Lecture-10:

Basic Biosafety
Principles

Scientific Ethics and

Safety
ENGR / SCI 304 Sherif H. El-Gohary , Phd
Assistant Professor,Biomedical Engineering
shamdy@zewailcity.edu.eg
Course Requirement
Assessment Type Percentage

Attendance & Participation 10%

Assignments 10%

Midterms 30%

Project 10%

Final Exam 30%

Poster 10%

Total 100%
 Learning objectives

At the end of the presentation, you

should understand:
• Safety and Rules of the Lab
• Principles of biosafety
• How to transport dangerous pathogens
• Biosafety levels in a laboratory
• General disinfection principles
Safety and Rules of
the Lab
What’s Wrong With This Picture?
What’s Wrong With This Picture?
What’s Wrong With This Picture?
WHY IS LABORATORY
SAFETY IMPORTANT?

• TO PREVENT:
• adverse health effects from exposure to
chemicals
• exposure to organisms, diseases, etc. in
laboratories
• laboratory equipment hazards - if not
maintained properly
Lab safety must be taught
to :

all employees, including service

groups
undergraduate students
graduate students
professors
visitors
Safety Symbols

• Know safety symbols

• They appear in your
laboratory activities
• They will alert you
to possible dangers
• They will remind
you to work carefully
Laboratory Policies
and Procedures
Must be:
written and available
General Procedures or Rules:

• No food or drink allowed in the

laboratory
• No mouth pipetting
• Individuals not allowed to work in the
laboratory alone
• Wear required personal protective
equipment
• No smoking in laboratory
• Maintain good housekeeping habits
 Protect Yourself
Eye Safety
• Wear safety goggles when
working with chemicals,
flames, or heating devices
• or if possibility of flying debris

• If you wear contact

lenses let your
teacher know
 Protect Yourself
Proper Attire
• Keep all long hair
tied back
• Do not wear loose
clothing that could
catch on fire

• Foot wear that

completely covers
the foot is required
Safety Symbols
Chemical Safety
• Read all labels twice before removing a chemical from
the container.
• Never touch, taste, or smell a chemical unless
instructed by the teacher.
• Transfer chemicals carefully!
Hand Safety
• If a chemical spills on your skin, notify immediatly
and rinse with water for 15 minutes.
• Carry glassware carefully.
Plant Safety
• Do not eat any plants in lab.
• Wash your hands after handling plants.
Handling animals and
plants
• Procedures for caring for animals and plants including
feeding and watering
• Procedures for cleaning cages
• Procedures for cleaning and/or decontaminating rooms or
locations
• Procedures for entering & leaving contaminated areas
• Procedures for handling the animals or plants
• Procedures for animal bites or scratches
• Procedures for disposal to prevent spread of disease
 Handling of diseases or
organisms
• Allow only authorized individuals in infectious disease
laboratories
• Do not allow individuals to work alone
• Procedures for proper use of equipment and maintenance
• Use proper containers for transportation, incubation, and
storage
• Labeling of laboratories and cultures
• Proper disinfecting procedures
• Hygiene procedures
• Procedures for exposure or release of material
Principles of biosafety
To protect:
•the patient
•yourself
•the environment
Biosafety in Various
Disciplines
Biosafety is related to several
fields
• ECOLOGY: referring to imported life
forms not indigenous to the region
(Reggie the alligator)
• AGRICULTURE: reducing the risk of
alien viral or transgenic genes, or
prions such as BSE/"MadCow“;
reducing the risk of food bacterial
contamination
• MEDICINE: referring to organs
or tissues from biological origin,
or genetic therapy products, virus;
levels of lab containment
protocols BSL-1, 2, 3, 4 in rising
order of danger
• CHEMISTRY: i.e., nitrates in
water, PCB levels affecting
fertility
• EXOBIOLOGY: i.e., NASA's
policy for containing alien
microbes that may exist on space
samples - sometimes called
"biosafety level 5"
 Biosafety in Academic
Research
• Research Universities:
Promoting safe laboratory
practices, and procedures;
proper use of containment
equipment and facilities;
provides advice on
laboratory design and risk
assesment of experiments
involving infectious agents,
rDNA in-vitro and in-vivo.

Bottom Line: Risk & Containment

Biohazard Symbol
• Charles Baldwin at National
Cancer Institute at NIH.
• Symbol to be “memorable
but meaningless” so it could
be learned.
• Blaze orange – most visible
under harsh conditions
 Biosafety Issues

Laboratory Safety
Bloodborne pathogens (BBP)
Recombinant DNA (rDNA)
Biological waste disposal
Infectious substance and
diagnostic specimen shipping
 Biosafety Issues
(cont..)
Respiratory Protection
Bioterrorism and Select agents
Mold and indoor air quality
Occupational safety and health in the use of
research animals
Biohazards used in animal models
 Transport regulations (1)
• Transport of infectious substances is subject to
strict national and international regulations:
• proper use of packaging materials
• proper labelling, notification

• Compliance:
• reduces likelihood of damaging packages
• minimizes exposure
• improves carrier’s efficiency and confidence in package
delivery
 Transport regulations (2)
• Subject to regular amendments
• shippers refer to latest issuances of national
and international regulations for regulations

• International regulations not intended to

supersede local or national requirements
• where national requirements do not exist,
international regulations should be followed
 Category B, “650
package”
UN 3373
No biohazard label
 Category A “602 package”
Labels: UN 2814 UN 2900 Biohazard
 Triple packages
Category B infectious substances may be
shipped in "602" packages, as long as the
correct marking and labelling is provided on
the outer package

Category A infectious substances cannot be

shipped in "650" packages

Category A Category B
(“602”) (“650”)
 Biosafety Concepts

Principles of Biosafety
• Practice and Procedures
• Standard Practices
• Special Practices & Considerations
• Safety Equipment
• Facility Design and Construction
• Increasing levels of protection
Principles of Biosafety

Biosafety Levels 1-4 (BSL)

• Increasing levels of employee and environmental
protection
• Guidelines for working safely in research & medical
laboratory facilities

Animal Biosafety Levels 1- 4

(ABSL)
• Laboratory animal facilities
• Animal models that support research
• Guidelines for working safely in animal research facilities
Biosafety Concepts
Biosafety in Microbiological and
Biomedical Laboratories (BMBL)

(1) Standard Microbiological Practices

• Most important concept / Strict adherence
• Aware of potential hazard
• Trained & proficient in techniques
• Supervisors responsible for:
• Appropriate Laboratory facilities
• Personnel & Training
• Special practices & precautions
• Occupational Health Programs
Biosafety Issues
The BMBL

(2) Safety Equipment

• Primary Containment Barrier
• Minimize exposure to hazard
• Prevent contact / Contain aerosols
• Engineering controls/ equipment
• Personal Protective Equipment (PPE)
• Gloves, gowns, Respirator, Face shield, Booties
• Biological Safety Cabinets
• Covered or ventilated animal cage systems
Biosafety Concepts
The BMBL
(3) Facility Design and
Construction
• Secondary Barrier/ Engineering
controls
• Contributes to worker protection
• Protects outside the laboratory
• Environment & Neighborhood
• Ex. Building & Lab design,
Ventilation, Autoclaves, Cage wash
facilities, etc.
 Discussion

• What are some of

the negatives and
positives of this open
lab concept?
 Biosafety Level-1
Concepts of Biosafety
Biosafety Level-1 (BSL-1 or ABSL-1)
• Well characterized agents
• Agents not known to cause disease (in healthy
human adults; now healthy immunocompetent adults)
• Prophylactic treatment available
• Open bench procedures
• Animals in open cage system or open
environment (outdoors)
• Good laboratory practices
 Risk Group 1 Agents
• E.coli K-12
• Transgenic Plants
• Plasmids
• Fungi
• Mold
• Yeast
 BSL-1 Practices

• Bench-top work allowed

• Daily Decontamination
• Manual pipetting
• Required Handwashing
• Red bag waste
• Bio cabinet not required (unless
creating aerosols)
 Risk Group 2 Agents

• Human or Primate Cells

• Herpes Simplex Virus
• Replication Incompetent
Attenuated Human
Immunodeficiency Virus
• Patient specimens
 BSL-2 Practices
Concepts of Biosafety

Practices & Procedures

• Agents associated w/ human disease
• Treatment for disease available
• Agent poses moderate hazard to personnel and
environment
• Direct contact or exposure
• Percutaneous exposure
• Scratch, Puncture, Needle stick
• Mucus membrane exposure
• Eyes, Mouth, open cut
 BSL-2 Practices

• Limited access to lab when work in

progress
• Daily decontamination
• Mechanical pipetting
• Labcoat, safety glasses and gloves
required
• Red bag & sharps containers required
 BSL-2 Practices
(cont.…)
• Biohaz. Sign posted at
entrance to lab
• Label all equipment
(incubators, freezers, etc.)
• Tissue culture (TC) room –
negative air flow
• Documented training
• Baseline serology or pre-
vaccination may be required
 Risk Group 3 Agents

• Human Immunodeficiency Virus

• Mycobacterium tuberculosis

• Coxiella burnetii
 Biosafety Level 3
Working in High Containment
Biosafety Level-3 (BSL-3 or
ABSL-3)
• Indigenous or exotic agents
• Aerosol transmission
• Serious health effects
• Treatment may or may not exist
BSL-3 Practices

• Public access NOT permitted

• Daily decontamination after spill and
upon completion of experiment
• Autoclave required and waste is
disposed at the end of day
• Required foot activated handwashing
sink and controls
• No sharps unless absolutely necessary
BSL-3 Practices (con’t)

• Aerosol minimization procedures required

• Wrap around disposable clothing is
required. Specialized equipment may be
required depending upon procedures
• Biohaz. Signs and labels posted
• Air flow from low hazard to high hazard
“Pressure Mapping”
BSL-3 Practices
(cont.…)
• Bench top work not permitted
• Documented training and personnel
competency certification (for BSL-3
procedures)
• Baseline serology
• Spills – report immediately and treat
accordingly
• Vaccinations/post exposure protocols and
SOP’s, Biosafety Manual, Biosafety Officer
BSL-3 Examples
 Biosafety Level-4
Working in High Containment

Biosafety Level-4
• Builds on BSL-3/ ABSL-3 practices
• Maximum containment facilities
• Pressurized Containment Suite
• BSL-3 + Class III Biosafety Cabinet
• Chemical decontamination showers
• Liquid effluent collection / decontamination
• No BSL-4 labs exist at UCSD
 Biosafety Level 4
• Lassa Fever Virus
• Ebola Hemmorrhagic Fever
Virus
• Marburg Virus
• Herpes B Virus
Biosafety Concepts
Working in High Containment

Biosafety Level-4 (BSL-4 or ABSL-4)

• Dangerous/exotic agents
• Life threatening disease
• Aerosol transmission
• Agents of unknown risk
of transmission or health affects
• No known treatment
Animal Biosafety Level-4
Working in High Containment
Brief :BioSafety Level 1

• Well characterized, non-pathogenic

organisms or agents

• Open bench- no containment

• Use good laboratory practices, waste

disposal, and aseptic techniques

• Example: E. coli K-12 strains

 Brief :BioSafety
Level 2
• Agents of moderate hazard to personnel or environment

• Basic lab, but restricted access, containment during

certain processes (i.e. aerosols, large volumes, etc.)

• Autoclave and Biological Safety Cabinet desired

• Use good laboratory practices, waste disposal, and aseptic

techniques

• Example: most non-respiratory, non lethal, agents

 Brief :BioSafety
Level 3
• Agents of high hazard to personnel or environment

• Respiratory exotic or indigenous agents which are easily

transmissible causing serious or lethal disease

• All work is contained, engineering controls and controlled

environments we currently do not have the facilities to
handle.

Example: Mycobacterium tuberculosis, SARS, etc.

Brief :BioSafety Level 4

• FORGET ABOUT IT!!!

• Hemorrhagic fever, deadly viruses, etc.

• Total containment, airtight labs, “submarine” doors,

air pumps, water treatment, HEPA filtration, etc.

• Positive pressure “moonsuits”