BIOSAFETY TRAINING

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BIOSAFETYTRAINING

• Tina Preseau
• Rita Toussaint-Archambault?
• Office of Risk Management
• ?Human Resources - Occupational Health
  Disability Leave

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COURSE OUTLINE

• Introduction
• Laboratory Associated Infections
• Blood-borne Pathogens
• Classification of Biohazards
• Infection/Biohazard Control
• Spill Response
• Biomedical Waste
• Regulations

BIOSAFETY
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INTRODUCTION
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WHAT IS A BIOHAZARD?

• A potential hazard to humans, animals or the
  environment caused by a biological organism, or
  by material produced by such an organism
• Examples
• Viruses, bacteria, fungi, and parasites and their
  product.
• Blood and body fluids, as well as tissues from
  humans and animals.
• Transformed cell lines and certain types of
  nucleic acids .

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WHAT IS BIOSAFETY?

• Measures employed when handling biohazardous
  materials to avoid infecting oneself, others or
  the environment.
• Achieved through
• Administrative Controls
• Engineering Controls
• Personal Protective Equipment
• Practices and Procedures

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WHAT IS BIOSECURITY?

• Measures employed to protect biohazardous
  materials, or critical relevant information,
  against theft or diversion by those who intend to
  pursue intentional misuse.
• Achieved through
• Physical barriers
• Psychological barriers
• Monitoring Activities
• Personnel Clearance

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WHO ARE THE STAKEHOLDERS?

• INTERNALLY
• Vice-President (Research)
• Committees
• University Services (ORM, HR, PRS, PS)
• Deans, Chairs, Principal Investigators,
  Employees, Students
• Manager of Biological Containment Suite

• EXTERNALLY
• Public Health Agency of Canada
• Canadian Food Inspection Agency
• Environment Canada
• Transport Canada
• Ontario Ministry of Labour
• Emergency Response Personnel
• Suppliers Contractors
• Community

ACCOUNTABILITY
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KEY SERVICES

• Office of Risk Management
• Training
• Interface with Regulatory Bodies
• Biosafety Program

certifications training procedures
inspections contingency planning
accident/incident follow-up
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KEY SERVICES

• HR (Occupational Health, Disability and Leave)
• Medical surveillance
• Immunizations
• Medical Follow-up
• Interface with Workplace Safety and Insurance
  Board

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WHY ARE WE CONCERNED?

• Potential for acquiring a laboratory-associated
  infection (LAI)
• Contamination of the environment
• Contamination of research
• Public perception

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LABORATORY ASSOCIATED INFECTIONS
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LABORATORY ASSOCIATED INFECTIONS
Infection Source

• Cultures and stocks
• Research animals
• Specimens
• Items contaminated with above

Route of Transmission
Susceptible Host

• Percutaneous inoculation
• Inhalation of aerosols
• Contact of mucous membranes
• Ingestion

• Immune system
• Vaccination status
• Age

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LAIS

• Only 20 causative or defined event
• 80 of which are caused by human factors
• 20 are caused by equipment failure
• Top 4 accidents resulting in infection
• Spillages splashes
• Needle and syringe
• Sharp object, broken glass
• Bite or scratch from animals or ectoparasites

http//www.weizmann.ac.il/safety/bio2.html
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LAIS
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BLOOD-BORNE PATHOGENS
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BLOODBORNE PATHOGENS (BBP)

• Sources
• Blood
• Semen
• Vaginal Secretions
• Other Bodily Fluids
• Cerebrospinal
• Amniotic
• Synovial
• Tissue Cultures
• Organ Cultures
• Infected Experimental Animals

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RISK OF EXPOSURE

• Pathogen involved
• Type of body fluid
• Route of exposure
• Duration of exposure
• Volume of blood involved in exposure
• Concentration of virus at time of exposure
• PPE worn

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SPECIFIC EXAMPLES OF BBPS

• Hepatitis B
• Hepatitis C
• HIV

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ISSUES TO CONSIDER

• Symptoms
• Mode of transmission
• Incubation period
• Survival outside host
• Communicability
• Immunization
• Prophylaxis / Treatment

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IF AN EXPOSURE OCCURS

• Initiate first aid
• Notify your supervisor / designated person
• Report to hospital emergency department or
  Universitys Health Services
• Report incident to OHDL
  
• Occupational Health, Disability and Leave Office
  telephone ext. 1472 http//www.rh.uottawa.ca/00_
  main/index_f.asp

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UNIVERSAL PRECAUTIONS

• Minimum standard of practice for preventing the
  transmission of BBP includes
• Education
• Hand washing
• Wearing protective barriers
• Use safe work practices

If samples cannot be guaranteed non-infective
treat as infectious!
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BIOHAZARD CLASSIFICATION
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BIOHAZARD CLASSIFICATION

• Conventional Agents
• Recombinant DNA
• Tissue Culture
• Animal Work
• Anatomical Specimens
• Unconventional Agents

Class D, division 3 of WHMIS (Poisonous and
Infectious Material - Biohazardous Infectious
Material)
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BIOHAZARD CLASSIFICATION

• Organisms are categorized into a group base on
  the particular characteristics of each organism,
  such as
• Pathogenicity
• Infectious dose
• Mode of transmission
• Host Range
• Availability of effective preventive measures
• Availability of effective treatment

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BIOHAZARD CLASSIFICATION

• Organisms are categorized base on the measures
  required for handling each organism safely in a
  laboratory setting, such as
• Engineering Requirements
• Operational Requirements
• Technical Requirements
• Physical Requirements

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CONVENTIONAL AGENTS
Unlikely to cause disease in healthy workers or
animals
Rarely cause serious human or animal disease
May cause serious disease
Likely to cause very serious disease
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RECOMBINANT DNA

• Genetic Engineering in vitro incorporation of
  genetic material from one cell into another
• In Canada the level of risk depends on source of
  DNA, vector and host.
• The Office of Risk Management will assist the
  investigator in this determination.

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TISSUE CULTURE

• Have the potential to contain pathogenic
  organisms
• In general
• Human non-human primate, and mycoplasma-containi
  ng cell lines Level 2
• Others Level 1

A detailed risk assessment should be undertaken
when using a new cell line.
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ANIMAL WORK

• Animals can harbour infectious organisms
  (naturally or introduced)
• Level dependent on type of work being conducted.
• Special Animal Care training is required for all
  personnel working with animals.
• All work involving animal use must receive prior
  approval from the Animal Care Committee

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ANATOMICAL SPECIMENS

• All specimens should be considered infectious due
  to potential presence of infectious agents
• Important to consider the type of specimen
• blood, organs, tissues
• Spinal sample, brain tissue
• From infectious patient
• In general Level 2 but it depends on the nature
  of the work.

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UNCONVENTIONAL PATHOGENS

• TSE prion diseases lethal transmissible
  neurodegenerative conditions
• Creutzfeld-Jakob disease, Variant C-J Disease,
  Mad Cow Disease, Scrapie, Chronic Wasting Disease
• Resistant to destruction by procedures that
  normally inactivate viruses.
• Contact ORM to assess requirements (containment,
  procedures, waste disposal, etc.)

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WHERE CAN WE FIND THESE?
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INFECTION/BIOHAZARD CONTROL
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INFECTION/BIOHAZARD CONTROL

• Administrative Controls
• Engineering Controls
• Personal Protective Equipment
• Practices and Procedures

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ADMINISTRATIVE CONTROLS

• Program based, information and methods to
  minimize risk of exposure
• Risk assessment
• Medical Surveillance
• Training/Education
• Resources
• Inspections
• Signs Labeling

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ADMINISTRATIVE CONTROLS

• Risk Assessment
• Will determine type of containment, procedures,
  and safety equipment required
• Responsibility of users, additional assistance is
  available from the ORM
• Consider areas such as experimental design,
  procedures to be employed and personal
  experience/knowledge, etc.
• Know and understand the various characteristics
  of the agent(s) you are working with. (Material
  Safety Data Sheets and suppliers or
  manufacturers)

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ADMINISTRATIVE CONTROLS

• Medical Surveillance
• Training Education
• WHMIS
• Lab specific policies and procedures
• Biosafety training
• Resources
• ORM web site, Biosafety page
• Faculty web sites
• Biosafety Manual
• Training Videos

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ADMINISTRATIVE CONTROLS

• Inspections
• Routine self-inspections
• Biosafety Inspection Checklist available on-line
• In addition, ORM, EHSOs and OHS will inspect
  labs to ensure compliance with regulations/
  guidelines and provide feedback.

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ADMINISTRATIVE CONTROLS

• Signs Labeling
• Biohazard warning signs must be posted on doors
  to rooms where biohazardous materials are used.
• Biohazard labels should be placed on containers,
  equipment and storage units used with biological
  agents.

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ENGINEERING CONTROLS

• Technology based, reduce or eliminate exposure to
  hazards by changes at the source of the hazard.
• Containment
• Types Primary and Secondary
• Containment levels

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PRIMARY CONTAINMENT

• First line of defence.
• Ensures protection of personnel and immediate
  environment from exposure to the infectious
  agent.
• Protective envelope that encapsulates the
  infectious agent or animal.
• Petrie dish, vial, stoppered bottle.
• Biological safety cabinets, glove boxes and
  animal caging equipment, etc.
• Effectiveness of control is based on the
  integrity of the containment.

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SECONDARY CONTAINMENT

• Protects the environment external to the
  laboratory from exposure
• Includes facility design and operational practices

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CONTAINMENT LEVEL 1

• Basic laboratory
• Requires no special design features
• Biosafety cabinets are not required and work may
  be performed on the open bench.

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CONTAINMENT LEVEL 2

• Clinical, diagnostic, research and teaching
  facilities with level 2 agents.
• Requires a class I or class II biological safety
  cabinet if any potential for aerosol or splash
  exists.
• An emergency plan for handling spills must be
  developed.
• Access should be controlled.

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CONTAINMENT LEVEL 3

• Specialized design and construction
• primary barriers to protect the individual
• secondary barriers to protect the environment
• All staff must undergo special training on the
  agents being used, PPE, equipment, waste
  management as well as practices and procedures
  above and beyond the scope of this course.

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CONTAINMENT LEVEL 4

• Only one level 4 facility in Canada (Canadian
  Centre for Human and Animal Health in Winnipeg,
  Man.)
• Design specifications are extremely stringent,
  worker is completely isolated from infectious
  material.

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BIOLOGICAL SAFETY CABINETS

• Effective means of primary physical containment
  for biological agents, especially when aerosols
  are generated.
• HEPA filters remove particles (min 0.3 microns)
  with 99.97 efficiency.
• There are 3 main classes of cabinets (I, II,
  III) which provide various levels of protection.

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BIOLOGICAL SAFETY CABINETS

• Laminar flow hoods
• Vertical or horizontal laminar flow
• HEPA filtered air (intake only)
• Product protection only

• Biological Safety Cabinet
• HEPA filtered laminar air flow
• Exhaust
• Personnel, environment product protection

VS
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WORKING SAFELY IN A BSC

• Before using the cabinet
• Ensure BSC is certified
• Turn off UV lamp turn on fluorescent lamp
• Disinfect work surfaces with appropriate
  disinfectant
• Place essential items inside cabinet
• Allow the blower to run for 5-10 min before work

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WORKING SAFELY IN A BSC

• While using the cabinet
• Ensure material and equipment is placed near the
  back of the hood, especially aerosol-generating
  equipment. Do not block any vents
• Use techniques that reduce splatter and aerosols.
• General work flow should be from clean to
  contaminated areas
• Minimize movement so as not to impede air flow
• Open flame in BSCs is controversial

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WORKING SAFELY IN A BSC
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WORKING SAFELY IN A BSC

• After using the cabinet
• Leave blower on at least 5 minutes to purge
  cabinet
• Remove and decontaminate equipment and materials
• Disinfect cabinet surfaces
• Turn off blower and fluorescent lamp, turn on UV
  lamp

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WORKING SAFELY IN A BSC

• Maintenance
• Before and after each use - Work surfaces wiped
  down
• Weekly - UV lamp should be wiped clean
• Monthly - All vertical surfaces wiped down
• Annually - UV lamp intensity verified
• - Decontamination with formaldehyde gas (ORM)
• - Certification (ORM)

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PERSONAL PROTECTIVE EQUIPMENT (PPE)

• PPE can become an important line of defence (last
  line of defense)
• Responsibility of both the user and the
  supervisor to ensure that PPE is worn

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PPE

• Criteria for consideration
• Routes of exposure that need to be blocked
• Degree of protection offered
• Ease of use
• Only effective if correctly selected, fitted,
  used and cared for, and the individual is trained
• Ensure PPE is removed before leaving the lab

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PPE

• Footwear
• Closed toed shoes should always be worn. Booties
  are worn in some higher containment labs and
  animal facilities.
• Lab Coats/Gowns
• Long-sleeved, knee length with snaps
• Elastic cuffs
• Back-closing gowns
• Periodic cleaning required

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PPE

• Gloves
• Latex, nitrile vinyl for work with biological
  agents.
• Exam gloves should not be reused, change
  frequently. Utility gloves can be disinfected and
  reused if they show no sign of degradation.
• Consider tensile characteristics, length of cuff.
• Double gloving.
• ORM can provide assistance finding an alternative
  for people with allergies.

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PPE

• Eye Face Protection
• Goggles, safety glasses to protect the eyes
• Full face shield to protect facial skin.
• Respirators
• Only personnel who have been fit-tested and
  trained should wear respirators.

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PRACTICES AND PROCEDURES

• General Safety Guidelines
• Good Microbiological Practice
• Handwashing
• Suspicious Packages
• Specific Procedures
• Centrifuges
• Needles Syringes and other sharps
• Pipettes
• Blenders, Grinders, Sonicators Lyophilizers
• Inoculation Loops
• Cryostats

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GENERAL LABORATORY SAFETY GUIDELINES

• Mostly common sense, but you must understand the
  hazards you face in the laboratory and be
  adequately trained to deal with them.
• Basic must be known for all labs.

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GOOD MICROBIOLOGICAL PRACTICE (GMP)

• Basic code of practice that should be applied to
  all types of work involving microorganisms.
• Objectives
• prevent contamination of laboratory workers and
  the environment
• prevent contamination of the experiment/samples
• Application of aseptic technique, minimization of
  aerosols, contamination control, personal
  protection, emergency response

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HANDWASHING

• One of the single effective means of preventing
  infections if done properly and frequently
• When to wash?
• Before starting any manipulations
• Before leaving the lab
• When hands are obviously soiled
• Before and after completing any task in a BSC
• Every time gloves are removed
• Before contact with ones face or mouth
• At the end of the day

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SUSPICIOUS PACKAGES

• Unopened
• Do not open and do not shake
• Place in secondary container or cover
• Inform others of the situation
• Clear the room and section off the area
• All individuals who may have come into contact
  with the material must wash their hands
• Call Protection Services and wait for their
  arrival
• List all the individuals present in the room or
  area when the package arrived. Give this list to
  Protection Services for follow-up

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SUSPICIOUS PACKAGES

• Opened
• Contents Intact
• Do not manipulate contents further
• Cover the package
• Inform others of the situation
• Clear the room and section off the area
• All individuals who may have come into contact
  with the material must wash their hands
• Call Protection Services and wait for their
  arrival
• List all the individuals present in the room or
  area when the package arrived. Give this list to
  Protection Services for follow-up

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SUSPICIOUS PACKAGES

• Contents not intact (spilled)
• Do not try to clean up the spill
• Gently cover the spill
• Inform others of the situation
• All individuals who may have come into contact
  with the material must wash their hands
• Call Protection Services
• Remove heavily contaminated clothing (place in
  bag) and shower using soap and water
• List all the individuals present in the room or
  area when the package arrived. Give this list to
  Protection Services for follow-up

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SAFE USE OF CENTRIFUGES

• Before use
• Stress lines? Overfilled? Balanced?
• Caps or stoppers properly in place?
• Run conditions achieved?
• Use sealable buckets (safety cups) or sealed
  rotors
• After run
• Centrifuge completely stopped?
• Spills or leaks?
• Allow aerosols to settle (30 min) or open in a
  BSC.

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NEEDLES AND SYRINGES

• Avoid use whenever possible
• Use a BSC for all operations with infectious
  material
• Fill syringes carefully
• Shield needles when withdrawing from stoppers
• Do not bend, shear or recap needles.
• Dispose of all used needles/syringes in yellow
  sharps containers

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PIPETTES

• Mouth pipetting is prohibited.
• Never force fluids out.
• To avoid splashes, allow discharge to run down
  dispense the receiving container wall.
• Never mix material by suction and expulsion.
• Reusable pipettes should be placed horizontally
  in a disinfectant filled pan.

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BLENDERS, GRINDERS, SONICATORS, AND LYOPHILIZERS

• Operate in a BSC whenever possible. Allow
  aerosols to settle for 5 minutes before opening.
• Safety Blender
• Do not use glass blender jars
• Decontaminate immediately after use
• Lyophilizers
• Use glassware designed for vacuum work, ensure
  there is no damage before using
• All surfaces should be disinfected after use
• Use vapour traps whenever possible

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INOCULATION LOOPS

• Sterilization in an open flame may create
  aerosols which may contain viable microorganisms.
• Use a shielded electric incinerator
• Shorter handles minimize vibrations
• Disposable plastic loops are good alternatives

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CRYOSTATS

• Wear gloves during preparation of frozen sections
  and heavy gloves when accessing the cryostat.
• Decontaminate frequently (100 or 70 Ethanol)

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SPILL RESPONSE
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SPILLS

• Spill response will vary depending on
• What was spilled?
• How much was spilled?
• Where was the spill?
• What is the potential for release to the
  environment?
• Spills should be cleaned up immediately (unless
  an aerosol was generated), to ensure proper
  decontamination.
• Ensure appropriate PPE is worn and clean-up
  equipment is readily available.

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SPILLS-GENERAL CLEAN-UP

• Cover spill area with absorbent material
• Soak the spill area with an appropriate
  disinfectant (i.e. 10 bleach)
• Pour disinfectant from the outside of the
  absorbent material towards the inside
• Ensure any broken glass is picked up (with
  forceps!) and placed in a sharps container
• Leave on for 20 to 30 minutes
• Wipe up with absorbent material
• Waste should be disposed in appropriate biohazard
  bags and where possible autoclaved

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SPILLS-SPECIAL CASES

• Within a Centrifuge
• Within a BSC
• Open Areas (lab, during transport)
• The spill response plan template is available at
  (http//www.uottawa.ca/services/ehss/SPILLRESPONSE
  PLAN.pdf)

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SPILLS

• All users of biological materials should be
  familiar with the spill clean-up procedures.
• All spills are to be reported ASAP to the lab
  supervisor and ORM.
• Additional assistance is available from
• ORM x 5892
• Your departmental safety officer
• ERT x 5411 (through Protection Services)

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BIOMEDICAL WASTE
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DECONTAMINATION, DISINFECTION, AND STERILIZATION

• Decontamination Free of contamination, the
  destruction of microorganisms to a lower level
  such that it removes danger of infection to
  individuals.
• Sterilization The complete destruction of all
  viable microorganisms.
• Disinfection Use of agents (physical or
  chemical) to destroy harmful organisms on
  inanimate objects (not necessarily all organisms)

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DECONTAMINATION PHYSICAL

• Heat
• Autoclaving (most practical and recommended)
• Incineration (for disposal of sharps and tissues)
• Irradiation
• UV light (wavelength of 253 nm is germicidal)
• Gamma (disrupts DNA and RNA)
• Filtration
• HEPA (biological safety cabinets, ventilation)

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AUTOCLAVES

• Items that CAN be autoclaved
• Cultures and stocks of infectious material
• Culture dishes and related devices
• Discarded live and attenuated vaccines
• Contaminated solid items (petrie dishes,
  eppendorf tips, pipettes, gloves, paper towels)

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AUTOCLAVES

• Items that CAN NOT be autoclaved
• chemicals (flammables, oxidizers, phenols, acids,
  alkalides)
• chemotherapeutic or radioactive waste
• Bleach (or other chlorinated products)
• certain kinds of plastics
• sharps (not at the University of Ottawa)

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AUTOCLAVES

• Preparation of waste
• Use only approved autoclave bags
• Do not overfill autoclave bags
• Separate material for re-use from that which will
  be disposed and dry from liquid material
• If outside of bag is contaminated, double bag
• All flasks containing biological material should
  be capped with aluminum foil
• Ensure items are labeled with contact information

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SAFE USE OF AUTOCLAVES

• Many autoclaves are now run by dedicated staff,
  however, if you are operating an autoclave
• Learn how to use it!
• Ensure PPE is worn
• Recognize acceptable material and packaging
• Proper loading and unloading
• All users/operators must take the autoclave
  training

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DECONTAMINATION CHEMICAL

• Generally for disinfection rather than
  sterilization
• Choice depends on
• Type of material to be disinfected
• Organic load
• Chemical characteristics
• Most common are chlorine compounds and alcohols
  (broad range)

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WHAT TO USE FOR MY AGENT?

• Vegetative bacteria (E.coli, Staph)
• 2 domestic bleach
• 75 Ethanol
• Quaternary ammonia
• 6 formulated Hydrogen peroxide
• Mycobacteria and fungi
• 10 domestic bleach
• 75 Ethanol
• Phenolic compounds
• 6 formulated Hydrogen peroxide
• Spore forming bacteria (Bacillus)
• 10 domestic bleach
• Gluteraldehyde
• Formaldehyde
• 6 formulated Hydrogen peroxide

• Viruses
• Enveloped (HIV, Herpes)
• 2 domestic bleach
• 75 Ethanol
• Quaternary ammonia
• 6 formulated Hydrogen peroxide
• Non enveloped (Hepatitis, Adenovirus)
• 10 domestic bleach
• 6 formulated Hydrogen peroxide
• Gluteraldehyde
• Formaldehyde

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WASTE MANAGEMENT

• Discarded biological material from teaching,
  clinical and research laboratories and operations
  is biomedical waste. Biomedical waste includes
  but is not limited to
• Animal waste
• Biological laboratory waste
• Human anatomical waste
• Human blood and body fluid waste
• Sharps

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WASTE MANAGEMENT

• All biological waste should be decontaminated
  prior to disposal (including level 1 agents).
• Treated waste is no longer considered
  biomedical (i.e. microbiological waste, blood
  and bodily fluid waste) and can be disposed in
  the regular waste stream.
• Any waste that cannot be treated (i.e. sharps,
  carcasses, tissues and body parts) remains
  biomedical waste and must be incinerated off site.

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WASTE DISPOSAL
Biomedical Waste (untreated)
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WASTE DISPOSAL
Biomedical Waste (treated)

In compliance with Sewer use by-laws
With H2O 110
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SPECIAL WASTE

• EtBr
• Toxins
• Recombinant DNA
• Contact ORM

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REGULATIONS
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KEY REGULATED ACTIVITIES

• Purchasing Receiving of Biological Agents
• PHAC, CFIA, Environment Canada
• Inventory Records
• Transportation/Transfer
• Transport Canada- TDG
• All Agencies (provincial and federal) emphasize
  and expect Biosecurity

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PURCHASING

• Importation permits required by Public Health
  Agency Canada or CFIA for certain agents
• US restrictions
• Ensure you meet all criteria and have all
  pertinent documentation

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INVENTORY

• What material is presently being used and/or
  stored
• Location
• Expiry date
• Use log book for remaining amount
• MSDSs
• Mandatory

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SHIPPING AND RECEIVING

• Transportation of Dangerous Goods Act Class 6.2
  of (Infectious Substances)
• PHAC/CFIA restrictions
• Ensure
• Proper classification
• Proper packaging
• Proper labeling
• Proper documentation
• Import/Export Permits

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TRANSPORTATION OF DANGEROUS GOODS

• Pre-approved
• Authorized Individuals
• Lead time (International Regulations.)
• Appropriate Scheduling (Holidays, Weekends)
• Transportation within the building
• Between lab to lab
• Colleague to Colleague
• Between Institutions

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TRANSPORTATION

• Important Considerations
• does material need to be transported at all
• packaging requirements
• means and route of transportation
• regulatory requirements
• Between lab transfers - 4 sided cart, sealed
  primary container, secondary container, low
  traffic route.
• Off Campus transfers consult ORM

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THE BOTTOM LINE

• If you are not careful and diligent with
  biological agents you risk
• Infecting yourself, others or the environment
• Contaminating your research
• Having Public Health Agency of Canada, Canadian
  Food Inspection Agency, Ministry of the
  Environment or Transport Canada after you

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CONCLUSIONS

• Biohazards - microorganisms, blood and body
  fluids, tissues and tissue culture
• Biosafety - ensuring that individuals and the
  environment are not infected
• Biosecurity - used in the context of protecting
  dangerous pathogens and toxins against
  intentional removal
• Everyone within the University community is
  responsible
• With proper knowledge, planning and care, a
  biological exposure is avoidable.

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BIOSAFETY WEBSITE

• http//www.uottawa.ca/services/ehss/biosafety.htm