Biosafety Training: Containment Level 2

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Biosafety Training Containment Level 2

• Cynthia Schwartz, Ph.D.
• Laboratory Safety Manager, SLRI

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Biosafety Training Topics

• What is Biosafety?
• Why Biosafety?
• What is a biohazardous material?
• Types of biological hazards
• Risk Assessment
• Protection
• Waste
• Emergency Procedures Biological Spill

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Biosafety Training

• What is Biosafety?
• The combination of laboratory practice and
  procedures, laboratory facilities and safety
  equipment used when working with potentially
  infectious microorganisms (CDC)

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Biosafety Training

• What are biohazardous materials?
• Agents of biological origin that have the
  capacity to produce deleterious effects on humans
  and animals.

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Biosafety Training

• Why Biosafety?
• US Survey registered 4,000 laboratory-associated
  infections between 1951-1996 60 from Research
  activities bacterial and viral agents accounted
  for one third of these
• About 20 of laboratory-associated infections
  are due to recognized accidents, 80 are
  attributed to unknown or unrecognized causes
  (Lawrence Berkeley National Laboratory, USA)

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Biosafety Training

• Biohazardous materials include
• (1) all infectious organisms (e.g. bacteria,
  viruses, fungi, parasites, prions, rickettesias,
  etc) that can cause disease in humans, animals or
  plants
• (2) certain types of recombinant DNA, transgenic
  animals or plants
• (3) human blood, tissues, fluids, cells and/or
  cell lines
• (4) potentially infectious animals or animal
  tissue
• (5) biologically active agents (i.e. toxins,
  allergens, venoms) that may cause disease in
  other living organisms or cause significant
  impact to the environment or community.

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Biosafety Training

• Classification of biological hazards
• Biohazards are classified into Risk Groups that
  are determined by the potential impact of the
  exposure to the agents upon the individuals that
  work with the agents, their colleagues and the
  potential effect upon the surrounding community

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Biosafety Training

• Classification of Organisms into Risk Groups is
  based on
• Pathogenicity
• Infectious dose
• Mode of transmission
• Host range
• Availability of effective preventative measures
• Availability of effective treatment

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Biosafety Training

• Classification of Biohazards
• Canadian Legislation
• WHMIS
• Class D Poisonous and Infectious Materials
• Division 3 Biological or infectious materials

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Classification of Organisms according to Risk
Groups

• Risk Group 1
• Low individual risk and community risk
• Risk Group 2
• Moderate individual risk, low community risk
• Risk Group 3
• High individual risk, low community risk
• Risk Group 4
• High individual risk, high and community risk

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Biosafety Training

• Risk Group 1
• It is unlikely that the agent will cause disease
  in healthy adults or animals
• Risk Group 2
• The agent can cause human disease but under
  normal circumstances it is unlikely to be a
  serious hazard to lab workers, the community,
  livestock or the environment
• Risk Group 3
• The agent usually causes serious human disease
  but can not be spread by casual contact or the
  disease is treatable
• Risk Group 4
• The agent causes very serious human disease,
  often untreatable, may be readily passed from one
  to another by casual contact

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Biosafety Training

• Containment Levels
• Containment levels describes safe methods for
  handling and managing infectious agents in the
  laboratory
• The purpose of containment is top reduce or
  eliminate the exposure to potentially hazardous
  agents
• Four containment levels are outlined in the
  Health Canada guidelines the highest safety
  standards (Containment Level 4) are reserved for
  the most hazardous pathogens (Risk Group 4), and
  the least stringent (Containment Level 1) for
  those which have minimal impact on health (Risk
  Group 1).

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Biosafety Training

• Containment levels take into account
• Risk group of the biohazardous material
• Handling, procedures, manipulations
• Scale

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Biosafety Training

• Containment Levels
• Containment is achieved through the use of
  appropriate safety equipment, facility design and
  lab procedures and practices

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Biosafety Training Containment Features

• Containment Level 1
• No special design features containment achieved
  through lab practices
• Containment Level 2
• Primary containment barriers are Biosafety
  Cabinets (BSCs), centrifuges with sealed rotors
  or safety cups, personal protective equipment and
  lab practices
• Containment Level 3
• Additional primary and secondary barriers are
  used to minimize the potential for release of
  infectious organisms respiratory protection,
  HEPA filtration of exhausted air from the
  laboratory and strictly controlled access to the
  lab
• Containment Level 4
• Maximum containment of infectious diseases
  positive pressure suits, Class III BSC,
  decontamination of the air and all other
  effluents produced in the lab

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Biosafety Training

• Risk Assessment
• Process used to identify the potential risk
  posed to human health and the environment as a
  result of exposure to hazardous materials
• Helps to determine the type of containment,
  procedures and safety equipment required

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Biosafety Training

• Protective Measures
• Equipment
• Personal Protective Equipment
• Work practices

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Biosafety TrainingProtective Measures Biosafety
Cabinets

• Class I Biosafety Cabinet
• Vented cabinet with inward flow of non-filtered
  air
• HEPA-filtered exhaust air
• Does not provide product protection

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Biosafety TrainingProtective Measures

• Class II Type Biosafety Cabinet
• Vented cabinet with HEPA- filtered supply air
  flow
• HEPA-filtered exhaust air
• Provide personnel, product and environmental
  protection
• May not be used with volatile, toxic or volatile
  radioactive materials

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Biosafety TrainingProtective Measures

• Class III Biosafety Cabinet
• Totally enclosed, gas-tight cabinet with HEPA-
  filtered supply and exhaust air
• Exhaust air flows though dedicated exterior
  exhaust system
• Provide maximum personnel, product and
  environmental protection

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Biosafety TrainingProtective Measures

• Personal Protective Equipment (PPE)
• Lab coat long sleeved and closed
• Gloves
• Closed-toe shoes
• Face or eye protection as needed

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Biosafety Training

• Decontamination
• Decontamination is the reduction of contaminants
  there are two levels of decontamination
  disinfection and sterilization
• Disinfection reduces the number of micoorganisms
  to a level no longer expected to cause disease
  Sterilization is the complete killing of the
  organisms/spores
• Biohazardous materials vary in their resistance
  to decontamination a disinfectant that destroys
  bacteria may be ineffective against viruses or
  fungi gram-negative and gram-positive bacteria
  respond differently bacterial spores are more
  resistant than vegetative forms, and
  non-enveloped, non-lipid-containing viruses
  respond differently than do viruses which have a
  lipid coating

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Biosafety Training

• Decontamination
• Information on the susceptibility of a particular
  microorganism to decontamination can be found in
  the Material Safety Data Sheet (MSDS) for that
  agent
• MSDSs provide additional details such as health
  hazards associated with the microorganism, mode
  of transmission, containment requirements and
  spill response procedures
• MSDSs for biohazardous agents are available on
  the Public Health Agency of Canada website

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Biosafety Training

• Types of Decontamination
• Physical Decontamination
• Heat types of radiation filtration
• Chemical Decontamination
• Liquid Chemicals
• Vapors and Gases

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Biosafety Training

• Types of Chemical Disinfectants
• Choice of a chemical disinfectant depends upon
  type of microorganisms (e.g., spores or
  vegetative cells, bacteria or viruses)
• purpose of treatment (e.g., disinfection or
  sterilization)
• possible interaction with other active chemicals
• presence of organic matter (e.g., blood) that may
  inactivate the disinfectant
• contact time required for disinfection
• toxicity to individuals, culture systems,
  environment, residual toxicity on items
• pH, temperature, hardness of available dilution
  water
• cost

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Biosafety Training

• Classes of Disinfectants
• Chlorine
• Iodine
• Alcohol
• Phenolics
• Quaternary Ammonium
• Glutaraldehyde
• Hydrogen peroxide

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Biosafety Training

• Resistance of Biohazardous Agents to
  Disinfectants
• Prions
• Bacterial spores
• Coccidia (Cryptosporidium)
• Mycobacterium
• Nonlipid viruses (Hep A, Polio)
• Fungi
• Rickettsiae, Chlamydiae
• Vegetative bacteria
• Lipid-containing viruses

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Biosafety Training

• Emergency Procedures Spills
• Spill in a BSC
• Spill outside of a BSC

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Biosafety Training Spills

• Cleaning a Spill in Biosafety Cabinet (BSC)
• Wait at least five minutes to allow the BSC to
  contain aerosols
• Don appropriate PPE laboratory coat, safety
  glasses, nitrile gloves
• With the BSC operating, apply generous amounts of
  Virox to the spill
• Allow a minimum of 20 minutes contact time
• Wipe with disposable, absorbent pads and place in
  a biohazardous waste bag. Wipe the walls, work
  surfaces and any equipment in the cabinet with
  Virox-soaked absorbent towels and place in
  biohazardous waste bag
• Place the biohazardous waste bag in a clean
  biohazardous waste bag for disposal
• Spray entire interior surface BSC with 70
  ethanol and wipe down to ensure that the area has
  been decontaminated
• Run the BSC for an additional 10 minutes after
  the clean-up before resuming work or turning off
  the BSC

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Biosafety Training Spills

• Cleaning a Spill outside of the Biosafety Cabinet
• Evacuate the room and close the door
• Remove any contaminated garments and place in a
  biohazardous waste bag. Even if you dont
  suspect that your skin was exposed, wash your
  face and hands with soap and water
• Warn others not to enter the area, contact your
  Laboratory Manager
• Wait 30 minutes to allow the aerosols created by
  the spill to settle
• Don appropriate PPE laboratory coat, safety
  glasses, nitrile gloves
• Remove broken glass with a dustpan and place
  glass in a sharps container dispose of the
  dustpan in the biohazardous waste.
• Place disposable, absorbent pads or pillows on
  top of the spill.
• Pour Virox around the edges of the spill and then
  onto the pads or pillow. Allow a minimum of 20
  minutes contact time.
• Using additional absorbent materials, wipe up the
  area
• Clean spill area again with Virox-soaked
  absorbent pads
• Place all of the waste in a biohazardous waste
  bag
• Place the waste bag in a clean biohazardous waste
  bag
• Remove PPE and place in biohazardous waste bag
• Wash your hands with soap and water prior to
  leaving the lab

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Biosafety Training Spills

• Cleaning a Spill inside a centrifuge
• Clear the area of all Staff
• Wait 30 minutes to allow the aerosols created to
  settle
• Don appropriate PPE laboratory coat, safety
  glasses, Nitrile gloves
• Remove rotors and bucket and place in the sink
• Thoroughly disinfect the inside of the centrifuge

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