Description and Prescription of BSL2+ Containment

Biosafety Level "2+" or "2 enhanced" training is the name is given to the scenario whereby a pathogen that is deemed to be BSL2 requires additional operational protocols in order to be handled safely. These operational protocols are prescribed as BSL3, however the pathogen is handled in a BSL2 physical space with BSL2 equipment.

The government-issued document which describes this type of handling is entitled "Containment Level 3 Operational Practices for use with specified RG2 organisms" and can be found here.

The decision for a given BSL2 pathogen to be handled with BSL3 operational protocols will come from one of three sources:

(1) The import permit under which the pathogen was imported requires it (see import permit restrictions)

(2) A formal Risk Assessment by the Presidential Biosafety Advisory committee has found that these precautions are warranted

(3) The above Risk Assessment was inconclusive and a query was forwarded directly to the Office of Laboratory Biosafety, PHAC and they have deemed that these precautions are warranted

BSL2+ is often required for those manipulating certain kinds of lentiviral vectors or those manipulating Macaque tissues. BSL2+ is also required when the scale of production of a BSL2 pathogen warrants extra precautions.

Lentiviral vectors - Information and Biosafety Considerations

Biosafety Considerations for Work With Lentiviral Vectors

Macaque tissue question on PHAC CL2 Checklist (Q 117)

 

Getting Your BSL2 Laboratory Ready for BSL2+ Work

Physical Space

Your laboratory must be suitable for BLS2 work. In practice these features are iterated by the Laboratory Biosafety Guidelines, Chapter 4 - Laboratory Design and Physical Requirements. The items appropriate for a BSL2 laboratory have been summarized in the document used for the annual audit of your BSL2 laboratory space, the Laboratory Containment Level 2 Audit Checklist. To ensure that your laboratory complies with this checklist, please contact the Biosafety Auditor, Carol Carte for help and suggestions.

If the work must be conducted in a shared space, it is imperative that only those workers that are BSL2+ trained be present in the work area when BSL2+ work is being conducted. Signage must be prominant.

Paperwork

Working under BSL3 operational protocols requires a complete set of BSL2+ SOPs to be written. The SOPs required include but are not limited to:

a) Layout of the BSL2+ area when in BSL2+ mode, Laboratory Access, Biosecurity

b) PPE Required, Entry and Exit Protocols, Decontamination, Housekeeping

c) Emergency Exit Protocols

d) Spill Protocols

e) Lab Specific protocols (experimental procedures done at BSL2+)

f) Use of a Biological Safety Cabinet

At any time during this process, you may contact the Carol Carte in the Biosafety Office for SOP review and comments. When completed, the set of SOPs are submitted to the Presidential Biosafety Advisory Committee for review.

Solid Front, Rear Tying, Tight Wristed Gowns

Options for these types of gowns are disposable or rental. For rental gowns, you must create an "on demand" account at Mohawk Linen Services by contacting customer service representative Randy Spencer. The account allows you to receive clean, nonsterile gowns in bulk which you then send back in bulk for laundering. The type of gowns to order are called "Polygown Class B" (one size fits all) and you will require a "nylon bag" for return of dirty gowns. Gowns from Mohawk are delivered to the MUMC Receiving docks in the Health Sciences Centre. Contact Mike Mans, Hamilton Health Sciences Customer Support Services Leader (x73399), for manual dropoff and pickup locations or to create an account for trucking options. ** if a gown becomes contaminated due to a spill, please bag the gown in an autoclave bag and contact the Biosafety Office. Do not bleach. Do not bag with uncontaminated gowns.

 

Receiving Training for BSL2+ Work

Site and work specific training are required by the Laboratory Biosafety Guidelines and these are re-iterated in the above referenced document "Containment Level 3 Operational Practices for use with specified RG2 organisms". Every worker must have completed BSL2 training, WHMIS and Fire Safety and all the associated updates. There are five additional tasks that need to be completed:

1. The new worker must be familiar with the physical space that is to be designated the BSL2+ work area. The supervisor should assure this.

2. The new worker must have read and understood the SOPs required for their BSL2+ work, both the general SOPs and the project-specific SOPs. The supervisor should assure this.

3. The new worker should understand the nature of the pathogen with which they are working. This information should be provided by the supervisor.

4. Once #1, #2 and #3 are completed, the new worker should contact the Biosafety Office to arrange for a training session and to receive the BSL2+ quiz. The quiz includes questions that relate to the physical space and the SOPs. The training session is approximately 1 hour in length depending on the number of participants.

5. The new worker should contact the FHS Safety Office (x24956) to arrange for repirator training and N95 mask fit testing.

6. Once the quiz is successful, the new worker should contact the Biosafety Office to schedule a practical spill test.

When the new worker has completed all 5 tasks, they are deemed competent to work safely at BSL2+.