Module 1A

1
Module 1A
CDC/UNMC General Biosafety Training Overview
2
General BiosafetyModule 1 Outline

• Overview
• Module 1A
• Biosafety Levels 1 and 2
• Module 1B
• Safety Equipment
• Module 1C
• Decontamination and Biohazardous Waste Handling
• Module 1D

3
General BiosafetyModule 1A Synopsis

• This module gives an overview of the federal
  guidelines concerning biosafety practices and how
  these practices are applicable to the research
  laboratory.

4
Who needs this training?

• Individuals who work with or have access to
  biohazardous materials.

5
Definition

• Biohazardous materials
• Materials of biological origin that have the
  capacity to produce deleterious effects on humans
  or animals.

6
Examples of Biohazardous Materials

• Recombinant DNA molecules
• Microorganisms containing recombinant DNA
  molecules
• Microorganisms classified as risk group 2 (RG-2),
  RG-3, or RG-4 (defined in the next series of
  slides)
• Biological products derived from RG-2, RG-3, or
  RG-4 organisms
• Diagnostic specimens known or reasonably expected
  to contain RG-2, RG-3, or RG-4 organisms.
• Clinical/ medical waste derived from the medical
  treatment of humans or animals or from biomedical
  research.

7
Definition

• Biosafety
• The combined application of laboratory practice
  and procedure, laboratory facilities, and safety
  equipment when working with potentially
  infectious microorganisms.

8
NIH Guidelines

• Federal guidelines that must be followed when
  performing research using biohazardous agents
• The Guidelines purpose is to specify practices
  for constructing and handling rDNA molecules and
  organisms containing these molecules.
• UNMC/ UNO also use these guidelines to include
  the handling of other biohazardous agents (not
  associated with rDNA) as a part of the biosafety
  program.

9
Institutional Biosafety Committee

• Members appointed by the University with the
  following duties
• Review research for compliance with NIH
  Guidelines
• Notify PI of the results of an IBC review
• Setting containment levels for experiments
• Periodic review of research to ensure adherence
  to NIH Guidelines
• Adoption of an emergency plan covering accidental
  spills and personnel contamination and
• Report significant problems with or violations of
  the NIH Guidelines.
• (NIH Guidelines, Section IV-B-2b)

10
Biosafety Officer

• Appointed by the Institution with duties to
  include
• Periodic inspections to ensure that laboratory
  standards are rigorously followed
• Reporting to the IBC any significant problems,
  violations of the NIH Guidelines, and significant
  research-related accidents or illnesses
• Developing an emergency plan for handling
  accidental spills and personnel contamination
• Providing advice on laboratory security and
• Providing technical advice to PIs and the IBC on
  research safety procedures.
• (NIH Guidelines, Section IV-B-3)

11
Principal Investigators Responsibility

• Make available to all laboratory staff the
  protocols that describe the potential biohazards
  and the precautions to be taken
• Instruct and train laboratory staff in the
  practices and techniques required to ensure
  safety and the procedures dealing with accidents
  
• Inform the lab staff of the reasons and
  provisions for any precautionary medical
  practices advised or requested
• Supervise the safety performance of the
  laboratory staff
• Investigate and report any significant problems
  pertaining to the operation and implementation of
  containment practices to the IBC
• Correct work errors and conditions that may
  result in a release of a biohazardous agent and
• Ensure the integrity of the physical and
  biological containment.
• (NIH Guidelines, Section IV-B-7-c,d,e)

12
NIH Committee Overviews

• OBA
• Office of Biotechnology Activities
• Office within the NIH that is responsible for
  review and coordinating all actions of the NIH
  Guidelines
• RAC
• Recombinant DNA Advisory Committee
• Public advisory committee that advises the
  Department of Health and Human Services and NIH
  Director on recombinant DNA research issues.

13
Experiments Requiring IBC Approval

• Six major categories
• IBC and NIH Director approval and RAC Review
  before initiation
• IBC and NIH/OBA approval before initiation
• IBC and IRB approval and RAC review before
  initiation
• IBC approval before initiation
• IBC notice simultaneous with initiation
• (NIH Guidelines, Section III)

14
IBC and NIH Director Approval and RAC Review

• Example
• Experiments involving the deliberate transfer of
  a drug resistant trait to microorganisms not
  known to acquire that trait naturally
• NIH Guidelines, Section III-A

15
IBC and NIH/OBA Approval

• Example
• Experiments involving the deliberate formation of
  rDNA containing genes for the biosynthesis of
  toxin molecules lethal for vertebrates at an LD50
  of less than 100 ng/kg body weight
• NIH Guidelines, Section III-B

16
IBC and IRB Approval and RAC Review

• Example
• Experiments involving the deliberate transfer of
  rDNA into human research participants
• Also called human gene transfer experiments
• NIH Guidelines, Section III-C

17
IBC Approval Before Initiation

• Examples
• Experiments involving the introduction of rDNA
  into risk group 2, RG-3, or RG-4 agents
• Experiments involving the use of infectious or
  defective viruses in the presence of helper virus
  in tissue culture
• Experiments involving whole animals in which the
  animal has been altered by the introduction of
  DNA or RNA (transgenic animals)
• Experiments to genetically engineer plants by
  rDNA methods
• Experiments involving more that 10 liters of
  culture material
• Experiments in which DNA for RG-2, RG-3, or RG-4
  agents is cloned into nonpathogenic prokaryotic
  or lower eukaryotic host vector systems
• NIH Guidelines, Section III-D

18
IBC Approval Simultaneous with Initiation

• Example
• Experiments in which all components are derived
  from non-pathogenic prokaryotes and
  non-pathogenic lower eukaryotes.
• NIH Guidelines, Section III-E

19
Why biosafety practices?

• To protect
• workers
• products
• co-workers
• environment
• students
• visitors

20
Risk Assessment Process

• To determine the level of containment to handle a
  biohazardous agent
• Subjective process
• Based on the following
• Virulence
• Pathogenicity
• Infectious dose
• Environmental stability
• Route of spread
• Communicability
• Operations
• Quantity
• Availability of vaccine or treatment
• Gene product effects
• Toxicity, physiological activity, and
  allergenicity
• The IBC gives final approval to the biosafety
  level of containment
• (NIH Guidelines, Section II)

21
Chain of Infection and Means of Protection
22
Laboratory Requirements

• Knowledgeable supervisor
• Knowledgeable personnel
• Aware of potential hazards
• Proficient in lab practices techniques
• Availability of a lab specific biosafety manual

23
Biosafety Lab Manual Components

• Contact information
• Copies of IBC, IRB, and/or IACUC protocols
• Standard Operating Procedures
• For unique procedures
• Laboratory inspection checklist
• Emergency plan for handling spills and personnel
  contamination
• UNMCs IBC spill procedure
• Relevant reference materials
• Information on the biology of the organism(s)
  used in the laboratory
• UNMC Biosafety Manual
• UNMCs IBC policies pertaining to biosafety

24
Classification of Infectious Agents

• Classified into risk groups on the basis of risk
  to the individual and to the community.
• Currently, 4 risk group levels have been
  designated.
• the least risk (RG-1) to the most risk (RG-4).
• NIH Guidelines, Appendix B

25
RG-1 Agents

• Not associated with disease in healthy adult
  humans.
• Generally only require a laboratory with minimal
  containment
• Biosafety Level 1
• Declaration of Dangerous Goods is not generally
  required for transportation by air.
• Classified as a diagnostic specimen

26
RG-2 Agents

• Associated with human disease which is rarely
  serious.
• Preventative and therapeutic interventions are
  available.
• Generally require a laboratory with moderate
  containment.
• Biosafety Level 2
• Declaration of Dangerous Goods required for
  shipment.

27
RG-3 Agents

• Associated with serious or lethal disease.
• High individual risk but low community risk
• Preventative or therapeutic interventions may be
  available.
• Generally require a laboratory with high level
  containments
• Biosafety level 3
• Declaration of Dangerous Goods required for
  shipment.

28
RG-4 Agents

• Likely to cause serious or lethal human disease
• High individual and community risk
• Preventative or therapeutic interventions are not
  usually available.
• Require a laboratory with extensive high-level
  containment
• Biosafety level 4
• Research on these agents are not allowed on
  campus since a BSL-4 containment lab is not
  currently available.

29
Biosafety Levels of Containment

• Combination of laboratory practices, safety
  equipment, and laboratory design to achieve
  levels of physical containment.
• Currently, 4 biosafety levels have been
  designated.
• The least restrictive (BSL-1) to the most
  restrictive (BSL-4).
• Specifics of these containment types will be
  discussed in more detail in Module 1B.
• NIH Guidelines, Appendix G

30
BSL-1 Containment Overview

• RG-1 Agents
• Not known to cause disease in healthy adult
  humans
• Practices
• Standard microbiological practices
• Safety equipment
• Minimal requirements
• Facilities
• Open bench top

31
BSL-2 Containment Overview

• RG-2 Agents
• Associated with mild to moderate disease in
  humans
• Practices
• BSL-1 plus limited access.
• Safety equipment
• Biological Safety Cabinet and personal protective
  equipment as needed.
• Facilities
• BSL-1 plus the availability of a mechanism for
  decontamination.

32
BSL-3 Containment Overview

• RG-3 Agents
• Associated with serious or potentially lethal
  disease in humans
• Practices
• BSL-2 plus controlled access.
• Safety equipment
• Biological Safety Cabinet and personal protective
  equipment required.
• Facilities
• BSL-2 with self-closing double door access and
  single-pass negative directional airflow.

33
BSL-4 Containment Overview

• RG-4 Agents
• Associated with high risk of life-threatening
  disease in humans and/or animals
• Practices
• BSL-3 plus controlled access
• Safety equipment
• Biological Safety Cabinet
• Full-body air-supplied, positive pressure
  personnel suit
• Facilities
• BSL-3 plus dedicated air and exhaust,
  decontamination procedures for exit, separate
  building, etc.
• (This containment lab is not available on the
  UNMC/UNO campuses.)

34
Risk Group vs Biosafety Level

• Biosafety risk assessment
• To determine the risk group of a biological agent
• RG-2 organisms are generally handled in BSL-2
  containment and RG-3 in BSL-3.
• Exceptions
• RG-2 agents used in large quantities may require
  BSL-3 containment
• RG-3 agents under some circumstances may be
  manipulated at BSL-2 containment.
• Institutional Biosafety Committee approves the
  level of laboratory containment required
  following an IBC protocol review.

35
This completes Module 1A. Continue with
Module 1B.