Biotechnology

1. BIOSAFETY CONCEPTS ANDBIOSAFETY CONCEPTS AND
ISSUESISSUES -AMRUTHA K.H
II MSc Zoology

2. BIOSAFETY
• Refers to the effort that ensure safety in
using,transporting,transferring,handling,relea
sing and disposing of biological organisms
including genetically modifies
organisms(GMO)which are capable of
harming human,animal,plants or environment

3. GOALS
• Provide the highest practical protection and lowest
practical exposure
• To determine in advance the hazards to human health
and natural system if any GMO is released
• To anticipate when GMO or any of its product will be
harmful if consumed in food
• Also defined as:
Avoidance of risk to human health and safety, and
conservation of environment, as a result of the use for
research and commerce of infectious or GMO
(Zaid,2001)

4. CONCEPTS AND ISSUES
1. Containment
• Safety measure which prevent the
escaping of GEOs from the laboratory
• 2 types
 Physical
 Biological

5. Physical
• To prevent escape of GEO to the
environment
• It include
Air filtration
Sterilization light
Waste disposal
Protective handling

6. Biological
• Makes the organism unable to survive in
the outside environment
• Prevents the spreading of vector DNA
outside the lab by using
conguation,transformation or transduction
• Some bacteria induce gene mutation
• Survive only in the culture

7. BIOSAFETY LEVEL
• set of biocontainment precautions required to isolate
dangerous biological agents in an enclosed laboratory
facility. The levels of containment range from the lowest
biosafety level 1 (BSL-1) to the highest at level 4 (BSL-4).

8. Biosafety level 1
• these agents should pose minimal potential
hazard to laboratory personnel and the
environment.
• Agents: Characterized strains of microorganisms
known to cause no disease in healthy adults. eg.
E. coli, S. cerevesiae, B. subtilis etc.
• Recombinant DNA based research activities
involving non-pathogenic micro-organisms for
expression of genes using plasmid vectors or
low risk viral vectors.

9. • Work practice: Standard aseptic microbiological
techniques.
• Safety equipment requirement: Lab coats and
eye protection recommended.
• Facilities: Bench top, sink etc.

11. Biosafety level 2
• Agents: Handling of micro-organisms which
possess moderate hazard to personal and
environment.
• rDNA based research activities in micro-
organisms using non-viral or viral vectors.
• Work practice: Standard BSL-I practices with
addition of limited access, biohazard sign,
defined procedure for disposal of “Regulated
Medical Waste”, proper training to lab personal
and medical surveillance.

12. • Safety equipment: Class-II biological safety
cabinet, lab coats, gloves, eye/face protection,
physical containment equipment to reduce
infectious aerosol exposure or splashes.
• Facility: BSL-I facility with addition of autoclave,
decontamination facility and proper airflow.

14. Biosafety level 3
• Agents: Handling of micro-organisms which are
designated as hazardous or potentially lethal
agents to personal and environment.
• Laboratory personnel must have specific training
in handling infectious micro-organisms and
should be supervised by scientist competent in
handling infectious agents.
• Work practices: BSL-2 practices, with the
addition of: controlled access, on-site
decontamination of all waste and lab clothing
and medical surveillance.

15. • Safety equipment: Class-III biological safety cabinet, lab
coats, gloves, eye/face protection, respiratory protection,
physical containment equipment to reduce infectious
aerosol exposure or splashes.
• Facility: BSL-III facility has specific criteria to meet.
• Lab should have double door entry with physical
separation of working area from the access corridors,
directional airflow in lab, and no recirculation of exhaust
air in the lab, sufficient decontamination facility, in lab
autoclave etc.

16. Researcher at US Centers for Disease Control, Atlanta, Georgia, working with influenza
virus under biosafety level 3 conditions, with respirator inside a biosafety cabinet (BSC

17. Biosafety level 4
• highest level of biosafety precautions, and is
appropriate for work with agents that could
easily be aerosol-transmitted within the
laboratory and cause severe to fatal disease in
humans for which there are no available
vaccines or treatments.
• BSL-4 laboratories are generally set up to be
either cabinet laboratories or protective-suit
laboratories.

18. • Agents: Hazardous and potentially lethal
organisms that posses high individual risk of
laboratory transmitted disease for which there is
no vaccine or treatment, or a related agent with
unknown risk of transmission.
• Laboratory personnel must have specialized
training in handling BSL-IV agents and should
be supervised by scientist competent in handling
infectious agents.

19. • Safety equipment: Class-IV biological safety
cabinet, lab coats, gloves, eye/face protection,
respiratory protection, physical and containment
equipment to reduce infectious aerosol exposure
or splashes.
• Facility: BSL-IV facility requires specialized
design to minimize the exposure to risk and only
the authorized entry should be permitted in
laboratory area in BSL-IV labs.

20. Scientists working inside positive pressure personnel suit at biosafety level 4
(BSL-4) laboratory of the NIAID Integrated Research Facility (IRF) in Frederick,
Maryland

21. Biosafety cabinet
• also called a biological safety cabinet or
microbiological safety cabinet—is an
enclosed, ventilated laboratory workspace for
safely working with materials contaminated with
(or potentially contaminated with) pathogens
requiring a defined biosafety level.
• Several different types of BSC exist,
differentiated by the degree of biocontainment
required.
• BSCs first became commercially available in
1950

22. Purposes
• The primary purpose of a BSC is to serve as a
means to protect the laboratory worker and the
surrounding environment from pathogens.
• All exhaust air is HEPA-filtered as it exits the
biosafety cabinet, removing harmful bacteria and
viruses.
• This is in contrast to a laminar flow clean bench,
which blows unfiltered exhaust air towards the
user and is not safe for work with pathogenic
agents.

23. • Neither are most BSCs safe for use as
fume hoods.
• Likewise, a fume hood fails to provide the
environmental protection that HEPA filtration in a
BSC would provide.
• However, most classes of BSCs have a
secondary purpose to maintain the sterility of
materials inside (the "product").

24. • BSCs into three classes.
• These classes and the types of BSCs within
them are distinguished in two ways:
 the level of personnel
 environmental protection provided and the level
of product protection provided.

25. Class I
• Class I cabinets provide personnel and
environmental protection but no product
protection.
• inward flow of air can contribute to
contamination of samples
• Inward airflow is maintained at a minimum
velocity of 75 ft/min(0.38 m/s).
• These BSCs are commonly used to enclose
specific equipment (e.g. centrifuges) or
procedures (e.g. aerating cultures) that
potentially generate aerosols.

26. • BSCs of this class are either ducted (connected
to the building exhaust system) or unducted
(recirculating filtered exhaust back into the
laboratory.

27. Class II
• Class II cabinets provide both kinds of
protection (of the samples and of the
environment) since makeup air is also
HEPA-filtered.
• There are five types: Type A1 (formerly A),
Type A2 (formerly A/B3), Type B1, Type
B2 and Type C1.

29. Class III
• The Class III cabinet, generally only installed in
maximum containment laboratories, is
specifically designed for work with BSL-4
pathogenic agents, providing maximum
protection.
• The enclosure is gas-tight, and all materials
enter and leave through a dunk tank or double-
door autoclave.

30. • Gloves attached to the front prevent direct contact with
hazardous materials (Class III cabinets are sometimes
called glove box).
• These custom-built cabinets often attach into a line, and
the lab equipment installed inside is usually custom-built
as well.

32. Risk assessment
I. Pathogenicity
II. Virulence
III. Proliferation
IV. Transmission route
V. Infectious dose
VI. Antibiotic resistance

33. Biosafety Issues
1. Risk of animal and human health
2. Risk for the environment
3. Horizontal gene transfer
4. Risk for agriculture
5. General concern

34. General guidelines for rDNA
research activity
• General
i. Harmonization of approaches to rDNA techniques can be
facilitated by exchanging principles or guidelines for
national regulations;
ii. There is no scientific basis for specific legislation for the
implementation of rDNA techniques and applications.
iii. Any approach to implement guidelines should not
impede future developments in rDNA techniques.
iv. To facilitate data exchange and minimize trade barriers
between countries, further developments such as testing
methods, equipment design, and knowledge of microbial
taxonomy should be considered at both national and
international levels.

35. • Industry
i. The large-scale industrial application of rDNA techniques
wherever possible should utilize microorganisms that are
intrinsically of low risk.
ii. If a recombinant microorganism cannot be handled
merely by GILSP, measures of containment
corresponding to the risk assessment should be used in
addition to GILSP.
iii. Further, research to improve techniques for monitoring
and controlling non-intentional release of rDNA
organisms should be encouraged in large-scale
industrial applications requiring physical containment.

36. • Environmental and Agricultural Considerations:
A. Ecological traits relating to the donor and recipient
environment.
B. Properties of environment where the engineered
organism are being applied.
C. Survival, multiplication and dissemination of the
engineered organism in the environment.
D. Interactions of engineered organism(s) with biological
systems (target and non-target populations, stability, and
routes of dissemination).
E. Potential environmental impacts (Effect on target and
non-target organisms and ecosystems).

37. Mechanism of implementation of
guidelines
1. RDAC
2. IBSC
3. RCGM
4. GEAC

38. BIOSAFETY PROTOCOL 2000
• The Cartagena Protocol on Biosafety to the
Convention on Biological Diversity is an
international agreement on biosafety as a
supplement to the Convention on Biological
Diversity effective since 2003.
• The Biosafety Protocol seeks to protect
biological diversity from the potential risks posed
by genetically modified organisms resulting from
modern biotechnology

39. • makes clear that products from new
technologies must be based on the
precautionary principle and allow developing
nations to balance public health against
economic benefits.
• It will for example let countries ban imports of
genetically modified organisms if they feel there
is not enough scientific evidence that the
product is safe and requires exporters to label
shipments containing genetically altered
commodities such as corn or cotton.

40. • Objective of the Protocol is to contribute to
ensuring an adequate level of protection in the
field of the safe transfer, handling and use of
'living modified organisms resulting from modern
biotechnology' that may have adverse effects on
the conservation and sustainable use of
biological diversity, taking also into account risks
to human health, and specifically focusing on
transboundary movements (Article 1 of the
Protocol, SCBD 2000).

41. Living modified organisms
(LMOs)
• The protocol defines a 'living modified organism'
as any living organism that possesses a novel
combination of genetic material obtained
through the use of modern biotechnology, and
'living organism' means any biological entity
capable of transferring or replicating genetic
material, including sterile organisms, viruses and
viroids.

42. • LMO Products' are defined as processed material that
are of living modified organism origin, containing
detectable novel combinations of replicable genetic
material obtained through the use of modern
biotechnology.
• Common LMOs include agricultural crops that have
been genetically modified for greater productivity or for
resistance to pests or diseases.
• Examples of modified crops include tomatoes, cassava,
corn, cotton
• Living modified organism intended for direct use as food
or feed, or for processing (LMO-FFP)' are agricultural
commodities from GM crops.

43. Procedures for moving LMOs
across borders
1. Advance Informed Agreement
2. LMOs intended for food or feed, or for processing
3. Handling, transport, packaging and identification
4. Biosafety Clearing-House

44. THANK YOU