted in the laboratory.
All procedures involving the manipulation of infectious materials that may generate an aerosol should be conducted within a BSC or other physical containment devices.
BSL-2 Standard and Special Practices, Safety Equipment, and Facility Requirements
Safety Equipment (Primary Barriers and Personal Protective Equipment):
Properly maintained BSCs, PPE, or other physical containment devices to be used whenever:
Procedures with a potential to create infectious aerosols or splashes are conducted via…
Pipetting, centrifuging, grinding, blending, shaking, mixing, sonicating, opening containers of infectious materials, inoculating animals, or harvesting infected tissues
Or, high concentrations or large volumes of infectious agents are used
Lab coats, gowns, smocks, or uniforms designated for laboratory use must be worn while working with hazardous materials
Protective clothing must be removed before leaving for non-lab areas i.e. cafeteria, library, administrative offices, or disposed of/deposited for laundering properly.
BSL-2 Standard and Special Practices, Safety Equipment, and Facility Requirements
Safety Equipment Continued… Eye and face protection i.e. goggles, mask, or face shield, shall be used for anticipated splashes or sprays of
infectious or other hazardous materials when the work is being handled outside of a BSC or containment device.
Dispose of properly or clean before reuse. Safety glasses are advised at all times in the labs.
Gloves must be worn when handling hazardous materials
Change gloves when contaminated or integrity compromised.
Remove gloves and wash hands after completion of work with hazardous materials, and before leaving the lab.
No washing or reusing disposable gloves. Dispose of in appropriate containers.
Hand washing protocols must be rigorously followed.
Eye, face and respiratory protection should be worn in rooms with infected animals, as determined by a risk assessment.
BSL-2 Standard and Special Practices, Safety Equipment, and Facility Requirements
BSL-2 Standard and Special Practices, Safety Equipment, and Facility Requirements
Laboratory Facilities (secondary barriers): Self closing lab doors with locks
Sinks for hand washing, located near the exit
Designed for easy cleaning and decontamination… no rugs or carpets
Benchtops impervious to water, and resistant to heat, organic solvents, acids, alkalis, and other chemicals
Chairs in the lab must be covered of non-porous material, which can be easily cleaned and decontaminated with appropriate disinfectant
Lab windows that can open are not recommended, however must have screens if so
BSCs strategically placed so as to not sustain airflow disruptions
Vacuum lines protected with liquid disinfectant traps
BSL-2 Standard and Special Practices, Safety Equipment, and Facility Requirements
Laboratory Facilities (secondary barriers) Continued… Eyewash station available and inspected per BWH policy (at least once a month)
Always consider inward flow mechanical ventilation systems without recirculation to space outside the lab
Certain classes of BSCs with HEPA filtration are recommended if recirculation back into the lab is necessary… discuss with Env Affairs
Available methods for waste decontamination are available i.e. autoclave or other committee approved options
BSL-2 Standard and Special Practices, Safety Equipment, and Facility Requirements
A sign incorporating the universal biohazard symbol must be posted at the entrance to the laboratory when infectious agents are present. Posted information must include: the laboratory’s biosafety level, the supervisor’s name (or other responsible personnel), telephone number, and required procedures for entering and exiting the laboratory. Agent information should be posted in accordance with the institutional policy.
BSL-2 Laboratory Signage
Quiz Time!
Who is responsible for ensuring training is completed for lab
personnel, and when is it required?