Biological Safety

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Biological Safety

• Biological Safety Cabinets

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Principal Investigator Responsibilities

• It is the responsibility of the P.I. to ensure
  that all workers in their research group who will
  be handling biological agents are registered with
  the EHS Office and have completed required
  biological safety training.
• It is the responsibility of the P.I. to ensure
  that biosafety cabinets are serviced and
  certified annually.
• Workers within the research group who carry out
  their duties in a registered biological lab must
  receive instruction relating to the biological
  hazards even if they will not be handling
  biological agents themselves.

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Canadian Regulations

• Federal Regulatory Authorities
• ? Public Health Agency of Canada (PHAC)
• ? Canadian Food Inspection Agency (CFIA)
• ? Transport Canada
• ? Environment Canada/Health Canada
• ? Department of Foreign Affairs and
  International Trade

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Laboratory Biosafety Guidelines 3rd ed. 2004

• This document is a guideline when working with
  indigenous species but is regulatory for all
  imported agents.
• This guideline covers facility design and
  operational requirements for work with human
  pathogens, containment levels 1-4

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Containment Standards for Veterinary Facilities,
1st ed. 1996

• This standard covers the design and operational
  requirements for facilities working with animal
  pathogens, containment levels 1-4 laboratories
  handling non-indigenous animal pathogens small
  and large animal containment facilities

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Principles of Biosafety

• Personal safety is the most important element
  when working with biological agents. It is
  estimated that there have been thousands of
  laboratory acquired infections. Hepatitis is the
  most commonly reported viral infection in lab
  workers.

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Principles of Biosafety routes of exposure

• Inhalation
• Ingestion
• Inoculation
• Absorption

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What is a Biological Safety Cabinet?

• A ventilated cabinet or enclosure
• Uses directional airflow and HEPA (high
  efficiency particulate air) filters to provide
• ? personnel protection
• ? environmental protection
• ? varying degrees of product protection

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Containment Hood

• Containment Cabinet designed to protect both
  the specimen and the worker. A laminar flow of
  HEPA filtered air is passed down from the top of
  the hood and across the work surface and is
  exhausted or re-circulated without entering the
  workers breathing space. The air is re-filtered
  before being exhausted.

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Biosafety Cabinet Classifications

• Class I
• Class II
• Class III

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Class I BSC

• Suitable for work with low to moderate risk
  agents where there is need for containment but
  NOT product protection
• Protects environment by filtering air before it
  is exhausted
• Movement of air into cabinet, away from user

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Class II BSC

• Class II Type A1
• Class II Type A2
• Class II Type B1
• Class II Type B2

• Protection for personnel, product, environment
• Front access opening with carefully maintained
  inward airflow
• HEPA filtered, vertical, unidirectional airflow
  with the work area
• HEPA filtered exhaust air to the room or to a
  facility exhaust system

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Class II BSC
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Class III BSC

• Offers the highest level of protection and must
  be used for Risk Group 4 agents
• All penetrations are sealed gas tight
• Air flow is maintained by a dedicated exhaust
  system exterior to the cabinet
• Access to the work area is by means of heavy duty
  rubber gloves attached to ports in the cabinet
• The cabinet has an attached sterilizable
  pass-through box
• The cabinet may be connected to a double door
  autoclave

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Biosafety Cabinets
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Clean Bench

• Clean Bench designed to protect the specimen by
  bathing the work area with a laminar flow air
  free of particulate contamination. The clean
  bench forces air out from the back of the hood,
  across the work surface and toward the worker.
  The specimen is protected not the worker.

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Working in the BSC start up procedures

• Ensure that the cabinet is certified
• Turn off UV lights if used
• Turn on fluorescent light and blower (if off)
• Allow the blower to run for 5 10 minutes
• Check air intake exhaust grilles for
  obstruction
• If BSC has an alarm, check switch on
• Disinfect all interior surfaces with 70 ethanol
  or suitable disinfectant

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Working in the BSC Standard Operating Procedure
(SOP)

• Place items and aerosol generating equipment near
  the back of the hood
• Do not obstruct grilles
• Don proper personal protective equipment (PPE)
• Position yourself with arm pits level with sash
  opening.

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Working in the BSC Standard Operating Procedure
(SOP)

• Do not hang spray bottles on the outside grille
• Perform operations as far to the rear of the work
  surface as possible
• Keep discarded, contaminated material to the rear
  of the BSC, do not discard in containers outside
  the BSC
• NO OPEN FLAMES (turbulence, damage to HEPA
  filter, fire hazard)
• Work from clean to dirty
• If a spill occurs surface decon objects and work
  surface with the BSC ON

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Work from clean to dirty
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Personal Protective Equipment
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Working in the BSC

• Upon completion of work
• ? close open containers
• ? allow BSC to run for 5 minutes with no
  activity
• ? surface disinfect all objects before removal
  from cabinet
• ? spray off or remove first pair of gloves to
  the biohazard bag within the cabinet
• ? wipe down work surfaces with 70 ethanol or a
  suitable disinfectant
• ? turn off fluorescent light cabinet blower
  (if applicable)

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UV lights

• Intended to destroy microorganisms in air or
  exposed surfaces
• Have a limited penetrating power and only
  effective when properly maintained and cleaned
• Have a limited germicidal life span even if you
  see the blue glow the lamp may not be effective
  for its intended purpose
• May damage eyes, skin and laboratory equipment
• Must always be turned off when the lab is
  occupied one does not need a direct exposure,
  exposure can occur from the reflective surfaces
  within the cabinet
• Though exposure decreases with distance (inverse
  square law), a study reported in a position paper
  by the American Biological Association notes that
  it is possible to reach permissible UV exposure
  limits within 32 minutes to 1.4 hours at general
  eye level in the center of standard research lab

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BSC Maintenance

• Twice daily wipe down work surfaces
• Weekly clean UV lamp
• Monthly wipe down all vertical surfaces within
  cabinet, clean underside of work surface as well
  as area underneath the work surface
• Annually cabinet certification

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Proper Glove Removal

• With both hands gloved, peel one glove off from
  top to bottom and hold it in the gloved hand
• With the exposed hand, peel the second glove off
  from the inside, tucking the first glove inside
  the second
• Dispose of the gloves promptly
• Never touch the outside of the glove with bare
  skin
• Wash hands as soon as possible

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Proper Glove Removal
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Basic hand washing guidelines

• Wash hands regularly
• Wet, soap and lather for at least 20 sec.
• Wash and scrub under the nails and cuticle
• Rinse thoroughly and dry
• Turn off tap using paper towel

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Recommended Hand Washing