1. Research & Teaching

2. 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

3. 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.

4. 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

5. 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.

6. 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

7. Institutional Biosafety Committee Training

8. 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).

9. 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

10. 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

11. 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

12. 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

13. Per 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) Appendix CNIH Guidelineshttp://oba.od.nih.gov/oba/rac/Guidelines/APPENDIX_C.htm

14. Per 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.htmhttp://oba.od.nih.gov/oba/rac/Guidelines/NIH_Guidelines.htm

15. 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

16. 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.

17. 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.

18. 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

19. 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

21. 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

22. 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

23. 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.

24. 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”

25.  Safety Considerations  Risk assessments: (Appendix B) - Appendix B lists human etiologic agents and zoonotic agents based on RG RG 1RG 2RG 3RG 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)

26. 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

27. BSLAgentsPracticesBarriers & Safety Equipment Facilities 1Not known to cause disease in healthy adults Standard microbiological practicesNone requiredLab 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

28. 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/http://www.selectagents.gov/

29. 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

30. 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.

31. 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).

32. Level of ReviewExample of r/sNA ResearchRelevant 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 initiationCreating 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 initiationCreating 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 GuidelinesPurchase or transfer of transgenic rodentsIII-F

33. Life 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.

35. 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

36. 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

37. 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

38. 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

39. IACUC ReviewIBC Review Animal Welfare o 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 Health o Transfer of genetically altered material, viral vectors, etc Risks to the Environment o Escape & establishment in the wild o Interbreeding with wild stock o Consumption by other animals

40. 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

41. 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

43. 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

44. 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

45. 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.

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

48. 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

49. 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

50. 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

51. 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

52. 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

53. 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

54. 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

55. 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

56. 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

57. “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.”

59. 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

60. With lab safety issues Personal protective equipment for personnel Disposal of waste Decontamination of laboratory and equipment Containment facilities Accidents (emergency plans and response)

61. 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)

62. 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

63.  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: ____________________________________________