1. Place Bunsen burner away from overhead shelving or equipment
2. Remove all paper, combustible materials and excess chemicals from area
3. Tie back long hair, loose clothing
4. Inspect hose for cracks, holes, pinch points or other defects and ensure hose fits securely on gas valve and Bunsen burner
5. Have a sparker with extended nozzle available before turning on the gas
6. Turn on the gas slowly and do not open completely
7. Adjust the flame to produce an appropriate flame for the experiment
8. Do not leave flames unattended
9. Shut off gas when flame is no longer needed
10. Allow burner to cool before handling

1. Only move compressed gas cylinders with a proper cart, with the chain or strap secured
2. Store upright in dry, ventilated areas, away from heat sources or electricity
3. Store strapped into appropriate cylinder holder, with straps at 2/3 of cylinder height
4. Store capped with valves closed when not in use
5. When in use, only use appropriate pressure regulator
6. All cylinders must be properly labelled
7. Inspect cylinders regularly for damage or corrosion

This SOP applies to EVERYONE working in the laboratory.

Review BEFORE working.

1. Access to the laboratory is limited to staff, or other persons with permission of the Principal Investigator, when work with BSL-2 pathogens is being conducted.
2. Laboratory doors must be kept closed, but locking is at the discretion of the Principle Investigator.
3. Laboratory staff and all other persons working with infectious substances must wear gloves. Contaminated gloves must be changed immediately.
4. Laboratory staff must wash hands after handling infectious materials, after removing gloves, and always before leaving the lab.
5. Laboratory staff must not eat, smoke, handle contact lenses, or apply cosmetics in the lab.
6. Food for human consumption must be stored outside laboratory work area in refrigerators designated for that purpose.
7. Only mechanical pipetting devices must be used in the laboratory.
8. All sharps must be place in red sharps containers.
9. All non-liquid contaminated cultures, stocks, plates, and other biologically contaminated waste must be placed in biohazard waste containers for pickup by Stericycle.
10. All infectious liquids must be autoclaved or disinfected with 1: 10 bleach for one day prior to being poured into drain.
11. All procedures must be performed to minimize creation of splashes or aerosols.
12. As much work as possible with pathogens should be performed in the Biological Safety Cabinet.
13. All work surfaces must be decontaminated at the completion of work, at the end of the day, or after any spill or splash of viable material.
14. All laboratory personnel must be advised of potential hazards, and read the laboratory safety manual.
15. All support personnel must be made aware of hazards before working in the laboratory.
16. Spills and accidents that result in overt exposures to infectious materials must be immediately reported to the Principle Investigator and appropriate medical evaluation must be provided.
17. Laboratory equipment, surfaces, and other contaminated materials must be decontaminated with an effective disinfectant on a routine basis. Decontamination must also be completed after overt spills, splashes, or other contamination according to the Spill Control Procedures.
18. Contaminated equipment must be decontaminated before the equipment is repaired in the laboratory or sent out for repair, maintenance, or other purposes.
19. Lab coats must be removed and left in the laboratory before going to non-laboratory areas.
20. Lab coats must be laundered by the department, and should never be taken home.
21. Open-toed shoes must not be worn in the lab.
22. Laboratory doors must be locked when the laboratory is not occupied.
23. Bench tops should be kept neat, orderly, and accessible for cleaning.
24. Paper work stations are located away from hazardous materials and never within the level 2 laboratory zone.

BEFORE STARTING WORK

• Wear gloves and a lab coat
• Make sure UV lights are off
• Ensure sash is in correct position
• Check that air grilles are unobstructed
• Make sure air is flowing inward
• Disinfect all working surfaces

DURING WORK

• Don’t move your hands and arms in and out of the BSC excessively
• Keep contaminated material towards the back of the BSC
• If you’ve used it in the BSC, throw it away in the BSC
• Don’t use open flames

AFTER FINISHING WORK

• Leave BSC unused and running for 5 minutes
• Make sure all containers are closed before taking them out of the BSC
• Disinfect all contaminated material with 70% ethanol or 1% NaClO (1:6 bleach solution) for 15 minutes before taking them out of the BSC
• Disinfect all work surfaces in the BSC while it is still running
• Remove and dispose of gloves in the BSC
• Wash hands before leaving the lab

PHYSICAL PROTECTION

The lab will be locked at night when there is no one working in the lab.  Unknown people entering the lab will be questioned and asked to leave.

Keys are issued only to Guttman lab members.  Do not share your lab key with anyone.

All risk group 2 biologicals must be stored in spill proof, impact resistant containers in equipment that is locked and labelled (door signage)

BIOSAFETY LEVEL 2 ORGANISMS that can be used in the Guttman lab:

Pseudomonas aeruginosa

Burkholderia cepacia

Aspergillus fumigatus

Candida albicans

Clavibacter michiganensis (only for DNA extraction and sequencing)

Neisseria subflava, N. polysaccharea, N. bergeri (only for DNA extraction and sequencing)

PERSONNEL SUITABILITY / RELIABILITY

All lab members working with any biosafety level 2 organisms must be trained in biosafety protocols.  Documentation of completion of training courses must be kept on file in the lab.

PATHOGEN ACCOUNTABILITY

The use, addition of new stocks, and disposal of any biosafety level 2 organisms must be documented in a log, stored in the lab.

Any discrepancies in the pathogen log must be reported to the Responsible Official.  The Responsible Official will account for any discrepancy and report justification.

EMERGENCY RESPONSE

Unknown people entering the lab must be questioned and asked to leave.  If they resist, campus police must be called to ensure their removal.

Intentional events like bomb threats will be reported to the campus police.  For fire alarms, the lab doors will be locked and everyone will leave the building by the fastest route.

For unintentional events like spills, follow the SOP in the biosafety manual.

All samples containing biosafety level 2 organisms must be stored in equipment that is plugged into the emergency (red) power outlets, so power failures won’t impact storage / growth.

BSC failure (alarm, loss of power):

1. Don’t panic
2. Stop work immediately
3. Follow shut down procedures
4. Report to PI

Emergency evacuation (fire alarm):

1. Turn off any direct sources of heat (ie bunson burners, hot plates)
2. Close any open containers of biologicals or chemicals.
3. If there is time, wash hands before leaving.
4. Take personal belongings (and coat).
5. Get out – shut doors behind you.  Doors should be locked.