Biological Safety Training

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Biological Safety Training

• modified slightly from REM 2008
• Rachael DeRudder

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Biohazards

• BIOHAZARDS
• A biological hazard or biohazard is a
  self-replicating organism, or substance derived
  from an organism, that poses a threat to
  (primarily) human health. This can include
  medical waste or samples of a microorganism,
  virus or toxin (from a biological source) that
  can impact human health. It can also include
  substances harmful to animals and plants.
• Human body fluids, unfixed tissues, cell lines,
  or genetic oncogenes
• Can also be a fungus, bacteria, prion, parasite,
  or DNA component.

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Exposure

• EXPOSURE to biohazards can occur by
• Injection (contact with open wound or abraded
  skin)
• Inhalation (absorption through respiratory tract
• Ingestion (e.g. contam.fingers in mouth)
• Absorption through mucous membranes (e.g. contam.
  finger in eye or nose)
• Intact skin is regarded as an effective barrier.

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NIH Biohazard Risk Groups

• RG 1 Agents of no or minimal hazard under
  ordinary conditions or handling
• RG 2 Includes agents which may produce disease
  of varying degrees of severity from accidental
  inoculation or injection or other means of
  cutaneous penetration but which are contained by
  ordinary laboratory techniques.

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NIH Biohazard Risk Groups

• RG 3 Includes pathogens that require special
  conditions for containment.
• RG 4 Require the most stringent conditions for
  their containment because they are extremely
  hazardous to laboratory personnel or may cause
  serious epidemic disease.

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Biohazard Classifications
Biosafety levels corresponds to the rating of the
biohazard, i.e., BSL2 is required for Risk Group
(RG) 2 biohazards. BSL2 is the most common type
of biohazard project that occurs on campus.
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BSL2

• Limited access,
• Door posts warnings,
• Surface decontamination methods,
• Waste handling,
• No eating or drinking,
• Hand washing,
• Appropriate personal protective equipment, and
• A Class 2 Biosafety Cabinet
• (if aerosol production is anticipated).

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Bio-Awareness Training

• Specific handling techniques for each RG 2 or
  higher pathogen that is used must be reviewed and
  explained by the Principal Investigator.
• Signs and symptoms for the onset of disease
  caused by the organism must be reviewed.

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Bio-Awareness Training

• Special hazards, i.e., complications for
  pregnancies and immuno-compromised lab staff,
  must be explained and posted.
• Know the proper Personal Protective Equipment
  (PPE).
• Appropriate decontamination procedures must be
  reviewed.
• Any required security steps must be taken.

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PPE

• Personal
• Protective
• Equipment

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• PPE can include the PROPER
• Goggles
• Gloves
• Footwear
• Lab Coats
• For the Task

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Respiratory Protection comes in many varieties.
Choose the appropriate mask for the task.
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Waste Handling
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Waste Handling Biohazard Bags

• BIO-RELATED WASTE MATERIALS
• Dispose of in properly labeled water impervious
  autoclavable bags (usually red or orange bags).
• Do not over fill bags.
• Utilize proper personal protective equipment and
  wash hands after completion.
• Keep bags closed until they are ready to be
  decontaminated.
• Do not dispose of a bio waste bag in the regular
  trash.
• Do not place these bags in public areas such as
  hallways. Contact REM for removal of deconned and
  autoclaved materials.

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Protect yourself from the contents. Hold Bio bags
away from your body. Dont squeeze the bag so
aerosols are released into your breathing space.
Take biowaste to the autoclave room as soon as
possible.
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Waste Handling - Autoclave

• Autoclaves are very effective decontamination
  devices that use high pressure and steam (270?F)
  to eliminate biological activity on equipment,
  wastes, or growth mediums.
• Extreme heat and high pressure produced in
  autoclaves can cause serious injury. Always read
  the operations manual and review usage procedures
  with the lab manager. Users should understand and
  respect the hazards these can create and use the
  proper personal protective equipment, i.e.,
  heat-resistant gloves, lab coat, and face shield.
  
• Do not put the following items in the autoclave
• - Sealed containers - Aerosol cans
• - Volatiles - Uncontained
  sharps

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Waste Handling - SHARPS

• SHARPS are items such as glass and plastic
  pipettes, broken glass, test tubes, petri dishes,
  razor blades, needles, syringes, etc. that are
  capable of puncturing, cutting, or abrading the
  skin.
• Properly handle, decontaminate, and dispose of
  sharps in order to prevent injury and potential
  disease transmission.

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DO NOT PLACE SHARPS CONTAINERS IN THE REGULAR
TRASH! CALL REM FOR PICKUP
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ALWAYS place sharps in puncture resistant
containers for disposal.
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In the event of a spill Use the proper PPE Use
the proper disinfectant solution. Dispose of
clean-up materials in biohazard bags.
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Security

• EVERYONE is responsible for security in the
  laboratory.
• If you are using RG 2 biohazards make sure these
  materials are secured.
• Keep your lab doors locked as well as the doors
  to areas outside your lab where RG 2 organisms
  are incubated, stored, or otherwise processed.
• Ask for identification from unknown individuals
  who enter your lab. Do not allow access to
  unauthorized people.
• Secure biohazardous waste and autoclave as soon
  as possible.
• Report the disappearance of any biohazardous
  material to your Supervisor.
• Special security measures apply to users of
  Select Agents. See the Biosafety Officer for
  more specific details.

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HEPA Filtered Cabinets

• Biological Safety Cabinet Laminar Flow Clean
  Bench
• HEPA stands for
• High
• Efficiency
• Particulate
• Air

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Biological Safety Cabinets
Laminar Flow Clean Benches

• Protects operator
• Protects the environment
• Protects the product
• Reduces the risk of airborne infection by
  reducing the escape of aerosols

• Protects the product
• Blows aerosols into the operators face.
• Does not protect the operator
• Should not be used with potential pathogens
• Is not a Biological Safety Cabinet

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BIOSAFETY CABINET
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BSC Safe for Biohazard Use
Visual Indicator
Sash
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BIOSAFETY CABINET AIRFLOW
(Courtesy of Eagleson Institute)
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BIOSAFETY CABINETS

• Class II Biosafety Cabinets, or BSCs, use
  filtered air to isolate materials from the
  operator and work environment.
• BSCs use multiple filtration systems, fans, and
  air-flow paths that must be carefully controlled,
  maintained and checked to ensure operator safety.
  Most BSCs will not be vented to the outside.
• BSCs protect both sample and workers from
  contaminating each otherdesigned to contain
  biological hazards (particles) not chemical fumes
  vapors.
• There are other Classes of Biosafety Cabinets.
  Not to be confused with Clean Bench or Chemical
  Fume Hood.

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Working Safely in a BSC

• Turn the blower on at least 5 minutes BEFORE
  beginning work to allow the BSC to remove any
  particulates in the cabinet.
• Check the Magnehelic gauge before using every
  time . Reading should be equal to approximately
  0.5 inches to assure proper operation of the
  cabinet before placing any materials into it.
  Higher readings may indicate filter clogging.
  Zero readings may indicate loss of filter
  integrity.

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Working Safely in a BSC

• Use the APPROPRIATE Personal Protective Equipment
  (PPE) while working at the BSC.
• Place necessary materials in the BSC BEFORE
  beginning work to minimize the number of arm
  movement disruptions to the air barrier of the
  cabinet.
• When working in a BSC, work with arms raised
  slightly above the front grille, NOT resting
  flatly on it. This alleviates the problem of room
  air flowing directly into the work area.

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Working Safely in a BSC

• DONT place objects directly on the grillwork at
  the opening or back of the cabinet. This greatly
  interferes with the laminar airflow curtain.
• After placing your equipment inside the cabinet,
  close the sash to the proper operating height.
• AVOID working in and operating the cabinet with
  the sash in any other position than the
  manufacturers recommended sash height.

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Working Safely in a BSC

• The workers face should be ABOVE the front
  opening of the BSC.
• All work should be performed at least 4 inches
  from the front grille on the work surface.

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Completing Work in the BSC

• All equipment that has come in contact with a
  biological agent should be decontaminated. 
• The cabinet should be allowed to run for at least
  3 minutes with no activity.

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Completing Work in the BSC

• Wipe down all surfaces with the appropriate
  disinfectant, including the cabinet sides, back,
  and the interior of the glass.
• Decontaminate or, where appropriate, autoclave
  all disposable materials and wastes before
  removal from the laboratory.
• Call REM for pickup.

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More Errors to Avoid

• NEVER disengage the alarm. Alarm indicates
  improper airflow that could effect performance
  and endanger the researcher or the experiment.
• Never completely close the window sash with the
  motor running.  This could damage the motor.

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CLEAN BENCH UNSAFE for Biohazard Use
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CLEAN BENCH AIRFLOW
POTENTIALLY CONTAMINATED AIR
HEPA FILTERED AIR
ROOM AIR
Courtesy of Eagleson Institute
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CLEAN BENCH

• Clean Benches typically provide only product
  protection by creating a unidirectional airflow
  through a HEPA filter.
• The discharged air goes directly into the
  workroom.
• These cabinets are not safe for work with
  biohazards, chemicals, or radioactivity.

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REMEMBER

• A Biosafety Cabinet is only as safe as the person
  using it.
• Use the APPROPRIATE cabinet type for the
  materials being used.
• Make sure the cabinet is working properly BEFORE
  beginning work.
• Consult the supervisor about questions or
  concerns regarding working in a Biosafety Cabinet.

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Select Agents

• Agents that could be used for terrorist acts
• Highly regulated
• Stiff penalties
• FBI background checks
• High security
• No one at Purdue has been approved for Select
  Agents at this time.

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Resources

• Center For Disease Control and Prevention
•  http//www.cdc.gov/
•  Select Agent Program
•  http//www.cdc.gov/od/sap/
•  Biosafety in Microbiological and Biomedical
  Laboratories 5th Edition, January 20007
•  http//www.cdc.gov/od/ohs/biosfty/bmbl5/BMBL_5th_
  Edition.pdf/
•  

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Resourcescontinued

• Material Safety Data Sheets For Research
  Pathogens
• http//www.phac-aspc.gc.ca/msds-ftss/index.html
• Purdue University Biosafety Manual
•  http//www.purdue.edu/rem/home/booklets/bioman.pd
  f
• Guidelines for Research Involving Recombinant DNA
  Molecules (NIH Guidelines)
• http//www4.od.nih.gov/oba/rac/guidelines/guidelin
  es.html