Complying with NIH Guidelines at CalvinResearch & Teaching

What this training covers• Various sections of the NIH Guidelines including:• NIH Overview• Calvin’s Institutional Biosafety Committee (IBC)• Recombinant DNA (rDNA) & Synthetic Nucleic Acid Molecule

guidelines• Animal guidelines• Plant guidelines• Roles & Responsibilities• Resources

• For the purposes of this training, Principal Investigator (PI) includes any faculty or researchers conducting research or teaching coursework at the college

NIH Guidelines for Recombinant DNA (rDNA) & Synthetic Nucleic Acid Molecules (SNA)

r/sNA

• Details procedures and practices for the containment and safe conduct of various forms of rDNA & SNA research.

• First drafted in 1976 as an outcome of a meeting of scientists concerned about addressing the potential public health and environmental risks associated with rDNA.

• Frequently amended to reflect evolving scientific understanding of rDNA and its applications.

The NIH Office of Biotechnology Activities (OBA)

• A core responsibility of OBA is to promote awareness of, and adherence to, the standards and practices set forth in the NIH Guidelines.

• Provide guidance to IBCs in the oversight of r/sNA research

• Disseminate information on technical and policy matters concerning r/sNA research

• Develop and contribute to public policy on r/sNA research

Training Requirements

• According to the NIH Guidelines • “The institution is responsible for ensuring that the Principal

Investigator (PI) has sufficient training” regarding laboratory safety and implementation of the NIH Guidelines.• “The PI is responsible for ensuring that laboratory staff are

appropriately trained.”

• We have developed this online training course to fulfill the training requirements for PIs; this will be tracked by the Biosafety Officer (BSO, Lori Keen) in the IBC database.

• PIs will be responsible for training their staff and will have access to a downloadable ppt.

Course Objectives• Upon completion of this course, participants will

have a better understanding of:• The IBC review and approval processes that apply to

various forms of research and teaching• The biosafety levels of the NIH Guidelines• The various types of research & teaching covered by the

NIH Guidelines• The roles and responsibilities of investigators under the

NIH Guidelines

The IBC at Calvin CollegeInstitutional Biosafety Committee Training

The Institutional Biosafety Committee (IBC)• IBCs were established under the NIH Guidelines to

provide local review and oversight of research using r/sNA.

• They ensure that r/sNA research conducted at or sponsored by the institution is in compliance with the NIH Guidelines.

• Many institutions, including Calvin, have chosen to assign their IBCs the responsibility of reviewing other research & teaching involving other biohazardous risks (e.g. infectious agents and human samples).

Calvin’s IBC• Made up of faculty, staff and community members• Members include representatives from:• Biology and/or Biochemistry- Dave Benson, Curt

Blankespoor, John Ubels• Biology Lab Manager - Lori Keen• Environmental Health & Safety – Heather Chapman• 2 community members – David LaGrand, Bob Leunk

Calvin College IBC Staff• IBC Coordinator, Lori Keen• Liaison between the investigators & the IBC• Assists faculty with the protocol review process• Notifies investigators of IBC review outcome• Ensures committee operations comply with federal and

state regulations

IBC Notification Process

PIs must notify the IBC prior to commencing your research or teaching, if you plan to use:• Recombinant DNA or Synthetic Nucleic Acid molecules• Infectious agents (BSL2)• Human body fluids or tissues

IBC Approval Process• If your research or teaching is non-exempt, you

must complete the Biosafety Application• If your research or teaching is exempt, you must

complete the IBC First Assessment Form (keep in mind that you may have already completed this)• See next slide for definitions of exempt vs non-exempt

• Both forms are to be submitted to the IBC Coordinator, Lori Keen who will ensure the appropriate review process

Exempt vs. Non-ExemptPer Section III-F of the NIH Guidelines, experiments are exempt when they involve recombinant DNA or SNA that:• Is not in organisms or viruses;• Uses a host vector system such as: E. coli K 12, Saccharomyces cerevisiae,

Saccharomyces uvarum or Bacillus subtilis; and their plasmids;• Uses rDNA molecules containing less than one-half of any eukaryotic genome that

are propagated and maintained in cells in tissue culture;• Consist entirely of DNA segments from a single nonchromosomal or viral DNA

source; though one or more of the segments may be a synthetic equivalent;• Consist entirely of DNA from a prokaryotic host including its indigenous plasmids

or viruses when propagated only in that host (or a closely related strain of the same species); or when transferred to another host by well-established physiological means;

• Consist entirely of DNA from a eukaryotic host including its chloroplasts, mitochondria, or plasmids (but excluding viruses) when propagated only in that host (or a closely related strain of the same species).

• Details on certain other experiments that may be exempt, as well as exceptions, can be found in Appendix C of the NIH Guidelines (http://oba.od.nih.gov/oba/rac/Guidelines/APPENDIX_C.htm)

Exempt vs. Non-ExemptPer Section III of the NIH Guidelines , research or teaching that meet any of the criteria below must submit a proposal to the IBC and receive IBC approval:

• Experiments involving the introduction of rDNA or SNA into risk group 2 organisms (includes adenovirus & murine retroviral vectors) or higher

• Experiments in which DNA from human or animal pathogens (risk group 2 or higher) is introduced into a non-pathogenic host vector system

• Experiments involving the use of infectious DNA or RNA viruses or defective DNA or RNA viruses (risk group 2 or higher) in the presence of helper virus in tissue culture systems

• Experiments involving formation of rDNA molecules containing no more than 2/3 of the genome of only eukaryotic virus (with some restrictions). It must be demonstrated that cells lack helper virus

• The deliberate transfer of a drug resistance trait to microorganisms that are not known to acquire the trait naturally, if such acquisition could compromise the use of the drug to control disease agents in humans, veterinary medicine or agriculture.

• Cloning of toxic molecules with LD50 of less than 100 nanograms/kg body weight• Experiments involving cultures of more than 10 liters• Whole animals in which the animal’s genome has been altered by the stable introduction of rDNA, or

RNA derived therefrom, into the germline (transgenic animals)• Experiments involving whole plants• Gene transfer experiments in humans

• For more information, you may view Section III at: http://oba.od.nih.gov/oba/rac/Guidelines/NIH_Guidelines.htm

IBC Approval Process• The Biosafety Officer can review and approve

research or teaching involving:• BSL1 agents, <10 L• Human blood and/or tissue (requires consult with the BSO)• Already approved infectious agents• Environmental swabbing for culturing

• If any of the following criteria are met, the research or teaching must be reviewed by the full IBC at a convened meeting:• BSL 2 or higher agents• > 10 L of an agent• Recombinant DNA that is NOT exempt

IBC Approval Process• Four possible outcomes:• Approved as is• Approved with conditions• Minor corrections required

• Tabled• More information required

• Denied• Major corrections and/or information needed. PI

generally instructed to consult BSO and re-submit application.

IBC Submission• Due to the low volume of Non-exempt work at the

College, the IBC meets as necessary, but at least annually to review protocols and annual reports. • If you submit an application, the IBC will convene within 2

weeks to review your application

• Applications that do not require full committee review may be submitted to the IBC Coordinator or the IBC Chairperson at any time.

IBC Renewal Process• PIs must submit annual reports for projects falling

within the NIH Guidelines• The PI will receive a reminder email at least 30

days prior to the report deadline• Approved non-exempt long-term projects must be

renewed every 3 years

IBC Renewal Process• The BSO can review and approve:• Closures• Renewals without change• Renewals with minor change• Such as personnel changes or changing project titles

• The IBC reviews and approves:• Renewals with major changes• Such as an adding an agent to a project, going from in

vitro to in vivo• 3-year renewals

The NIH Guidelines for Research Involving Recombinant DNA & Synthetic Nucleic Acid Molecules

Contents of the NIH Guidelines• Section I –Scope of the Guidelines• Section II –Safety Considerations for r/sNA

research• Section III –Types of Experiments Covered by the

NIH Guidelines and the levels of review these experiments require

• Section IV –Roles and Responsibilities of individuals and entities involved in the conduct and oversight of r/sNA research

• Appendices –large number of appendices covering particular topics in greater detail

Purpose of the Guidelines• To specify the practices for constructing and

handling: • (i) recombinant nucleic acid molecules, • (ii) synthetic nucleic acid molecules, including those that

are chemically or otherwise modified but can base pair with naturally occurring nucleic acid molecules, and • (iii) cells, organisms, and viruses containing such

molecules

Section I - Scope• In the context of the NIH Guidelines, recombinant

and synthetic nucleic acids are defined as: • (i) molecules that a) are constructed by joining nucleic

acid molecules and b) that can replicate in a living cell, i.e., recombinant nucleic acids; • (ii) nucleic acid molecules that are chemically or by other

means synthesized or amplified, including those that are chemically or otherwise modified but can base pair with naturally occurring nucleic acid molecules, i.e., synthetic nucleic acids, or • (iii) molecules that result from the replication of those

described in (i) or (ii) above.

Section I –Scope Compliance with the NIH Guidelines

• All institutions that receive NIH funding for r/sD research must comply with the NIH Guidelines.

• All research & teaching at an institution that are subject to the NIH Guidelines must comply with the requirements even if their individual projects are not funded by NIH.

• Compliance with the NIH Guidelinesis mandatory as a condition of receiving NIH funding.

• “Guidelines” does not mean “optional”

NIH Guidelines – Section II Safety Considerations

Risk assessments: (Appendix B) - Appendix B lists human etiologic agents and zoonotic agents based on RG RG 1 RG 2 RG 3 RG 4

Agents that are not associated with disease in healthy adult humans

Agents that are associated with human disease which is rarely serious and for which preventive or therapeutic interventions are often available

Agents that are associated with serious or lethal human disease for which preventive or therapeutic interventions may be available (high individual risk but low community risk)

Agents that are likely to cause serious or lethal human disease for which preventive or therapeutic interventions are not usually available (high individual risk and high community risk)

Section II –Safety Considerations

• Physical Containment• Defined by 4 Biosafety Levels • BSL1, the least stringent; BSL4, the most stringent• These biosafety levels consist of a combination of • Lab practices and techniques,

• Safety equipment, and• Lab facilities appropriate for the operations being

performed

• Consistent with the CDC Biosafety in Microbiological and Biomedical Laboratories

• Described in detail in Appendix G

Summary of Recommended Biosafety Levels for Infectious AgentsBSL

Agents Practices Barriers & Safety Equipment

Facilities

1 Not known to cause disease in healthy adults

Standard microbiological practices

None required Lab bench and sink required

2 • Associated with human disease

• Routes of transmission include percutaneous injury, ingestion, mucous membrane exposure

BSL-1 practice plus:• Limited access• Biohazard warning signs• Sharps precautions• Biosafety manual

defining any needed waste decontamination or medical surveillance policies

Primary barriers:• Class I or II BSCs or

other physical containment devices used for manipulations of agents that cause splashes or aerosols of infectious materials

• PPE:• Lab coat, gloves,

respirator as needed

BSL-1 plus:• Autoclave

available

3 • Indigenous or exotic agents with potential for aerosol transmission

• Disease may have serious or lethal consequences

BSL-2 practices plus:• Controlled access• Decontamination of all

waste• Decontamination of lab

clothing before laundering

• Baseline serum

Primary barriers:• Class I or II BSCs or

other physical containment devices used for manipulations of agents that cause splashes or aerosols of infectious materials

• PPE:• Lab coat, gloves,

respirator as needed

BSl-2 plus:• Physical

separation from access corridors

• Self-closing, double-door access

• Exhaust air not recirculated

• Negative air flow into lab

4 • Dangerous/exotic agents which pose high risk of life threatening disease

• Aerosol-transmitted lab infections have occurred; or related agents with unknown risk of transmission

BSL -3 practices plus:• Clothing change before

entering• Shower on exit• All material

decontaminated on exit from facility

Primary barriers:• All procedures

conducted in class III BSCs or Class I or II BSCs in combination with full-body, air-supplied positive pressure personnel suit

BSL-3 plus:• Separate building

or isolated zone• Dedicated supply

and exhaust vacuum, and decon systems

• Other requirements may apply

Select Agents• Select Agents are pathogens or toxins considered

to have potential as bioweapons. They are regulated by CDC and/or USDA and require approval from those organizations to possess, use, receive or ship them.

• A list of Select Agents, and information on the Select Agent Program is available at http://www.selectagents.gov/

Section II –Safety Considerations

• Biological Containment• Refers to the application of highly specific biological

barriers (appendix I)• Limit infectivity of a vector for specific hosts• Limits dissemination and survival in the environment• Example – genetically designed to be replication

defective outside of host

Risk Group and Biosafety Level are NOT the same

• Risk Group (RG) is a stable comparative descriptor of the inherent pathogenic nature of a given microorganism. RG designation is based on how a particular organism is associated with disease in a “healthy” adult and whether prevention or intervention exists for that organism. RG does not change based on how or where the agent is used.

• Biosafety Level (BSL) is a variable comparative descriptor of the facility, equipment, practices that serve to “contain” a microorganism, and to ensure the safe use of that organism, while it is being handled. BSL is based on risk assessment and technical judgment and may vary with the use of the agent.

Section III –Experiments Covered

• Six categories of experiments involving r/sNA based on the level of review required (III-A, III-B, III-C, III-D, III-E, III-F)

• Experiments which pose great risk to human health require the highest level of review (category III-A), such as the deliberate transfer of antibiotic resistance into a pathogen, that would then compromise the use of that drug to treat the disease.

• Experiments that are not considered to pose a risk to human health or the environment are exempt from the Guidelines(category III-F) and do not require review (e.g., the purchase of transgenic animals that can be housed at BSL1).

Six Levels of ReviewLevel of Review Example of r/sNA Research Relevant

section(s) of the NIH Guidelines

IBC, RAC review, & NIH Director review and approval – Major Action

Experiments that compromise the control of disease agents in medicine through deliberate transfer of a drug resistance trait

III-A

IBC approval & NIH review for containment determinations

Experiments conducted with a rDNA modified restricted agent in a whole animal

III-B

IBC & IRB approval and NIH review before research participant enrollment

Experiments involving the deliberate transfer of rDNA into a human research participant

III-C

IBC approval before initiation Creating stable germline alterations of an animal’s genome, or testing viable rDNA modified microorganisms on whole animals; experiments in which whole plants are genetically engineered or used with rDNA modified insects; generally require BSL-2 containment or greater

III-D

IBC notice at initiation Creating stable germline alterations of rodents using rDNA; experiments involving rDNA modified whole plants that are not noxious weeds; require only BSL 1 containment

III-E

Exempt from the NIH Guidelines

Purchase or transfer of transgenic rodents III-F

Dual Use ResearchLife sciences research that yields information or technologies with the potential to be misused to threaten public health or national security.

• The National Science Advisory Board for Biosecurity (NSABB) has identified seven categories of experiments that have the potential to fall under Dual Use Research of Concern:• Enhance the harmful consequences of a biological agent or toxin. • Disrupt immunity or the effectiveness of an immunization without clinical and/or

agricultural justification. • Confer resistance to prophylactic or therapeutic interventions, or facilitate the

ability of an agent or toxin to evade detection methodologies. • Increase the stability, transmissibility, or the ability to disseminate a biological

agent or toxin. • Alter the host range or tropism of a biological agent or toxin. • Enhance the susceptibility of a host population. • Generate a novel pathogenic agent or toxin or reconstitute an eradicated or

extinct biological agent.

Recombinant and Synthetic Nucleic Acid Research Involving Animals and the NIH Guidelines

Section III-D-4 Experiments Involving Animals

• IBC and Institutional Animal Care & Use Committee (IACUC) Approval Before Initiation

• Includes Experiments in which:• the animal’s genome has been altered by stable

introduction of r/sNA into germline (i.e., transgenic animals)• applies to both the generation and use of transgenic

animals (including arthropods)• r/sNA modified microorganisms are tested on whole

animals• BSL2 or BSL2-N or greater containment

Section III-E-3 Experiments Involving Transgenic Rodents

• IBC and IACUC Approval Before Initiation at Calvin• Includes Experiments in which:• Rodent’s genome has been altered by stable introduction

of recombinant or synthetic nucleic acid into germline• applies only to the generation of transgenic rodents

• BSL1 containment is appropriate

Appendix C-VI Exempt Animal Research

• The purchase or transfer of transgenic rodents that require BSL-1 containment

• Further manipulations of these animals with r/sNA are not necessarily exempt from the NIH Guidelines

IBCs and IACUCs and Animal Research

• Joint purview and collaborative review over certain types of research • Transgenic or cloned animals• Use of recombinant or synthetic nucleic acid molecules in

animals• Pre-clinical studies and data assessment for human gene

transfer protocols

• At Calvin, the EHS Officer is a member of both committees

IBC and IACUC Review of Animal Research Utilizing r/sNA

IACUC Review IBC Review

Animal Welfareo Pain & distress from

adverse phenotypes (behavioral, anatomical & physiological abnormalities)

o Risks to other animals in the facility from the inadvertent spread of vectors

Risks to Human Healtho Transfer of genetically

altered material, viral vectors, etc

Risks to the Environmento Escape & establishment in

the wildo Interbreeding with wild

stocko Consumption by other

animals

Points to Consider for Animal Research with r/sNA

• Containment procedures (SOPs)• Physical and biological• Plans for recapture of escapees• Consequences should containment fail

• Procedures for transfer and transportation of animals

• Disposal and destruction methods• Breeding SOPs• Occupational biosafety concerns• Personal protective equipment• Decontamination

Appendix Q –Physical and Biological Containment for Large Animals

• Applies when research animals are of a size or have growth requirements that preclude laboratory containment• For example, cattle, swine, sheep, goats, horses, poultry,

etc.

• Addresses containment and confinement practices in animal facilities (BL1-N to BL4-N)

• Applies to animals:• In which genome is altered by stable introduction of rDNA

or SNA; or• On which rDNA or SNA-modified microorganisms are being

tested

Recombinant & Synthetic Nucleic Acid Research Involving Plants and the NIH Guidelines

Section III-D-5 Experiments Involving Plants

• IBC Approval Before Initiation• Includes Experiments in which:• Plants are genetically engineered by r/sNA or synthetic

biology methods, or• Plants are used with r/sNA modified insects• Generally BL1-P through BL4-P, depending on risk

• IBC Approval Before Initiation • Includes Experiments:• Involving r/sNA-modified whole plants• That are not noxious weeds and will not hybridize with

noxious weeds, and • For which BL1-P containment is appropriate

Appendix P –Physical and Biological Containment for Plants

• Defines physical containment levels for r/sNA research involving plants• Describes the four plant Biosafety Levels BL1-P through

BL4-P and provides details on the standard practices and any special procedures that should be followed

• Describes biological containment practices for r/sNA-containing plants, plant-associated microorganisms, and plant-associated small animals.

Section IV of the Guide

Section IV –Roles and Responsibilities

• Institution• Institutional Biosafety Committee (IBC)• Biological Safety Officer (BSO)• Principal Investigator (PI)• NIH

Institutional Responsibilities• Establish and implement policies for the safe

conduct of recombinant or synthetic nucleic acid research

• Establish an IBC• Ensure compliance with the NIH Guidelines by

investigators • Ensure appropriate training for IBC members and

staff, PIs, and laboratory staff• Determine necessity for health surveillance of

personnel• Report any significant incidents or violations to the

NIH Guidelines to OBA within 30 days

IBC Responsibilities• Assessment of r/sNA research

• Conformity with the NIH Guidelines• Potential risk to environment and public health

• Containment levels per NIH Guidelines• Adequacy of facilities, SOPs, PI and lab personnel training• Institutional and investigator compliance; e.g., adverse event reports

• Periodically review r/sNA use for compliance• This may include periodic unannounced inspections

• Adopt emergency plans covering spills, contamination, other accidents

• The IBC may not authorize initiation of r/sNA experiments not explicitly covered by the NIH Guidelines until NIH establishes the containment requirement

IBC Membership• Membership requirements• At least 5 individuals• Appropriate r/sNA expertise• Plant and animal experts if needed, biosafety officer as

appropriate

• At least two members not affiliated with the institution• represent the interest of the surrounding community with

respect to health and protection of the environment

• Laboratory technical staff (recommended)• Expertise in assessment of risk to environment and

public health • Knowledge biological safety and physical containment

BSO Responsibilities• The Biosafety Officer’s (BSO) duties include:• Reporting to the IBC and institution of any problems,

violations, research-related accidents or illnesses• Advice on lab security• Technical advice to PIs and IBCs on research safety

procedures

• The BSO shares certain responsibilities including:• Periodic inspection of labs• Developing emergency plans for handling accidental spills

and personnel contamination

PI Responsibilities• Full compliance with the NIH Guidelines

• PIs must:• Be familiar with institutional policies which may go above

and beyond the NIH Guidelines• Not initiate or modify r/sNA research prior to IBC approval• Be adequately trained in good microbiological techniques• Inform and train laboratory staff• Risk assessment• Containment Practices• Procedures for dealing with accidents• Occupational health

PI Responsibilities• Supervise r/sNA research• Adhere to all approval conditions imposed by the IBC• Correct work errors and conditions that may result in the

release of rDNA materials• Ensure the integrity of containment

• Comply with permit and shipping requirements• Adhere to IBC-approved emergency plans• Report any significant problems, violations of the

NIH Guidelines, or any significant research-related accidents and illnesses to the BSO • Know the Calvin IBC Policy on reporting incidents involving

r/sNA to the NIH OBA

NIH Responsibilities• Recombinant and Synthetic nucleic acid

experiments requiring review by NIH OBA• Deliberate transfer of drug resistance to microorganisms, if it

could compromise disease control• Cloning of toxin molecules with LD50 <100 ng/Kg bodyweight• DNA from restricted agents transferred to nonpathogenic

prokaryotes or lower eukaryotes• The term restricted agent in the NIH Guidelines is used

specifically to refer to the pox viruses, small pox and white pox, and should not be confused with “select agents”

• DNA from nonpathogenic prokaryotes or lower eukaryotes transferred to restricted agents

• Use of infectious or defective restricted poxviruses in presence of helper virus

NIH Guidelines -Appendices

• There are 15 appendices. Those highlighted are the ones you are most likely to consult.

• Appendix A –Exemptions: Natural Exchangers• Appendix B –Classification of Etiologic Agents• Appendix C –Exemptions under III-F• Appendix D –Major Actions• Appendix E –Certified Host-Vector Systems• Appendix F –Biosynthesis of Toxic Molecules• Appendix G –Physical Containment• Appendix H –Shipment

NIH Guidelines -Appendices• Appendix I –Biological Containment• Appendix J –Biotechnology Research

Subcommittee• Appendix K –Large Scale Physical Containment• Appendix L –Gene Therapy Policy Conferences• Appendix M –Points to Consider in Human Gene

Transfer Research• Appendix P –Physical and Biological Containment:

Plants• Appendix Q –Physical and Biological Containment:

Animals

Good Judgment is Key!

“The NIH Guidelines will never be complete or final since all conceivable experiments involving recombinant DNA cannot be foreseen. Therefore, it is the responsibility of the institution and those associated with it to adhere to the intent of the NIH Guidelines as well as to the specifics.”

Resources for Investigators

The IBC Can Help You• Ensure that you are working with infectious

agents, human samples, and rDNA/SNA safely• Meet all compliance requirements for use of rDNA

& SNA• Meet all compliance requirements • Avoid preventable accidents and incidents that

might cause harm or undermine public confidence in your research/teaching activities

The BSO & EHS Can Help You• With lab safety issues• Personal protective equipment for personnel• Disposal of waste• Decontamination of laboratory and equipment• Containment facilities• Accidents (emergency plans and response)

Web Resources

• NIH Guidelines for Research Involving Recombinant DNA Molecules (http://oba.od.nih.gov/rdna/nih_guidelines_oba.html)

• Biosafety in Microbiological and Biomedical Laboratories(BMBL) 5th Edition (http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm)

• National Select Agent Registry (http://www.selectagents.gov/) Select Agent and Toxins list (http://www.selectagents.gov/Select%20Agents%20and%20Toxins%20List.html)

• American Biological Safety Association (ABSA) Resources (http://absa.org/resmenu.html)

Additional Information• Contact• IBC Coordinator & BSO, Lori Keen• 526.6080• keel@calvin.edu

• IBC Chairperson, John Ubels• 526.6219• jubels@calvin.edu

• Environmental Health & Safety Officer, Heather Chapman• 526.8591• hlc5@calvin.edu

PI Verification of Training

I acknowledge that I have read and understand the IBC training module.

I didn’t quite get it and would like the BSO to contact me to discuss further

• Please select a box above and then print and send to the BSO.

NAME: ___________________________________________DATE: ____________________________________________