Module 1A

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Module 1A
UNMC General Biosafety Training Overview
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General BiosafetyModule 1 Outline
Note Module 1 is composed of 5 subsections (1A,
1B, 1C, 1D, and 1E)

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

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General BiosafetyModule 1A Synopsis

• Information about this training module
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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

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Who needs this training?

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

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Definition

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

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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.

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Definition

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

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NIH Guidelines

• Federal guidelines that must be followed when
  performing research using biohazardous agents
• The Guidelines purpose
• 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 a total
  biosafety program.

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Institutional Biosafety Committee

• UNMC IBC membership
• appointed by the Associate Vice Chancellors for
  Academic Affairs
• Duties include
• Review research for compliance with NIH
  Guidelines
• Notify Primary Investigator of the results of an
  IBC review
• Set containment levels for experiments
• Review research periodically to ensure adherence
  to NIH Guidelines
• Adopt 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)

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Biosafety Officer

• Appointed by the Vice Chancellor for Academic
  Affairs
• Duties
• Periodic inspections to ensure that laboratory
  standards are rigorously followed
• Report to the IBC any significant problems,
  violations of the NIH Guidelines, and significant
  research-related accidents or illnesses
• Develop an emergency plan for handling accidental
  spills and personnel contamination
• Provide advice on laboratory security and
• Providr technical advice to PIs and the IBC on
  research safety procedures.
• (NIH Guidelines, Section IV-B-3)

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Principal Investigators Responsibility

• Make available to all laboratory staff
• 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)

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NIH Committee Oversight

• OBA
• Office of Biotechnology Activities
• Office within the NIH that is responsible for
  reviewing 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 the
  NIH Director on rDNA research-related issues.

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Experiments Requiring IBC Approval

• Five major categories of the review process
  dependent on the type of experiments conducted
• 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)

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Review Process 1IBC and NIH Director Approval
and RAC Review Before Initiation

• 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

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Review Process 2IBC and NIH/OBA Approval
Before Initiation

• 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

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Review Process 3IBC and IRB Approval and RAC
Review Before Initiation

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

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Review Process 4IBC 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

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Review Process 5IBC 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

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Why biosafety practices?

• To protect
• Workers,
• Products,
• Co-workers,
• Environment,
• Students, and
• Visitors

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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
• All IBC applications include a section describing
  this risk assessment process
• The IBC gives final approval to the biosafety
  level of containment
• (NIH Guidelines, Section II)

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Chain of Infection and Means of Protection
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Laboratory Requirements

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

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Biosafety Lab Manual Components
Note All laboratories conducting biohazardous
agent research are required to have a biosafety
manual available

• 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

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Classification of Infectious Agents

• Classified into risk groups on the basis of
• Risk to the individual and
• Risk to the community
• Four risk group levels have been designated
• Minimal risk (RG-1)
• Moderate risk (RG-2)
• High risk (RG-3)
• Extreme risk (RG-4)
• NIH Guidelines, Appendix B

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RG-1 Agent

• Not associated with disease in healthy adult
  humans
• Generally only require a laboratory with minimal
  containment
• Biosafety Level 1
• Shipment by air
• Biological Substance Category B
• UN 3373

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RG-2 Agent

• Associated with human disease
• But rarely serious
• Preventative and therapeutic interventions are
  available
• Generally require a laboratory with moderate
  containment
• Biosafety Level 2
• Shipment by air
• Biological Substance Category B
• UN 3373

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RG-3 Agent

• 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
  containment
• Biosafety level 3
• Shipment by air
• Category A agent
• Declaration of Dangerous Goods statement
  required
• Label with
• UN2814 Infectious Substance Affecting Humans
• Infectious Substance 6

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RG-4 Agent

• 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
• No BSL-4 containment lab is available

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Biosafety Levels of Containment

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

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BSL-1 Containment Overview

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

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BSL-2 Containment Overview

• Agents used
• RG-2 Agents
• Associated with mild to moderate disease in
  humans
• Practices
• BSL-1 plus
• Limited access
• Safety equipment
• Biological safety cabinet
• Personal protective equipment as needed
• Facilities
• BSL-1 plus
• the availability of a mechanism for
  decontamination

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BSL-3 Containment Overview

• Agents used
• RG-3 Agents
• Associated with serious or potentially lethal
  disease in humans
• Practices
• BSL-2 plus
• Controlled access
• Safety equipment
• Biological safety cabinet
• Personal protective equipment as required
• Facilities
• BSL-2 with self-closing double door access and
• Single-pass negative directional airflow

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BSL-4 Containment Overview

• Agents used
• 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.)

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Risk Group vs Biosafety Level

• Biosafety risk assessment
• To determine the risk group of a biological agent
• Risk group and biosafety level generally
  correspond
• RG-1 used in BSL-1
• RG-2 used in BSL-2
• etc.
• 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
• Through an IBC protocol review.

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This completes Module 1A

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