BSL 1 Biosafety Training FHSc. Safety Office Health Sciences Center 1J11, ext. 24956 Print out the accompanying test and return (in handwriting) to 1J11.

BSL 1 Biosafety Training

FHSc. Safety Office

Health Sciences Center 1J11, ext. 24956

Print out the accompanying test and return (in handwriting) to 1J11 HSC or

fax to 905 528 8539

How to find us HSC 1J11

[Printable

Information for undergrad courses

• You will be contacted only if you have failed your quiz, <75%. Your instructional assistant will be contacted at the same time.

• You will not be permitted to attend the biological labs of your course until you pass.

Biosafety in Canada: Standards Public Health Agency of Canada (PHAC)

through the “Laboratory Biosafety Guidelines”

Canadian Food Inspection Agency (CFIA) has “Veterinary Standards for Animal Facilities”

Human Pathogens and Toxins Act C-11

Ministry of Environment Canada (MOE) for waste disposal

PHAC has “Laboratory Biosafety Guidelines”

• Sets the standards for work with biological agents such as bacteria, viruses, fungi parasites, prions and hazardous cell lines.

• The health risk associated with an organism will determine the protocols which must be followed to work with it.

Level of Hazard: BSL 1,2,3,4

Internationally and in Canada, there are four levels of biosafety, each associated with a type of lab and specific equipment.

The personal protection required increases with the level of hazard.

Labs, techniques and methods required increased safety with the level of hazard.

Risk evaluation to determine level of use

Risk evaluation of an agent is part of the responsibility of the supervisor for the project and is reviewed by the University Presidential Biosafety Advisory Committee.

All individuals working with an agent must comply with the level of risk.

Factors affecting risk

• agent in use eg. specific bacterium

• pathogenicity eg. degree of disease

• infectious dose eg. # organisms to cause disease

• mode of transmission eg. directly by blood or indirectly through the air

Factors affecting risk

• techniques in use

• host range eg. does it infect animals as well as humans

• effective treatment or prevention eg. is there a vaccine?

• Potential for aerosol creation eg. sonication, vortexing or homogenization

Factors affecting risk

Volume of material eg > 10 liters

• Concentration of material

• Stability in the environment eg. will it survive on a benchtop?

• Use of re-combinant DNA eg. oncogenicity, replication ability, host range, ability to revert to wild type

Biosafety Level 1 (BSL 1)

• unlikely to cause disease in healthy workers or animals

• treatment / prevention is available

Low individual risk

Low community risk

Biosafety Level 2 (BSL 2)

• causes human or animal disease • under appropriate use, is unlikely to cause death

to healthy laboratory workers or animals • treatment / prevention is available

Moderate individual risk

Limited community risk

Biosafety Level 3 (BSL 3)

• usually causes serious human or animal disease including death. Usually spreads by direct contact of blood or body fluids from one individual to another

• generally, no cure / prevention available

High individual risk

Moderate community risk

Biosafety Level 4 (BSL 4)

• human disease including death and may be readily transmitted through casual contact

• no treatment / prevention available

High individual risk

High community risk

Which level of organisms are being used in your lab ? • Ask to see the biological equivalent of an

“MSDS” for the agents you will be using.• Or look up the information on the Internet

site for the supplier of the product. • Or reference a large database such as

ATCC http://www.atcc.org/ or

PHAC http://www.phac-aspc.gc.ca/index-eng.php/

Basic Safety Practices - BSL 1

These requirements must be followed in all labs working with biological agents.

Know and understand them.

Basic Safety Practices

Longer hair is to be restrained so that it cannot come into contact with hands, specimens, containers or equipment.

Oral pipetting of any substance is prohibited in any lab, use only mechanical fluid transfer devices.

Basic Safety Practices

No eating or storage of food or drink inthe labs…dispose of any food wrapping or containers BEFORE you enter the lab, including water bottles and gum.

Keep personal items out of the working area of the lab.

Basic Safety Practices It is wise to report any increased susceptibility to infection to your supervisor or medical advisor.

Ask the lab supervisor if you are unclear about any hazard or process.

Dispose of all biological waste in the designated container only.

Waste disposal

• Biological waste is to be disposed of into the yellow or red bags in the biohazard labeled cardboard boxes

• Sharps (scalpel blades, razor blades, needles on syringes) must be disposed into a sharps container and then disposed as biological waste when ¾ full

Basic Safety Practices

Open wounds, cuts, scratches and grazes should be covered with waterproof dressings BEFORE working with any biological material.

Limit use of sharps and glass.

DO NOT RECAP NEEDLES

Basic Safety Practices

Minimize aerosol creation

Label all biological material

Where possible and appropriate, protect yourself by immunization esp Hep B when working with blood or body fluids.

Personal protection for BSL 1

Protective laboratory clothing must be worn in the lab but not worn out of the lab

- closed toed footwear

- wear lab coats at all times

- goggles – if using contact lenses

- if required by the protocol

Personal protection for BSL 1

Gloves…latex for water based materials

…nitrile or vinyl for organics

Change gloves when contaminated.

Wash your hands frequently, particularly when leaving the laboratory.

Emergencies

Staff must understand all hazards with which they work, including emergency response.

- read and know the emergency and spill response protocol posted in your lab

- report any spills or injuries to your supervisor- know the location of spill kits and first aid kits- know the location of eyewash and shower

Training

personnel must receive training on biohazards and safe handling protocols

- pass for biotraining is 75%

- local lab training to be signed by trainee and trainer

- retraining program includes annual update

Physical lab requirements for BSL 1

Physical requirements are a functional and well designed lab

Doors to the laboratory must not be left open

Working requirements for BSL 1

• Basic microbiology lab practices, keep the lab organized and clean

• Open bench is suitable

• Routine decontamination techniques to maintain sterility - contamination of cultures is a risk in an unorganized lab

Supervisor responsibilities for BSL 1 Access to the laboratory is restricted to

authorized personnel

ie. those who have been advised of the potential hazards, trained and approved by the supervisor

Supervisor responsibilities for BSL 1 Post the lab with a biohazard sign

indicating the biosafety Level and organism(s) in use

Contaminated materials or equipment leaving the lab must be appropriately decontaminated first

Supervisor responsibilities for BSL 1

All materials must be contained so that they will not be released to the environment.

Disinfectants effective against the material in use, must be available at all times.

Have available leak proof containers for transporting materials between labs.

Supervisor responsibilities for BSL 1

Maintain an effective insect and rodent control program.

Investigate spills and accidents, keep records, use results in future training.

Supervisor responsibilities for BSL 1 Biological safety information, such as a

manual, must be available

- with procedures for the lab

- it must be read and followed by all

- it must be updated regularly

Supervisor responsibilities for BSL 1Provide MSDSs or equivalent.

Ensure compliance within lab.

Any use of animals must comply with the CCAC and any training requirements of the CAF and follow the SOP.

Additional biosafety lab information 1) Bunsen burners

2) Ultraviolet light

3) Causes of contamination

4) airflow diagram of a biological cabinet

5) airflow diagram of a fume hood

6) airflow diagram of a clean air bench

Bunsen Burners• Do not use combustibles or flammables in a

biological cabinet – 70% recirculated will allow buildup of leaking gases

• Upward hot air flow disrupts downward laminar flow

• May create holes in the HEPA filters• Splashes may occur (when using loops) and

could cause contamination

Substitute:• Micro incinerator• Alcohol burner

Ultra Violet Light

UV lights are not recommended:

• they provide a false sense of security - not giving off UV light (just prior to the end life of the bulb)

• only disinfects the surface – could contact a layer of media protein covering the pathogen

• casts a shadow, leaving areas that remain unexposed to the UV wavelengths

• could cause burns / damage eyes – turn off when working in the room

Causes of Contamination1) YOU!

hands not clean, bacteria/spore under your nailsgloves – not sterile from the box dirty lab coatslab coat sleeves should be tapedplace all materials (cleaned) required into hood BEFORE work

2) Incubator Avoid dip in line from CO2 tank Clean humidity source regularly Clean all surfaces after spill, including holes on trays If at ground level – tape bench paper

onto floor

Class II Biological Safety Cabinetuse with BSL 2 agents

yes -personnel protection

yes - product protection

yes - environmental protection

Chemical Fume Hooddo not use with biological agents

yes - personnel protection

no - product protection

no - environmental protection

Clean Air Benchuse only with BSL 1

no - personnel protection

yes - product protection

no - environmental protection

Information for McMaster staff/students • Do not ship any samples, or take samples

with you on a trip. Check with Mike McGuire to ensure it does not require Transportation of Dangerous Goods compliance

• File a Materials Transfer Agreement with Bertha Monrose before you share material with another institution

Information for McMaster staff/students • Do not transform your own body cells.

The immune protection you would normally have has now been bypassed

• Obtain a biosafety approval number from the Biosafety office, HSC 1J11 ext 24956 before importing material or working with biological material

Information for McMaster staff/students • Cell lines can be BSL 1 or BSL 2. Check

with your supplier or your supervisor.

• If you are isolating primary cell lines from ill humans or animals, they may be BSL 2 until they are well characterized. Check with the Biosafety office first.

Use of a biological cabinet

• Some labs may chose to use Class II biological cabinets to maintain sterility for their BSL 1 agents.

• The following slides cover information for the use of biological cabinets.

Before you begin

• Check to ensure you are using the right cabinet for your work.

• Check to see it has been certified in the last 12 months.

• Ensure the UV light is off anytime anyone is in the room.

• To start, let the cabinet run for 3 minutes to purge air.

Before you begin

• Use a disinfectant that will kill any organism in the cabinet

• Check to see that the window is in the correct position

• Do not place pens or paper inside the cabinet

• Never block the grilles

• Segregate clean items from dirty ones

Before you begin

• Wear a lab coat and tape the coat at the wrist

• Pull the gloves over the taped coat

• Sterilize the gloves with a 70% alcohol wash

• As you place items into the hood, spray them down with alcohol to sterilize

Before you begin

• Place sterile items on the left side

• Place items from the incubator at center back

• You may place a work pad soaked in disinfectant on the center front work area

• On the right side, place a waste bucket for fluids, one for sharps, one for solids

Working in the cabinet

• Keep lab doors closed and keep others out of the room

• Only one person working in the cabinet

• Use good aseptic technique

• Slow movements directly in or out

• Keep sterile and non-sterile items separated

• Do not use Bunsen burners in cabinet

Working in the cabinet

• Have posted spill protocol nearby

• No gum, food or drink

• See the two powerpoint presentations on Good Culture Practices and Misidentification and Contamination, to learn more about good practices and why they are important.

When you are finished

• Spray each item with a disinfectant as you take it out of the cabinet

• Fill the liquid waste to the max fill line to achieve appropriate concentration. Mark on the time when it can be disposed

• Seal the top of the solid waste bag, spray it, put it into the biohazard cardboard box

• Close the top of the sharps container, leave it in the hood

When you are finished

• Spray all interior surfaces with the disinfectant, remember to spray the plastic window facing you.

• If possible, leave the cabinet running.

• If not, let it purge for 3-4 minutes.

• If there is anything unusual, tell your supervisor and leave a note for the next person who will be using the hood.

Clean ----------------------- Dirty

Anew materials

B materials

from incubator

C

garbage

D materials

to go back into

incubator

E working

surface

F wet material

solution (bleach)

Suggested layout for working in a Class II biohoodSuggested layout for working in a Class II biohood

Contact information

For additional training videos, biosafety approval, import permits, reporting of spills

or lab acquired infections, or general questions

FHSc. Safety Office

Health Sciences Center 1J11, ext. 24956

Carol Carte or Karin Cassidy