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Biosafety Level 1 & 2 Training Colorado State University Biosafety Office Environmental Health Services 141 General Services Building 970-491-6729
Purpose To conform to University, Federal, and Funding Regulations Colorado State University Biosafety Policy http://ricro.colostate.edu/IBC/policy.htm Biosafety in Microbiological and Biomedical Laboratories (BMBL 5) http://www.cdc.gov/biosafety/publications/bmbl5/ NIH recombinant DNA and Synthetic Nucleic Acid Guidelines http://oba.od.nih.gov/rdna/nih_guidelines_oba.html To inform the CSU Community of the expectations of BSL-1 and
BSL-2 safety practices To increase awareness of biosafety issues and, thereby, increase safety To prepare labs for site visits that will be conducted by Biosafety Officers
CSU Biosafety PolicyResponsibilities Principal Investigators
It is the responsibility of the Principal Investigator to ensure that personnel have been appropriately trained in biosafety procedures and to maintain documentation of site – specific employee training records.
CSU Biosafety PolicyResponsibilities Principal Investigators
Regulatory: Submit Agent and Project protocols for review by Institutional Biosafety Committee (IBC) prior to beginning experiment http://web.research.colostate.edu/ricro/ibc/ibc.aspx Ensure no project is modified without IBC approval Occupational Health: Ensure personnel are enrolled in the Occupational Health Program https://wsnet.colostate.edu/cwis86/WOHSP/RiskAssessment/ad d/AddRisAssessment.aspx Incidents: Report accidents, exposures or violations to Biosafety http://www.ehs.colostate.edu/WOHSP/PDF/IncidentReport Form.pdf Report all incidents to Risk Management http://www.ehs.colostate.edu/WWorkComp/Home.aspx
CSU Biosafety PolicyResponsibilities Personnel
Follow approved IBC protocols Follow all safety policies, guidelines, and procedures Be familiar with workplace hazards Consult with supervisors to receive appropriate training and instruction Help improve the safety of your work environment:
If you are unsure, ASK! Report all accidents, incidents, near misses, and areas of needed improvement http://www.ehs.colostate.edu/WBiosafety/PDF/IncidentReportF orm.pdf
Training and Requirements All Employees
Need to enroll in the Occupational Health Program online
Fill out risk assessment at: http://www.ehs.colostate.edu/WOHSP/Home.aspx File assessment annually and when your job/ duties/ health changes Based on certain job risks, you may be asked to enroll in medical surveillance or to take additional training
Training and Requirements Provided by Environmental Health
Services:
Biosafety Level (BSL) 1 and 2 Online Training If you will be using a Biosafety Cabinet (BSC), contact Heather Blair (
[email protected]) or Claudia Gentry-Weeks (
[email protected]) to schedule hands-on training
Training and Requirements Provided by your Department/ Principal
Investigator (PI)/ Supervisor
Building safety and emergency action plans Site specific training (e.g. Lab Standard and Special Practices Training ) Information about site specific risks and protocols to reduce risk
Before you Start Work Get information about the biological agents
you work with and the associated risks. Be aware of symptoms that may indicate work related illness.
Helpful links: Public Health Agency of Canada: http://www.phacaspc.gc.ca/lab-bio/res/psds-ftss/index-eng.php Iowa State University Center for Food Security and Public Health: http://www.cfsph.iastate.edu/DiseaseInfo/?transmis sion[]=006&lang=en CSU Factsheets: http://www.ehs.colostate.edu/WOHSP/Bsl3Packets .aspx
Important Resource: NIH/CDC Biosafety in Microbiological and Biomedical Laboratories (BMBL)
The BMBL serves as the guide for training and
practices for working with microbiological agents. The BMBL provides an overview of
General risk assessment Description of Biosafety Levels 1-4 Facility, equipment and safety practices Agent specific information
The most recent edition is available at http://www.cdc.gov/OD/OHS/biosfty/bmbl5/BMBL_5th _Edition.pdf
BMBL- Risk Groups versus Biosafety Levels Risk group (RG):
Is a comparative descriptor of the inherent pathogenic nature of a given microorganism. Is based on how a particular organism is transmitted, how it is associated with disease in a “healthy” adult and whether prevention or intervention exists for that organism. Does not change based on how or where the agent is used.
Biosafety Level (BSL):
Is a variable comparative descriptor of the facility, equipment and practices that serve to “contain” a microorganism, and to ensure the safe use of that organism while it is being handled. Is based on risk assessment and technical judgment and may vary with the use of the agent.
BMBL- Human Risk Group Classifications http://www.absa.org/riskgroups/ Risk Group (RG)
Description
Risk Group 1 (RG1)
A microorganism that is unlikely to cause human disease or animal disease
Risk Group 2 (RG2)
A pathogen that can cause human or animal disease but is unlikely to be a serious hazard to laboratory workers, the community, livestock or the environment. Laboratory exposures may cause serious infection, but effective treatment and preventative measures are available and the risk of spread of infection is limited
Risk Group 3 (RG3)
A pathogen that usually causes serious human or animal disease but does not ordinarily spread from one individual to another. Effective treatment and preventative measures are available.
Risk Group 4 (RG4)
A pathogen that usually causes serious human or animal disease and that can be readily transmitted from one individual to another, directly, indirectly. Effective treatment and preventive measures are not usually available
Biosafety Levels (BSL) Biosafety levels are designed and constructed to provide a barrier of protection for individuals and animals outside the lab environment. The recommended biosafety level depends on the risk of the microorganism being worked with.
Biosafety Level
Agent Characteristics
Biosafety Level 1
Well characterized agents that are not known to cause disease in immunocompetent adult humans, and present minimal potential hazard to personnel and the environment (e.g. E. coliK12, Bacillus subtilis, canine hepatitis virus, murine cell lines)
Biosafety Level 2
Agents associated with human disease and pose moderate hazards to personnel and the environment. It also addresses hazards from ingestion as well as from percutaneous and mucous membrane exposure (e.g. Measles virus, human cell lines, Toxoplasma, Salmonella, Adenovirus, Lentiviral vectors)
Biosafety Level 3
Indigenous or exotic agents, agents that present a potential for aerosol transmission and agents causing serious or potentially lethal disease (e.g. B. anthracis, SARS coronavirus, Mycobacterium tuberculosis, St. Louis encephalitis virus)
Biosafety Level 4
Dangerous and exotic agents that pose a high individual risk of life-threatening disease, aerosol transmission, or related agent with unknown risk of transmission (e.g. Marburg Virus, Congo-Crimean hemorrhagic fever, Ebola)
Biosafety Controls Incorporate the following safety practices: • • • •
Facility design and safety equipment Good laboratory/ microbiological practices Appropriate training, awareness and guidance Personal protective equipment
Standard Microbiological Practices Wash hands after work
with biological material and before leaving laboratory or work space No eating, drinking,
smoking, handling contact lenses, applying cosmetics, and storing food in laboratory
16
Standard Microbiological Practices Wear appropriate personal protective equipment
(PPE), such as gloves, lab coat, safety glasses Do not mouth pipette Work to avoid or minimize aerosols YES!!!
NO!!!
Standard Microbiological Practices Follow precautions when working with sharps
CSU Sharps Policy: http://www.ehs.colostate.edu/WBiosafety/Home.a spx Decontaminate work surfaces at least daily and after spills Use bench tops that are impervious to water, resistant to heat and chemicals; no carpet or rugs; no fabric furniture or chairs Decontaminate all infectious materials before disposal using an effective method
Summary of Biosafety Practices: Biosafety Level 1
2
Practices Standard Microbiological Practices: o Wash hands o No food, drink, or applying cosmetics o Biohazard warning signs o Sharps policy o Appropriate training BSL1 Practices PLUS: Limited access and advice on hazards Biosafety manual defining lab specific guidelines Proficiency training Incident evaluation No outside animals or plants
Primary Barriers and Safety Equipment PPE: o Gloves o Protective eyewear as needed
Facilities (Secondary Barriers) Laboratory bench Easily cleaned surfaces Sink
Class II biosafety cabinet for manipulations that cause aerosolization PPE: o Laboratory coat o Gloves o Closed toe shoes o Eye/Face protection as needed o Removal of PPE before leaving for non-lab areas
BSL1 PLUS: o Self-closing doors with locks o Eyewash available o Mechanical ventilation of inward airflow o Method of decontaminating waste
Requirements Personal Protective Equipment (PPE) Biohazard Symbol Caution Sign
Lab Specific Biosafety Manual
Contains
Risk assessment of microorganisms used in lab Lab specific biosafety procedures Training records PPE required IBC-approved protocols
Personal Protective Equipment (PPE) PPE is protective clothing or equipment that
prevents direct contact with infectious agents. MINIMUM PPE while working with biologicals/ chemicals: Close toed shoes Laboratory coat Gloves Additional PPE is determined by risk assessment and may include: Eye protection Respirators Apron
The Biohazard Symbol The biohazard symbol is a universal warning
against materials that could be infectious. Biohazardous waste must be labeled with this symbol and the word “Biohazard” and be decontaminated prior to disposal.
Biohazard Caution Signs Posted at entrance to laboratory Needs to include: Biosafety Level (1 or 2) Name of the infectious agent(s) Special entry requirements (minimum of lab coat when work is in-process) Supervisor’s name and phone number Secondary contact name and phone number National Fire Protection Association (NFPA) ratings Lab specific caution stickers to reflect hazards
BSL2
Supervisor Info special requirements Name of agents
Lab Specific Biosafety Manual Information about the University Biosafety Manual and
Bloodborne Pathogen Exposure Control Plan (if applicable) The link to the current CSU Biosafety manual is: http://www.ehs.colostate.edu/WBiosafety/PDF/Bios afetyHanbookCSU.pdf The link to the current CSU Bloodborne Pathogen Exposure Control Plan is: http://www.ehs.colostate.edu/WBiosafety/PDF/Expo sure_Control_Plan.pdf Lab Specific Biosafety Procedures Instructions on PPE and how to use equipment Special practices specific to the lab or work space
Lab Specific Biosafety Manual Emergency Contact Information and Protocols
CSU Emergency Contact information can be found online at www.ehs.colostate.edu/WBiosafety/Home.aspx Spill Cleanup Procedures -See Appendix A in CSU Biosafety manual for generic spill cleanup protocol Incident reporting Procedure and link to Biosafety Incident report form: http://www.ehs.colostate.edu/WBiosafety/PDF/I ncidentReportForm.pdf
Lab Specific Biosafety Manual Agent specific fact sheets and risk assessments – Can be
acquired through the the following websites:
Public Health Agency of Canada Pathogen Safety Data Sheets and Risk Assessment American Association of Zoo Veterinarians: Transmissible Disease Fact Sheets Animal Disease Information, Iowa State University IACUC (Institutional Animal Care and Use Committee) Resources, Washington State University Technical disease cards, The World Organization for Animal Health (OIE)
IBC protocols (PARF, AARF and and IACUC protocols-if animals
used)
Note: employees MUST review the protocols.
The protocols can be found by logging in at:
https://protocols.research.colostate.edu/rco/
Precautions for Handling Sharps CSU Sharps Procedures http://www.ehs.colostate.edu/WBiosafety/Home.aspx
Sharps
Any item having corners, edges or projections capable of piercing the skin: Needles Blades (razors, scalpels, X-acto, etc.) Broken glassware Glass slides Glass Pasteur pipettes and glass capillary tubes
Precautions for Handling Sharps Use plastic lab ware when possible Don’t bend, break, recap, or remove
needles from syringes Always use forceps, tongs or dust pan and broom when picking up broken glass Use a puncture resistant, properly labeled, sharps container for disposal of sharps
Place items into sharps disposal container immediately after use Never reach into the sharps disposal container Never overfill a sharps disposal container Never force materials into a sharps disposal container
Biological Sharps Injury Contain the sharp object Remove gloves or contaminated clothing
immediately Cleanse all exposed skin with soap and water for 3 -5 minutes Rinse exposed mucous membranes or eyes with water Apply antiseptic and a band-aid Report the incident to Biosafety (491-0270)
and your supervisor
Biological Sharps Injury Seek medical care at University’s Authorized
Treating Physician, if needed
During business hours: Go to a CSU Authorized Treating Physician (see list at http://www.ehs.colostate.edu/WWorkComp/Home.aspx)
After hours: Go to Emergency Room or Urgent Care
Follow up:
Fill out a Worker’s Compensation form: http://www.ehs.colostate.edu/WWorkComp/Home.aspx
within 4 days
Fill out a Biosafety Incident Report Form http://www.ehs.colostate.edu/WBiosafety/PDF/Incid entReportForm.pdf
Safety Equipment- Biological Safety Cabinet (BSC) A biosafety cabinet (BSC) is an enclosed
workspace
It is able to protect personnel, product, and the environment from exposure to biohazards and cross contamination, if used properly.
Safety Equipment- Biological Safety Cabinet (BSC) Aerosol generating activities at BSL-2 should be
performed in a Class II BSC Do not block BSC air vent or the function will be compromised Avoid storing equipment and supplies in the BSC, only use what is needed for task Do NOT use bunsen burners in the BSC Contact Heather Blair or Claudia Gentry-Weeks for BSC training:
[email protected] [email protected]
Biosafety Cabinet Certification All BSCs on campus must be certified annually
to:
Certify integrity of HEPA filters Certify airflow rates
Certification is performed by Technical Safety
Service, Ed Canfield, 720-981-4965
Safety EquipmentMicrocentrifuge Place in BSC, if possible
Use safety cups with gaskets or sealed rotors when using high
concentrations or large volumes of biohazardous materials Ensure that the rotor is properly balanced Know the speed ratings for rotor and tubes If unattended, post a warning sign with contact information (recommended) Have an emergency plan in place - many lab associated infections have been linked to centrifuge use
Disinfection vs Sterilization Both are processes that clean objects or substances to remove contaminants (such as disease causing micro organisms). Differences include: Disinfect
Sterilize
Definition:
To disinfect means to eliminate most harmful microorganisms (not including spores) from surfaces or objects; inactivate viruses.
To sterilize means to kill ALL microbes whether harmful or not - and spores present on a surface or object.
Methods:
Heating and pasteurization; phenolic compounds, heavy metals, halogens (eg chlorine), bleach, alcohols, hydrogen peroxide, detergents.
Heat, chemical, irradiation, high pressure, and filtration.
Types:
Surface disinfectants, alcohols, aldehydes, oxidizing agents, phenolics.
Steam, heating, chemical sterilization, radiation sterilization, sterile filtration.
Application:
Disinfection is used mostly to decontaminate surfaces and air.
Sterilization is used for food, medicine and surgical instruments.
Chemical Disinfectants Different micro-
organisms have different susceptibilities to disinfectants and require different methods for decontamination
Chemical Disinfectants When choosing a disinfectant, consider:
Concentration – Disinfectants MUST BE USED at the concentrations recommended by the manufacturer – MORE is not always better!
Contact time – Disinfectants must be in contact with objects for the time recommended by the manufacturer.
Stability – Some disinfectants are not stable when diluted and must be made fresh for use, or can only be stored for a specific period of time after dilution.
Toxicity – Disinfectants are made to destroy living things – avoid direct contact or inhalation.
Corrosiveness – Many disinfectants are corrosive to metals and other surfaces.
Flammability
Equipment- Autoclaves Autoclaves use
pressurized steam at specific temperatures to sterilize substances, and inactivate infectious materials prior to disposal Users MUST be trained Verify load is sterilized
Autoclave Monitoring Autoclave tape Operators must ensure minimum temperature
of 250°F / 121°C for at least ½ hour.
( > time/temp. may be required) Autoclave thermometer calibration at least annually
Structural Testing
Must be inspected by a qualified pressure vessel inspector at least
every two years.
Autoclave Monitoring Autoclave tape
Use on every item placed in the autoclave
Indicates increase in temperature, and is an indication that items have been subjected to heat, but:
Autoclave tape does NOT verify sterilization time and temperature
Untreated/failed
autoclaved
Autoclave Record Keeping Chemical Indicators
Designed to monitor exposure to steam conditions. A visual to validate that sufficient exposure conditions have occurred.
Biological Indicators
(Geobacillus sterothermophilus)
Performed at least every 6 months. Preferably monthly
Autoclave Record Keeping Maintain and post an Autoclave user log: Name Date Time Temperature Pressure Type of waste
Biohazardous Waste Needs to have the biohazard symbol and
the word “Biohazard” on the bag. Designate a space for biohazardous waste
and avoid contamination of other lab equipment. Inspect biohazard bags and containers for
protruding sharps or lab glass. Items with protruding sharps or lab glass
must be placed in a secondary, leak-proof container.
Transportation of Biological Materials within the Building
Use a durable, leak proof, secondary container.
Plastic bin Cooler
Label with infectious substance sticker with contact information on container. Decontaminate outer container prior to leaving laboratory.
Transportation off Campus When transporting biological material on
commercial roads or via air transport, proper training in Shipping Infectious Agents MUST be completed (Class 6.2 IATA/ DOT Dangerous Goods Training).
https://wsnet.colostate.edu/cwis86/WTrainReg/ ClassSignUp.aspx?TabID=Biosafety
Biological Spill Cleanup PPE
Risk dependent Minimum Closed toed shoes Lab coat Gloves Eye protection Disposal Dispose of all cleanup supplies in appropriate container (e.g. biohazard bag, sharps container, etc.) All biohazardous materials must be autoclaved prior to disposal Sharps: Use tongs/ dust pan and broom Place sharp objects in puncture resistant containers Refer to CSU Sharps Policy (http://www.ehs.colostate.edu/WBiosafety/Home.aspx)
Biological Spill Cleanup Isolate the area; put up a sign (if available) Get the biological spill kit Put on PPE, if not already on: Lab coat or closed front surgical gown 2 pairs of disposable gloves (double-glove) Eye protection/ face mask Respiratory protection may be advisable. Gently put down absorbent towels over the spill Pour disinfectant around the spill area so the disinfectant soaks into
the spill zone Give the disinfectant its contact time Wipe up spill area and dispose of towels in a biohazard bag Mop surrounding area with pathogen specific disinfectant (10 feet radius) Seal the biohazard bag with tape Autoclave
Biological Spill Cleanup General Protocols are found in the CSU
Biosafety Manual:
http://www.ehs.colostate.edu/WBiosafety/PDF/POST ED_CSU_Biosafety_Manual.pdf)
Contact your supervisor for work site specific
protocols. For large spills, evacuate area and call Biosafety: 491-0270.
Incident Response and Reporting Potential Biological Exposure Stop what you were doing. Remove gloves or contaminated clothing
immediately. Cleanse all exposed skin with soap and water for 3 5 minutes. Rinse exposed mucous membranes or eyes with water. Apply antiseptic and band-aid Report the incident to Biosafety (491-0270) and your supervisor.
Incident Response and Reporting Potential Biological Exposure Seek medical care at University’s Authorized
Treating Physician, if needed. During business hours: Go to a CSU Authorized Treating Physician (see list at http://www.ehs.colostate.edu/WWorkComp/Home .aspx After hours- go to Emergency Room or Urgent Care. Follow up: Fill out a Worker’s Compensation form: (http://www.ehs.colostate.edu/WWorkComp/Hom e.aspx) within 4 days Fill out a Biosafety Incident Report Form http://www.ehs.colostate.edu/WBiosafety/PDF/Inc identReportForm.pdf
Biosafety Laboratory Site Visits How to Prepare: Request a BSL-1 or BSL-2 Audit Form from the Biosafety Office Have a Lab Specific Biosafety Manual available Review this training
What to expect: Assessment of lab facility Assessment of laboratory practices Discussion of research performed Action items to complete E-mail of results following site visit
Biosafety Office Contact Information Dr. Robert Ellis – Biosafety Officer
[email protected] Dr. Claudia Gentry-Weeks – Associate Biosafety Officer
[email protected] Heather Blair – Assistant Biosafety Officer
[email protected] Joni Triantis Van Sickle– Occupational Health Coordinator and
Assistant Biosafety Officer
[email protected]
http://www.ehs.colostate.edu/WBiosafety/Home .aspx
CSU BSL-3 Facility Concepts Training Contact Information: Biosafety Office Environmental Health Services 141 General Services Building
[email protected] 970-980-9335
Table of Contents Section 1: BSL3 Hazards and Risks Section 2: Requirements Prior to Entering
BSL3 Section 3: Material Entry, Storage and Exit Section 4: Personnel Barrier Entry Section 5: Select Agent Work Requirements Section 6: Personnel Barrier Exit Section 7: Emergency Procedures
Section 1 BSL3 Hazards and Risks
What are the Hazards and Risks in our BSL3/ ABSL3 labs?
Biological Infectious agents including volumes, concentrations, routes of infection. These are described in detail in the Pathogen Characteristics and Symptoms training.
Chemical Exposure to chemical agents including disinfectants, biological reagents (DMSO, Trizol) and other common laboratory chemicals. Many of these must be treated has hazardous waste and are further described in Hazardous Waste Generator Training. Specific chemical hazards are available on safety data sheets (SDS) which must be made accessible in the laboratory.
Animal Animals pose an inherent risk to allergens, bites and scratches. Those that are infected with a pathogen may shed the organism in urine, feces or respiratory secretion resulting in a risk to the users. Proper handling is critical and covered in detail in Animal Care and Use Trainings offered through LAR, RICRO and the University Veterinarian.
What are the Hazards and Risks in our BSL3/ ABSL3 labs? Motion
Repetitive motion and heavy lifting can result in ergonomic risks.
Equipment
Using equipment in the BSL3 laboratory poses a risk of exposure particularly centrifuges and shaker incubators. Know how to properly use equipment safely. Take efforts to reduce the aerosolization of materials and use a biological safety cabinet. Know how to properly handle and dispose of sharps.
People
Be aware of your health; Do you get tired/ sick/ agitated? Breach protocol? How about those you work with? Does what happens outside the lab affect what happens in the lab? Be aware of how others are working in the laboratory to assure safe practices and reduce security threats.
Section 2 Requirements Prior to Entering BSL3
BSL3 Requirements/ Documents Occupational Health Risk Assessment Form
https://wsnet.colostate.edu/cwis86/WOHSP/RiskAssessment/add/A ddRisAssessment.aspx Mycobacterium tuberculosis work in Area: Medical Surveillance (PPD skin test) Virology Phase III: Yellow Fever Vaccination N95 Respirator Medical Clearance Discovery: Seasonal Influenza vaccine Facility Orientation Tour This training N95 Respirator Initial Training
Required to wear an N95 Respirator: https://wsnet.colostate.edu/cwis86/WTrainReg/OnlineClass/N95_Train/Train .aspx
Mock BSL3 Training PI/ Supervisor requests this training for you
BSL3 Requirements/ Documents Online Pathogen Characteristics and Symptoms training Select Agent and some BSL3 Viruses: https://wsnet.colostate.edu/cwis86/WTrainReg/OnlineClass/Unit4a/Train.as px M. tuberculosis: https://wsnet.colostate.edu/cwis86/WTrainReg/OnlineClass/Bio_Unit4MBT/ Train.aspx Online Select Agent Rules, Regulations and Insider Threat Training For
individuals working in Select agent areas: https://wsnet.colostate.edu/cwis86/WTrainReg/OnlineClass/Bio_Unit6/Train.asp x
HPAIV Self Quarantine Form (Contact PI or Heather Blair for the form) For Pod 1 and DMC suites Animal BSL3 training Achieved through the PIs or LAR training Lab Specific In Barrier Training Contact PI or Supervisor
Occupational Health Program Occupational Health services provided by
PVHS Occupational Health Services. Information related to these services is posted at:
www.ehs.colostate.edu/WOHSP/PDF/OCCU PATIONAL_HEALTH_PROVIDER_INFORMA TION_WEB_POST.pdf.
Contact Joni Triantis Van Sickle with
questions:
[email protected]; 970‐491‐3102 or 970‐420‐8172.
Keycard Access Requirements You need to complete and maintain all required training and
documentation (refer to slides 2 and 3) before keycard can be activated for BSL3 areas. “All access points, including emergency exits, must be secured”.
If a gate is opened by you or your car/ bike, make sure it closes before leaving the area. Do not let others enter with you or let them use your keycardunless you have been given approval by Biosafety
Escorting a visitor
If you notice a door propped open, CLOSE IT!
Propping doors open to the building or into hallways leading to BSL3 areas is NOT allowedunless you are there to watch it!!!
BSL3 Visitor Requirements We must have documentation of the visit Inform Biosafety of the visitor WELL in advance due to requirements that need to be met: We need to know WHO they are, WHY they are requesting a visit and WHEN they expect to be on site. The visitor needs to be informed of hazards and have proper training. Medical documentation may be needed depending on the visit location: Occupational Health Risk Assessment PPD surveillance HPAIV Policy Required vaccination(s) N95 clearance/ waiver Documentation of entry and exit times is also required.
Section 3 Material Entry, Storage and Exit
Materials Requiring Approval Prior to Entry into BSL3 Biological Agents
Institutional Biosafety Committee (IBC)/ Biosafety Approval
http://ricro.colostate.edu/IBC/IBC.htm
Animals
Lab Animal Resources (LAR) and Animal Care and Use Committee (ACUC) Approval
http://ricro.colostate.edu/IACUC/IACUC.htm http://web.research.colostate.edu/LAR/
Ionizing Radiation
EHS Radiation Office
http://www.ehs.colostate.edu/
Instruments/Equipment
Contact Building Operations Group Contact Biosafety if the equipment has the potential to generate a biological hazard
http://www.ehs.colostate.edu/WBiosafety/Home.aspx
Material Entry into the BSL-3 Materials that can be held in one hand can be taken
through the locker room with the researcher.
Uncrate materials from primary shipping container/ cardboard or from any extraneous cardboard and transfer the materials into the BSL3 barrier.
Materials that require two hands need to be placed in
the Material Transfer Area (MTA)
Place materials on cart(s) by inner anteroom door. Do not leave materials on these carts for extended periods of time; they need to be available for other groups/materials. Material should be easily retrieved on the “dirty side” with clean gloved hands. Avoid contact with the clean side of the door or cart when retrieving from the “dirty side”.
Material Storage in the BSL-3 Material should be stored off the floor so it
does not impede traffic, workflow, and cleaning. Biological materials are labeled with:
Genus, species, researcher’s first initial and complete last name, date, and PI last name
No biological agents are allowed to be poured
down the drain.
Chemical Storage in the BSL-3 Chemicals need to be labeled with
Chemical name, concentration, storage/accumulation date, PI’s last name and researcher’s first initial and complete last name
Safety Data Sheets (SDS) need to be accessible Complete Hazardous Waste Training (EHS)
Make sure all chemical types and quantities are accurate and labeled appropriately on all container(s) with the EHS eRFD. When Hazardous Waste accumulation date is close to 70 days, or the container(s) is full, you can do a surface wipe down using pathogen specific disinfectant to remove container(s) from the BSL3. Request a pickup from the EHS-Hazardous Waste group.
Biohazardous Waste Disposal Users must complete training before
operating the autoclave(s) Never abort an autoclave cycle! Report all alarms immediately to the BSL-3
Facility Manager/Section Head/ Operations Manager
Biohazardous Waste Disposal Biohazard waste must be labeled with the biohazard
symbol, the word “Biohazard” and be properly decontaminated prior to disposal. Do not overfill bags For autoclavable biohazardous waste:
Use autoclavable biohazard bags Add disinfectant to all bags before sealing Seal the bag with autoclave tape
Use a piece of autoclave tape and a black thick sharpie to label the bag with:
First initial, complete last name Date generated waste Principal Investigator’s last name “Save” or “Trash”
Instruments & Equipment in the BSL-3 Employees must be properly trained by their
PI/ Supervisor/ Lab Manager before operating equipment independently. Report any non-functioning equipment
immediately to the BSL-3 Operations Manager/ Principal Investigator. Label any broken or out of use equipment as
such.
Material Exit from the BSL-3 One of the following must happen for materials to exit
the BSL-3:
Shower/ Wash Personnel/ hands
Autoclave Trash, laundry Whatever can be
Chemical Decontamination (VHP/ Pathogen Specific Disinfectant)
Keycard and glasses Non-autoclavable/ sensitive material
Data transfer (e-mail, fax)
Paper
SECTION 4 Personnel Barrier Entry
Locker Room Entry Hang photo ID onto
Personnel Locator Boards OR sign into logbook Read all posted signs Check photohelic and/or monitors Swipe keycard Enter locker room
No gum, candy, food or beverages allowed beyond this point
Photohelic Gauge
Green light indicates appropriate air flow and safe entry
Red light indicates disrupted air flow. Do NOT enter.
General BSL3 PPE Remove street clothes, non-permanent jewelry,
hair ties, lanyards… Put on
Don’t forget to put applicable.
Put
corridor.
in shirt scrub pocket, where
on in De-gowning room or BSL3
BSL3 Additional PPE Additional PPE may be required for necropsy,
aerosolizations, animal handling or other specific procedures.
BSL3 Lab Entry Confirm directional
airflow monitoring device(s) throughout barrier. Read all posted signs. Movement (deliberate and focused). One door open at a time.
Section 5 Select Agent Work Requirements
Required for Select Agent Areas Required: SURGICAL GOWN OR TYVEK SUIT when performing necropsies and other animal handling work Increased risk of organism shed in feces and urine. EYE PROTECTION if wearing contacts
Section 6 Personnel Barrier Exit
General Barrier Exit- What to Remove First, Second, Third… 1.
5.
2.
6. 3. 7. 4. 8.
9.
“Special” Barrier Exit
Showering out is required if: • •
There is a biological spill outside of the BSC HPAIV is an agent used or stored in the suite
Section 7 Emergency Evacuation
Emergency Equipment Locations Spill Kits/ Rescue Blanket Stretchers and
Emergency PAPRS
Janitor closets or hallways
Flashlight/ First Aid Kits/ Soap/ Eye Wash
Station
Near sink area(s)
Fire Extinguishers
By doors and in hallways
Fire Alarm The fire alarm will be audible (“ping ping”, “BEEEERMP”) and
visual (strobe lights) Fire alarms are located in each area, as well as throughout the building Evacuate Immediately! Working inside a biosafety cabinet:
Seal all open work Remove outer gloves Leave sash open
Working outside a biosafety cabinet:
Turn off centrifuges, if in use.
Fire Alarm Remove
Surgical gown and PAPR, if applicable
Shoes and all gloves
Step onto the “clean side” Put on tyvek suit
Exit facility to rally point
Fire Alarm Rally Points
Tornado Evacuation Notifications may come electronically via email
through CSU or by phone. Evacuate as per facility procedures.
BARRIER INTEGRITY ALARM IDRC: Police Siren Painter Center: ALL CALL ADL: Loud alarm DMC: Eerily quiet/ or alarm sounds Close sash and turn off BSC
NOTE for the Painter Center BSCs: If you cannot close the sash, then leave the BSC on and running. Exit the barrier following normal barrier exit procedures Meet at designated area for information
POWER OUTAGE: GENERATOR ON Generator turns on (lights
back on):
Treat LIKE BARRIER INTEGRITY ALARM
Go to designated meeting area for information
Blackout Happens when the generator does not
turn on:
Exit lab following barrier exit procedure.
Contain cultures and animals (use touch light or emergency lighting) Close the sash of the BSC and turn the switch to the off position Find flashlight and use it to exit Disinfect keycard and other materials (glasses) using provided disinfectant. Use hand sanitizer and a towel to “wash” your hands.
Go to designation meeting area for further instructions.
BLACK OUT for People Who Shower Out Generator does not turn on: Treat like Fire alarm, BUT CLOSE THE BSC SASH
Contain cultures and animals (use touch light or emergency lighting) Remove outer gloves before exiting BSC Close BSC sash and turn switch to off position Find flashlight and use it to exit Follow emergency evacuation procedures - Tyvek suit
Meet at designated location
Spill Outside the Biosafety Cabinet (BSC)
EXIT! SHOUT and let everyone else know to EXIT! POST sign on lab door to inform others Now go shower out! POST sign on locker room door CONTACT Biosafety 491-0270
Illness Procedure If you are feeling sick, don’t go into BSL3 areas! Know the symptoms associated with the organisms you work
with. Know your baseline body temperature. CSU Authorized Treating Physician (Occ. Health Provider) should be sought: If you are concerned you have a work related illness/injury If you develop a fever (2 degrees above your normal body temperature) within 5 days of being in the BSL3 barrier and fever is a symptom associated with the pathogen/s that you work with: Notify Occupational Health Coordinator/Biosafety
491-0270
Person Down
ASSESS
CALL
DISINFECT
EXIT
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