dedicated to all zoologists

1. Biosafety LeVeLs &
Biosafety CaBinets

2. Topics
Bio safety Levels
Air Filtration in Biosafety cabinets
Standards
Hood , Laminar flow & Bio safety Cabinets
Classification –Bio safety Cabinets

3. Biosafety Levels

4. Biosafety Levels
Bio Safety Level (BSL)-I:
Well-characterized agents not known to consistently cause disease in healthy adult
humans, and of minimal potential hazard to laboratory personnel and the
Environment.
Standard practices required:
frequent hand washing
door that can be kept closed when working
limits on access to the lab space when working
no eating, drinking, storage of food in laboratory
care to minimize splashes and actions that may create aerosols (tiny droplets)
decontamination of work surfaces after every use
Primary Safety Barriers- Not Required

5. Biosafety Levels
Bio Safety Level (BSL)-II:
• Suitable for work involving agents of moderate potential hazard to personnel
and the environment.
• Generally associated with human disease
Safety Equipment (Primary barriers):
• Class I or II Bio safety Cabinets or other physical containment devices (eg….
Aerosol tight lid in Centrifuges) are used
• Personnel Protection Equipment: Laboratory coats, gloves, face protection as
needed
Facilities (Secondary barriers):
• Provide lockable doors for facilities that house restricted agents
• Locating new laboratories away from public areas.
• Furniture used in laboratory work should be covered with a non-fabric material that
can be easily be decontaminated.
• An eye wash station is readily available.

6. Biosafety Levels
Bio Safety Level (BSL)-III:
Applicable where work is done with indigenous or exotic agents which may cause
serious or potentially lethal disease as a result of exposure by the inhalation
route.
Safety Equipment (Primary barriers):
• Class I or II Bio safety Cabinets
• Personnel Protection Equipment: Laboratory coats; gloves; face protection as
needed
• Facilities (Secondary barriers):
• The laboratory has special engineering and design feature . The exhaust air from the
laboratory room is discharged to the outdoors
• The ventilation to the laboratory is balanced to provide directional airflow into the
room

7. Biosafety Levels
Bio Safety Level (BSL)-IV:
Dangerous/exotic agents which pose high risk of life-threatening disease, aerosol-
transmitted lab infections or related agents with unknown risk of transmission.
Safety Equipment (Primary barriers):
All procedures conducted in Class III Bio safety Cabinets
or Class I or II Bio Safety Cabinets in combination with full-body, air-supplied, positive
pressure personnel suit.
Facilities (Secondary barriers):
BSL-3 plus:
• Separate building or isolated zone
• Dedicated supply and exhaust and decontamination systems

8. Air Filtration in Biosafety cabinets

9. Air Filtration in Biosafety cabinets
•High efficiency particulate air (HEPA) filters
•Ultra low particulate air (ULPA) filters
Variations in HEPA
Depending upon its quality, a HEPA
filter is able to trap from 9,995 to 9,999
of every 10,000 particles
filter efficiency from
99.95 % to 99.99 % are
usually due to
manufacturing
techniques.
An ULPA filter is able to
trap from 99,995 to 99,999
every 100,000 particles
ULPA - this is at least 10 times
better than HEPA filtration !!!

10. International Standards on Biosafety Cabinet

11. Why a standard ?

12. International Standards on Biosafety Cabinet
International Standards are mainly issued by two Organisations:
International Standard Organisation (ISO), based in Geneva
(Switzerland),
Comitee Europeen de Normalisation (CEN) based in Brussels
(Belgium).
These standards replace all other local standards on the same
subject.
The aim of the standards is to construct a mechanism to provide
manufacturer and user of a common site to specify technical
specifications and features.
Both construction and testing of the cabinet are covered

13. International Standards on Biosafety Cabinet
Standards:
BS-5726 (U.K.)
NF X44-201 (France)
DIN 12950 (Germany)
Now Replaced by EN 12469
EN 12469
European Standards on Design, Construction & Testing of Bio safety Cabinets
In addition to comply DIN 12980 for safety handling of cytotoxic drugs
NSF-49 (National Sanitation Foundation)
In the USA the NSF-49, was one of the first standards on this subject and was the
reference point for a lot of manufacturers. NSF is an independent organisation that
works as neutral agency to help on the problems that could affect to public health
and environment protection.
Was published in 1976.

14. Difference between Hood and Safety Cabinet
Hood Safety Cabinet
Exhaust of
vapors and
aerosols from
Laboratory to
outside
Filters and
blower(s)
incorporated
Validatable
personnel, product
& environmental
protection
No validatable
product or
personnel
protection

15. Fume Hood
A fume hood is a ventilation device that is designed to
limit exposure to hazardous fumes vapors or dust. A fume
hood is typically a large piece of equipment enclosing
five sides of a work area, the bottom of which is most
commonly located at a standing work height.
Two main types exist, ducted and recalculating. The
principle is the same for both types: air is drawn in from
the front (open) side of the cabinet, and either expelled
outside the building or made safe through filtration and
fed back into the room.
Fume Hood

16. Laminar Flow
Only for Product Protection
No Personnel or Environment Protection
A. front opening
B. sash
C. supply HEPA filter
D. blower
Vertical Laminar Flow

17. Laminar Flow
A. front opening
B. supply grille
C. supply HEPA filter
D. supply plenum
E. blower
F. Grille
Horizontal Laminar Flow

18. Classification of Bio Safety Cabinet
Biological Safety Cabinet
Class
Class I
Type A
Class II
III
Type B
Type A
I
Type A
II
Type B
I
Type B
II

19. Classification of Biosafety Cabinet
Class I:
Personnel and Environment protection.
Product is not protected.
Class II:
Personnel, Product and Environment are protected.
Class III:
Maximum protection to Personnel, Product and Environment.

20. Classification of Biosafety Cabinet
Class I Biosafety Cabinets
A. Front opening
B. Sash
C. Exhaust HEPA
Filter
D. Exhaust plenum
This is a negative-pressure,
ventilated cabinet usually
operated with an open front.
All of the air from the cabinet is
exhausted through a HEPA filter
either into the laboratory or to
the outside.
The Class I BSC is designed for
general microbiological research
with low- and moderate-risk
agents.

21. Classification of Biosafety Cabinet
Class II Type A-I Biosafety Cabinets
A. front opening
B. sash
C. exhaust HEPA filter
D. rear plenum
E. supply HEPA filter
F. blower
It is designed with inward air
flow to protect personnel,
Product & Environment
HEPA-filtered downward
vertical laminar airflow for
product protection, and HEPA-
filtered exhaust air for
environmental protection

22. Classification of Biosafety Cabinet
Class II Type A-I Biosafety Cabinets
Face Velocity 75 fpm
70% re circulated through HEPA Filter
30% Exhausted Through HEPA Filter
Cannot be used for Radio Nucleotides and Toxic Chemicals
Biosafety Levels 2 & 3
User, Product, & Environment Protection

23. Classification of Biosafety Cabinet
Class II Type A-II Biosafety Cabinets
A. front opening
B. sash
C. exhaust HEPA filter
D. supply HEPA filter
E. positive pressure plenum
F. negative pressure plenum
Note: The cabinet exhaust
needs to be connected to the
building exhaust system.

24. Classification of Biosafety Cabinet
Class II Type A-II Biosafety Cabinets
Downward Velocity 100 fpm
70% re circulated through HEPA
Filter30% Exhausted Through HEPA Filter
Can be used for Radio Nucleotides and Toxic Chemicals
Biosafety Levels 2 & 3User, Product, & Environment Protection
Under Negative Pressure to room
Exhaust Air is Hard Ducted

25. Classification of Biosafety Cabinet
Class II Type B-I Biosafety Cabinets
A. front opening
B. Sash
C. exhaust HEPA filter
D. supply plenum
E. supply HEPA filter
F. Blower
G. negative pressure exhaust plenum
Note: The cabinet exhaust needs to be
connected to the building exhaust
system.

26. Classification of Biosafety Cabinet
Class II Type B-I Biosafety Cabinets
Downward Velocity 100 fpm
30% re circulated through HEPA Filter
70% Exhausted Through HEPA Filter
Can be used for Radio Nucleotides and Toxic Chemicals
(Low Levels / Volatility)
Biosafety Levels 2 & 3User, Product, & Environment
Protection

27. Classification of Biosafety Cabinet
Class II Type B-II Biosafety Cabinets
A. front opening
B. Sash
C. exhaust HEPA filter
D. supply HEPA filter
E. negative pressure exhaust plenum
F. supply blower
G. filter screen
Note: The carbon filter in the building
exhaust is not shown. The cabinet
exhaust needs to be connected to
the building exhaust system

28. Classification of Biosafety Cabinet
Class II Type B-II Biosafety Cabinets
Downward Velocity 100 fpm
No Re-circulation100% Exhausted Through HEPA Filter
Can be used for Radio Nucleotides and Toxic Chemicals
Biosafety Levels 2 & 3User, Product, & Environment
Protection
Exhaust HEPA Filtered
Exhaust Hard Ducted out.

29. Classification of Biosafety Cabinet
Class III Biosafety Cabinets
A. glove ports with O-ring for attaching
arm-length gloves to cabinet
B. Sash
C. exhaust HEPA filter
D. supply HEPA filter
E. double-ended autoclave or pass-
through box
Note: A chemical dunk tank may be
installed which would be located
beneath the work surface of the BSC
with access from above. The cabinet
exhaust needs to be connected to the
building exhaust system.

30. Classification of Biosafety Cabinet
Class III Biosafety Cabinets
This is a totally enclosed, ventilated cabinet
Gas-tight construction
Offers the highest degree of personnel and environmental protection
from infectious aerosols as well as protection of research materials
from microbiological contaminants.

31. Cytotoxic Drug Safety Cabinets
Carbon filter exhaust #3 (option)
Dedicated exhaust flow
blower
Hepa filter exhaust #2
Laminar flow filter
All contaminated areas under
NEGATIVE pressure
Dedicated Laminar flow
blower
70% re circulated HEPA
filtered laminar flow
Provides sample
protection
30% inflow provides operator
Safety barrier
Hepa filter (pre-filter) #1

32. Can you see the problems ?
Ceiling clearance
or wall clearance
Air vent
Window
Front
Venting
Exhaust
BEWAR
E

33. Location In Lab
Location B is often
nfluenced from air
ows in the Room.
Location A, E and F
are well selected.
As well influencing
air flows through
the Laboratory Location C is
influenced from
personnel traffic
and Room air
flows.
should only be
aced in quiet areas!personnel traffic is
avoided.
Location D is also very often influenced
from personnel traffic and air flows.

34. THANK YOU