BIOSAFETY
TRAINING
Uottawa2k9
Human Resources Occupational Health
Disability & Leave
Pierre Laflamme
May 16, 2012
Office of Risk Management,
Environmental Health
and Safety
COURSE OUTLINE
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Introduction
Laboratory Associated Infections
Blood-borne Pathogens
Classification of Biohazards
Infection/Biohazard Control
Spill Response
BIOSAFETY
Biomedical Waste
Regulations
INTRODUCTION
What is a BIOHAZARD?
Any organism or its toxin that is known to cause disease
in humans or animals or that is a potential hazard to
humans, animals or the environment.
Examples:
Microorganisms such as viruses, bacteria, fungi, and parasites
and their toxins.
Blood, body fluids and tissues from humans and animals.
Transformed cell lines
What is BIOSAFETY?
 The combination of measures employed when
handling biohazardous materials to:
 Protect personnel from exposure to infectious agents
 Prevent environmental contamination
 Provide an environment for high quality research
while maintaining a safe work place
 Comply with applicable federal, provincial and
municipal requirements
How is BIOSAFETY achieved?
 Administrative controls
 Training, Inspections, Permits and Certificates
 Engineering Controls
 Biological Safety Cabinets, Ventilation
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Personal Protective Equipment
Practices and Procedures
Medical Surveillance
Immunization when necessary
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.
How is BIOSECURITY achieved?
 Physical barriers
 Buildings, doors, locks, key card access
 Psychological barriers
 Security personnel, cameras
 Monitoring Activities
 Patrols, monitoring by support staff
 Personnel Clearance
 Access to authorized personnel only
Who are the STAKEHOLDERS?
INTERNALLY
EXTERNALLY
 Vice-President (Research)
 Committees
 University Services (ORM, HR,
PRS, PS)
 Deans, Chairs, Principal
Investigators
 Employees, Students
 Manager of Biological
Containment Suite
 Public Health Agency of
Canada
 Canadian Food Inspection
Agency
 Environment Canada
 Transport Canada
 Ontario Ministry of Labour
 Emergency Response
Personnel
 Suppliers & Contractors
 Community
University KEY SERVICES
 Office of Risk Management, Environmental Health
and Safety
Certificates and Permits
Training
Procedures (Waste disposal)
Risk Identification (Inspections)
Emergency plans
Accident/Incident follow-up
University KEY SERVICES
 HR (Occupational Health, Disability and Leave)
 Medical surveillance
 Immunizations
 Medical Follow-up
 Interface with Workplace Safety and Insurance Board
WHY ARE WE CONCERNED?
 Potential for acquiring a
laboratory-associated infection
(LAI)
 Contamination of the environment
 Contamination of research
LABORATORY ASSOCIATED
INFECTIONS
LABORATORY ASSOCIATED INFECTIONS
Infection Source
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Microorganisms
Cells and tissues
Blood and body fluids
Any items contaminated
with the above
Susceptible Host
Route of Transmission
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Immune system
Vaccination status
Age
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Percutaneous inoculation (needles
and bites)
Inhalation of aerosols
Contact of mucous membranes
Ingestion
LAIs
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Only 20% of LAIs are related
to a causative or defined event
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80% are caused by human errors
20% are caused by equipment failure
Types of accidents causing LAIs
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Spills and sprays
Needles
Sharp objects and broken glass
Bites or scratches from animals
http://www.weizmann.ac.il/safety/bio2.html
BLOOD-BORNE PATHOGENS
Human Resources
Occupational Health Disability & Leave
BLOODBORNE PATHOGENS (BBP)
 Sources
 Blood
 Semen
 Vaginal Secretions
 Other Bodily Fluids:
Cerebrospinal
Amniotic
Synovial
 Tissue Cultures
 Organ Cultures
 Infected Experimental
Animals
RISK OF EXPOSURE
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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
SPECIFIC EXAMPLES OF BBPS
Hepatitis B
Hepatitis C
HIV
FACTS ABOUT SOME BBPs
Pathogen
Hepatitis B
Hepatitis C
HIV
Pathogenicity
2 major forms:
• asymptomatic
• symptomatic
• asymptomatic
• symptomatic
• non-specific symptoms
• acute infection: non-specific “flu-like”,
“mono-like” symptoms
Mode of
transmission
• percutaneous/ permucosal
exposure to body fluids, organs
• indirect contact with
contaminated lab items (e.g.
needles, syringes)
• Percutaneous exposure to
contaminated blood (102 –
103 infectious particles/
mL)
• intravascular inoculation (e.g. transfusion)
of contaminated blood products
• direct exposure of virus to mucosa (oral,
rectal, vaginal)
Incubation
period
• usually: 24 - 180 days
• average: 60-90 days
• 2 weeks - 6 months
• most commonly 7 – 10
weeks
• chronic infection may
persist up to 20y before
onset of cirrhosis
• variable
• generally 1 - 3 months between time of
infection to development of detectable Ab’s
• time from HIV infection to diagnosis of
AIDS ranges from < 1y to 15y or more
Survival
outside host
• Survives in dried blood for long
periods (weeks)
• stable on environmental services
for at least 7 days at 25 °C
• not known
• similar to hep B (survives
in dried blood for long
periods…weeks)
• viable in blood in syringes @ RT for 42d
• Cell-free HIV dried on glass coverslips in
10% serum can survive forlonger than 7d,
depending on initial titre
Laboratoryacquired
infections
(LAIs)
• MOST FREQUENTLY
occurring LAI
• lab workers incident rate: 7X >
general population
• health care workers
handling blood at higher
risk to infection
• low (e.g. as of 2001, total of 57 cases of
documented occupationally acquired HIV
among US health care workers
[Source: http://www.phac-aspc.gc.ca/lab-bio/res/psds-ftss/index-eng.php]
ISSUES TO CONSIDER
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Symptoms
Mode of transmission
Incubation period
Survival outside host
Communicability
Immunization
Prophylaxis / Treatment
IF AN EXPOSURE OCCURS
 Initiate first aid
 Notify your supervisor / designated person
 Report to hospital emergency department or
University’s Health Services
 Report incident to OHDL
Occupational Health, Disability and Leave Office, ext. 1472
http://www.rh.uottawa.ca/00_main/index_f.asp
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 noninfective …… treat as infectious!
BIOHAZARD
CLASSIFICATION
BIOHAZARD CLASSIFICATION
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Conventional Agents – Risk Groups 1 to 4
Recombinant DNA
Tissue Culture
Animal Work
Anatomical Specimens
Unconventional Agents
Class D, division 3 of WHMIS
(Poisonous and Infectious Material - Biohazardous
Infectious Material)
BIOHAZARD CLASSIFICATION
 Conventional agents are categorized into risk groups
based on their particular characteristics such as:
 Pathogenicity (Infectivity of the agent - disease,
severity, mortality)
 Infectious dose
 Mode of transmission (Airborne, Ingestion, Parenteral)
 Host Range (Animal or human pathogen
 Availability of effective preventive measures (PPE)
 Availability of effective treatment
BIOHAZARD CLASSIFICATION
 Conventional agents are categorized based on the
measures required for handling each organism safely
in a laboratory setting, such as:
 Operational Requirements (Protocols, Biological safety
cabinets, Lab safety practices)
 Engineering Requirements (Maintenance, certification,
repairs)
 Physical Requirements (PPE)
CONVENTIONAL AGENTS
Risk
Group
Individual
Risk
Community
Risk
Containment
Level
Examples
1
Low
Low
Level 1
Escherichia Coli
Level 2
Bacteria: Streptococcus and
Salmonella
Viruses: Adenovirus, Hepatitis
A, B & C, Influenza
Level 3
Bacteria: Bacillus anthracis and,
Mycobacterium tuberculosis
Virus: HIV
Level 4
Viruses: Ebola virus and Lassa
virus
Unlikely to cause disease in
healthy workers or animals
2
Moderate
Limited
Rarely cause serious
human or animal disease
3
High
Low
May cause serious disease
4
High
High
Likely to cause very serious
disease
RECOMBINANT DNA
 Recombinant DNA technology or genetic engineering:
 in vitro incorporation of genetic material from one cell into
another or from one organism to another
 In Canada the level of risk depends on
 the source of DNA being transferred
 the vector
 the host
 The Office of Risk Management will assist the investigator
in this determination.
TISSUE CULTURE
 Mammalian cell lines have to be considered
infectious as they may contain infectious agents
 Untransformed mammalian cell lines - Risk Group 1
 MCF-7 (Human breast carcinoma cell line)
 NIH 3T3 (Mouse fibroblast cell line)
 Transformed mammalian cell lines – Risk Group 2
 HeLa (Human - contains papovavirus)
 All mammalian cell lines should be handled in a
Level 2 Containment.
ANIMAL WORK
 Animals can harbour infectious agents (naturally or
introduced) which can be transmitted to humans
 Scratches, bites, aerosols (needles and litter changes),
body fluids and excrements
 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
ANATOMICAL SPECIMENS
 All specimens should be considered infectious due to
potential presence of infectious agents
 It’s 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.
UNCONVENTIONAL PATHOGENS
 Includes unconventional agents, slow viruses and
prions causing progressive neurological diseases
 Creutzfeld-Jakob disease in humans, Mad Cow Disease,
Scrapie in sheeps and goats
 Resistant to destruction by chemical and physical
procedures that normally inactivate viruses
 Precautions:
 Handle tissues as Risk Group 2 or higher
 Handle formalin-fixed tissues and paraffin-embedded
blocks as if still infectious
 Follow up-to-date disinfection protocols.
WHERE ARE BIOHAZARDS FOUND?
Biology
Earth
Sciences
Bio-Engineering
University
Engineering
Civil
Chemical
Human
Kinetics
Chemistry
Medicine/
Nursing
INFECTION/BIOHAZARD
CONTROL
INFECTION/BIOHAZARD CONTROL
1. Administrative Controls
2. Engineering Controls
3. Personal Protective Equipment
4. Practices and Procedures
INFECTION/BIOHAZARD
CONTROL
1. ADMINISTRATIVE
CONTROLS
ADMINISTRATIVE CONTROLS
 Administrative procedures to minimize the risk of
exposure:
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Risk assessment
Training/Education
Resources
Inspections
Permits and Certificates
Medical Surveillance
Signage
ADMINISTRATIVE CONTROLS
Risk Assessment
 Will determine for each biohazard:
 Risk group
 Containment level
 Operational practices
 Safety measures
 Responsibility of users
 Know and understand the various characteristics of the
agent(s) you are working with.
 (Material Safety Data Sheets and suppliers or
manufacturers information sheets)
ADMINISTRATIVE CONTROLS
 Medical Surveillance
 Training & Education
 WHMIS
 Lab specific policies and procedures
 Biosafety training, Laboratory safety training
 Resources
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ORM web site, Biosafety page
Faculty web sites
Biosafety Manual
Training Videos
National and International Biosafety Guidelines
ADMINISTRATIVE CONTROLS
Inspections
 Routine self-inspections
 Biosafety Inspection Checklist available online
 In addition, ORM and Health, Safety and Risk
Officers will inspect labs to ensure
compliance with regulations/ guidelines and
provide feedback.
ADMINISTRATIVE CONTROLS
Signs & Labeling
 Biohazard warning signs must be posted on
doors to rooms where biohazardous materials
are used and/or stored.
 Biohazard labels should be placed on
containers, equipment and storage units used
with biological agents.
INFECTION/BIOHAZARD
CONTROL
2. ENGINEERING
CONTROLS
ENGINEERING CONTROLS
 Technology based
 Reduce or eliminate exposure to hazards
 Containment:
 Types: Primary and Secondary
 Levels: 1, 2, 3 and 4
 For effective containment, users must
 be aware of the potential hazards
 be trained
 Handle the material safely by adhering to standard
microbiological practices and techniques
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.
 Petri dish, vial
 Biological safety cabinets
 animal caging equipment
SECONDARY CONTAINMENT
 Protects the environment external to the laboratory
from exposure
 Includes facility design and operational practices
CONTAINMENT LEVEL 1
 Basic laboratory
 Requires no special design features
 Biosafety cabinets are not required and
work may be performed on the open
bench.
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.
CONTAINMENT LEVEL 3
 Specialized design and construction with
 primary barriers to protect the individual
 secondary barriers to protect the environment
 Requires type II or type III biosafety cabinets
 All staff must undergo specific training on the agents
used, PPE, equipment, waste management as well as
practices and procedures.
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
 The worker is completely isolated from infectious
material.
BIOLOGICAL SAFETY CABINETS
 Primary containment
 Minimize contact between operator and the
infectious agent by the use of directional airflows
 There are 3 main classes of cabinets (I, II, III) which
provide various levels of protection.
 Class II and III BSC contain HEPA filters which
remove particles (min 0.3 microns) from supply and
exhaust air with 99.97% efficiency .
 BSC should be located away from doors and high
traffic areas
BIOLOGICAL SAFETY CABINETS
Biological Safety Cabinet
Laminar air flow and HEPA filtered
exhaust air
Personnel
and
environment
protection
HEPA filtered supply air & product
protection with Class II + III
To be used
with biohazards
Laminar Flow Hoods or
Clean Air Benches
Vertical or horizontal laminar flow
HEPA filtered supply air only
Provide product protection only
Not to be used with biohazards
VS
WORKING SAFELY IN A BSC
Step 1
Before using the cabinet:
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Turn off UV lamp; turn on fluorescent lamp
Ensure BSC is certified
Disinfect work surfaces with appropriate disinfectant
Place essential items inside cabinet
Allow the blower to run for 5-10 min before work
WORKING SAFELY IN A BSC
Step 2
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
WORKING SAFELY IN A BSC
Step 3
After using the cabinet:
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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
WORKING SAFELY IN A BSC
Maintenance:
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Before and after each use - Wipe down work surfaces
Weekly - Clean UV lamp
Monthly - Wipe down all vertical surfaces
Annually - VerifyUV lamp intensity
- Decontamination with formaldehyde gas
(by ORM)
- Certification (by ORM)
INFECTION/BIOHAZARD
CONTROL
3. PERSONAL PROTECTIVE
EQUIPMENT
PERSONAL PROTECTIVE EQUIPMENT
 PPE is an important line of defence
 Responsibility of both the user and
the supervisor to ensure that PPE is
worn
 PPE is specific to each containment
level
 Examples of PPE?
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
 the individual is well trained
 Ensure PPE is removed before leaving the lab
PPE
Footwear
 Closed toe and heel shoes only.
 No sandals!
 Shoe coverings are worn in some
higher containment labs and animal
facilities.
PPE
Lab Coats/Gowns
 Protect street clothing from spills
 Offer additional body protection
 Long-sleeved, knee length with snaps
 Elastic cuffs
 Back-closing gowns
 Periodic cleaning required
PPE
Gloves
 Offer protection against a variety of hazards (heat,
cold, chemical agents, biological agents,
radioisotopes…)
 Latex, nitrile, rubber & vinyl for work with biological
agents.
 Gloves should not be reused and should be changed
frequently.
 Utility gloves can be disinfected and reused if they
show no sign of degradation.
PPE
Eye & Face Protection
 Goggles, safety glasses to protect the eyes
 Full face shield to protect facial skin
 Offer protection against chemical and biological
splashes
 No contact lenses
Respirators
 Only personnel who have been fit-tested and trained
should wear respirators.
 Worn in atmospheres that pose an infectious or
toxic hazard
MOVIE
BREAK
INFECTION/BIOHAZARD
CONTROL
4. PRACTICES AND
PROCEDURES
PRACTICES AND PROCEDURES
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General Safety Guidelines
Good Microbiological Practice
Handwashing
Receipt of Packages
Opening Packages
Specific Procedures
 Centrifuges
 Needles & Syringes and other sharps
 Pipettes
 Blenders, Grinders, Sonicators & Lyophilizers
 Inoculation Loops
 Cryostats
GENERAL LABORATORY SAFETY GUIDELINES
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Understand the hazards you face in the laboratory
Be adequately trained
Appropriate PPE must be worn
The lab should be kept clean and in order
Long hair must be tied back
Work surfaces must be cleaned and decontaminated
daily
 The use of needles should be limited
 Lab doors have to be closed
 Access to the lab has to be restricted
b i o s a f e t y
1. GOOD MICROBIOLOGICAL PRACTICE
(GMP)
Mitigates the risk of:
1. Personnel exposure
2. Contamination
a) sample
b) environment
What are we talking about ?
2. GOOD MICROBIOLOGICAL PRACTICE
(GMP)
Universal Precautions:
• More knowledge about the organism being used =
easier to take the necessary precautions
• Appropriate PPE greatly minimizes risk of
exposure
• Engineered controls (BSC’s) prevent release of
aerosols outside cabinet (and helps protect user!)
• Frequent hand washing = avoid infections
3. GOOD MICROBIOLOGICAL PRACTICE
(GMP)
• Prepare yourself for the work:
o Know what you will be doing
o Structure the work in a logical fashion (work flow)
• Prepare the work area
• Ensure all material that needs to be in the BSC is sterile
before placing it there
• Ensure waste containers are at hand’s reach and are not
overflowing and likely to collapse/ fall over
• Use aseptic technique
• Consult web [http://www.protocol-online.org] for SOPs &
techniques
• Properly trained to use equipment accordingly and when
in doubt…ASK!
• Clean up and decontaminate
4. GOOD MICROBIOLOGICAL PRACTICE
(GMP)
• Disinfect work surfaces with suitable disinfectant before and after
• Clean spills immediately and disinfect area thoroughly
• Keep bench top uncluttered
• Minimize traffic and unnecessary movements around work area
• all work with infectious material should be carried out in a specific area
• Material should not be carried throughout, or out of lab, unless in a
closed or capped container
• Minimize aerosol generation; if unavoidable, carry out activities in a BSC
5. GOOD MICROBIOLOGICAL PRACTICE
(GMP)
• Keep sterile and non-sterile objects separate
• Minimize exposure to outside air
• Avoid contact with non-sterile surfaces and items
• Hold open containers at an angle whenever possible
• Identify and properly dispose of different types of waste
HANDWASHING
 One of the single effective means of preventing
infections if done properly and frequently
 When to wash hands?
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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 one’s face or mouth
At the end of the day
RECEIPT OF PACKAGES
At Shipping & Receiving
 Verify shipment is yours, and expected.
 Inspect the integrity of container.
 If damage and breakage possible, transfer the package into a
secondary container lined with absorbent paper (absorbent side
up)
 Transfer to a cart with 4 sides for transfer to lab.
 Decontaminate all the areas in S&R where the package came
into contact with.
 All individuals who may have come into contact with the
material must wash their hands
 REMEMBER AT THIS POINT YOU DO NOT KNOW IF THE
SAMPLE HAS BEEN BE BREACHED !
OPENING PACKAGES – IN LAB
Scenario 1: Package appears damaged.
• Transfer the sample to a biological cabinet and open
and inspect each layer of packaging confronted with for
signs which would indicate the sample integrity.
• If damaged, inform your supervisor and ORM (x. 3153)
• Dispose of sample in the appropriate manner
• Package must be sterilized or sent for incineration.
OPENING PACKAGES – IN LAB
Scenario 2: Package is intact.
• Open package in the containment level required by
the sample
• Add sample to inventory
• Read and file MSDS or supplier information sheet
• Deface all markings on the package prior to disposal
SAFE USE OF CENTRIFUGES
 Before use
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Check centrifuge tubes for cracks
Avoid Overfilling
Place caps or stoppers properly
Balance loads
Use sealed buckets (safety cups) or sealed rotors
 Before leaving: ensure centrifuge achieves run conditions
 After run
 Centrifuge has to be completely stopped before opening the lid
 Check for spills or leaks before removing samples. Clean spills
 Allow aerosols to settle (30 min) or open in a BSC
NEEDLES AND SYRINGES
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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
PIPETTES
 Mouth pipetting is prohibited.
 Never force fluids out.
 To avoid splashes, discharge the liquid down the
receiving container wall.
 Never mix material by suction and expulsion.
 Reusable pipettes should be placed horizontally in a
disinfectant filled pan.
BLENDERS, GRINDERS, SONICATORS,
AND LYOPHILIZERS
• Operate in a BSC whenever possible. Allow aerosols
to settle for 5 minutes before opening.
• Decontaminate after use
• Blender
 Do not use glass blender jars
 Use safety blenders which can be autoclave
• Lyophilizers (used for dehydration process)
 Use glassware designed for vacuum work, ensure
there is no damage before using
 Use vapour traps whenever possible
INOCULATION LOOPS
 Sterilization in an open flame may
create aerosols which may contain
viable microorganisms.
 Shorter handles minimize vibrations
 Disposable plastic loops are good
alternatives
CRYOSTATS
 Wear gloves during preparation of frozen
sections and heavy gloves when accessing the
cryostat.
 Decontaminate frequently (70% Ethanol)
SPILL RESPONSE
SPILLS
 Spill response will vary depending on:
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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.
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
 Leave on for 20 to 30 minutes
 Pick up any broken glass (with forceps!) and place in a sharps
container
 Wipe up with absorbent material
 Waste should be disposed in appropriate biohazardous waste
container
SPILLS-SPECIAL CASES
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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/d
ocs/SPILLRESPONSEPLAN.pdf)
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)
BIOMEDICAL WASTE
DECONTAMINATION, DISINFECTION,
AND STERILIZATION
 Decontamination: 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
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)
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 (petri dishes, eppendorf
tips, pipettes, gloves, paper towels)
AUTOCLAVES
Items that CANNOT 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)
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
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
DISINFECTION: 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)
WHAT TO USE FOR MY AGENT?
Vegetative bacteria (E.coli, Staph)
Viruses




Enveloped (HIV, Herpes)
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




2% domestic bleach
75% Ethanol
Quaternary ammonia
6% formulated Hydrogen peroxide*
Non enveloped (Hepatitis, Adenovirus)




10% domestic bleach
6% formulated Hydrogen peroxide*
Gluteraldehyde
Formaldehyde
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
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 of 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.
 LABEL YOUR WASTE 
 IDENTIFY CONTENTS 
WASTE DISPOSAL
Biomedical Waste (untreated)
WASTE DISPOSAL
Biomedical Waste (treated)
*in compliance with
sewer use by-laws
with H2O (1:10)
REGULATIONS
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
PURCHASING
 Importation permits required by Public
Health Agency Canada (PHAC) or Canadaian
Food Inspection Agency (CFIA) for certain
agents
 Material Transfer Agreements (MTAs)
between importer & exporter
 US restrictions
 Ensure you meet all criteria and have all
pertinent documentation
BIOMATERIAL ACQUISITION/ MTAs
“How soon do you need it?”
“You want it when?”
 In order to facilitate a quick turnaround, provide
ORM with:
 copies of MSDS’s
 references (hardcopies)
 as much background information re:
product as possible
INVENTORY
 What material is presently being used
and/or stored
Location
Expiry date
Use log book for remaining amount
MSDS’s
 Mandatory
SHIPPING AND RECEIVING
 Transportation of Dangerous Goods Act: Class 6.2
(Infectious Substances)
 PHAC/CFIA restrictions
 Ensure:
 Proper classification
 Proper packaging
 Proper labeling
 Proper documentation
 Import/Export Permits
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
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
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
BIOSAFETY WEBSITE
http://www.uottawa.ca/services/ehss.biosafety.htm
Biohazardous Materials
User Registration
Biosafety Health
Assessment Survey
Practical Training Form
MOVIE