▪ Containment principles, technologies

and practices implemented to prevent

unintentional exposure to pathogens
and toxins, or their unintentional
release
▪ Protection, control and accountability
for valuable biological materials within
laboratories in order to prevent
unauthorized access, loss, theft,
misuse, diversion or intentional release
4
5
Laboratory
Acquired
Infections
▪ Infections (symptomatic /
asymptomatic)
▪ Acquired through laboratory related
activities as a result of working with
infectious agents
7
▪ Causative or defined event
□ 20%  equipment failure
□ 80%  human factors

8
9
▪ Top “4” accidents resulting in
infection

10
▪ INGESTION
□ Consumption of a substance by an
organism
▪ INOCULATION
□ Act of introduction of a substance
into the body 11
▪ CONTAMINATION
□ Presence of a minor and unwanted
substance or impurity in the skin or
mucous membrane
▪ INHALATION
□ Act of drawing air or other substances
into the lungs 12
HISTORY OF
BIOSAFETY

13
A.G. Wedum, MD
• U.S. Biological Research Laboratories at Fort Detrick
• one of the pioneers in developing biosafety
measures after the Second World War
• evaluated the risks of handling hazardous biological
agents and developed practices, equipment, and
facility safeguards for their control
A.G. Wedum
Asilomar Conference in 1975
• general principles for dealing with potential
biohazards related to GMOs were drafted
• It was suggested that containment should be an
essential consideration in the experimental design
and that the effectiveness of the containment should
match the estimated risk.
mid-1970s
• designation of 4 levels of biosafety
1976
• first edition of the National Institutes of Health
(NIH) guidelines for research involving DNA
molecules was published
1984
• first edition of a guidebook, called Biosafety in
Microbiological and Biomedical Laboratories was
produced
• U.S. NIH and Centers for Disease Control and
Prevention (CDC)
• now considered a major reference text for biosafety
1990
• Directive on the protection of workers from risks
related to exposure to biological agents at work
(European Commission)
• Directive on the contained use of genetically
modified microorganisms(European Commission)
ORGANIZATIONS
 provides oversight of public
health and safety, including
the laboratory

Develops and enforces workplace standards to

protect employees’ safety and health.
Recommendations include guidelines
addressing blood-borne pathogens, chemical
safety, phlebotomies, latex gloves, ergonomics,
and any other potentially hazardous situation
22
that may be found in the workplace
International Federation of Biosafety
Associations

23
24
FUNDAMENTAL
CONCEPTS
▪ Laboratory safety
▪ Blood borne pathogens
▪ Recombinant DNA
▪ Biological waste disposal
▪ Transport of biological materials

26
▪ Respiratory protection
▪ Bioterrorism and select agents
▪ Mold and indoor air quality
▪ Occupational safety
▪ Health in the use of research animals

27
▪ The principle of holding or be capable of
holding or including within a fixed limit
or area
▪ Biocontainment: preventing the release
of biological agents
28
 PRIMARY BARRIERS
(control hazard at source)
BSC
HEPA filter

BSC

SECONDARY BARRIERS
(structure surrounding primary barrier)
Sealed perimeter
Exhaust HEPA filters 29
▪ Primary containment equipment
□ BSC
□ Animal enclosures
□ Sealed centrifuge rotors

30
▪ HEPA filters
▪ Liquid effluent treatment
▪ Sealed laboratory walls and floors

31
▪ Practice and procedures
□ Standard practices
□ Special practices and considerations
▪ Safety equipment
▪ Facility design and construction
▪ Increasing levels of protection
32
 1

▪ Most important concept / strict

adherence
▪ Aware of potential hazard
▪ Trained and proficient in techniques
33
 2
▪ Primary containment barrier
▪ Minimize exposure to hazard
□ Engineering controls / equipment
□ PPE
□ Biological safety cabinets
34
35
▪ Lab Coats and gowns
□ used to protect from
infectious fluids
□ Don’t wear lab coats
outside of the lab or take
them home
□ Cuffed sleeves can protect
the wrists and lower arms 36
▪ Gloves
□ Wear disposable vinyl, synthetic or N-
DEX nitrile gloves when working with
biohazardous materials

37
▪ Gloves
□ Avoid latex gloves (may cause allergies)
□ Do not reuse gloves
□ Do not wear gloves outside of the
laboratory
□ Wash hands after removing gloves

38
▪ Eye and Face Protection
□ protect mucous membranes and
prevent ingestion whenever there is
potential for splash to eyes/face

39
▪ Foot/Skin Protection
□ Open toed shoes,
sandals and other open
footwear should be
prohibited
□ Shorts and other
garments that leave skin
unprotected are not
appropriate 40
▪ Respiratory Protection
□ Two types: air supplying and air
purifying
□ Full face, half face, PAPR (Powered Air
Purifying Respirator)
□ Special considerations: fit testing; facial
hair; comfort; care and maintenance
41
▪ Respiratory Protection
□ N95 respirators

□ N100 respirators

42
▪ BSCs provide effective primary
containment for work with infectious
material or toxins when they are
properly maintained and used in
conjunction with good laboratory
techniques
43
▪ Fume Hood
▪ Laminar Flow Cabinet (LFC)
▪ Biohazard Safety Cabinet (BSC)

44
45
46
47
48
49
▪ Removes a broad –Soot
range of airborne –Pollen
contaminants: –Radioactive particles
–Fine dust –Impurity ion can
–Smoke affect Integrated
–Bacteria (typical Circuit speed
size: 500 to 0.3μm
50
51
52
53
54
55
56
57
58
 

59
60
61
62
63
▪ HEPA filter leak test (LT)
□ This test is conducted to check the HEPA
impulsion/downflow and exhaust filters,
the filter housing, and the mounting
frames for possible leakage.
64
▪ Inflow velocity test (IV)
□ This test is conducted to
determine the nominal
value of the inflow velocity
(air entered to the BSC)

65
▪ Downflow velocity test
□ This test is conducted to check the
nominal value of the downflow
velocity (displacement airflow) in
the work area of the safety
cabinet.
66
▪ Downflow velocity test

67
▪ Airflow pattern test
□ This test is conducted to check the behavior of
the airflows in the sample chamber.
□ Check to see if the displacement airflow
passes along the entire work area, if the
vertical passage is correct, and of air escapes
through joints or seals of the housing and of
the front window.
68
69
 3
▪ Secondary barriers / engineering
controls
□ Contributes to worker protection
□ Protects outside the laboratory

70
 3
▪ Example:
□ Building and lab design
□ Ventilation
□ Autoclave
□ Cage wash facilities
71
3

72
 4
▪ Biosafety levels
□ Increasing levels of employee and
environmental protection
□ Guidelines for working safely in
research and clinical laboratory
73
Classification of
infective microorganisms
by risk group
▪ Assignment of microorganisms based on:
□ PATHOGENICITY
□ MODE OF TRANSMISSION AND HOST RANGE
□ Local availability of effective PREVENTATIVE
MEASURES
□ Local availability of EFFECTIVE TREATMENT
Risk A microorganism that is unlikely to cause human or
Group 1 animal disease.

BSL 1
Typical Open bench
Work
Area

Example S. cerevisiae (yeast), Lactobacillus, B. subtilis

76
Risk A pathogen that can cause human or animal disease
Group 2 but is unlikely to be a serious hazard to laboratory
workers, the community, livestock or the
environment.
Laboratory exposures may cause serious infection,
but effective treatment and preventive measures are
available and the risk of spread of infection is limited.
BSL 2
Typical Biosafety cabinet / laminar flow hood
Work
Area 77
Risk
Group 2

BSL 2
Example Hepatitis B virus, HIV, the salmonellae and
Toxoplasma spp.
78
Risk A pathogen that usually causes serious human or
Group 3 animal disease but does not ordinarily spread
from one infected individual to another. Effective
treatment and preventive measures are available.
BSL 3
Typical Class 3 biosafety cabinet
Work
Area

Example M. tuberculosis, St. Louis encephalitis virus and

Coxiella burnetti, Yersinia pestis, SARS virus 79
Risk
Group 4

80
Risk A pathogen that usually causes serious human or
Group 4 animal disease and that can be readily
transmitted from one individual to another, directly
or indirectly. Effective treatment and preventive
measures are not usually available.
BSL 4
Typical Full isolation suits
Work
Area
Example Ebola, Marburg or Congo-Crimean hemorrhagic fever
viruses 81
82
▪ Combination of laboratory practices
and procedures, safety equipment
(PRIMARY BARRIERS) and laboratory
facilities (SECONDARY BARRIERS)
□ Somewhat related to risk groups

83
▪ For Risk Group 1
▪ Safety practices:
□ Restrict or limit access when working.
□ Prohibit eating, drinking, and smoking.
□ Prohibit mouth pipetting.
□ Needles and sharps precautions
▪ Basic level of containment
84
85
▪ For Risk Group 2
▪ Safety practices:
□ BSL-1 practices plus:
□ Use Biological Safety Cabinet (BSC)-Class II
□ Use leakproof containers
▪ Clinical, diagnostic, teaching and other
laboratories
▪ Splashes or aerosols is low 86
87
▪ For Risk Group 3
▪ Safety practices:
□ BSL-2 practices plus:
□ Use BSC-Class II 100% air exhausted to outside
through double HEPA filtration or HEPA plus
incineration.
□ Cabinet is gas tight and sealed, with operation
performed through rubber gloves.
□ Use complete PPE (personal protective
equipment). 88
▪ For agents that pose an increased risk of
aerosol spread
▪ More emphasis on primary and secondary
barriers
▪ Controlled access to the laboratory and
ventilation requirements
▪ All laboratory manipulations are performed
in a BSC 89
90
▪ For Risk Group 4
▪ Dangerous and exotic agents
▪ Transmitted via aerosol route
▪ Maximum containment
▪ Class III BSC
▪ Suit laboratory
91
92
BIOSECURITY
93
▪ Biological materials
▪ No devices to detect pathogens being
removed from facility
▪ Easy to hide small vials, filter paper
▪ Present in clinical labs, research labs,
private labs and government labs
1. Physical security
2. Personnel security
3. Pathogen security
4. Transport security
5. Information security
 Physical security
• Access control
• Intrusion detection
• Alarm assessment and
response

Information security Personnel Security

• Confidentiality, integrity and • Background check
availability of info
• Periodic investigations
• Passwords, back-ups
• Personnel reliability program

Transport security Pathogen security

• 3 way packaging system • Detailed inventory
• “From – How – Where” • Internal / external transfer
• Knowledge IATA certified personnel • Inactivation and disposal records
of pathogens 96
Effectiveness of biosecurity will depend on
the integrity of the individuals with access
to the pathogen

97
98