Biological Safety

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Biological SafetyIts in Your Hands
University of Kentucky Department of Biological
Safety
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Biological Safety Training

• Module 1 Basics of Biosafety
• Module 2 Risk Assessment
• Module 3 Working with Infectious Agents
• Module 4 The Biosafety Levels
• Module 5 The Biosafety Audit

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Biosafety Levels

• Do you know your labs biosafety level?
• What are the proper procedures and practices for
  your biosafety level?
• How does waste need to be decontaminated at your
  biosafety level?

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Biosafety Practices and Procedures

• The University of Kentucky adheres to the
  guidelines presented in the CDC publication,
  Biosafety in Microbiological and Biomedical
  Laboratories
• Each laboratory is required to have a copy
  available as a paper version or a webpage link on
  the laboratory computer
• http//www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.h
  tm

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Can you recognize safe practices?Whats wrong
with these pictures?
Biological Safety Levels in the Lab
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Biosafety Levels at UK

• Registration with the Department of Biological
  Safety is required for BSL-1 projects
• Approval from the Institutional Biosafety
  Committee (IBC) is required for BSL-2 and BSL-2
  projects
• Approval from the IBC and possibly other entities
  is required for BSL-3 projects
• BSL-4 projects are not allowed at UK

Biosafety Levels Build on Each Other
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Procedures and Practices for BSL-1 Labs

• Wash hands often
• Know location of safety showers and eyewashes
• Wipe down work surfaces with disinfectant daily
  and after spills
• No eating, drinking, smoking, applying cosmetics,
  handling contact lenses

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Procedures and Practices for BSL-1 Labs

• Label biohazard use and storage areas
• Decontaminate waste prior to disposal
• Keep the laboratory neat and organized
• Proper sharps and broken glass disposal is in
  place

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Procedures and Practices for BSL-1 Labs

• Mouth pipetting is prohibited
• Access to the laboratory is limited during
  experiments
• Doors are locked when the laboratory is
  unoccupied
• Work is performed carefully to minimize splashes
  and aerosols

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Personal Protective Equipment for BSL-1 Labs

• No Sandals, No Shorts
• Lab Coats worn only in lab
• Gloves as needed for procedures
• Protective eyewear if splashes are anticipated

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Procedures and Practices for BSL-2 Labs

• All BSL-1 practices are followed, plus
• Access is restricted and laboratory doors are
  labeled when work with infectious agents is
  performed
• All personnel working in the lab must have
  adequate lab-specific training
• Appropriate immunizations are received if
  available

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Procedures and Practices for BSL-2 Labs

• Nothing stored in the biosafety cabinet and
  anything used in the biosafety cabinet properly
  decontaminated before removal.
• Sharps and glassware should be avoided if
  possible
• Substitute plastic for glass
• Use safer sharps
• Self-sheathing needles

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Procedures and Practices in a BSL-2 Lab

• Approved secondary container utilized
• for transport.
• Sealed centrifuge buckets or rotors preferred.
• Aspiration used with proper trap and filter setup.

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Safety Equipment in a BSL-2 Lab

• Biological Safety Cabinets are utilized
• Nothing stored in the biosafety cabinet and
  anything used in the biosafety cabinet properly
  decontaminated before removal.
• Personal Protective Equipment
• Gown
• Gloves
• Face and Eye Protection

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Practices and ProceduresBSL-2 Lab

• Sharps not used to harvest virus solution or for
  culture of infected cells.
• Aspiration not used with infected cultures or
  proper trap and filter setup utilized.
• Properly dispose of sharps
• Call Environmental Management (3-6280) for
  disposal if laboratory located outside of the
  Hospital or Medical Sciences Building

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Practices and ProceduresBSL-2 Lab

• The door to the culture room will remain closed
  at all times and only necessary personnel will be
  permitted to use the room when experiments with
  infectious agents are being performed.

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Practices and ProceduresBSL-2 Lab

• One-glove rule utilized for transport of
  materials to centrifuge.
• Approved secondary container utilized for
  transport of infectious agent.
• Centrifuge buckets or sealed rotors opened/closed
  only in BSC and the outside is decontaminated in
  BSC.
• Cultures housed in a dedicated and labeled
  incubator.

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Personal Protective Equipment in BSL2 Lab

• Face Shield/Safety Goggles and Face Mask for
  Splash Protection Outside BSC
• Respiratory Protection as needed
• Disposable Wraparound Lab Gown
• Double Gloves
• Standard lab safety dress code will be followed
  (no shorts or open toed shoes, etc.) as required
  by OSHA.

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Practices and ProceduresBSL-3 Lab

• A lab specific biosafety manual addressing all
  precautions and procedures must be in place
• Negative pressure, HEPA filtered exhaust air is
  utilized in the facility
• All penetrations in outer walls sealed
• Entry is through double self-closing doors
• BSL-3 facilities must be tested and verified for
  containment

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Personal Protective Equipment in BSL-3 Lab

• Lab-specific risk assessments can determine
  appropriate PPE
• Additional protective equipment may be necessary
  in a BSL-3 facility
• Detailed standard operating procedures are
  written for entering and exiting the facility and
  PPE required

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Personal Protective Equipment in BSL-3 Lab

• Respiratory protection may be necessary depending
  upon risk assessment
• Extra layer of PPE required for work with
  infected animals

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Decontamination and Waste Disposal Procedures

• In BSC, all disposables are either inactivated
  with 0.5 sodium hypochlorite or placed in an
  autoclavable cardboard box when working with
  infectious materials.
• The box placed in an autoclave bag, the outside
  of the autoclave bag decontaminated, bag removed
  from the biosafety cabinet and put into another
  autoclave bag to be autoclaved under appropriate
  conditions.

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Decontamination and Waste Disposal Procedures

• BSC surfaces thoroughly wiped with disinfectant.
• Biohazard Waste Containers
• hard-sided
• foot operated lid
• easily decontaminated with liquid disinfectant
• only the inside of the autoclave bag is visible

Fisher Scientific https//www1.fishersci.com/index
.jsp
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Types of waste
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What is Medical Waste?

• Definition Medical waste is generally defined
  under state regulations. Medical waste is often
  described as any solid waste that is generated in
  the diagnosis, treatment, or immunization of
  human beings or animals, in research pertaining
  thereto, or in the production or testing of
  biologicals, including but not limited to
• blood-soaked bandages
• discarded surgical instruments - scalpels
• needles - used to give shots or draw blood
• removed body organs - tonsils, appendices, limbs,
  etc.
• lancets
• intentionally infected animal carcasses

Environmental Protection Agency
http//www.epa.gov/epaoswer/other/medical/
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Autoclaving Biohazardous Waste

• Wear buttoned lab coat, eye protection,
  closed-toed shoes, heat resistant gloves.
• Add 1 liter of water to properly vented clear or
  orange autoclave bag by pouring carefully down
  the side of the bag
• Use a solid secondary container or pan approved
  for autoclave use
• Promptly remove cooled autoclaved biohazardous
  waste to trash
• Record use in user log

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Review of Training Objectives

• Equipment for Biological Safety in the Lab
• Biological Safety Cabinet
• Closed Doors
• Personal Protective Equipment (PPE)
• Appropriate PPE and Practices
• No Shorts, No Sandals, Right PPE for the Job
• Decontamination of Waste and Work Surfaces

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Whats wrong with these pictures?
Food or Drink in Lab
No Gloves Worn While Working in BSC
Overfilled Autoclave Bag
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Need More Information?

• For more information on the topics covered in
  this training check out these websites
• UK Department of Biological Safety
• http//ehs.uky.edu/biosafety/
• Biosafety in Microbiological and Biomedical
  Laboratories
• http//www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.h
  tm
• NIH Guidelines for Recombinant DNA
• http//www4.od.nih.gov/oba/rac/guidelines/guidelin
  es.html

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Questions?
Marcia Finucane Biological Safety
Officer Department of Biological Safety 505
Oldham court Lexington, KY 40502 Phone 
859-257-1049 Cell  859-699-1553 mfinu2_at_email.uky.
edu http//ehs.uky.edu/biosafety