BIOSAFETY LEVELS AND RISK ASSESSMENT POLICY

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1 A. Purpose Research in the life sciences frequently requires the use of biological agents that may pose an infection risk to laboratory workers. Any work with biological agents should be preceded by a comprehensive risk assessment to account for the characteristics of the agent being used, the procedures to be performed, the possible routes of exposure, and the consequences of any potential exposure. These considerations will assist the laboratory in determining the appropriate biosafety level for the work to be performed, and thus the procedures and precautions to be used when working with the agent. The risk assessment process must be carried out thoroughly and carefully to ensure that the appropriate biosafety level is chosen for the task at hand; underestimation of risk can lead to serious health or environmental hazard, while overestimation can result in excessive burden to the laboratory. B. Applicability/scope This policy applies to all laboratories at Columbia University conducting work with biological materials. While some materials inherently present very little risk and may be safely manipulated with minimal risk of infection, a risk assessment is required in all cases to initially establish the appropriate biocontainment requirements. C. Responsibilities 1. Principal Investigators Risk Assessment - Principal Investigators (PIs) at Columbia University working with biological materials are responsible for ensuring that these materials are managed in a way that minimizes the risk of laboratory acquired infections (LAI) or exposure to personnel. A risk assessment must be performed prior to the start of any work with a new agent or new procedure. Any significant changes to the agents being used or the procedures performed must also be considered. Environmental Health & Safety (EH&S) and the Institutional Biosafety Committee (IBC) are available to assist with risk assessments. The outcome of a risk assessment should serve to determine the appropriate biosafety level for handling a given agent, which in turn dictates the engineering controls, work practices, and personal protective equipment (PPE) that must be employed. The PI is responsible for ensuring that the laboratory and all of its personnel operate at the appropriate biosafety level for all activities. EH&S and the IBC are available to advise laboratories on how to meet the requisite biosafety level. Training - The PI is required to complete all safety training pertaining to the materials and activities present in the laboratory in accordance with the guidelines described here: Environmental Health & Safety Page 1 of 10

2 The PI must also ensure that all laboratory personnel complete training relevant to their respective job duties. 2. Laboratory personnel Training - All laboratory workers are responsible for complying with the requirements of the biosafety level established by the PI for any given material and procedure, and for completing all applicable safety training. All laboratory accidents and potential exposures to infectious material must be reported to the PI and EH&S Biosafety Officers. 3. Institutional Biosafety Committee (IBC) Protocol and research review - The IBC reviews all work at Columbia University involving the use of recombinant DNA, in accordance with the NIH Guidelines, and provides high level review for other significant biosafety concerns such as work with agents that can cause disease in humans. The IBC determines whether experiments that have been submitted to the Committee for review have been planned with the appropriate biosafety level for the agents and procedures to be used. 4. Environmental Health & Safety (EH&S) D. Definitions EH&S provides technical guidance to PIs, laboratory workers, and the IBC through its Biosafety Program. Biosafety Officers are available to conduct or assist with risk assessments, and to advise laboratories on how to meet the requirements of the biosafety level that applies to their activities. ABSL Vertebrate animal biosafety level (ABSL) criteria are established for the use of experimentally infected animals in animal care facilities, as well as for animals that may harbor zoonotic infectious agents. In general, the animal biosafety level for working with any given infectious agent in vivo is comparable to the biosafety level (BSL) for that agent in vitro, though exceptions may occur based on the transmissibility of the agent, the route of administration, and/or the potential for the animal to shed the agent into its environment. Criteria for each animal biosafety level include standard animal husbandry practices, safety equipment (including caging, primary containment devices and PPE), facility design criteria (including autoclaves and secondary barriers, if any), and any special practices. These criteria are well-established in the BMBL. ABSL-1 The lowest level of infectious risk and corresponding protective measures for work with animals, which serves as the default for any animals that are not administered infectious materials and are not known to naturally harbor zoonotic agents affecting humans. ABSL-1 is suitable for work with well-characterized agents that are not known to Environmental Health & Safety Page 2 of 10

3 cause disease in healthy human adults and present minimal risk to laboratory personnel, the environment, or for transmission between animals. ABSL-2 This level of infectious risk and protective measures is suitable for work with vertebrates involving agents of moderate hazard to personnel and the environment where 1) laboratory personnel have specific training in animal facility procedures and handling pathogenic agents and infected animals; 2) personnel are supervised by individuals with adequate knowledge of potential hazards, microbial agents, animal manipulations and husbandry procedures; 3) access to the laboratory is restricted; 4) all procedures in which infectious aerosols or splashes may be created are conducted in BSCs or other physical containment equipment; 5) the facility meets design and containment requirements as specified in the BMBL. ABSL-3 The highest level of infectious risk and corresponding protective measures for work with animals allowed at Columbia University, for agents that may cause serious or potentially lethal disease through the inhalation route of exposure. This differs from ABSL-2 in that all procedures involving the manipulation of infectious materials and infected animals must be conducted within BSCs or other physical containment devices, and the laboratory requires special engineering and design features. Special safety practices, equipment, and facility requirements are necessary for work at ABSL-3. Biocontainment The practice of physically isolating potentially hazardous biological agents in order to prevent infection of workers or environmental release. BMBL Biosafety in Microbiological and Biomedical Laboratories, 5 th Edition is the standard biosafety reference document promulgated by the CDC and NIH. See BSC Biological Safety Cabinet BSL The biosafety level designated for any agent or activity is both a risk designation and the level of biocontainment precautions required to prevent infection or release of biological agents. Criteria for each biosafety level include standard microbiological practices, safety equipment (including primary containment devices and PPE), facility design criteria (including secondary barriers, if any), and any special practices. These criteria are well-established in the BMBL. BSL-1 The lowest level of infectious risk and corresponding protective measures, suitable for work with well-characterized agents that are not known to cause disease in healthy human adults and present minimal risk to laboratory personnel or the environment. BSL-2 This level of infectious risk and corresponding protective measures is suitable for work involving agents of moderate hazard to personnel and the environment where 1) laboratory personnel have specific training in handling pathogenic agents and are Environmental Health & Safety Page 3 of 10

4 supervised by scientists competent in handling infectious agents and associated procedures; 2) access to the laboratory is restricted when work is being conducted; and 3) all procedures in which infectious aerosols or splashes may be created are conducted in BSCs or other physical containment equipment. BSL-2+ - Also known as BSL-2 with enhanced procedures, work under this level of infectious risk and corresponding protective measures may be conducted in facilities that are designed for BSL-2 operations, but requires several practices that are normally used for BSL-3 operations. BSL-2+ criteria are not strictly defined in the BMBL and other standard biosafety references, though common themes have emerged in the biosafety field. Examples of BSL-2+ criteria can be found in Appendix 1. BSL-3 The highest level of infectious risk and corresponding protective measures allowed at Columbia University, for agents that may cause serious or potentially lethal disease through the inhalation route of exposure. This differs from BSL-2 in that all procedures involving the manipulation of infectious materials must be conducted within BSCs or other physical containment devices, and the laboratory requires special engineering and design features. Special safety practices, equipment, and facility requirements are necessary for work at BSL-3. CDC Centers for Disease Control and Prevention, US Department of Health and Human Services EH&S Environmental Health and Safety FACS Fluorescence-Activated Cell Sorting IBC Columbia University Institutional Biosafety Committee LAI Laboratory acquired infection LV Lentivirus NIH National Institutes of Health, US Department of Health and Human Services NIH Guidelines The NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acids is the regulatory standard governing institutional oversight of work with recombinant nucleic acids, which mandates the establishment of an Institutional Biosafety Committee. See PI Principal Investigator PPE Personal protective equipment Environmental Health & Safety Page 4 of 10

5 RMW Regulated medical waste WHO World Health Organization Risk Group Classification Risk Group 1 Risk Group 2 Risk Group 3 Risk Group 4 E. Risk Assessment Procedures 1. Determine Agent Characteristics The first consideration for a biological risk assessment should involve the characterization of the inherent hazard of the agent. This level of hazard is defined by several properties, including its ability to infect humans or animals, the severity of disease that results, and the availability of effective prophylaxis or treatment. For wellcharacterized agents, risk group classifications have been established by both the WHO and NIH, which roughly correlate to biosafety levels in most cases, though a risk assessment is still necessary to determine whether a particular procedure or other factors may elevate or lower the biosafety level necessary to safely handle an agent. Table 1: Classification of Infectious Microorganisms by Risk Group* NIH Guidelines for Research Involving World Health Organization Laboratory Biosafety Recombinant or Synthetic Nucleic Acid Manual Molecules Agents not associated with disease in healthy adult humans. Agents associated with human disease that is rarely serious and for which preventive or therapeutic interventions are often available. Agents associated with serious or lethal human disease for which preventive or therapeutic interventions may be available (high individual risk; low community risk). Agents likely to cause serious or lethal human disease for which preventive or therapeutic interventions are not usually available (high individual risk; high community risk). *Taken from BMBL 5 th edition. A microorganism unlikely to cause human or animal disease (no or low individual and community risk). A pathogen that can cause human or animal disease but is unlikely to be a serious hazard to laboratory workers, the community, livestock, or the environment. Laboratory exposures may cause serious infection, but effective treatment and preventive measures are available and the risk of spread of infection is limited (moderate individual risk; low community risk). A pathogen that usually causes serious human or animal disease but does not ordinarily spread from one infected individual to another. Effective treatment and preventive measures are available (high individual risk; low community risk). A pathogen that usually causes serious human or animal disease and can be readily transmitted from one individual to another, directly or indirectly. Effective treatment and preventive measures are not usually available (high individual risk; high community risk). Environmental Health & Safety Page 5 of 10

6 2. Laboratory Procedure Hazards In addition to the inherent risks posed by the agents manipulated in a laboratory, researchers must always consider the activities and procedures that are to be performed, and whether these activities could open a potential route of exposure, when conducting a risk assessment to determine appropriate safety precautions. Possible routes of exposure may include percutaneous injuries such as needle sticks, animal bites and scratches, direct contact by a splash to the skin or mucous membranes, accidental injection, or inhalation of aerosols. In particular, procedures that can potentially generate aerosols are of paramount concern. In some cases, such as with fluorescence activated cell sorting (FACS), the biosafety level required for working with a given material may be higher than that required for other more routine manipulations (see Columbia University s FACS policy: While aerosols may present the most common route of exposure in laboratory acquired infections, other potential routes of exposure must always be considered and minimized where possible. In addition to the use of appropriate engineering controls and personal protective equipment, appropriate laboratory technique employed by the researcher can make a large difference in the potential for exposure. 3. Safety Equipment and Facility Safeguards The use of appropriate engineering controls and biosafety facilities should be considered the top line of defense for the conscientious researcher. Without appropriate engineering controls and facilities, many agents simply cannot be handled safely. Biological safety cabinets (BSCs) serve as the primary engineering control in most biomedical research settings, and should always be used for any open manipulation of infectious materials or any procedures that may generate aerosols (See Columbia University s BSC policy: Similarly, centrifuge safety cups with gasket-sealed lids are excellent examples of engineering controls that must be used whenever centrifugation of a potentially infectious sample is needed. The use of engineered safe sharps devices should be another consideration for any manipulation of infectious materials requiring a syringe, and is mandated by OSHA for work with any materials that may contain bloodborne pathogens, including human cell lines (see the Columbia University Bloodborne Pathogens and Exposure Control Plan Environmental Health & Safety Page 6 of 10

7 Personal protective equipment is another consideration that must not be overlooked. While for many BSL-1 and BSL-2 activities the common PPE ensemble of a lab coat, safety glasses, and gloves may suffice, some activities may warrant the use of additional precautions such as sleeve covers, rear-closing impermeable lab coats, face shields, and/or respiratory protection. Additionally, appropriate laboratory facilities must be used for activities involving infectious materials. Laboratory and animal care facilities are designed to meet the specifications required for the biosafety level of the work that will take place in them. BSL-2 activities should always be confined to areas approved for BSL-2 work. BSL-3 activities require elaborate engineering controls and must never take place outside of laboratories designed to meet BSL-3 standards. Likewise, in animal care, animals that are administered infectious materials and pose a risk of shedding infectious agents may require special ABSL-2 or ABSL-3 housing arrangements, and must not be kept in general housing. 4. Determine Appropriate Biosafety Level and Precautions By considering the agents involved in an experiment and the procedures that are planned, a risk assessment should help researchers to determine the appropriate biosafety level for a given activity. High risk activities involving high concentrations of agents and procedures that may generate aerosols may warrant elevating the biosafety level above what would normally be dictated by a given agent s risk group. On the other hand, low risk activities performed in animals, such as stereotactic intracranial administration of certain replication-deficient viral vectors, may not result in a risk of viral shedding by the animal, and could allow for the animals to be housed at ABSL-1 following administration even if the agent requires in vitro manipulation at BSL-2. Once the biosafety level for an activity is chosen, the standard precautions for conducting work at that biosafety level are generally well-established (BMBL, 5 th edition). There may be some cases, such as with work that warrants BSL-2+ precautions, that special procedures may need to be adopted in consultation with EH&S. 5. Evaluate Proficiencies of Personnel In addition to generic biosafety training provided in a classroom or online setting, laboratory personnel must be trained by skilled advisors for any procedures involving infectious materials. For high risk procedures in particular, it is critical that junior researchers are thoroughly trained by their seniors and observed for competency in laboratory techniques before being allowed to manipulate infectious materials. Environmental Health & Safety Page 7 of 10

8 6. Review Risk Assessment with Biosafety Professionals Many routine procedures with common agents require only a simple risk assessment on the part of the investigator, after which they may be safely conducted at the appropriate biosafety level. Researchers working with novel, high-risk, or exotic agents, or who plan to perform any high-risk procedures, should consult with EH&S before initiating work, as special precautions may be necessary. In such cases, the researcher may be required to submit a protocol to the Institutional Biosafety Committee for review. F. Emergency contacts Public Safety Morningside: , Emergency: Public Safety Medical Center: , Emergency: Facilities: EH&S Morningside: EH&S Medical Center: Locations and contact info for health care providers: G. Medical Surveillance Personal exposure takes priority over clean up. If medical follow-up is warranted it should be sought immediately. For details on health care providers see: For more information on post exposure response, review the Bloodborne Pathogens Exposure Control Plan ( H. Recordkeeping N/A Environmental Health & Safety Page 8 of 10

9 I. Appendices Appendix 1: Examples of BSL-2+ precautions. The decision on which precautions to employ is dependent on a risk assessment. Type of control Training Administrative Engineering PPE Procedural Practice Personnel must have demonstrated proficiency in microbiological practices and techniques. An apprentice program may be established for personnel where they shadow more experienced personnel and are not allowed to work independently until they demonstrate proficiency. A laboratory member was appointed to serve as the BSL 2+ Manager and oversees daily operations in the lab including, ensuring that adequate PPE and supplies such as disinfectants are available, monitoring conditions in the lab including PPE usage, and reporting issues that may require retraining. Dedicated BSL2+ rooms. Access restricted to those performing BSL2+ work. Door signs are posted while BSL2+ work is taking place. The containment laboratory must be kept locked. A protocol specific to the operation of the laboratory developed and read by personnel; employees must certify in writing that they have understood the material in the protocol. General protocols must be supplemented with protocols specific to each project in progress. Procedures in the event of an accident are clearly posted in the laboratory. A hands free sink and soap are present in the lab for hand washing. Two pairs of gloves are worn during procedures. The outer pair of gloves should be removed inside the biosafety cabinet to prevent contamination of equipment and surfaces outside of the cabinet. Disposable solid gown with cuffed sleeves that ties in the back. Gowns are identified with "owners" name and may be reused a limited number of times, unless known to be soiled. Alternatively, a cotton lab coat lab that is kept in the room and used in conjunction with disposable sleeves. Safety glasses with side shields are worn at all times. Sealed tubes are wiped down with a bleach wipe prior to leaving the BSC. Infected cultures should be in a dedicated incubator or held within secondary containment in a common incubator. Solid waste is decontaminated daily by autoclave. No notebooks or other paperwork associated with lentiviral experiments can be taken in and out of the laboratory. Keep all required equipment within the BSL2+ animal room; no sharing between rooms. Animal cages are not changed for 48 or 72 hours. All cages are sealed in autoclavable bags prior to leaving the room and are autoclaved prior to going through the cage wash. Environmental Health & Safety Page 9 of 10

10 Procedural Substitution All waste generated in the room is considered biohazardous and collected as RMW. There are no regular trash cans in the room. Ready mix bleach spray is used for decontamination of BSC interior and objects leaving the BSC. Create a Spill Kit and store within the lab. Transport of LV or LV transduced cells within the facility must occur in a double packaging. Transport of LV or LV transduced cells outside the facility must occur in triple packaging according to regulations regarding the transport of Dangerous Goods. Plastic substituted for glass where possible (e.g. glass Pasteur pipettes substituted with plastic transfer pipettes). Use a non pseudotyped ecotropic virus. J. Forms N/A K. References Biosafety in Microbiological and Biomedical Laboratories, 5 th Ed; (2009, December); U.S. Department of Health and Human Services. Bloodborne Pathogens Exposure Control Plan, Columbia University, NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules; (2013, March); Office of Biotechnology Activities, National Institutes of Health; Policy for Fluorescence-Activated Cell Sorting (FACS), Columbia University, Policy for the Use of Biosafety Cabinets, Columbia University, Regulated Medical Waste Policy, Columbia University, Environmental Health & Safety Page 10 of 10

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