Chapter 26
Biosafety

Contents

Approved by Bruce King
New 1/07

26.1 General Biosafety Policy
26.2 Biological Risk and Requirements

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26.1 General Biosafety Policy

Work with biological material at Berkeley Lab is conducted in a safe, ethical, environmentally sound, and compliant manner using the principles and functions of Integrated Safety Management (ISM) and work authorization (PUB-3000, Chapter 6).  For the purpose of this policy, biological materials encompass a broad range of organisms, cells, viruses, and other materials of biological origin that pose differing levels of risks to plants, animals, or humans.  Line management and researchers are expected to define their biological work, evaluate the biological hazards, determine the risk, and implement required biosafety containment controls (i.e., establish a Biosafety Level).  This is accomplished with the assistance and oversight of the Institutional Biosafety Committee (IBC), the Environment, Health, and Safety Division (EH&S), and other Berkeley Lab ES&H functions.  The Biological Use Notification, Registration, and Authorization process is used to facilitate and document the above process and ensure compliance.

Berkeley Lab’s policy incorporates required federal and state government, contract, and funding regulations and standards.  This chapter provides an overview of biosafety policy to assist in identifying and understanding the requirements for evaluation and control of biological work in research.  LBNL’s policies and requirements for work with biological material are further detailed in the online Biosafety Program and Biosafety Manual.

26.2 Biological Risk and Requirements

Identification of biological risk and requirements associated with an operation must consider three primary factors:  the inherent hazard (perceived or real) posed by the biological material or agent, the susceptible hosts (i.e., receptors) that may be affected by the material or agent, and the exposure pathways between the threat hazard and the susceptible host.  This section discusses these factors and provides an overview of biological risk and hazard categories, biosafety standards, and associated requirements.

26.2.1 Risk Assessment and Groups

The term “biohazardous” is used to describe biological materials that present potential risk to the health of humans or other organisms, either directly through infection or indirectly through damage to the environment. 

To define the risk associated with a biological agent, the researcher must make an initial risk assessment based on the Risk Group (RG) of the agent.  Four Risk Groups (RGs) have been defined by the World Health Organization (WHO) and adopted by the Centers of Disease Control and Prevention (CDC).  These Risk Group categories are defined by the National Institutes of Health (NIH) in Appendix B of the NIH Guidelines for Research Involving Recombinant DNA Molecules.  These NIH Risk Group definitions are based on risk of disease in humans and are shown in Table 26-1.  Assignments of specific agents to Risk Groups can also be found in Appendix B of the same document or in other sources such as the American Biological Safety Association (ABSA) risk group database.  Risk Group definitions and classification may vary slightly between countries.  Berkeley Lab uses the NIH Risk Group definitions and agent classifications. 

Table 26-1  Risk Groups and Definitions

When deciding the appropriate containment for the biological work, consideration should be given to the Risk Group assignment, agent factors, how the agent is to be manipulated, and if medical surveillance is required.  Agent factors that should be considered include: virulence, pathogenicity, infectious dose, environmental stability, route of spread, and communicability.  Manipulations that should be assessed include: handling operations (e.g., use of sharps), quantity of agent, and aerosol generation.  Medical surveillance factors may include: availability of vaccine or treatment, physiological activity, and allergenicity.  Any recombinant biological material that is known to be more hazardous than the parent (wild-type) may require handling at a higher containment level.

26.2.2 Bloodborne Pathogens and Human Materials

Bloodborne pathogens (BBPs) are infectious agents that are capable of causing human disease and are transmitted through human blood and tissues.  Examples include the hepatitis B virus (HBV) and the human immunodeficiency virus (HIV).  According to the Federal Occupational Safety and Health Administration (OSHA) BBPs Standard, materials that are regulated based on their potential to contain BBPs include human blood, human blood components, products made from human blood, and other potentially infectious materials (OPIM) listed in Table 26-2.  Work that may involve exposure to these human materials is conducted at Biosafety Level 2 containment (see Section 26.3) and requires compliance with the specific requirements of the Fed/OSHA BBPs Standard.  Dried blood and some human secretions (e.g., urine, feces, vomit, tears, sweat, sputum, nasal secretions, and saliva) that do not contain blood are not considered OPIM.

Table 26-2  Human and Other Materials Covered by the OSHA Bloodborne Pathogens Standard*

26.2.3  Recombinant Organisms and Genomic Materials

Work with recombinant organisms or specific recombinant genomic materials and nucleic acids may involve risks to humans, animals, plants, or the environment.  Requirements for work with recombinant DNA molecules are specified in the NIH Guidelines for Research Involving Recombinant DNA Molecules (NIH Guidelines), and recombinant DNA molecules are defined in Section I-B.  Institutions such as LBNL that receive NIH funding are required to comply with the NIH Guidelines.  As a condition for NIH funding, LBNL and researchers must ensure that all recombinant DNA research conducted at or sponsored by LBNL, irrespective of the source of funding, complies with the NIH Guidelines.

26.2.4  Pathogenic and Etiologic Agents

26.2.4.1 Pathogens and Virulence

Pathogens are infectious microbial (e.g., bacteria, protozoa, fungi, viruses, etc.) or other agents that can cause disease in healthy host organisms such as humans, animals, or plants.  “Opportunistic pathogens” may become pathogenic on their own or by infecting immuno-compromised hosts.  Avirulent or attenuated agents are pathogens that were physiologically modified or genetically altered in a way that they are several orders of magnitudes less likely to produce disease in a healthy host organism.  Pathogens may be regulated by a variety of agencies (e.g., OSHA, NIH, CDC, USDA) depending on the potential hosts and impacts (e.g., humans or livestock).

26.2.4.2 Biological Etiologic Agents

A “biological etiologic agent” is an agent of biological origin (e.g., bacterium, fungus, parasite, virus, etc.) that causes disease in humans (i.e., pathogenic to humans).  See Appendix B of the NIH Guidelines for a list and Risk Group categorization of human etiologic agents.

This section describes specific requirements and programs for biological etiologic agents that are included in LBNL’s biosafety program, as required by the DOE Worker Safety and Health Program (10 CFR 851, Appendix A, Section 7).  Line management is responsible for reporting work with biological etiologic agents.  The Institutional Biosafety Committee (IBC) and EH&S Division have specific oversight and reporting responsibilities listed in the following sections.

26.2.4.2.1  Institutional Biosafety Committee

Berkeley Lab has an established IBC that has the following responsibilities related to biological etiologic agents:

• The IBC reviews work with biological etiologic agents for compliance with applicable CDC, NIH, WHO, and other international, federal, state, and local guidelines, and assesses the containment level, facilities, procedures, practices, training, and expertise of personnel using these agents.  Applicable guidelines include the NIH Guidelines for Research Involving Recombinant DNA Molecules, the CDC-NIH Biosafety in Microbiological and Biomedical Laboratories (BMBL), and the WHO Laboratory Biosafety Manual.
• The IBC reviews LBNL’s security, safeguards, and emergency management plans and procedures to ensure they adequately consider work involving biological etiologic agents. 
• The IBC (which includes the LBNL Medical Director and Biosafety Officer) assesses potential vaccines and the need for immunizations when it reviews work to be conducted with biological etiologic agents.  Any immunization requirements will then be incorporated into the operation’s biosafety documentation.

26.2.4.2.2  EH&S Division

The EH&S Division has the following responsibilities related to biological etiologic agents:

• The EH&S Industrial Hygiene Group maintains an inventory and status of biological etiologic agents, and provides to the DOE Berkeley Site Office (through the LBNL IBC) an annual status report describing the status and inventory of biological etiologic agents and their biological safety program.  The inventory includes the agent(s)’ names, PI’s or owner’s name, and location(s) of the researcher’s laboratory operation.  The annual report provides specific inventory information for select agents, and information on how and where inventory information for non-select agents is maintained.
• The EH&S Security and Emergency Operations and Industrial Hygiene Groups confirm that the site safeguards and security plans and emergency management programs address biological etiologic agents and biological select agents.

26.2.5  Select Agents and Toxins

The term “select agents and toxins” is used to describe the list of specific pathogenic agents and toxins that are strictly regulated by the CDC and USDA, because they may potentially be used as biological weapons or pose a severe threat to human, animal, and plant health.  Specific genetic elements, recombinant nucleic acids, and recombinant organisms that may pose a similar threat are also regulated. 

26.2.5.1  Select Agent and Toxin Requirements

Possession, use, or transfer of select agents and toxins must be conducted in compliance with the CDC and USDA regulations related to human, plant, and animal select agents and toxins.

26.2.5.2  Select Agent Reporting to DOE

Select agents are defined in Section 26.2.5.  The LBNL EH&S Responsible Official or Alternate Responsible Official for select agents submits to the DOE Berkeley Site Office (BSO) the following select agent notifications as required under the DOE Worker Safety and Health Program (10 CFR 851, Appendix A, Section 7):

• Each Laboratory Registration/Select Agent Program registration application package requesting registration of a new laboratory facility for the purpose of transferring, receiving, or handling select agents.  DOE’s review and concurrence is required before transmittal to the CDC.
• A copy of each CDC Transfer of Select Agents form: (a) upon initial submission of the form to a vendor or other supplier requesting or ordering a select agent for transfer, receipt, and handling in a registered facility, and (b) within ten days upon completion of the form documenting final disposition of the select agent.
• Documentation of the destruction of a complete stock of a select agent within ten days following the destruction.

26.2.6  Animals, Plants, and Soils

Some organisms, agents, or materials may significantly harm domestic or native animals and plants, or U.S. agricultural and natural resources.  These biological materials are regulated by the U.S. Department of Agriculture (USDA) and require permitting (e.g., soil or import permit) because they may cause harm directly (e.g., predator or pathogen) or indirectly (e.g., vector).  General examples include specific animals, plants, animal pathogens, plant pathogens, materials that may contain such pathogens (e.g., soil), and agents that pose a severe threat.

26.3 Biosafety Containment Levels

Based on the evaluation of biological hazards related to the work, a biosafety containment level and specific work practices shall be established and implemented for the operation.  Biosafety containment levels consist of combinations of standard microbiological practices, safety equipment, and facilities needed to properly contain the biological work.  Biosafety Levels (BLs) are defined in the LBNL Biosafety Manual and are based on the Biosafety Level requirements in the NIH Guidelines (Section II-B, Appendix G, and Appendix I) and the Biosafety Level guidelines in the CDC/NIH Biosafety in Microbiological and Biomedical Laboratories.  Agents requiring implementation of BL3 or BL4 currently are not used or stored at Berkeley Lab.

26.4 Biological Work Review, Documentation, and Authorization

Researchers and line management are expected to define their biological work, evaluate the biological hazards, implement biosafety containment controls, and ensure their work is authorized.  This is accomplished with the assistance and oversight of the Institutional Biosafety Committee (IBC), EH&S Division (e.g., biosafety and hazardous waste management programs), and other Berkeley Lab ES&H functions.  See Table 26-3 for a summary of the review, documentation, and authorization requirements for work with biological materials.  The paragraphs below provide additional explanation.

26.4.1  General Process

The Principal Investigator (PI) and line management are responsible for ensuring that research with biological material is documented, reviewed by the IBC, and authorized when and as required.  Submission and authorization of documentation results in a Biological Use Notification, Registration, or Authorization as detailed in Table 26-3.  LBNL forms are available for this purpose.  Other documentation may also be required.  EH&S Biosafety Program personnel assists the PI in screening the work documentation, determining the applicable requirements and documentation, and coordinating the IBC review process.

The PI is responsible for keeping the work documentation updated, and submitting it for review to the EH&S Biosafety Program either according to the renewal cycles shown in Table 26-3 or when there are significant changes in the work (e.g., addition of recombinant DNA work, addition of pathogens, new types of biohazardous materials, change of location, or change of responsible personnel).  If the changes in work are not significant and the IBC reviewed the document less than three years prior, the EH&S Biosafety Program may approve and distribute the updated document without IBC review. 

26.4.2  Biological Use Notification

A Biological Use Notification is used for work that involves only Risk Group 1 biological material or recombinant material that is exempted by NIH, and does not involve other conditions that require review as listed in Table 26-3.  The PI-authorized Biological Use Notification must be submitted to the IBC simultaneously with experiment initiation, and must receive concurrence by the Biosafety Officer or IBC Chair.  This notification process at the start of a project serves to define and assess the work, establish containment levels, and ensure that the requirements and hazards for the project are correctly identified and addressed.

26.4.3  Biological Use Registration

A Biological Use Registration is required for work with Risk Group 1 recombinant biological materials that are covered by the NIH Guidelines (i.e., recombinant DNA molecules or Risk Group 1 organisms and viruses containing recombinant DNA molecules).  The PI-authorized Biological Use Registration must be submitted to the IBC simultaneously with experiment initiation.  The Biological Use Registration must be reviewed and approved by the IBC.

26.4.4  Biological Use Authorization

A Biological Use Authorization (BUA) is required for biological work that (a) involves biohazardous materials (i.e., Risk Group 2 or higher materials, or the use of Biosafety Level 2 containment for safety), or (b) requires a regulatory registration or permit for the operation (i.e., USDA soil or plant pest permit, select agent registration, etc.).  A BUA is a type of LBNL “Formal Authorization” as defined in Chapter 6 of PUB-3000.  PI-approved Biological Use Authorizations require full IBC review and approval, and authorization by the PI’s supervisor prior to commencing work.

Table 26-3     Berkeley Lab Review, Documentation, and Authorization for Work with Biological Materials

26.5     Biological Waste

Guidance for the disposal of hazardous waste is provided in Chapter 20 (Hazardous Waste Disposal) of PUB-3000.  Specific guidance for the disposal of medical or biohazardous waste is provided in the Medical and Biohazardous Waste Generator’s Guide (PUB-3095).  Generally, when biohazardous waste is to be rendered noninfectious, the process is accomplished either by the addition of bleach (i.e., sodium hypochlorite) to biological liquids, or the treatment of solid waste by the LBNL biohazardous waste subcontractor.  Refer to the Solid Medical Waste Disposal Procedures chart for a summary of solid waste disposal.

26.6     Biosafety and Waste Training

Supervisors and principal investigators (PIs) are responsible for ensuring their workers have sufficient skills, knowledge, and ability to do their work.  In addition, supervisors and PIs must ensure workers have sufficient job-specific training on the work tasks to be performed, hazards of the specific biological materials that may be present, and biosafety containment to be used.

Specific biosafety or biohazardous waste courses are also required.  At a minimum, anyone working with biological material of any risk level (e.g., microorganisms, cells, cell lines, tissue cultures, recombinant nucleic acids, blood, body fluids or tissues, or animals) must attend EH&S course EHS 739 (General Biosafety Training).  In addition, EHS 730 (Medical/Biohazardous Waste Training) is recommended for anyone working with biological material, and required for anyone generating medical or biohazardous waste. EHS 730 is available as online training or classroom training.  Anyone working with human blood or blood products, or human materials (e.g., cells, tissues, or fluids) considered “Other Potentially Infectious Materials” under the OSHA BBP Standard (see Section 26.2.2) must also take EHS 735 (Bloodborne Pathogen Training).  This latter course requires annual refresher training, termed EHS 738.  EHS 739 and 738 are also available as Web-based challenge exams.

26.7     Biosafety or Waste Questions

For concerns regarding the handling and use of biohazardous materials, contact the EH&S Biosafety Officer at extension 2768 (495-2768).  Questions regarding biohazardous or medical waste may be directed to the Medical Waste Coordinator, extension 7663.  For related medical information, call the Health Services Group, extension 6266.

26.8     Glossary

This section lists and/or defines terms and acronyms as used in this chapter.
 
American Biological Safety Association (ABSA) is a professional association that promotes biosafety as a scientific discipline and serves the growing needs of biosafety professionals throughout the world.

Biohazardous is an adjective used to describe biological materials that present potential risk to the health of humans or other organisms, either directly through infection or indirectly through damage to the environment.

Biohazardous Waste is waste that requires biological inactivation in an approved manner prior to disposal, but is not regulated by the California Department of Health Services as regulated medical waste.  Note PUB-3095, Medical and Biohazardous Waste Generator Guidelines, for additional information.

A Biological Etiologic Agent is an agent of biological origin (e.g., bacterium, fungus, parasite, virus, etc.) that causes disease in humans (i.e., pathogenic to humans).

A biological material is a broad range of organisms, cells, viruses, and other materials of biological origin that pose differing levels of risks to plants, animals, or humans.

Biosafety Level (BL) is a combination of practices and techniques, safety equipment, and facilities that are specified by the applicable agency as being appropriate to safely contain the biohazardous materials or agents to be used in the work. 

Bloodborne pathogens (BBPs) are infectious agents that are capable of causing human disease and are transmitted through human blood and tissues.

Containment is the method(s) used to reduce or eliminate exposure of workers and the environment to biohazardous materials or agents.

Centers for Disease Control and Prevention (CDC) is one of the thirteen major operating components of the U.S. Department of Health and Human Services.

Etiologic is an adjective that means disease-causing.

Medical Waste is waste that is generated or produced as a result of the following:  diagnosis, treatment, or immunization of human beings or animals; research pertaining to the diagnosis, treatment, or immunization of human beings or animals; or the production or testing of biologicals.  Note PUB-3095, Medical and Biohazardous Waste Generator Guidelines, for additional information.

National Institutes of Health (NIH) is one of eight health agencies of the U.S. Public Health Service (PHS), and the PHS  is part of the U.S. Department of Health and Human Services.

NIH Guidelines is an abbreviated title used by NIH for the document titled NIH Guidelines for Research Involving Recombinant DNA molecules

Occupational Safety and Health Administration (OSHA) is an agency of the U.S. government that assures the safety and health of U.S. workers (e.g., by setting and enforcing standards).

Other Potentially Infectious Materials (OPIM) are materials that are regulated by OSHA under the Bloodborne Pathogens Standard based on their potential to contain BBPs.  These materials do not include bloodborne pathogens, human blood, human blood components, or products made from human blood.

 A pathogen is an infectious microbial (e.g., bacteria, protozoa, fungi, viruses, etc.) or other agent that can cause disease in healthy host organisms such as humans, animals, or plants.

Risk Group (RG) is a system (e.g., adopted by the CDC and NIH) for classifying biological agents by the degree of hazard.  There are four risk groups: A higher RG number indicates a higher level of hazard.

Select Agents and Toxins are:  a) specific pathogenic agents and toxins that are listed and strictly regulated by the CDC and USDA (i.e., under 7 CFR 331, 9 CFR 121, and 42 CFR 73) because they may potentially be used as agents of mass destruction or pose a severe threat to human, animal, and plant health, and b) specific genetic elements, recombinant nucleic acids, and recombinant organisms that are related to the list of select agents and toxins as described in the regulations.

United States Department of Agriculture (USDA) is an agency of the U.S. government with the following types of mission areas:  farm and foreign agricultural, food, food safety, nutrition, natural resources, environment, research, education, economics, and rural development.

World Health Organization (WHO) is an agency of the United Nations that specializes in the attainment by all peoples of the highest possible level of health.

26.9  Standards

• 7 CFR 331 and 9 CFR 121, Possession, Use, and Transfer of Biological Agents and Toxins, U.S. Department of Agriculture (USDA), Animal and Plant Health Inspection Service (APHIS)
• 7 CFR 330, Plant Pest Regulations; General; Plant Pests; Soil, Stone, and Quarry Products; Garbage.  Importation of Plant Pests, USDA/APHIS
• 9 CFR Parts 92, 94, 95 96, 122 and 130 (note especially Part 122, Organisms and Vectors).  Importation of Etiologic Agents of Livestock, Poultry, and Other Animal Diseases; USDA/APHIS
• 10 CFR 851, Worker Safety and Health Program, Depart of Energy (DOE)
• 29 CFR 1904.8, Recording criteria for needlestick and sharps injuries, Occupational Safety and Health Administration (OSHA)
• 29 CFR 1910.1030, Bloodborne Pathogens, OSHA
• 42 CFR 71, Foreign Quarantine, Part 71.54 Etiologic agents, hosts, and vectors;  Importation of Etiological Agents of Human Disease and Other Materials That May Contain These Agents; United States Public Health Service (USPHS)
• 42 CFR 73, Select Agents and Toxins, Department of Health and Human Services (HHS)
•  Biosafety in Microbiological and Biomedical Laboratories, 4th Edition, Centers for Disease Control (CDC) and National Institutes of Health (NIH), as applicable to biological etiologic agents and select agents
• Biosafety Program and Manual, LBNL
• California Health and Safety Code, Sections 117600 - 118360, California Medical Waste Management Act
• Guidelines for Research Involving Recombinant DNA Molecules, National Institutes of Health (NIH), Federal Register (current version)
• Laboratory Biosafety Manual, Second Edition (revised), Interim Guidelines, World Health Organization (WHO), Geneva 2003, as applicable to biological etiologic agents
• OSHA Standard Interpretation on Applicability of 1910.1030 to Establish Human Cell Lines

26.10   Related LBNL/PUB-3000 Chapters

• General Policy and Responsibilities (Chapter 1)
• Health Services (Chapter 3)
• Industrial Hygiene (Chapter 4)
• Safe Work Authorizations (Chapter 6)
• Personal Protective Equipment (Chapter 19)
• Hazardous Waste Disposal (Chapter 20)
• EH&S Training (Chapter 24)
• Research with Human and Animal Subjects (Chapter 22)

26.11   References

• American Biological Safety Association (ABSA) Web page
• Facility Safety Plan Requirements, United States Army Medical Research and Materiel Command (USAMRMC) webpage Biosafety, Security, and Incident Response Plan for Select Agents, LBNL, latest version
• General Emergency Response Guide (wall posting) and EH&S Emergency Preparedness Web page, LBNL
• How to Import Foreign Soil and How to Move Soil within the United States, Circular Q-330.300-1 Soil (10/2006), USDA/APHIS, Plant Protection and Quarantine (PPQ)
• National Sanitation Foundation (NSF)/ American National Standard (ANSI) Standard 49: Class II (laminar flow) biosafety cabinetry, March 19, 2002
• Primary Containment for Biohazards: Selection, Installation, and Use of Biological Safety Cabinets, 2nd Edition, 2000, CDC-NIH
• PUB-533, Master Emergency Program Plan for the Ernest Orlando Lawrence Berkeley National Laboratory, LBNL, latest version
• PUB-3140, Integrated Environment, Health & Safety Management Plan, LBNL, latest version
• PUB-3095, Medical and Biohazardous Waste Generator Guidelines, LBNL, latest revision
• Site Safeguards and Security Plan, LBNL, latest version

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