Biological Safety Training

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Biological Safety Training
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Introduction to Biological Safety

• Biological Hazards are divided into 4 Biosafety
  Levels
• BSL 1
• BSL 2
• BSL 3
• BSL4
• Biosafety levels define the lab requirements,
  protective clothing, and work practices

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BioSafety Definitions

• BSL 1 (BMBL)
• Not known to consequently cause disease in
  healthy human adults
• Pose minimal hazards under ordinary conditions of
  handling

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BioSafety Level 1

• Standard Practices
• Use Mechanical Pipetting devices
• No Eating, Drinking, Smoking in Lab
• Minimize splashes and aerosols
• Decontaminate work surfaces
• Safe handling of sharps
• Wash Hands before leaving lab

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BioSafety Level 1

• Protective Clothing
• Lab Coat
• Gloves
• Eye Protection

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BioSafety Definitions

• BSL 2 (BMBL)
• suitable for work involving well-characterized
  agents not known to cause disease in healthy
  adult humans, and of minimal potential hazard to
  laboratory personnel and the environment
• Examples Measles Virus, Salmonella,
• Hepatitis B Virus

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BioSafety Definitions

• Other Materials Handled at BSL 2
• (Requires compliance with OSHA BBP Standard)
• Human Blood
• Human Blood Components
• Human Tissues
• Human Cell Lines

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BioSafety Level 2

• Standard Practices
• All requirements for BSL 1 plus
• Access to laboratory is limited or restricted
  when work is being conducted
• Personnel have specific training in handling
  pathogenic agents
• Biohazard Sign posted on the door
• Extreme precautions are taken with contaminated
  sharp items

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BioSafety Level 2

• Standard Practices cont.
• Written Biosafety Procedures
• Report Spills
• Minimize aerosol generation
• Personnel receive appropriate immunizations or
  test (e.g. Hep B vaccine or TB skin Testing)

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BioSafety Level 2

• Written Biosafety Procedures
• Prepared specifically for lab by PI
• Incorporated into Lab Safety Plan
• Personnel are advised of special hazards
• Personnel are required to read and follow
  procedures

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BioSafety Level 2

• Personal Protective Equipment
• Lab Coats
• Gloves
• Double Gloving when appropriate
• Eye Protection - Safety Glasses/Shield

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BioSafety Level 3

• BSL 3 (BMBL)
• applicable to clinical, diagnostic, teaching,
  research, or production facilities in which work
  is done with indigenous or exotic agents which
  may cause serious or potentially lethal disease
  as a result of exposure by the inhalation route.

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BioSafety Level 3

• Standard Practices
• All requirements for BSL 2 plus
• Limited lab access
• 2 doors in a series to access lab
• Able to wash entire lab
• Special exhaust ventilation (Not re circulated,
  negative pressure)

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BioSafety Level 3

• Standard Practices cont.
• High level of training
• Personnel receive vaccinations if available
• Work in Biosafety cabinets

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BioSafety Level 3

• Examples
• Mycobacterium Tuberculosis
• Hanta virus
• SARS

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BioSafety Level 4

• BSL 4 (BMBL)
• required for work with dangerous and exotic
  agents which pose a high individual risk of
  aerosol-transmitted laboratory infections and
  life-threatening disease.

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BioSafety Level 4

• Requirements
• All requirements for BSL3 plus
• Class III Biosafety cabinet or positive pressure
  suits
• Shower/Change rooms
• Clothing Autoclaved before laundering
• Air Locks

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BioSafety Level 4

• Examples
• Ebola Virus
• Monkey B Virus
• Marburg Virus

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Containment

• Primary
• Protection of personnel and immediate lab from
  exposure
• Good Techniques
• Safety equipment
• Biological Safety Cabinet
• Engineering Controls
• PPE
• Serologic surveillance

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Containment

• Secondary
• Protection of environment external to lab
• Facility Design
• Operational practices

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Safe Work Practices for all Levels

• Wash hands after work when removing gloves
  before leaving lab
• No eating, drinking, applying cosmetics, handling
  contact lenses in the lab.
• Maintain labs in clean, orderly fashion.

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Safe Work Practices for all Levels

• Limit access to lab when work with organisms is
  in progress
• Use good microbiological techniques
• No mouth pipetting
• When possible use plastic instead of glass

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Sharps Safety

• Sharps include needles, syringes, razor blades,
  lancets, slides, scalpels, pipettes,
  micropipettes, pipette tips, broken plastic or
  glassware, and other devices capable of cutting
  or piercing the skin.

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Sharps Safety

• Contaminated needles shall not be bent, recapped,
  or removed unless there is no feasible
  alternative.
• If required, use a mechanical device or a one
  handed technique.

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Sharps Safety

• Safety devices or alternatives to needles should
  be used when available.

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Sharps Disposal

• Sharps containers for disposal of these items
  should be conveniently located and easily
  accessible in all work places where sharps are
  used.

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Sharps Disposal

• Syringes and syringes without a needle attached
  go into a sharps container
• Contaminated micropipettes, pipette tips, and
  Pasteur pipettes are discarded in a
  puncture-resistant container or a sharps
  container for disposal.

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Sharps Disposal

• Dont place needles or sharps in office waste
  containers

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Laminar Flow Equipment

• BioSafety Cabinets (BSCs)
• contain infectious agents to protect personnel
  and the environment
• Laminar Flow Clean Benches (LFBs)
• Non Hazardous work only
• Protect work from contamination

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BioSafety Cabinets

• 3 Classes
• All exhaust is HEPA filtered before leaving the
  cabinet
• Class I
• Do not protect the work from contamination
• Air entering cabinet is not filtered

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Biosafety Cabinets

• Class II (4 types A, B1, B2, and B3)
• Each type recirculates different amount of air
• Some are hard ducted, and some exhaust into the
  room.
• Class III
• Totally enclosed, ventilated cabinets
• Work through portals with attached gloves

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BioSafety Cabinets

• All equipment is laid out to not restrict airflow
  in the cabinet

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Laminar Flow Benches

• Do not protect personnel or the environment
• Discharges HEPA filtered air across the work
  surface toward the user.

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Use of Laminar Flow Equipment

• Minimize airflow disturbances (moving in/out of
  cabinet, people walking by, opening doors,
  blocking grills with equipment)
• Wipe down surfaces with approved disinfectant
  before and after work.

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Use of Laminar Flow Equipment

• Minimize use of flames in cabinet
• Try not to use chemicals in Class I or II hoods
  that are ducted
• Do not store excess equipment in cabinet
• All laminar flow equipment must be certified
  annually.

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Centrifuges
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Centrifuge Hazards

• Mechanical failure of Machine
• Lab equipment failure (tubes etc.)
• Aerosol generation
• Operator error

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Centrifuge Operation

1. Check tubes for cracks/chips
2. Use matched sets of tubes, buckets, etc. Make
   sure the rotor is balanced properly
3. Tightly seal all tubes and safety cups
4. Ensure that rotor is locked to spindle and the
   bucket is seated
5. Close lid during operation
6. Allow to come to complete stop before opening

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Centrifuge Safety

• Use sealed tubes, safety buckets or rotors when
  possible
• When possible fill and open centrifuge tubes or
  buckets in the biological safety cabinet.
• Lubricate O-rings and rotor threads weekly
• Disinfect weekly and after all spills or breaks

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Biohazardous Waste

• All cultures, stocks, and other regulated waste
  are decontaminated before disposal by an approved
  decontamination method such as autoclaving.

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Biohazardous Waste

• Examples
• stock cultures
• materials containing or contaminated with blood
• blood and body fluids
• sharps
• pipette tips
• animal waste
• other contaminated lab material

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Blood and Body Fluids Waste

• Serum,
• Plasma
• Other blood components
• Semen
• Vaginal secretions
• Cerebrospinal fluid
• Pleural fluid
• Peritoneal fluid
• Pericardial fluid
• Amniotic fluid
• Any other body fluid visibly contaminated with
  blood

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Infectious Sharps Waste

• Hypodermic needles
• Scalpels
• Pipettes /pipette tips
• Breakable containers
• Glass products (i.e., slides or cover skips)

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Biohazardous Waste

• Inappropriate materials
• Dispose of noninfectious waste in regular trash

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Biohazardous Waste

• Inappropriate materials
• These include aerosol cans, chemical waste,
  flammable material

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Biohazardous Waste

• Inappropriate packaging
• Use puncture resistant package if needed

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Biohazardous Waste Containers

• Biohazardous waste containers shall be clearly
  marked with the universal biohazard symbol.