biosafety

1. Basics of Biosafety
Dr Vikas D Dighe, M.V.Sc, Ph.D.
Scientist E and Head,
National Centre for Preclinical Reproductive and Genetic Toxicology
ICMR-National Institute for Research In Reproductive Health, Mumbai
Email:dighev@nirrh.res.in
Working Safely with Biological Materials

2.  Principles and practices employed to protect
laboratory personnel and the environment from
exposure or infection while working with living
organisms, biological materials, or agents.
 Included are any materials that may be potentially
infectious.
 Includes recombinant DNA research
What is Biosafety?

3.  The “agent” is the what creates risk
 Risks to the worker or environment are
often unknown
 Determining “acceptable risk”?
Agents and Risks

4. There is always risk!
The risk must be identified
The risk is evaluated
The risk must be measured
Plan to minimize the risk
Assessing Risk

5.  Assessment is conducted by a Biosafety
Professional in partnership with and based
on information provided by the Principal
Investigator
 The assessment is presented to the
Institutional Biosafety Committee (IBC) for
approval
Who Determines Acceptable Risk?

6.  Understand the biology of the agent
 Susceptibility and transmission within the
host
 Hazards associated with equipment and
procedures
 Goal:
Provide the highest practical protection and
the lowest practical exposure
Identifying Risk

7.  Worst case scenario -What might happen?
 Likelihood of an event
 Seriousness of the incident
 Actions needed to resolve the problems
Evaluating Risk Acceptability

8.  Since there is no such thing as “no risk”
 “Safe” means risk has been judged acceptable
 Judging risk is a subjective- humans make
decisions
 Measuring risk is objective- use available
guidelines, data, and documentation
 Keep records of how determinations were made
due to subjective nature of the process
What is Acceptable Risk?

9. Agents Assigned Risk Groups
 RG-1 Unlikely to cause disease in humans or animals
 low individual or community risk
 RG-2 May cause disease but typically not serious
 individual risk, low community risk, treatable
 RG-3 May cause serious disease, usually treatable
 High individual but low community risk, serious respiratory agents
 RG-4 Serious or fatal, often not treatable,
 Easy transmission, high individual and community risk
WHO-World Health Organization

10.  Different than the Risk Groups!!
 Risk groups used in risk assessment
 BSL are used in risk management
 BSL are ways to control the agent
 facilities, safety equipment, practices, PPE, etc.
 Once risk is assessed then the appropriate BSL
is determined
Biosafety Levels (BSL)

11.  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
BioSafety Level 1

12.  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
BioSafety Level 2

13.  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.
BioSafety Level 3

14.  FORGET ABOUT IT!!!
 Hemorrhagic fever, deadly viruses, etc.
 Total containment, airtight labs, “submarine”
doors, air pumps, water treatment, HEPA
filtration, etc.
 Positive pressure “moonsuits”
BioSafety Level 4

15. Bacterial:
76% from clinical labs
8% from research labs
Exposure:
60% acquired from inhalation
Other exposures include:
digestion, sharps, splashes, direct and indirect contact
Laboratory Acquired Infections (LAI)

16. Viral
 16% from clinical labs
 70% from research labs
32% from animal related activities
Laboratory Acquired Infections (LAI)

17. Biohazardous/Medical Waste
Waste that is potentially infectious to
humans, animals or plants.

18. Biohazardous Waste Categories
Cultures and stocks of infectious
agents and associated biologicals
laboratory waste
biological production waste
discarded live and attenuated vaccines
culture dishes and related materials
contaminated PPE

19. Biohazardous Waste Categories
Liquid human and animal waste
liquid or semi-liquid blood and blood
products and body fluids
contaminated items that would release
blood or items that are caked with blood
or other potentially infectious materials;
NOT including urine or materials stained
with blood or body fluids
infectious animal waste (research)

20. Biohazardous Waste Categories
Pathological waste
tissues
body parts other than teeth
products of conception
fluids removed by trauma or during
surgery or autopsy/necropsy or other
medical procedure and not chemically
fixed.

21. …And More Biohazardous
Waste Categories
 Animal and plant pathogen waste
 Recombinant DNA waste
 Sharps

22. Biowaste vs. Trash
3 basic questions to differentiate:
1. Is it contaminated with viable
biological material?
2. Can blood or other regulated
body or biological fluids be
released?
3. Is it a sharps hazard?

23. #1
Is it contaminated with
viable biological
material?
Examples:
• Contaminated lab waste
• Personal protective equipment
used for handling potentially
infectious materials (including
handling infected animals or their
products)
• Wastes from infectious disease
research (carcasses, body
fluids…)

24. #2
Can blood or other
(regulated) body fluids or
viable biological materials be
released?
Some Examples…
Tubes of blood
Vacuum flasks containing body
fluids or cell line waste

25. Managing Liquid Biohazardous Waste
Storage:
 Label and secure bulk vessels if
not disposed of immediately
Treatment:
 Chemical disinfection OR
 Autoclave
Disposal: THEN
 Flush to sewer
 Use proper PPE!

26.  10% bleach solution
 good for general disinfection
 High organics use 20%
 Needs to be made weekly
 Test contact time
 Ethanol
 Use 70% solution (most effective)
 Longer contact time and flammable
*Should research and know effectiveness and contact time for the
best disinfectant against your agent!
Disinfection

27. WRAPPERS/NON-
ABSORBENT MATERIALS
CONTAMINATED WITH
BLOOD

28. BANDAGES/OTHER
ABSORBENTS
SATURATED OR CRUSTED
WITH BLOOD

29. STAINED?….
or SATURATED?

30. Managing Non-Sharp Biohazardous Waste
 labeled
container
 lined with a
biohazardous
waste bag
 equipped with a
lid.

31. Managing Non-Sharp
Biohazardous Waste
 Securely tie bags for
transport to
treatment/collection
site.
 When moving wastes,
use secondary
containment; avoid
using public halls and
elevators.

32. “Breakable” Non-sharps Biowaste
Store in labeled containers
that are puncture-resistant,
closable and will capture
leakage, BUT….
…Do NOT use
SHARPS containers!

33. Effective Waste Autoclaving
 Leave bag open during
autoclaving or loosely closed
 Add water to bag prior to
autoclaving if primarily dry
materials
 Steam must contact
materials
 Place bag in autoclavable
tray with sides

34. Treated Waste Bag Disposal
 Allow waste bag to cool
 Use fume hood to reduce
odors
 Securely tie bag shut
 Place bag in a non-
transparent black bag for
regular disposal
Remember: NO ORANGE
BAGS IN DUMPSTER!

35. #3
Is it a sharps hazard?
Examples:
– needles
– syringes
– scalpels
– all biologically contaminated objects that
can easily penetrate skin (Pasteur pipettes,
razor blades, etc.)
Place sharps in approved sharps container
for disposal!

36. …Syringes in research settings should
be disposed of as a sharp to avoid
public relations concerns!

37. Sharps Containers
 Containers must be leak-proof,
puncture-resistant, closable & labeled
with the biohazard symbol.
 Proper sharps containers
must be used for
both clinic and
field work.

38. Proper Use of Sharps Containers
 Place tops on containers before use
on lab bench
 Don’t forget to date the container
when first put into use
 Remember: sharps
containers are a
one-way disposal
system

39. Proper Use of Sharps Containers
Use sharps containers for sharps
ONLY!
• No solid biohazardous waste (i.e.
gauze, un-broken pipettes, gloves)
• No mercury
thermometers

40. What’s wrong with this picture?

41. Sharps Container Disposal
 Containers must be permanently closed
and disposed of through the animal facility
manager:
Within 90 days
of first use
When ¾ full
 Disposal methods:
Landfill
Incineration

42. Safety Notes on Sharps Use
 Do not re-cap sharps
 Keep sharps container in
close proximity to point of
use (i.e. limit handling) for
easy disposal
 Do not leave needles in
pockets of coveralls or
smocks

43. Carcasses and Body Parts
 Human tissues
 Unfixed tissues are medical waste
 Make waste unrecognizable!
 Animal tissues, carcasses
 When generated in infectious disease or recombinant
DNA research, these are medical waste
 These items must be stored in biolabeled,
leakproof containers for incineration.
 Waste service- see Audrey Brown

44. Managing All That Other Waste…
 Drain bottles of non-hazardous
materials before disposal in trash
 <3% of volume is considered empty
 Higher volumes must not be thrown
in the trash

45. Managing All That Other Waste…
Do NOT discard
medications in the
trash.
Return to source for
disposal or seek
assistance from
your campus waste
group.
See Jaime Stock!

46. Recombinant DNA Safety Guidelines
Published by
(Department of Biotechnology,
Ministry of Science and Technology, Govt. of India)

47. Biotech Industry and rDNA Research
Indian Companies in modern biotechnology
Over 900 companies operating in all sectors of
biotechnology,
 Biopharmaceuticals- >49
 Transgenic Crops/Seeds- >60
 Industrial Products- 15 (Probiotics, Enzymes)
Indian Institutions in modern Biotech Research
Over 90 Institutions engaged in rDNA Research
 Public Funded Institutions- >57
 Private Institutions (Teaching & Research)- >37
 Universities- >115
Total number of organisations involved in rDNA Research- 256

48. A typical case of stakeholderds’ interaction
Shaping the future of transgenics
Central government - want to enforce EP Act through sate
governments as per biosafety guidelines
Politicians - want protection of public interests and safety of
environment with punishment to guilty as per Law
Public general – seeks information and are more concerned for
future with benefits and risks of rDNA products
Scientists - want to set an example by providing more products
with modern biotech research
Media – want report regularly and views of all without wrong
interpretations
Industry - request to protect their investment and enforce law at
the same time
Consensus is building on to protect public interest, punish guilty and
ensure maximum safety to environment with no (relatively low) risk !!!

49. Why Regulations are Necessary for Using GMOs
and Products Thereof?
GMOs and and their products are to play important role
including human and animal health care system, agriculture,
industrial products, environment management
Concurrently, there could be unintended hazards and risks
from the use of GMOs and products thereof, if the new
technology was not properly assessed before use
A GMO can be safe but this can be unsafe too depending
upon the trans-genes, the host organism and the environment
where the GMO is being tested
GMOs can be microorganisms, plants, and animals

51. Genetically Modified Organisms (GMOs) and r-
DNA Products Governed By
Environment (Protection) Act, 1986;
- Rules, 1989 of EPA
Industries (Development & Regulation) Act, 1951
- New Industrial Policy & Procedures, 1991
- EXIM Policy
Drugs & Cosmetics Act, 1940
- Rules 1945
Pharmaceutical Policy 2002

52. Indian EPA implementation structure for GMOs
(1989 RULES)
In order to contain possible hazards to
environment from the release of GMOs, the
Ministry of Environment and Forests has
notified in December 1989, the “Rules for the
manufacture, use, import, export and storage of
hazardous Micro-organisms/ Genetically
Engineered Organisms or Cells” under the
Environment (Protection) Act (EPA)1986.

53. APPLICATIONS OF 1989 RULES
Manufacture, import and storage of microorganisms
and gene technological products
Genetically engineered organisms/ microorganisms
and cells and correspondingly to any substance and
products and food stuffs, etc., of which such cells,
organisms or tissues form part
New gene technologies in addition to cell
hybridization and genetic engineering

54. Recombinant DNA Safety Guidelines
Scope:
1. Research
2. Large scale operations
3. Environmental risks
Research involving genetically engineered organism.
Genetic transformation of green plants,
rDNA technology in vaccine development
Large scale production of vaccine
Deliberate/ accidental release of organisms, plants and animals
Release of products derived by rDNA technology into the environment.
Not covered:
The issues relating to Genetic Engineering of human embryos,
use of embryos and foetuses in research,
human germ line gene therapy
• Indian Council of Medical Research, Ministry of Health and Family
Welfare

55. Guidelines for rDNA research activities
Category I: Exempt for the purpose of intimation and
approval of competent authority.
Category II: Require prior intimation of competent authority .
Category III: Require review and approval of competent
authority before commencement.

56. Category I: Exempt category
•The experiments involving self cloning, using strains and
also inter-species cloning belonging to organism in the
same exchanger group
•Organelle DNA including those from chloroplasts and
mitochondria.
•Host-vector systems consisting of cells in culture and
vectors, either non-viral or viral containing defective viral
genomes (except from cells known to harbour class III, IV
and special category etiologic agents

57. Category II: Intimation to competent authority
• Experiments falling under containment levels II, III and IV.
• Experiment wherein DNA or RNA molecules derived from
any source except for eukaryotic viral genome may be
transferred to any non-human vertebrate or any
invertebrate organisms and propagated under conditions of
physical containment PC1 and appropriate to organism
under study.
• Experiments involving non pathogen DNA vector systems
and regeneration from single cells.
• Large scale use of recombinants made by self cloning in
systems belonging to exempt category (e.g. E.coli,
Saccharomyces, and B. subtilis)

58. Category III: Review & approval of competent authority
• Toxin gene clonings .
• Cloning of genes for vaccine production
• Cloning of mosquito and tick DNA experiments
• Genes coding for antibiotic resistance into pathogenic organisms
• Introduction into cultured human cells of rDNA molecules containing
genes of potentially oncogenic viruses or transformed cellular genes.
• Introduction into animal cells of unidentified DNA molecules derived from
cancer cells or in vitro transformed cells.
• Experiments involving the use of infectious animal and plant viruses in
tissue culture systems.
• Experiments involving gene transfer to whole plants and animals.
• Cell fusion experiments of Animal cells.
• Transgenosis in animal experiments
• All experiments involving the genetic manipulation of plant pathogens
• Transfer of genes with known toxicity
• Experiments requiring field testing and release of rDNA engineered
microorganisms and plants
• Experiments involving engineered microbes with deletions and certain
rearrangements.
• Gene therapy for hereditary diseases of genetic disorders.

59. Containment
• Physical Containment:
Aim: to limit the spread of dangerous microorganisms
• Adopting good laboratory practices& techniques
• Using Safety equipments( primary barrier)
• Laboratory design and facilities (Secondary barrier)
• Biological Containment:
• Use of the combination of vector and host in such way that it
can limit the infectivity of vector to specific hosts
Control host vector survival in the environment
 Green house A
 Green house B
Purpose:
To reduce exposure of laboratory workers, other persons, and
outside environment to potentially hazardous agents

60. Recognition of facility
• Application for recognition of research facility to carry out
genetic manipulation should be made to the Department
of Environment before commence of work in the
prescribed Performa as per 7(2) of the rules of
hazardous microorganisms/ generically engineered
organisms notified under the EPA1986

61. STATUTORY BODIES
1. The Recombinant DNA Advisory Committee (RDAC):
2. Institutional Biosafety Committee (IBSC)
3. Review Committee on Genetic Manipulation (RCGM)
4. Genetic Engineering Approval Committee (GEAC)
5. State Biotechnology Coordination Committee (SBCC)
6. District Level Committee (DLC)

62. Recombinant DNA Advisory Committee (RDAC)
Keep updates of-
• developments at national and international levels in Biotechnology
• safety regulation for India on rDNA research use and applications.
Specific terms of reference for RDAC
• To evolve long term policy for research and development in
Recombinant DNA research.
• To formulate the safety guidelines for Recombinant DNA Research
to be followed in India.
• To recommended type of training programme for technicians and
research fellows for making them adequately aware of hazards and
risks involved in recombinant DNA research and methods of
avoiding it.

63. Institutional Biosafety Committee (IBSC)
IBSC constitutes of:
Head of the Institution or nominee
3 or more scientists
A member with medical qualifications - Biosafety Officer
One member nominated by DBT.
Functions :
• Nodal point for interaction within institution
• Project having biohazard potential needs to be notified
• Training of personnel on biosafety.
• Instituting health monitoring programme for laboratory personnel.
• On site emergency plan must be ready
• IBSC need to submit half yearly reports on ongoing projects to
RCGM
• Authorization for inter-state exchange of etiological agents,
diagnostic specimens and biological products

64. Review Committee on Genetic Manipulation (RCGM)
• RCGM constitutes of:
Department of Biotechnology
Indian Council of Medical Research
Indian Council of Agricultural Research
Council of Scientific & Industrial Research
Three Experts in Individual capacity
Department of Science & Technology
Functions :
• To review the reports in all approved ongoing research projects
involving high risk category and controlled field experiments
• Site visit of experimental facilities periodically where projects with
biohazard potential are being pursued.
• To issue authorization for import and receipt of etiological
agents, vectors, germ plasm, organelle etc
• To assist the Bureau of Indian Standards to evolve standards for
biologics produced by rDNA technology.
• To advise on intellectual property rights with respect to rDNA
technology on patents.

65. Genetic Engineering Approval Committee (GEAC):
Function under the Department of Environment
Gives approval for:
1. Import, export, transport, manufacture, process, selling of any
microorganisms or genetically engineered substances or cells
2. Discharge of Genetically engineered/classified organisms/cells
from Laboratory, hospitals and related areas into environment.
3. Large scale use of genetically engineered organisms/classified
microorganisms in industrial production and applications.
(Production shall not be commenced without approval).
4. Deliberate release of genetically engineered organisms. The
approval will be for a period of 4 years.

66. Recombinant DNA Safety Guidelines, 1990
Recombinant DNA Safety Guidelines and Regulations, 1990
Revised Guidelines for Safety in Biotechnology, 1994
Revised Guidelines for Research in Transgenic Plants, 1998
Guidelines for generating pre-clinical and clinical data for rDNA
vaccines, diagnostics and other Biologicals, 1999.
Guidelines and Standard Operating Procedures (SOPs) for
Confined Field Trials of Regulated, Genetically Engineered (GE)
Plants – 2008
Guidelines for the Safety Assessment of Foods Derived from
Genetically Engineered Plants – 2008
Protocols for Food and Feed Safety Assessment of GE crops -
2008
Revision of Guidelines is a continuous process
In Order To Evaluate Proposals, DBT has Issued
Following Guidelines:

67. Rules 1989 are applicable to the manufacture, import and storage of micro-organisms
and Gene-Technological products
Rules 1989
Para Deals with
7 Approvals to individuals on the import, export, transport, manufacture, process, use or sale of
GMOs and use of GMOs for research
8 Authorization for production of genetically modified microorganisms, plants and animals
9 Approval for deliberate or unintentional release of GMOs into the open environment
10 Approval for production, sale and import of substances and products which may contain
GMOs or cells.
11 Approval for production, sale and import of foodstuff, ingredients in foodstuff including
processing aid which may contain GMOs or cells
12 Procedures for obtaining approvals in different conditions
13 Conditions of approval of GMOs
14 Mechanism for supervising the implementation of term and conditions given with
authorization for commercial use
15 Penalties that can be levied for non compliance of measures for safe use of GMOs and
products thereof
19 Redress mechanism through National Environment Appellate Authority
Important paras of Rules, 1989

68. State Biotechnology Coordination Committee
(SBCC)
Inspect, investigate and take action against violation of
statutory provision through nodal dept and state pollution
control board/directorate of Health and medical services
Periodical review of safety and control measures in
various institutions and industries.
District level committee( DLC)
Monitor safety regulations
Coordidate activities in view of meeting any emergency
To prepare off-site emergency plan
Submit report to SBCC and GEAC

69. Large Scale Industrial Processes and Operations
Operation handing more than 20 liter are considered as large scale
Approval from GEAC is mandatory
Fundamental details of design, operation and methods of proper neutralization/
disposal
Regular monitoring by in-house and out side agency should be conducted of
control measures and safety equipments pertaining to the operation
Proper training to should be imparted to personnel for safe execution and disposal
of primary and by products
Stringent code of conduct should laid down and implemented
Regular monitoring of viable process organism in outside environment is necessary
Health status monitoring of personnel involved in process need to be done
regularly Pathogenic organism in use should be neutralized/ destroyed in proper
demonstrable way

70. A Biosafety System
Guidelines People
Biosafety Review
Process

71. GOI Government of India
DBT Department of Biotechnology
RDAC Recombinant DNA Advisory Committee
IBSC Institutional Biosafety Committee
RCGM Review Committee on Genetic Manipulation
DOEn Department of Environment
GEAC Genetic Engineering Approval Committee
SBCC State Biotechnology Coordination Committee
PI Principal Invstigator (R&D/Industry/Others)
FA Funding Agency (Govt./Private & Public Institutions)
GOI
DOen
Large
Scale
Production
&
Release
GEAC
SBCC
IBSC
PI
RCGM
RDAC
FA
rDNA Safety
Guidelines
DBT
Mechanism of Implementation of
Biosafety guidelines

72. The step-wise regulatory procedures /protocols for
five categories
Protocol-I:Indigenous product development, manufacture and
marketing of pharmaceutical products derived from LMOs but the
end product is not a LMO.
Protocol-II:Indigenous product development, manufacture and
marketing pharmaceutical products where the end product is a
LMO.
Protocol-III:Import and marketing of LMOs as Drugs/Pharmaceuticals in
finished formulations where end-product is a LMO.
Protocol-IV:Import and marketing of LMOs as Drugs/Pharmaceuticals in
bulk for making finished formulation where end product is a LMO.
Protocol-V:Import and marketing of products derived from LMOs as
Drugs/Pharmaceuticals and bought in bulk and/or finished
formulations where end product is not a LMO.

73. Protocol – I Indigenous product development derived from LMOS but end product
not a LMO.
Applicant
IBSC
RCGM approves pre-clinical trials
Pre-clinical trial conducted
RCGM recommends human CT to
DCGI and forwards views on
containment facilities to GEAC
DCGI approves human CT
Human CT conducted
GEAC examines
environmental risk v/s
benefit based on the
information on
containment facilities and
data on clinical trials
DCGI approves market
authorization under Drugs and
Cosmetic Rules based on the
clinical trials data
DCGI - Post release monitoring
Risk Group III and above Risk Group I & II
IBSC
RCGM approves pre-clinical trials
Pre-clinical trial conducted
RCGM recommends human CT
DCGI approves human CT
Human CT conducted
Environmental
Clearance
Rule1989
DCGI approves market
authorization under Drugs and
Cosmetic Rules based on the
clinical trials data
DCGI - Post release monitoring

74. Protocol – II Indigenous Product development where end Product is a
LMO
Applicant
IBSC
RCGM
(approves pre- clinical trials)
Pre-clinical trials conducted
RCGM
(evaluates toxicity and allergenicity data and
Containment facilities and recommends CT)
GEAC
(recommends Human CT)
DCGI
(approves Human CT and protocols)
A A

75. HUMAN
CT conducted
GEAC
(examines
environmental risk
versus benefits and
accords approval for
environmental release
under Rule 1989)
DCGI
(approves Market
Authorization under
Drugs & Cosmetics
Rules based on clinical
trials data)
DCGI
(Post Release Monitoring)
A
Protocol – II Contd.

76. Protocol – III Import and marketing of LMOs as drugs in finished
formulations
Applicant
GEAC
(examines data generated in the Country of origin and other countries where
the product has been tested and accords ‘In Principle’ approval for import
and conduct of clinical trials. GEAC recommends to DCGI)
DCGI
(approves Human CT and protocols)
HUMAN
CT conducted
GEAC
(examines environmental risk
versus benefits and accords
approval for environmental release
under Rule 1989).
DCGI
(approves Market Authorization under Drugs &
Cosmetics Rules based on clinical trials data)
DCGI
(Post Release Monitoring)

77. Protocol – IV Import of LMOs as drugs in bulk for making
finished formulations.
Applicant
GEAC
(examines data generated in the Country of origin and other countries where the
product has been tested and accords “in principal” approval for limited import for
conduct of clinical trials, GEAC informs DCGI and directs the applicant to setup
IBSC)
IBSC
RCGM
(approves activity, recommends to DCGI for clinical
trials and forward views to GEAC on containment
facilities)
GEAC
(recommends Human CT)
DCGI
(approves Human CT and protocols)
A A

78. Protocol – IV Contd.
HUMAN
CT conducted
GEAC
(examines environmental risk
versus benefits and accords
approval for environmental
release under Rule 1989)
DCGI
(approves Market Authorization under
Drugs & Cosmetics Rules based on clinical
trials data)
DCGI
(Post Release Monitoring)
A

79. Protocol – V Import and marketing of products derived from
LMOs as Drugs and bought in bulk and/or finished formulations.
Applicant
DCGI
(Examination of complete dossier including human clinical
trials protocols and trials if conducted and to accord
approval for Human CT and protocols after obtaining the
comments of RCGM)
HUMAN
CT conducted
DCGI
(approves Market Authorization under Drugs &
Cosmetics Rules based on clinical trials data)
DCGI
(Post Release Monitoring)

80. Guidelines for generating Pre-clinical and Clinical data
for r-DNA Based vaccines, diagnostics and other
biologicals, 1999
 Specification and characterization information
Details description
• of the method of r-DNA product
• sequence verification
• identity-Physical, Chemical, Immunological and Biological properties
• Potency (for recombinant vaccines & biologicals)
• General Safety Test
Data on sterility tests as per Indian Pharmacopia guidelines.
Data on purity of recombinant product
Description of constituent materials like preservatives etc.
Data on stability of finished formulation
Specific mandatory information required
Quality Control of Biological products produced by
rDNA technology

81. Preclinical Testing:
General Principles
Biological activity/ pharmacodynamics
Animal species/ model selection
Number/ gender of animals
Administration/ dose selection
Immunogenicity
Specific Considerations:
Safety pharmacology
Toxicology and pharmacokinetics(ADME)
Immunotoxicity
Reproductive performance and developmental toxicity
Genotoxicity studies
Carcinogenicity studies
Guidelines for generating Pre-clinical and Clinical data
for r-DNA Based vaccines, diagnostics and other
biologicals, 1999

82. In vitro diagnostic:
• recombinant reagents
• monoclonal antibodies
Guidelines for generating Pre-clinical and Clinical data
for r-DNA Based vaccines, diagnostics and other
biologicals, 1999
Clinical Trials:
• Human/ Clinical Pharmacology (Phase I):
Immunogenic Potency
• Exploratory Clinical Trials (Phase II):
Preventive/ Therapeutic Efficacy
• Confirmatory Trials (Phase III):
Newer vaccines
Established vaccines

83. Classification of micro-organisms: Bacteria
Risk Group I Risk Group II Risk Group III
Allbacterial
agents not
included in
higher classes
according to
"Basis for Agent
Classifications
Actinobacillus Actinobacillus mallei
Arizona hinshawii - Bartonella
Bacillus anthracis Brucella -
*Bordetella Clostridium botulium, Cl. tetani
Borrelia recurrentis, B. vincenti Francisella tularensis
Cl. chauvoei, Cl. difficle Cl. fallax, Cl. haemolyticum, Mycobacterium avium, M.bovis, M. tuberculosis, M. leprae.
Cl. histolyticum, Cl. novvi, Cl. perfringes, Cl. septicum, Cl.sordelbi Pasteurella multocida
Corynebacteriumdiptheriae*,C.equi, C.haemolyticum Pseudomonas pseudomallai
C.pseudotuberculosis,C.pyogenes, C.renale Yersinia pestis
Diplococcus (Streptococcus) pneumoniae
Erysipelothrix insidiosa
Escherichia coli-
Haemophilus ducreyi, H.influenzae, H. pneumoniae
Herellea vaginicola
Klebsiella-Letionella Leptospira interrogans ,Listeria, all species
Mima polymorpha Moraxella-all species
Mycobacteria-allspeciesincluding Mycobacterium avium, M.bovis,
M. tuberculosis, M.leprae.
Mycoplasma-all species except M.mycoides and M.agalactiae
Neisseria gonorrhoeae, N. meningitidis
Pasteurella . Salmonella,Shigella
Streptococcuspyogenes,S.equi, S.pneumonine
Treptonema carateum, T.pallidum and T. pertenue
Sphaerophorus neorophorus, Staphylococcus aureus,
Streptobacillus moniliformis
Vibrio foetus, V.comma
Streptomyces madurae pelleteri somaliensis

84. Classification of micro-organisms: Fungi
Risk Group I Risk Group II Risk Group IIi
All fungal
agents not
included in
higher classes
according to
"Basis for Agent
Classifications
Actinomycetes (including) Nocardia and
Actinomyces and Arachina propionica
Coccidioides immitis
Aspergillus fumigatus Histoplasma capsulatum
Blastomyces dermatitidis Histoplasma capsulatum var duboissi
Cryptococus neoformans C. fersiminosos
Epidermophyton madurella, E. microsporon
Paracoccidioides brasiliensis
(Sporothrix Trichoderma Trichophyton)

85. Classification of micro-organisms: Parasites
Risk Group I Risk Group II Risk Group III
All Parasitic
agents not
included in
higher classes
according to
"Basis for Agent
Classifications:.
Entamoeba histolytica Schisistosoma *mansomi
Leishmania species
Naegleria gruberia
Plasmodium thcilera
Plasmodium fabesia, P.falciparum
Schistosoma
Toxoplasma gondii
Toxocara canis
Trichinella spiralis
Trichomonas
Trypanosoma cruzi

86. Risk Group II Risk Group II Risk Group III
All viral, rickettsial
and chlamydial
agents not included in
higher classes
Adenoviruses - African Horse Sickness
Avian loukosis, Leucosis complex Alastrim, monkey pox and whotepox
Cache Valley virus Arboviruses, Blue Tongue virus, Epstein -
Barr viurs
CELO (avain adenovirus) Feline Leukemia, Feline
sarcoma
Coxsackio A and B viruses Gibbon Ape Lymphosarcoma
Corona viruses, Cytomegalo viruses Herpes simplex saimiri
Newcastle disease virus Dengue virus Herpes virus ateles
Rinderpest- attenuated virus strain (e.g. Kabatte-O Simian viruses ,Simian virus 40
Ad 7 SV 40 (defective)
Sindibis virus,Rensaw virus,Turlock virus
Vaccinia virus,Varicella virus
Vole rickettsia,Yellow fever virus, 17D vaccine strain
Foot-and-Mouth Disease virus (all serotypes
and subtypes)
Echo viruses Herpes simplex 2
Risk Group IV Encephalomyocarditis virus (EMC) HIV-1 & HIV-2 and strains of SIV
Alastrim, monkeypox, whitepox Flanders virus, Hart Park virus Infectious Equine Anaemia
Hemorrhagic fever &Crimean hemorrhagic hepatitis A and B viruses, non A and non B, HDV Herpes Lymphocytic choriomeningitis virus (LCM)
Korean hemorrhagic fever (Congo) Infectious Bovine Rhinotraechitis virus (IBR). Psittacosis-ornithosis-trachoma group of
agents
Herpes virus simae (monkey B viurs) Infectious bronchitis, Infectious Bursal diseases. Pseudorabies virus
Tick-borne encephalitis virus complex, including
Russian Spring
Infectious Laryngotraechitis (ILT) Rabies street virus, when used inoculations
of carnivores
Summer Encephalitis, Kyasanur Forest Disease, Omsk
hemorrhagic fever and Central European
Encephalitis viruses
Influenza virus- Langat virus Risckettsia
Lymphogranuloma venereum agent. Sheep pox (field strain)
Marek's Disease virus, Measles, virus Mumps virus Swine Fever virus
Newcastle disease Vesicular stomatitis virus
Parainfluenza viruses Wooly monkey Fibrosarcoma
Polio viruses, Poxviruses. Yaba pox virus
Rabies, Rubella virus Non-defective Adeno-2 SV-40 hybrids
Reoviruses, Respiratory syncytial virus
Rinderpest, Rhinoviruses
Classification of micro-organisms: Viruses

87. Inter-ministerial Standing Committee on Biotechnology
Regulation
Constitution of a standing inter-ministerial committee to redress and look into
various regulatory aspects and make issue-based recommendations on
case-by-case basis. Prior to any deviation from the proposed regulatory
mechanism, which when comes in vogue, the views of this inter-ministerial
committee should be obtained in the first instance.
The suggested composition of the committee is as follows:
Chairman -To be an Eminent Scientist
Chairman, GEAC -Member
Chairman, RCGM -Member
Member-Secretary, GEAC -Member
Member-Secretary, RCGM -Member
Joint Secretary (Seeds), MoA -Member
DDG, ICAR (Crop Sciences) -Member
Joint Secretary (MoEF) -Member
Joint Secretary (Food Processing) -Member
Adviser (Industry, DBT) -Member
DG/Representative (ICMR) -Member
DCG(I) -Member
Experts on Immunobiologicals, Biogenerics, Plant Breeding, Molecular Biology,
Environmental Sciences and other relevant areas may be co-opted from time to time.

88. National Biotechnology Regulatory Authority/
Commission
•Alternate models of a ‘National Biotechnology Regulatory
Authority’
•Amendment of EPA and harmonization Seeds Act/ Drugs &
Cosmetics Rules/ PFA Act may be necessary to obviate the
approvals required under these statues.
•Harmonization is an essential prerequisite for establishing
the national biotechnology regulatory authority.
•Recommended an inter-ministerial group to examine the
model proposed and make specific proposals with respect
to the implementation including the budgetary
requirements.

90. GOI
DOen
Large
Scale
Production
and
Release
GEAC
SBCC
IBSC
PI
RCGM
RDAC
FA
rDNA Safety
Guidelines
DBT