Biological Safety Manual

The purpose of this document is to increase awareness of biological hazards frequently encountered in research, clinical, and teaching laboratories at Northwestern University (Northwestern).  Additionally, this manual provides guidance on recommended practices when working with and disposing of biological hazards.  Biological hazards include infectious or toxic microorganisms (including viral vectors), biological toxins, manufactured and pre-assembled "kits" that may include any of these agents, and substances from which transmission of infectious agents or toxins could be reasonably anticipated such as tissues from humans and research animals.  Due to the diverse nature of biological hazards, they are often generically referred to as “agents” so that a discussion may encompass all known and unknown hazards.  The intrinsic danger associated with a particular biological hazard may be mitigated or compounded by the presence of recombinant or synthetic nucleic acids (rDNA). This document is supported by a separate document at Northwestern called the Exposure Control Plan required by the Occupational Safety and Health Administration (OSHA).

The objective of safety awareness and practice is to assure laboratory personnel that, with proper precaution, equipment, and facilities, most biohazardous materials can be handled without undue risk to themselves, their peers, their families, and the environment.

This document is intended not only for trained microbiologists, but also for individuals handling human clinical materials in other laboratory disciplines, such as biochemistry, genetics, oncology, immunology, and molecular biology.  Individuals with little or no microbiological training might not appreciate the potential hazard involved in handling the types of samples described above. It is important not to be complacent about certain agents due to their widespread use and availability.

The health and safety principles described in this document are based on sound safety practices, common sense, good housekeeping, personal hygiene, and emergency planning.  Many of the practices and procedures described herein have been adapted from the Biosafety in Microbiological and Biomedical Laboratories, 6th Edition (BMBL) and the National Institutes of Health Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules (NIH Guidelines).

All personnel are accountable for the establishment of a culture of responsibility in their labs and at their institutions.  Fundamental to this culture of responsibility are scientific integrity and adherence to ethical codes of conduct.  For the individual, an ethical code of conduct centers on personal integrity.  It embodies, above all, a commitment to intellectual honesty and personal liability for one’s actions, and to a range of practices that characterize the responsible conduct of research.  Such practices include:

• Intellectual honesty
• Accuracy
• Fairness
• Collegiality
• Transparency in conflicts of interest
• Protection of human subjects
• Humane care of animals

All individuals at Northwestern working with biological materials and potentially hazardous substances must demonstrate the following knowledge, skills, and abilities:

• Adherence to all safety protocols
• Adherence to spill and exposure protocols and reporting requirements
• Awareness of the reporting requirements of Northwestern and the various research oversight committees
• Awareness of emergency response protocols (e.g. fire, tornado, inclement weather)
• Completion of all training requirements
• Completion of and proficiency in all lab-specific training requirements
• Completion of all Occupational Heath requirements, including documentation of required physicals, medical clearances, and vaccinations, as applicable
• Immediate reporting to the Principal Investigator (PI) of any situation that compromises an individual’s ability to perform as required in a research laboratory, including physical or psychological issues
• Immediate reporting to the PI and Research Safety, where appropriate, of behavior or activities inconsistent with safety and security plans
• Adherence to security protocols

A. Wellbeing and Support

Northwestern has established various support mechanisms to help develop a culture of responsibility and support their researchers. Northwestern has resources available to help manage stress and significant life events.

To support faculty and staff Northwestern has created the Employee Assistance Program. The program is a network of services, including short-term counseling, to help you and your household members cope with everyday issues. Professional counselors can help you with everyday needs and life events, relationship/marital concerns, workplace concerns, anxiety and stress, family issues, coping with a serious illness, sleeping difficulties, loss of a loved one, emotional concerns, depression, and communication. The program provides counseling sessions with a local, licensed counselor and unlimited, 24/7 telephonic counseling. For assistance, contact (855) 547-1851 or visit https://www.northwestern.edu/hr/benefits/well-being/programs/employee-assistance-program/index.html.

Full-time matriculated students have opportunities to receive mental health counseling services at no charge through the Counseling and Psychological Services (CAPS) . CAPS provides mental health services for students at Northwestern, with offices on the Evanston and Chicago campuses. CAPS provides clinical services, referrals, educational workshops, and faculty, staff, and parent consultations. For more information, please call (847) 491-2151 or visit http://www.northwestern.edu/counseling/.

The Feinberg School of Medicine has an Ombudsperson who serves as an impartial, neutral, and confidential facilitator for students, residents, and fellows. The Ombudsperson will work with students, residents, and fellows to describe available options to address the issue and, if desired, help resolve conflicts. Student, resident, and fellow interactions with the Ombudsperson are handled as discreetly as possible. For more information, please visit https://www.feinberg.northwestern.edu/md-education/current-students/policies-services/ombuds.html.

B. Institutional Training Requirements

All personnel must take required laboratory and biological safety training based on the materials and agents handled. Information about what training is needed, how it is accessed, and the frequency of training can be found on the Research Safety website under the tab: “What Training Do I Need?” website. For individuals who work with animals, additional animal care safety information can be found on the IACUC website. Assigned training is mandatory; some training assignments are annual, while others only need to be completed once.

 C. Reporting Concerns

There are formal, confidential reporting mechanisms in instances of non-compliance with established, Northwestern University or lab-specific, safety and security policies.

There are multiple mechanisms for reporting non-compliance at Northwestern:

• PI/supervisor.
• Department Chair or Dean
• Research Safety at researchsafety@northwestern.edu or call (312) 503-8300 (Chicago) or (847) 491-5581 (Evanston)
• Research Integrity Office (https://researchintegrity.northwestern.edu/)
• Ethics Point is an anonymous reporting system through a third-party (https://secure.ethicspoint.com/domain/media/en/gui/7325/index.html) or call (866) 294-3545

Ensuring health and safety is a shared responsibility at Northwestern. Senior leadership at the University has entrusted the necessary authority to Research Safety to support the safe conduct of research and correct unsafe actions or conditions.

A. Senior Leadership

• Establishes a culture of safety at Northwestern University.
• Identifies staffing needs for oversight of biological research safety.
• Provides resources to ensure research safety.

B. Research Safety

• Establishes safety standards and policies to support an environment of safety and accountability for all university employees and to protect the health of university employees and minimize their exposure to potentially harmful biological materials.
• Evaluates the potential biological, chemical, radiological, and physical hazards within research labs and core facilities.
• Conducts audits of all research spaces as part of its laboratory safety review process to assess compliance, and, as necessary, non-compliance is reported to the appropriate University leadership for resolution.
• Suspends and prohibits unsafe acts and conditions in laboratories at Northwestern.
• Ensures compliance with various labor and environmental regulations that apply to research laboratories.
• Ensures that worker safety and health concerns are addressed during animal protocol review process.
• Supports the Institutional Biosafety Committee (IBC) by implementing policy and guidance established by the IBC.
• Provides staff support to the IBC.

C. Biosafety Officer (BSO)

• Develops efficient procedures to register biological research.
• Maintains an engaged IBC.
• Suspends and prohibits unsafe acts and conditions in laboratories at Northwestern.
• Oversees the evaluation and inspection of laboratory facilities for work with biological materials.
• Investigates laboratory accidents, incidents, and near-misses.
• Develops materials and training that promote safe microbiological practice.
• Maintains training records relating to biological safety, bloodborne pathogens, and shipment of biohazardous materials.
• Consults with Research Safety, the IBC, and the Northwestern University research community on biological research.
• Updates policies relating to work with biological materials.
• Ensures appropriate reporting to the National Institutes of Health (NIH) about Recombinant and Synthetic DNA (rDNA) research.
• Shares information about biological research safety through announcements, events, webinars, emails, and training courses.
• Notifies Principal Investigators of the results of their biological protocol reviews.
• Coordinates with Occupational Health Services for IBC protocols requiring medical surveillance.
• Supports Research Safety and the IBC in performing their duties.

D. Institutional Biosafety Committee

• Reviews and approves research involving rDNA.
• Reviews and approves research involving human, animal, and plant pathogens.
• Reviews and approves research involving animals associated with biological materials and rDNA.

E. Animal Care and Use Committee

• Ensures that worker safety and health concerns are addressed during animal protocol review process.
• Reviews and approves animal protocols.
• Ensures the humane care and use of animals in research.

F. Principal Investigators (PIs)

• Enforces institutional policies that control safety and access to the laboratories.
• Ensures lab personnel receive appropriate training based on their duties, potential hazards, and precautions to minimize exposure with annual updates and additional training when procedures or policies change.
• Reports all laboratory safety incidents to Research Safety within 5 days of their occurrence.
• Informs all personnel and visitors of hazards associated with their assigned facilities.
• Adopts engineering and work practice controls that reduce the risk of sharps injuries.
• Ensures laboratory personnel demonstrate proficiency in standard microbiological practices and techniques for working with agents requiring BSL-2 and greater containment.
• Develops laboratory specific biosafety standard operating procedures to minimize exposure risk to biohazardous agents.
• Verifies that all safety equipment in the laboratory (including BSC, PPE, etc.) are accessible for laboratory staff and functioning properly.

G. Center for Comparative Medicine (CCM)

• Enforces policies, procedures, and protocols for biosafety, biosecurity, and emergencies within the animal facility.
• Ensures animal facility personnel receive appropriate training based on their duties, potential hazards, and precautions to minimize exposure with annual updates and additional training when procedures or policies change.
• Informs all personnel and visitors of hazards associated with their assigned facilities.
• Adopts engineering and work practice controls that reduce the risk of sharps injuries.
• Animal care staff will report all accidents, spills, and exposures to the animal facility supervisor.

H. Laboratory Staff

• Follows all established biosafety practices to minimize exposure to biological materials and to avoid other incidents (such as personal injuries, chemical and radiological spills, laboratory fires, explosions, etc.).
• Completes all assigned biological and general research safety training.
• Reports all accidents, spills, and exposures to the laboratory supervisor.
• All personnel should be familiar with the agents used in their laboratory and the hazards they pose.
• All personnel should be familiar with the emergency spill protocols and the location of cleanup equipment.

A. Risk Assessment

A risk assessment should be conducted with all biological research. A risk assessment is mandatory for all potentially infectious materials.

A thorough risk assessment considers the following:

• The agent’s biological and physical nature
• The sources likely to harbor the agent
• Host susceptibility
• The procedures that may disseminate the agent
• The best method to effectively inactivate the agent

Globally, numerous government agencies have classified microorganisms pathogenic to humans into Risk Groups (RG). RG classification is based on transmissibility, invasiveness, virulence, or capability of causing disease or death.  It also considers the availability of vaccines or therapeutic interventions.

Pathogenic microorganisms include bacteria, viruses, fungi, parasites, and other infectious agents. The classification scheme ascends in order of increasing hazard from Risk Group 1 agents, which are non-pathogenic for otherwise healthy adults, to RG4 agents, which display high morbidity and mortality and for which treatments are not readily available. The complexity of controls required to contain and work with an organism in a particular Risk Group increases with the risk group. Microorganisms in RG1 require standard laboratory facilities and microbiological practices, whereas those in RG4 require elaborate procedures, engineering controls, and facilities for maximum containment.

In the United States, Risk Groups are assigned to pathogens that pose a risk to humans. Only human and animal pathogens capable of infecting humans are assigned a risk group. Pathogens that exclusively infect animals or plants are not assigned risk groups in the United States. The Risk Group listing in the NIH Guidelines is an accepted standard even when recombinant or synthetic nucleic acid molecules (rDNA) technology is not being used. A list of Risk Groups is found in the NIH Guidelines: https://osp.od.nih.gov/policies/biosafety-and-biosecurity-policy#tab2/. The American Biological Safety Association International (ABSA) also provides a comprehensive Risk Group database. This database also references other global agencies and their Risk Group classification. The database is accessible and can be found at https://my.absa.org/tiki-index.php?page=Riskgroups.

Health Canada has developed and maintains a website with detailed Pathogen Safety Data Sheets for many human and animal pathogens. These documents describe the hazardous properties of a specific pathogen and recommendations for work involving the agent in a laboratory setting. The Health Canada Pathogen Safety Data Sheets can be found at https://www.canada.ca/en/public-health/services/laboratory-biosafety-biosecurity/pathogen-safety-data-sheets-risk-assessment.html.

Microorganisms classified as RG2 or higher have been reported to cause infection and disease in otherwise healthy adults. Many have been associated with laboratory-acquired infections. Furthermore, lab-acquired infections with RG2 agents are the most common, much more than with RG3 or RG4 agents. At Northwestern University, RG2 or RG3 pathogens are designated as moderate and moderate to high hazard. These agents require more sophisticated engineering controls (such as facilities and equipment) than standard laboratories.

Risk groupings of infectious agents usually (but not always) correspond to Biosafety Levels (BSL or BL), which describe the recommended containment practices, safety equipment, and facility design features necessary to handle benign and pathogenic microorganisms safely.

 Table 1: Risk Group (RG) Classification Definitions and Examples

Definitions and examples for Risk Groups 1-4

	DEFINITION	EXAMPLES
Risk Group 1	Agents not associated with disease in healthy adult humans	E. coli K-12 derivatives, Saccharomyces cerevisiae
Risk Group 2	Agents associated with human disease that is rarely serious and for which preventative or therapeutic interventions are often available	E. coli O157:H7, Salmonella, Cryptosporidium, Hepatitis A, B, C, D, and E viruses
Risk Group 3	Agents associated with serious or lethal human disease for which preventative or therapeutic interventions may be available	Yersinia pestis, Brucella abortus, Mycobacterium tuberculosis, Human Immunodeficiency Virus (HIV), Transmissible spongiform encephalopathies (TSE) agents
Risk Group 4	Agents associated with serious or lethal human disease for which preventative or therapeutic interventions are not usually available	Ebola virus, Macacine herpesvirus (Monkey B virus)

 

B. Potential Routes of Infection

Pathogens may be transmitted via one or more routes of infection. The route(s) of infection depends on the characteristics of the pathogen. The most common routes of infection are inhalation of infectious aerosol, dust, or small droplets, exposure of mucous membranes to infectious droplets, ingestion from contaminated hands or utensils, or percutaneous inoculation (injection, incision, or animal bite). Appropriate precautions should be implemented to avoid such exposures.

The progression from entry into the body to infection depends upon several factors, including the (1) route of transmission, (2) form and infectious dose, (3) invasive characteristics of the agent, and (4) natural resistance of the person exposed. Not all contacts result in infection, and even fewer develop into clinical disease. Even when disease occurs, severity can vary considerably.

C. Potential Exposure in the Laboratory Environments

1. Teaching Laboratories

Whenever possible, teaching laboratories should use avirulent strains of infectious microorganisms. If pathogenic microorganisms must be used for teaching purposes, consult Research Safety and the Biosafety Officer.

While often considered safer (but never risk-free) attenuated microorganisms require good microbiological practice, especially with immunocompromised individuals. Students should be cautioned against and trained to prevent unnecessary exposure, as exposure to an avirulent strain may cause harm to an immunocompromised individual. Establishment of safety consciousness is integral to the conduct of good science.

2. Research and Animal Laboratories

Experiments involving high concentrations or large quantities of pathogen(s) increase the risk of exposure and infection. Using animals in research on infectious diseases also presents greater risk of exposure. Research and animal environments can be complex and sometimes unpredictable, requiring strict adherence to good lab practices and a conscious and continuous approach to hazard and risk analysis.

3. Clinical Laboratories

By design, clinical and diagnostic laboratories are responsible for helping to identify infectious materials in humans or animals. Clinical lab personnel must take precautions due to the increased risk of exposure to infectious agents. Keep in mind that the absence of an infectious disease diagnosis does not preclude the presence of a pathogen, and testing has limitations.  This is especially true of materials from patients who have received immunosuppressive therapy since such treatment may activate latent infections or make hosts more likely to harbor infectious agents. Standard precautions apply to all clinical specimens, with some activities requiring BSL-3 practices and procedures.

D. Health Status

Personal health status may affect an individual’s susceptibility to infection and ability to receive available immunizations or prophylactic interventions. It is recommended that all laboratory personnel discuss occupational risks with their personal healthcare provider. Do not hesitate to contact them or Corporate Health if you have questions or concerns.

Supervisors must ensure that laboratory and animal care staff are adequately informed about the potential occupational hazards associated with their work. Certain medical conditions may increase an individual’s risk of health problems when working with animals and pathogenic microorganisms.

Common examples include:

• Diabetes
• Pregnancy
• Autoimmune diseases
• Immunodeficiency/immunosuppression
• Animal-related allergies
• Chronic skin conditions
• Respiratory disorders
• Steroid and other drug therapies

At Northwestern University, research involving recombinant or synthetic nucleic acids must comply with the National Institute of Health’s Guidelines for Research Involving Recombinant and Synthetic Nucleic Acid Molecules (NIH Guidelines). 

The NIH Guidelines apply to all recombinant and synthetic nucleic acids research within the United States or its territories, which is conducted at or sponsored by an institution that receives any support for recombinant or synthetic nucleic acids research from the NIH. 

Federal funding agencies have similarly adopted the NIH Guidelines and require adherence to them as a condition of funding.  Any individual receiving support for research involving recombinant or synthetic nucleic acids must be associated with or sponsored by an institution that assumes the responsibilities assigned by the NIH Guidelines.

For the most up-to-date version of the NIH Guidelines, please visit https://osp.od.nih.gov/policies/biosafety-and-biosecurity-policy#tab2/.

A. Definition

The National Institutes of Health have defined recombinant and synthetic nucleic acids 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.

B.  National Institutes of Health (NIH) Guidelines

The purpose of the NIH Guidelines is 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.

The NIH Guidelines provide oversight of any nucleic acid molecule experiment, which according to the NIH Guidelines requires approval by NIH, must be submitted to the NIH or to another Federal agency that has authority for review and approval.  Once approvals, or other applicable clearances, have been obtained from a Federal agency other than NIH, the experiment may proceed without requiring NIH review or approval.

For experiments involving the deliberate transfer of recombinant or synthetic nucleic acid molecules, or DNA or RNA derived from recombinant or synthetic nucleic acid molecules, into human research participants (human gene transfer, HGT), no research participant shall be enrolled until (1) the NIH protocol registration process has been completed, (2) Institutional Biosafety Committee (IBC) approval (from the clinical trial site) has been obtained, (3) Institutional Review Board (IRB) approval has been obtained, and (4) all applicable regulatory authorizations have been obtained.

C. Institutional Biosafety Committee (IBC)

Section IV-B-2 of the NIH Guidelines mandates the formation of an Institutional Biosafety Committee (IBC) and the institution's appointment of a Biosafety Officer (BSO).  For more information regarding the NIH-mandated requirements for the IBC and the most up-to-date version of the NIH Guidelines, please visit https://osp.od.nih.gov/policies/biosafety-and-biosecurity-policy#tab2/.  The responsibilities of the IBC are described in the IBC Charter and the IBC Policies and Procedures Manual. If you have questions, do not hesitate to contact IBC Staff at ibc@northwestern.edu.

1. Functions of the IBC
As outlined by the NIH Guidelines, the IBC is responsible for reviewing recombinant or synthetic nucleic acid molecule research conducted at or sponsored by the institution for compliance with the NIH Guidelines. At the discretion of the IBC itself, the review includes the following:

• Assessment of the containment levels for the proposed research.
• Assessment of the facilities, procedures, practices, and training and expertise of personnel involved in the research.
• Assessment of awareness of all personnel to potentially hazardous agents used in the lab.

In addition to reviewing proposed research with recombinant or synthetic nucleic acid molecules, the Northwestern IBC also reviews and approves:

• Research with biohazardous materials including human and zoonotic pathogens both in the laboratory and in facilities that house vertebrate animals, regardless of the presence of recombinant or synthetic nucleic acids.
• Research with human blood, bodily fluids, human tissues, and animal tissues including non-human primates, regardless of recombinant or synthetic nucleic acids.
• Research with excluded strains or excluded amounts of select agents and toxins.
• Transportation of biological materials between Northwestern University, local medical facilities, and other locations upon the recommendation of the Biosafety Officer.

For Human Gene Transfer (HGT) studies, Northwestern works with two commercial Institutional Biosafety Committees to oversee such studies. These IBCs, with the support of the Northwestern Biosafety Officer, ensure that appropriate regulatory requirements are met, including:

• Ensuring compliance with the NIH Guidelines.
• Ensuring that no research participant is enrolled in an HGT experiment until the NIH protocol registration process has been completed, IBC approval has been obtained, IRB approval has been obtained, and all applicable regulatory authorizations have been obtained.
• Reviewing the recommendations of the Novel and Exceptional Technology and Research Advisory Committee (NExTRAC) for HGT studies, where applicable, and ensuring that the PI has appropriately responded to them.
• Ensuring final IBC approval is granted only after the NIH protocol registration process has been completed.
• Ensuring compliance with all surveillance, data reporting, and adverse event reporting requirements outlined in the NIH Guidelines.

Following review, it is the responsibility of the IBC to notify the PI of the result of the IBC’s review and subsequent approval, if applicable. The IBC is also responsible for setting the containment level of research involving recombinant or synthetic nucleic acids in organisms from Risk Groups 2, 3, and 4, as well as research involving live animals and plants. The IBC is granted the authority by the NIH to lower the containment level for specific experiments as specified in Section III-D-2-a of the NIH Guidelines.

The IBC is also responsible for periodically reviewing the recombinant and synthetic nucleic acids research conducted at the institution to ensure compliance with the NIH Guidelines.  Finally, the IBC must adopt and ensure the implementation of emergency plans covering accidental spills and personnel contamination resulting from recombinant or synthetic nucleic acids research.

2. The Relationship between the IBC and Research Safety

At Northwestern, Research Safety works closely with the IBC to manage many day-to-day operations of the IBC and to ensure IBC policy implementation. Several members of the Research Safety staff act as administrators of the IBC and maintain accurate records of the IBC meetings. Some staff also sit on the IBC and act as voting members. 

Research Safety representatives review biological IBC registrations to verify compliance with biosafety policies. The IBC has entrusted Research Safety with IBC policy implementation and compliance with IBC decisions. The IBC and Research Safety work together to ensure biological safety at Northwestern.

The Northwestern IBC has authorized the Biosafety Officer (or their designee) to approve and document exempt work as allowed by the NIH Guidelines.

D. Dual Use Research of Concern (DURC)

Broadly defined, “dual use” refers to the malevolent misapplication of technology or information initially developed for benevolent purposes. In life sciences, “dual use” refers to the potential misuse of microorganisms, toxins, recombinant/synthetic DNA technology, or research results to threaten public health or national security. “Dual Use Research of Concern,” referred to as DURC, is research that has the potential to be directly misapplied.

 Consistent with the September 2014 DURC Policy, and under Northwestern University Policy, research that uses one or more of the biological agents or toxins listed in this Policy and produces, aims to produce (or be anticipated to produce) one (or more) effects listed in this Policy will be evaluated for possible DURC potential.

Biological agents and toxins of concern:

1. Avian influenza virus (highly pathogenic)
2. Bacillus anthracis
3. Botulinum neurotoxin
4. Burkholderia mallei
5. Burkholderia pseudomallei
6. Ebola virus
7. Foot-and-mouth disease virus
8. Francisella tularensis
9. Marburg virus
10. Reconstructed 1918 influenza virus
11. Rinderpest virus
12. Toxin-producing strains of Clostridium botulinum
13. Variola major virus
14. Variola minor virus
15. Yersinia pestis

There are seven categories of experiments that require oversight under this policy.  Experiments that fall into one or more of these categories are not necessarily forbidden.  Instead, experiments within these categories require additional oversight. 

 These categories are:

1. Enhances the harmful consequences of the biological agent or toxin.
2. Disrupts immunity or the effectiveness of an immunization against the biological agent or toxin without clinical and/or agricultural justification.
3. Confers to the biological agent or toxin resistance to clinically and/or agriculturally useful prophylactic or therapeutic interventions against that agent or toxin or facilitates their ability to evade detection methodologies.
4. Increases the stability, transmissibility, or the ability to disseminate the biological agent or toxin.
5. Alters the host range or tropism for the biological agent or toxin.
6. Enhances the susceptibility of a host population to the agent or toxin.
7. Generates or reconstitutes an eradicated or extinct biological agent or toxin listed above.

 Northwestern University has developed and implemented a formal policy regarding DURC. For further information and questions regarding this Policy, please contact IBC Staff at ibc@northwestern.edu or call Research Safety at (312) 503-8300.

A. Microorganisms

A primary tenet of biological safety is to control, limit, or contain the generation of aerosols to reduce the potential for a lab-acquired infection when inhalation is a potential route of entry for the microorganisms. Cultures of infectious agents must be manipulated carefully to avoid the uncontrolled release of aerosols or the generation of droplets or spills. Droplets are larger particles that may travel a distance and fall onto surfaces whereas aerosols are smaller particles that may remain suspended in the air for an extended period. Accidental spilling liquid infectious cultures is an obvious hazard due to the generation of aerosols and/or small droplets.

Aerosols are generated when mixing, stirring, pipetting, plating, shaking, vortexing, or spill a liquid containing microorganisms. However, even routine manipulations of cultures may release microorganisms via aerosol and/or small droplet formation. Such manipulations include, but are not limited to:

• Removing stoppers from culture vessels
• Opening closed vessels after vigorous shaking/vortexing
• Spattering from flame-sterilized loops
• Expelling the final drop from a pipette (“blowing out”)
• Spinning centrifuge tubes without secondary containment
• Opening a container immediately following vortexing

The lyophilization of cells may disperse concentrated particles if the ampoules are not appropriately sealed, are mishandled, or break. Some pathogens can even survive in the liquid phase of Nitrogen.

The biosafety cabinet (BSC), covered in a later section, is a critical safety device for protection working with microorganisms that you must understand and consistently use properly. Modern equipment for manipulating of infectious materials, such as cell sorters and automated harvesting equipment, have safety features and requirements for proper use and decontamination. They still must be evaluated to determine the need for secondary containment. Maintain your equipment to the manufacturer's standards. Remember that costly equipment of this type is often operated as multi-user or core facilities; the inherent variability in risk from one project to another makes it imperative that operators and users of these facilities understand the possible risks and methods of control.

B. Human Cells and Tissues

Using well-established human or animal cell cultures in laboratories requires special consideration. Cell or tissue cultures in general present few biohazards, as evidenced by their extensive use and the low incidence of laboratory-acquired infections. When a cell culture is inoculated with or known to contain an etiologic agent, it should be classified and handled at the prescribed biosafety level for the agent. 

Biosafety Level 2 containment must be used for cell lines of human origin, even well-established ones like HeLa and HEK293, and all human clinical material (such as tissues and fluids obtained from surgery or autopsy). Manipulation of large volumes of human tissues or cell lines, or manipulations that have the potential to create aerosols, should all be performed within a certified biosafety cabinet.

Any specimen from human patients may contain infectious agents. Specimens most likely to harbor such microorganisms include blood, sputum, urine, semen, vaginal secretions, cerebrospinal fluid, synovial fluid, pleural fluid, pericardial fluid, peritoneal fluid, amniotic fluid, feces, and tissues.

C. Bloodborne Pathogens

Personnel in laboratories and clinical areas handling human blood, body fluids, and non-human primate material, and established human cell lines (even if screened for pathogens) must practice Standard Precautions (previously known as Universal Precautions).

Standard Precautions consider that all human blood and certain human body fluids contain infectious agents and bloodborne pathogens such as HIV, Hepatitis B Virus (HBV), and Hepatitis C Virus (HCV), regardless of what is known about the source. Hence, all the safety practices and procedures are standardized and applied in all situations. 

Persons with the potential for occupational exposure are required by law (OSHA 29 CFR 1910.1030) to complete bloodborne pathogen training. 

You can find details on Northwestern University’s Exposure Control Plan for Bloodborne Pathogens at https://researchsafety.northwestern.edu/safety-information/biological/bloodborne-pathogens-program.html.

D. Animals and Animal Cells and Tissues

Animals may harbor infectious pathogens that can affect humans, sometimes without showing signs or symptoms of disease in the host animal. Exposure can occur through bites, scratches, or excreted materials. When working with animals, use the same protection strategy as human blood, body fluids, and tissues—Standard or Universal precautions.

Assume that all animals, as well as their tissues, blood, and body fluid, are potentially infectious—follow appropriate biosafety practices and procedures. All non-human primate specimens (e.g., cell cultures, blood and body fluids, and tissues) should be handled with the expectation that they may contain infectious agents. Animals inoculated with human-derived materials including immortalized or established human cell lines must be housed in accordance with IACUC and CCM policies.

In the process of inoculating animals, an investigator can be exposed to infectious material by inadvertent self-inoculation or inhalation of infectious aerosols. Surgical procedures, necropsies, and the processing of tissues, can produce aerosols or droplets unintentionally. Needle sticks, cuts, and lacerations are often accidentally self-inflicted and may involve contaminated instruments. Research Safety provides cut-resistant gloves that may be useful in reducing the harm from cuts and lacerations. Animal excrement is a source of infectious microorganisms and investigators should take precautions to minimize aerosols and dust when changing bedding and cleaning cages. Use all required engineering controls in the vivarium.

All research staff must be on an approved protocol and complete all IACUC and the Center for Comparative Medicine (CCM) training requirements (protocol approval) as a prerequisite for entering and working in the CCM. Research personnel must first contact IACUC to begin the credentialing process.

E. Viral Vectors

Viral vectors are standard tools for molecular biologists. Their usefulness is attributed to the abilities these vectors retain from their parent virus: the ability to enter a host cell and transfer genetic material. The number of commercially available viral vector systems grows daily. Many viral vector systems are designed to allow expression in different cells, including human cells (tropism). 

 Standard viral vector systems commonly used in research include:

• Lentivirus vectors
• Gammaretrovirus vectors
• Adenovirus vectors
• Adeno-associated virus vectors
• Baculovirus vectors
• Herpesvirus vectors
• Rabies virus-based vectors
• Poxvirus vectors
• Sendai virus-based vectors

Although viral vectors can be purchased from commercial vendors, hazardous features of the original nature of the virus remain intact. Various mechanisms are used to enhance safety in the creation and manipulation of viral vectors. Many viral vectors have been rendered replication incompetent, meaning the virus cannot independently drive its replication within the host cell. Another safety feature is separation or reduction of the parent virus’s genome into multiple plasmids, which can reduce the likelihood of recombination into either a replication-competent and/or pathogenic virus. Additionally, some vectors have their host range limited based on the viral envelope included in the vector packaging. Regardless of the mechanism(s) used to enhance a viral vector’s safety, the agent is specifically designed to transduce a target host cell, which may be human. These agents are equally capable of transducing an incidental host during exposure. In some cases, these agents insert into host chromosomes and are, therefore, insertional mutagens that could promote oncogenesis in the long term. Viral vectors designed to express genes involved in growth control (e.g., proto-oncogenes) or to knock-down tumor suppressors should lack human-tropism whenever possible.

For these reasons, scientists using viral vectors must demonstrate knowledge of the products they buy or acquire in assessing the potential risks to individuals, public health, and the environment in their biological safety registration. Please contact the Biosafety group of Research Safety for more information.

F. Biological Toxins

Biological toxins are natural, toxic substances produced as by-products of microorganisms (exotoxins, endotoxins, and mycotoxins such as T-2 and aflatoxins), plants (plant toxins such as ricin and abrin), and animals (zootoxins such as marine toxins and snake venom). Unlike pathogenic microorganisms, including those that produce toxins, toxins themselves are not contagious and do not replicate. In this regard, toxins behave more like chemicals than infectious agents. However, unlike many chemical agents, biological toxins are not volatile and are odorless and tasteless. The stability of toxins varies widely, depending on the toxin structure (low molecular weight toxins are very stable).

Most biological toxins, except T-2 Mycotoxin, are not dermally active, making intact skin an excellent barrier against most toxins. In contrast, mucous membranes of the eyes, nose, mouth, and broken skin all serve as points of entry into the body. Aerosol transmission, ingestion, and percutaneous transmission are potential concerns for most biological toxins.

Bacterial toxins can be exotoxins (including enterotoxins or endotoxins). Exotoxins are cellular products excreted from certain Gram-positive and Gram–negative bacteria, highly toxic, and unstable. These toxins are lethal in microgram quantities.

Exotoxins are destroyed when heated above 60°C for at least ten minutes. Bacterial endotoxins are lipopolysaccharide complexes derived from the membrane of Gram-negative bacteria and endotoxins are released when bacteria die. Endotoxins are moderately toxic and relatively stable. Endotoxins are lethal at tens- to hundreds-micrograms quantities and can withstand heating at 60°C for hours without losing activity.

The modes of action of biological toxins vary but include damage to cell membranes or cell matrices (e.g., Staphylococcus aureus alpha toxin), inhibition of protein synthesis (e.g., Shiga toxin), or via activation of secondary messenger pathways (e.g., Clostridium botulinum and C. difficile toxins).

Most work with biological toxins requires BSL-2 containment. In some cases, such as large-scale production or the manipulation of large quantities of a toxin in powder form, manipulation at BSL-3 may be required, depending on the toxin in question and the quantities used. Please discuss this in advance with the Biosafety Officer.

The most hazardous form of any toxin is as a dry powder. Manipulating dry powder toxins must be performed in a biological safety cabinet or a fume hood. In some cases, the Biosafety Officer may recommend a glove box. Once a toxin is reconstituted in a liquid, BSL-2 practices and procedures are usually sufficient.

Biological toxins must be secured in locked refrigerator or freezer. Alternatively, a locked container within the refrigerator or freezer may be used. An inventory must also be maintained to document obtaining, use, and destruction of remaining toxins. A paper or electronic inventory may be used.

G. Select Agents and Toxins

Following the enactment of the Public Health Security and Bioterrorism Preparedness Act of 2002, the United States Department of Health and Human Services (HHS) established a list of biological agents and toxins that are highly regulated. Similarly, the Agricultural Bioterrorism Protection Act of 2002 requires that the United States Department of Agriculture (USDA) establishes and regulates a list of biological agents that have the potential to pose a severe threat to the health and safety of animals, plants, or animal and plant products. 

Combined, these Acts seek to ensure the health and safety of humans, livestock, and crops and forestall a bioterror attack. The Centers for Disease Control and Prevention (CDC) and the Animal and Plant Health Inspection Service (APHIS) enforce these regulations and share the responsibility of some agents due to the potential to threaten both humans and animals. The laws mentioned above require that HHS and USDA review and republish the lists of so-called Select Agents and Toxins lists on at least a biennial basis (https://www.selectagents.gov/sat/list.htm).

Specific Select Agents and Toxins strains have been excluded from regulation and listed. An excluded select agent strain or modified toxin will be subject to the regulation if there is a reintroduction of factor(s) associated with virulence, toxic activity, or other manipulations that modify the attenuation such that virulence or harmful activity is restored or enhanced. 

Northwestern University only allows work with excluded select agents and toxins (https://www.selectagents.gov/sat/exclusions/index.htm) or excluded, permissible amounts of toxins (https://www.selectagents.gov/sat/permissible.htm).

Biosafety Levels and Animal Biosafety Levels consist of combinations of laboratory practices and procedures, safety equipment and laboratory facility design features commensurate with laboratory operations performed and are based on the potential hazards imposed by the agents used and for the specific activity. It is the combination of practice, equipment, and facility that form the basis for physical containment strategies for infectious agents. There are four biosafety levels with Biosafety Level 1 (BSL-1 or BL-1) being the least stringent and Biosafety Level 4 (BSL-4 or BL-4) being the most stringent. Similarly, there are four animal biosafety levels with Animal Biosafety Level 1 (ABSL-1) being the least stringent and Animal Biosafety Level 4 (ABSL-4) being the most stringent. There are other specialized containment criteria for animals in less restrictive housing, penned in agricultural research settings, or the containment of insects and other arthropods. The general recommendations for the four Biosafety Levels are as follows:

• A/BSL-1 is recommended for non-pathogenic agents.
• A/BSL-2 is recommended for potentially pathogenic and pathogenic agents transmitted by direct contact (percutaneous inoculation, ingestion, or mucous membrane exposure).
• A/BSL-3 is recommended for pathogenic agents with the potential to be transmitted via aerosol.
• A/BSL-4 is recommended when the total separation between the pathogenic agent and investigator is critical.

Risk Group designations often, but not always, correlate with the physical containment level appropriate for a given research activity.  It is important to note that while Risk Group designations are inflexible and set by health and/or regulatory agencies, the local Institutional Biosafety Committee has some flexibility in assigning Biosafety and Animal Biosafety Level requirements. 

A brief description of the requirements (according to the BMBL) for each Biosafety Level is provided as well as a correlation between Risk Group and Biosafety Level is provided below in Tables 2-3 and Tables 4-5, for research and animal laboratory facilities respectively. It is important to note that this manual focuses on Biosafety Level 2 and Animal Biosafety Level 2.

Table 2: Summary of Biosafety Level Requirements

Biosafety Level Requirements, Levels 1-4

	BIOSAFETY LEVEL
	1	2	3	4
Restricted access1	Yes	Yes	Yes	NA
Controlled access2	No	Preferred	Yes	Yes
Isolation of laboratory	No	No	Preferred	Yes
Room sealable for decontamination	No	No	Yes	Yes
Inward airflow ventilation	No	Preferred	Yes	Yes
Mechanical ventilation via building system	No	Preferred	Preferred	No
Mechanical, independent ventilation	No	No	Preferred	Yes
Filtered air exhaust	No	No	Yes	Yes
Double door entry	No	No	Yes	Yes
Airlock	No	No	No	Yes
Airlock with shower	No	No	No	Yes
Effluent treatment	No	No	No	Yes
Autoclave on-site	Yes	Yes	Yes	Yes
Autoclave in laboratory room	No	No	Yes	Yes
Double-ended autoclave	No	No	Preferred	Yes
Class I or II BSC3	No	Yes	Yes	Preferred
Class II BSC	No	No	Preferred	Yes

1. Restricted access, laboratory doors are closed during work hours and locked after hours

2. Controlled access, only laboratory staff is allowed access at any time

3. BSC, Biological Safety Cabinet

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Table 3: Summary of Animal Biosafety Level Requirements

Animal Biosafety Level Requirements, Levels 1-4

	ANIMAL BIOSAFETY LEVEL
	1	2	3	4
Restricted access1	Yes	Yes	Yes	Yes
Controlled access2	No	Preferred	Yes	Yes
Separation from general traffic	Preferred	Preferred	Yes	Yes
Room sealable for decontamination	No	Preferred	Yes	Yes
Seamless floors and monolithic ceilings	Preferred *Yes	Yes	Yes	Yes
Inward airflow ventilation	Preferred	Preferred	Yes	Yes
Ducted exhaust air ventilation system	No *Yes	Yes	Yes	Yes
Exhaust air discharge without recirculation	No *Yes	Yes	Yes	Yes
Filtered air exhaust	No	No	Yes	Yes
Filtered air supply	No *Yes	No *Yes	No	Yes
Double door entry (swing inwards)	No	Yes	Yes	Yes
Anteroom entry	No	No	Yes	Yes
Personnel shower	No *Yes	Yes	Yes	Yes
Effluent treatment	No	No	No	Yes
Autoclave in animal facility	Yes	Yes	Yes	Yes
Pass-through autoclave	No	Preferred	Yes	Yes
Class I or II BSC3	No *Yes	Yes	Yes	Yes
Class II BSC	No *Yes	No *Yes	Preferred	Yes

1. Restricted access, animal facility doors are closed during work hours and locked after hours

2. Controlled access, only animal facility or laboratory staff is allowed access at any time

3. BSC, Biological Safety Cabinet

*Indicates additional features implemented at Northwestern University above standard practice

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Table 4: Relationship of Risk Groups to Biosafety Levels, Practices, and Equipment

Risk groups with biosafety levels, laboratory type, laboratory practices and safety equipment

RISK GROUP	BIOSAFETY LEVEL	EXAMPLES OF LABORATORY TYPE	LABORATORY PRACTICES	SAFETY EQUIPMENT
1	Basic – BSL-1	Basic teaching	SMP1	None; open bench work
2	Basic – BSL-2	Primary health services; primary level hospital; diagnostic, teaching, and public health; most biomedical research	SMP plus protective clothing; biohazard sign	Open bench plus BSC2 for potential aerosols
3	Containment – BSL-3	Special diagnostics; highly pathogenic biomedical research	BSL-2 Practices plus special clothing, respiratory protection, controlled access, directional airflow	BSC and/or other primary containment for all activities
4	Maximum Containment – BSL-4	Dangerous pathogens units; extremely pathogenic biomedical research	BSL-3 Practices plus airlock entry/exit, shower exit, special waste disposal	Class III BSC or positive pressure suits, pass-through autoclave, filtered air

1. SMP, Standard Microbiological Practices. Standard Microbiological Practices consists of aseptic techniques and other practices that are necessary to prevent contamination of the laboratory with the agents being handled and contamination of the work with agents from the environment. SMP is used to keep the agents being handled inside their primary containers without any other organisms getting in and contaminating the research materials.

2. BSC, Biological Safety Cabinet

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Table 5: Relationship of Risk Groups to Animal Biosafety Levels, Practices, and Equipment

Risk groups with animal biosafety levels, facility type, laboratory practices and safety equipment

RISK GROUP	BIOSAFETY LEVEL	EXAMPLES OF FACILITY TYPE	LABORATORY PRACTICES	SAFETY EQUIPMENT
1	Basic – ABSL-1	Conventional animal housing	Standard animal care and management practices, medical surveillance	Protective clothing and gloves
2	Basic – ABSL-2	Containment animal caging; special animal housing; diagnostic, teaching, and public health; most biomedical research	ABSL-1 plus limited access, proficiency demonstration, decontamination of wastes and cages, closed doors, sharps precautions	Animal containment equipment plus BSC1 for potential aerosols
3	Containment – ABSL-3	Special animal diagnostics; highly pathogenic biomedical research	ABSL-2 plus controlled access, decontamination of clothing before laundry, decontamination of bedding before removal from caging	Containment equipment for housing and cage dumping, BSC and/or other primary containment for all activities, respiratory protection
4	Maximum Containment – ABSL-4	Dangerous pathogens units; extremely pathogenic biomedical research	ABSL-3 practices plus clothing change, anteroom entry/exit, shower to exit, waste disposal before facility removal	Class III BSC or partial containment equipment with positive pressure suits

1. BSC, Biological Safety Cabinet

For more information regarding Biosafety Levels, consult the Biosafety in Microbiological and Biomedical Laboratories, 6^th Edition (BMBL) (https://www.cdc.gov/labs/BMBL.html).  Experiments involving rDNA are also governed by another method of providing containment called biological containment. For biological containment, highly specific biological barriers are considered in the risk assessment process. Specifically, biological containment considers natural barriers that limit either (1) the infectivity of a vector or vehicle (plasmid or virus) for specific hosts, or (2) its dissemination and survival in the environment. For additional information on biological containment, consult the NIH Guidelines (https://osp.od.nih.gov/policies/biosafety-and-biosecurity-policy#tab2/).

A. Laboratory Design

B. Laboratory Ventilation

To provide containment, it is important that laboratory air pressure be lower than (or “negative” to) that in the adjacent spaces. This negative air pressure differential ensures that air will enter the laboratory and not egress into the hallway. Labs with multiple rooms should have the airflow balanced such that the most negative room is where the most hazardous work takes place. To maintain negative room pressure, laboratory doors must be kept closed and should be self-closing with locks.

Exhaust air from biohazardous laboratories should not be recirculated in the building. It should be ducted to the outside and released from a stack remote from the building air intake. In certain special situations, including many BSL-3 labs, air exhausting from a containment facility may need to be filtered through High Efficiency Particulate Air (HEPA) filters.

C. Animal Facility Design and Ventilation

Animal facilities must be designed as described above for general BSL-2 laboratory spaces, but they have additional design and ventilation requirements detailed below.

• ABSL-2 facilities should be separated from the general traffic patterns of the building and restricted.
• Doors to areas where infectious materials and/or animals are housed must open inward, be self-closing, be kept closed when experimental animals are present, and never be propped open. Doors to cubicles inside an animal room may open outwards or slide either horizontally or vertically.
• Sink traps are filled with water and/or appropriate disinfectant to prevent the migration of vermin and gases. If open floor drains are provided, the traps are also filled as above.
• Interior surfaces of the animal facility are water-resistant. Floors are slip-resistant, impervious to liquids, and resistant to chemicals. Floors with drains are sloped towards the drains to facilitate cleaning.
• Penetrations in floors, walls, and ceiling surfaces are sealed, including openings around ducts, doors, doorframes, outlets, and switch plates to facilitate pest control and proper cleaning.
• Internal facility fixtures, such as light features, air ducts, and utility pipes, are designed and installed to minimize horizontal surface areas to facilitate cleaning and minimize the accumulation of debris or fomites.
• Equipment and furnishings are carefully evaluated to minimize exposure of personnel to pinch points and sharp edges and corners.
• If external windows are present, they are sealed and resistant to breakage.
• Ventilation for animal facilities is provided in accordance with the Guide for the Care and Use of Laboratory Animals. The ventilation system design considers the heat and high moisture load produced during the cleaning of animal rooms and during the cage washing process. The cage wash area is designed to accommodate high-pressure spraying systems, humidity, and chemical disinfectants.
• The cage wash procedure includes use of a mechanical cage washer where the final rinse temperature is at least 143-180^oF.
• The direction of airflow is into the animal facility such that the rooms are negative compared to adjoining hallways. Exhaust air from animal facilities is not recirculated in the building and is ducted to the outside and released from a stack remote from the building air intake.
• An autoclave is present in the animal facility to facilitate the decontamination of infectious materials and waste. A validated alternative process (alkaline hydrolysis digestion, incineration, etc.) may be used for the decontamination and disposal of animal carcasses.

D. Biological Safety Cabinets and Other Primary Containment Devices

Biological Safety Cabinets (also called Biosafety Cabinets or BSCs) are the primary means of containment developed for working safely with infectious organisms. When functioning correctly and in conjunction with Standard Microbiological Practice (SMP), BSCs are remarkably effective at controlling infectious aerosols. BSCs are laboratory safety equipment utilizing airflow to provide a barrier between the worker and the biohazardous materials. BSCs are distinct from other safety equipment, such as chemical fume hoods and clean benches. Class II BSCs are designed to provide personnel, environmental, and product protection when appropriate practices and procedures are followed.

BSCs and other primary containment barrier systems must be installed and operated in a manner to ensure their effectiveness. BSCs must be installed to ensure that general lab ventilation (air supply and exhaust) do not interfere with the ability of the cabinet to contain aerosols. 

BSCs must be located away from doors, windows that open, heavily trafficked areas, and other sources of airflow disruption. If biosafety cabinets are connected to the lab exhaust system, they must be either through a canopy connection (Class IIA type cabinets only) or hard ducted to the outside (Class II types B and C or Class III BSCs). 

The exhaust from Class IIA and IIC BSCs can be safely recirculated back into the laboratory if no volatile, toxic chemicals are used in the cabinet. BSCs and other primary containment units (including HEPA filters and filter housings of actively ventilated caging systems) must be certified at least annually, and if moved or repaired, to ensure appropriate performance.

Contact the Research Safety office for more information on the certification of BSCs. This information is also on the Research Safety website.

The following practices correspond to BSL-2 and ABSL-2 containment. They are essential for the prevention of laboratory infection and disease, as well as for reducing the potential for contamination of experimental material. If you are considering research with a risk group (RG) 2 or higher organism, or feel as though your research may require BSL-3 containment, contact Research Safety at (312) 503-8300 (Chicago) or (847) 491-5581 (Evanston) or researchsafety@northwestern.edu.

A. Prerequisites for Work

• The Principal Investigator and staff should a develop practices and procedures manual that includes safety. In most cases, the lab’s Northwestern Institutional Biosafety Committee (IBC) protocol, together with this Biosafety Manual, will provide the necessary information to work safely. This manual should be reviewed and updated as necessary.
• When working with rDNA and agents at A/BSL-2 or higher, the Principal Investigator must obtain IBC approval before the work can begin. Please note that, per the NIH Guidelines, the local IBC determines the Biosafety Level for a given research project. If you are uncertain of the Biosafety Level of your research, please get in touch with Research Safety at (312) 503-8300 (Chicago) or (847) 491-5581 (Evanston) or researchsafety@northwestern.edu.
• Post a biohazard sign at the laboratory or animal room entrance when infectious materials are present. This sign should list the biosafety or animal biosafety level, contact information for a responsible person, the Personal Protective Equipment (PPE) necessary for safe entry, any occupational health requirements, and procedures for entry and exit. Contact Research Safety to help update your door sign(s).
• Advise custodial staff of hazardous areas and locations that should be off-limits. Use appropriate signage, including biohazard signs.
• An effective integrated pest management program is implemented.
• Animals and plants not associated with the work performed are prohibited in the lab or animal areas.
• Limit access to laboratory spaces and animal rooms to those required for experimentation, husbandry, or support purposes.

B. Personal Hygiene

• Do not eat, drink, take medication or dietary supplements, chew gum, use tobacco, apply cosmetics or lotions, or handle contact lenses in the laboratory or animal areas.
• Do not store food and drink for human consumption in the laboratory or animal areas. This includes refrigerators and freezers as well as desks, cabinets, etc.
• Wash hands frequently including after handling infectious materials, after removing disposable gloves and protective clothing, and always before leaving the laboratory.
• Keep hands away from the mouth, nose, eyes, face, and hair while working.
• Do not store personal items such as coats, boots, bags, and books in the laboratory or animal areas.
• Long hair is restrained so it cannot contact hands, animals, specimens, containers, or equipment.

C. Mandatory Procedures for Handling Pontentially Infectious Materials at BSL-2

• Good housekeeping practices are essential. All working surfaces in the lab must be visibly clean.
• Disinfect equipment before beginning work and immediately upon the conclusion of an experiment or at the end of the day. Including the decontamination of all shared equipment and common areas.
• Plan and organize materials/equipment before starting work.
• Never mouth-pipette; always use mechanical pipetting devices.
• Perform all laboratory and animal procedures to minimize the generation of aerosols, splashes, and splatters of potentially infectious materials.
• Whenever possible, perform all aerosol-producing procedures such as shaking, grinding, sonicating, mixing, and blending in a properly operating biological safety cabinet. Note that some equipment may compromise the cabinet function by disturbing the air curtain. If performing a procedure within a BSC or another primary containment device is impossible, you must conduct a risk assessment to determine an appropriate combination of PPE and administrative and engineering controls.
• Centrifuge materials containing infectious agents in shatter-resistant, closable tubes. Use a centrifuge with a sealed rotor or screw-capped safety cups. Loading and unloading of these tubes into the rotors or centrifuge safety cups should be done in a biological safety cabinet or other containment devices.
• Cover countertops with plastic-backed disposable paper pads where hazardous materials are used to absorb spills and facilitate cleanup. Regularly dispose of these pads when visibly soiled or a spill occurs.
• Wipe work surfaces with an appropriate disinfectant after experiments and immediately after spills (following your established spill procedure) or splashes of potentially infectious materials. Decontaminate all potentially infectious materials before transport or disposal using an effective method. For help selecting the appropriate disinfectant, refer to the Chemical Disinfection section. For help with a spill response, refer to the Spill Protocols section.
• Decontaminate all contaminated or potentially contaminated materials including cultures and stocks by appropriate methods before disposal. Refer to the Biohazardous Waste Disposal and Methods of Decontamination section for more information about decontamination methods available for all lab wastes.
• Equipment is decontaminated routinely, after spills, splashes, or other potential contamination, and before maintenance, repair, or removal from the laboratory.

D. Procedures for Handling Sharps

• Policies for safely handling sharps (needles, scalpels, pipettes, broken glassware, etc.) are developed, implemented, and followed based on applicable federal, state, and local requirements.
• Plasticware is substituted for glassware whenever it is possible to do so. Broken glassware is not handled directly but removed using a brush, dustpan, tongs, or forceps.
• The use of sharps is limited and restricted to situations with no alternative. Active or passive needle-based safety devices are used whenever possible.
• Needles are uncapped to reduce the potential for recoil causing an accidental needlestick. Ensure other lab personnel are not within arm’s length when using sharps.
• Needles may not be bent, sheared, broken, recapped, removed from disposable syringes, or manipulated by hand before disposal. If necessary, a hands-free device or similar safety procedure must be used to remove a needle from a syringe or recap a needle.
• Used, disposable sharps devices are carefully placed into puncture-resistant sharps disposal containers immediately after use. Position these containers as close to the point of use as possible while sharps are in use.
• Non-disposable sharps are placed in hard-walled, puncture-resistant containers for transportation to the processing area for decontamination, including autoclaving.

E. Procedures for Handling Animals

• A risk assessment determines the type of physical containment devices used for animal work when a BSC may not be suitable. If procedures, especially those that may generate an aerosol, cannot be performed in a BSC or other physical containment device, a combination of appropriate PPE, administrative and engineering controls must be used.
• Whenever possible, restraint devices and practices are used to reduce the risk of exposure during animal manipulations.
• Equipment, cages, and racks are handled in a manner that minimizes contamination of other areas. Cages are decontaminated before washing using an appropriate disinfectant.
• When indicated by the risk assessment, animals are housed in primary biosafety containment equipment appropriate for the animal species such as micro-isolators or equivalent systems for larger animals. If used, actively ventilated caging systems are designed to contain microorganisms. Exhaust plenums for these systems are sealed and safety mechanisms are in place to prevent the cage and exhaust plenums from becoming positively pressurized. An alarm is included to indicate operational malfunction.
• Decontamination of an entire animal room is considered when there has been gross contamination of the space, significant changes in usage, and major renovations or maintenance shutdowns. A risk assessment is used to select the appropriate materials and methods for decontamination.
• Equipment is decontaminated routinely, after spills, splashes, or other potential contamination, and before maintenance, repair, or removal from the laboratory. A method for decontaminating routine husbandry and sensitive electronic or medical equipment is identified and implemented.
• Decontamination processes are verified on a routine basis.

F. Procedures for Handling Materials Requiring BSL-2 Enhanced Containment

Although not an official biosafety level, biosafety level 2 enhanced (also referred to as BSL-2 with BSL-3 practices or BSL-2+), may be assigned to specific IBC registrations. This level is a combination of the laboratory design and engineering controls of BSL-2 laboratories with some of the work practices and PPE used at BSL-3. At Northwestern the following practices and PPE apply to all BSL-2+ work:

• Access to the designated laboratory space or tissue culture room is restricted to laboratory personnel who have proper training and a need to access the space.
• The laboratory door displays signage indicating when BSL-2+ work is in progress and that BSL-2+ work is performed in that laboratory space.
• The dedicated BSL-2+ space has negative pressure (directional airflow) compared to other laboratory and administrative spaces.
• An emergency eyewash is located near the laboratory and a sink is available for handwashing in the room or nearby. If no sink for handwashing is present in the room or dedicated space, hand sanitizer is available and used before leaving the laboratory space. Individuals must proceed immediately to the nearest laboratory sink to perform handwashing after using hand sanitizer.
• Individuals receive specific training on BSL-2+ containment requiring tasks and must demonstrate proficiency before working. Standard Operating Procedures (SOPs) are developed specifically for this work. A BSL-2+ lab manager is assigned to oversee the additional requirements detailed for this work.
• The generation of aerosols is avoided. All work is performed in a properly maintained and certified BSC. Centrifuge safety cups and/or sealed rotors are used for all centrifugations. These cups or rotors are only opened in the BSC.
• All waste that is generated in the BSC is collected and secured in the BSC prior to removal for disposal. Although it is not common practice, the IBC may insist that solid waste is decontaminated (autoclaved) prior to disposal. Liquid biohazard waste should be disposed of in the space in which it is generated (e.g., in the BSL-2+ space) as described in this manual.
• Materials utilized in BSL-2+ laboratories must be transported in a secondary container that is hard-walled, puncture-proof, and effectively cleaned and disinfected. This secondary container must be secured so it would not come open if dropped and be labeled with the biohazard symbol and lab contact information. This material may only be transported to other approved BSL-2+ laboratory spaces.
• The use of sharps is avoided wherever possible. If there are no non-sharps alternatives that can be used, sharps must be used as described in the Procedures for Handling Sharps section above.
• A biohazardous materials spill kit is created and located in the BSL-2+ laboratory space.
• All PPE used in the BSL-2+ space is either single use or dedicated to the space. Eye and face protection is required for all work in BSL-2+ spaces. Two pairs of latex or nitrile gloves are required. A back closing disposable gown or dedicated lab coat must be worn. Consideration is given to wearing lab coats or gowns with cuffed sleeves. Tyvek sleeves that protect the wrists and lab coat or gown sleeves are recommended.

Additional practices may be required by the IBC depending on the proposed work including:

• Medical surveillance and available vaccinations
• Inventory of agents with an activity log
• Use of respiratory protection and enrollment in Northwestern’s respiratory protection program
• Restriction of BSL-2 work occurring simultaneously with BSL-2+ work in the same space

Selection of all Personal Protective Equipment (PPE) and protective clothing is based on an appropriate risk assessment. Purchase of PPE necessary to safely conduct your work is the responsibility of your PI. Research Safety supports this by offering many basic components of PPE at no cost.

PPE may include body coverings (lab coats, gowns, scrubs, or uniforms), eye and face protection (safety glasses, goggles, masks, face shields, or other splatter guards), hand protection (latex or nitrile gloves, bite resistant gloves, chemical resistant gloves), and respiratory protection (N95 respirator, powered air-purifying respirator [PAPR]). If research animals are present, the risk assessment will consider appropriate eye, face, and respiratory protection including the potential for exposure to animal allergens. Additional PPE is considered for individuals working with large animals.

All protective clothing and PPE must be contained, decontaminated, and/or disposed of properly. Reusable clothing is contained and decontaminated before laundering by the institution. No protective clothing or PPE may be taken home.

Eye and face protection must be used for manipulations or activities that may result in splashes or sprays of infectious or other hazardous materials in the laboratory and animal facility environment. Eye and face protection are disposed of with other contaminated facility waste or decontaminated after use if reusable. If non-human primates are used, the risk of mucous membrane exposure is assessed, and appropriate PPE is selected to protect this route of exposure.

Gloves are worn to protect hands from exposure to hazardous materials and when handling animals. Disposable gloves provide barrier protection against biohazardous materials. When handling animals, assess the need for a bite and scratch-resistant glove over a disposable one. Gloves must not be worn outside of laboratory or animal facility and gloves should be changed regularly, including when contaminated or when glove integrity is compromised. Disposable gloves may not be decontaminated, washed, or reused. Contaminated gloves must be disposed of with other biohazardous waste. 

Gloves and all other PPE are removed in a manner that minimizes personal contamination and transfer of infectious materials outside of the laboratory or animal facility where the materials or animals are handled.

Respiratory protection is considered based on a risk assessment of the work to be performed. Personnel using respirators based on the potential for inhalational hazard or animal allergy prevention are enrolled in an appropriately constituted respiratory protection program. Contact Research Safety if you have any questions or concerns.

Please refer to Appendix B of the BMBL (6th Edition) for more information.

The proper disposal of biohazardous waste protects the public, the environment, laboratory and custodial personnel, and transportation workers. Generators of biohazardous waste must ensure that waste labeling, packaging, and intermediate disposal conforms to federal, state, and local regulations as prescribed by Research Safety.

The definitions below are essential:

• Decontamination refers to removing disease-causing microorganisms and agents, rendering an object safe for general handling. Thoroughly cleaning surfaces and equipment is a prerequisite for good microbiological practice.
• Disinfection is a process that kills or destroys most disease causing microorganisms, except spores using a chemical agent.
• Sterilization is a process that destroys all forms of microbial life, including spores, viruses, and fungi.

A.  What is Infectious Waste?

The following are common examples of items usually considered to be an infectious waste:

• Microbiological laboratory wastes such as cultures derived from clinical specimens and pathogenic microorganisms.
• Laboratory equipment that may have encountered clinical specimens, pathogenic microorganisms, or cultures derived from them.
• Tissues, large quantities of blood, and/or body fluids from humans.
• Tissues, large quantities of blood, and/or body fluids from one or more animals carrying an infectious agent that can be transmitted to humans.
• Contaminated sharps (needles, broken glass, etc.)

The following items require decontamination prior to disposal even though they are not necessarily considered infectious waste:

• Regulated rDNA-containing organisms
• Exotic or virulent plant pathogens
• Exotic or virulent animal pathogens
• Recombinant arthropods

The following are usually not included in the definition of waste but should be placed in containers, such as plastic bags, prior to disposal to contain the waste.  If these items are mixed with infectious wastes, they must be managed as though they were infectious.  For this reason, segregate infectious waste from other waste.

• Items soiled or spotted, but not saturated, with human blood or bodily fluid (e.g. blood-spotted gloves, gowns, dressings, etc.).
• Containers, packages, waste glass, laboratory equipment, and other materials that have had no contact with blood, body fluids, clinical cultures, or infectious agents.
• Non-infectious animal waste (e.g. manure, bedding, tissue, blood, and body fluids) or cultures from an animal that is not known to be carrying an infectious agent that can be transmitted to humans.

B.  Packaging Waste

Laboratory materials used in experiments with potentially infectious microorganisms, such as discarded cultures, tissues, media, plastics, sharps, glassware, instruments, and laboratory coats, must either be handed off to a contractor licensed as an infectious waste treatment facility or be decontaminated before disposal or washing for reuse. 

Collect contaminated materials in durable, leak-proof containers labeled with the universal biohazard symbol. For infectious materials, the outer surface of the container must be disinfected and wiped dry before moving the materials. Autoclavable biohazard bags are recommended.

For researchers at Northwestern University, autoclaving biohazard waste prior to disposal is neither required nor recommended.  Since many autoclaves are not regularly validated for their effectiveness, all biohazard waste should be disposed of in the large collection bins located in designated research areas (refer to the Research Safety website for their locations).  A third-party vendor collects all biohazard waste at Northwestern, transports it to a disposal facility, and is responsible for decontamination and disposal. 

If you plan to autoclave biohazard waste prior to disposal, please contact Research Safety and the Biosafety Officer. After autoclaving biohazard waste, all items must still be disposed of in the large collection bins located in designated research areas.

Uncontaminated sharps (and other noninfectious items that may cause a cut or laceration) require special precautions to prevent injury, even if uncontaminated. Sharps must be collected in rigid, puncture-resistant containers to prevent injuries to scientists, custodial personnel, and waste technicians. Standardized sharps containers and other waste collection containers are available through the Scientific Stockrooms.

For more information, please refer to the Research Safety Hazardous Waste Information webpage (https://researchsafety.northwestern.edu/safety-information/hazardous-waste.html).

C. Methods of Decontamination

Choosing the appropriate method to eliminate or inactivate a biohazardous agent can be complicated. The method depends on the treatment equipment available, the infectious agent, and the presence of interfering substances (e.g., media, high organic content, tissues). Various treatment techniques are available, but practicality and effectiveness determine the most appropriate method.

Ideally, decontaminate your biohazardous waste before the end of each day unless it is to be collected for treatment off-site. In the latter case, the waste should be packaged and stored (frozen, if pathological waste) until the scheduled pickup by Research Safety. 

Biohazardous waste should never be compacted, and ordinary lab wastes should be disposed of as routinely as possible to reduce the amount requiring special handling.

Regardless of its infectious nature, all organisms containing recombinant or synthetic nucleic acids (rDNA) must be decontaminated before final disposal. This includes non-pathogenic strains used for cloning, protein expression, diagnostics, etc. 

For help determining the best method of decontamination and disposal, consult Research Safety at researchsafety@northwestern.edu, or call (312) 503-8300 (Chicago) or (847) 491-5581 (Evanston).

1. Steam Sterilization

Decontamination by autoclave of biohazardous waste is prohibited at Northwestern except in special situations with written approval of the Biosafety Officer. All solid biological waste is collected and disposed of by third-party contractors. 

In some cases, steam sterilization using an autoclave is required before collecting and disposing of certain biological wastes. The autoclave should be properly functioning and routinely monitored for efficacy with a biological indicator, such as spores of Geobacillus stearothermophilus.

When autoclaving biohazardous waste, it is recommended that the cycle be at least one hour. However, the nature of the batch should determine the cycle duration. For example, if the batch contains a dense organic substrate such as animal bedding or manure, more than one hour may be needed to inactivate pathogens within the material. Some volume of liquid is necessary within the bag and the bag must not be closed in an airtight fashion to ensure appropriate steam sterilization, consult Research Safety for assistance. 

Use caution when treating waste co-contaminated with volatile, toxic, or carcinogenic chemicals, radioisotopes, or explosive substances. Autoclaving this type of waste may release dangerous gasses into the air (e.g., autoclaving waste previously treated with bleach may release chlorine) and damage the autoclave.  Anticipate the creation of mixed waste before starting your experiment. Contact Research Safety for assistance.

2. Drain Disposal of Liquid Biohazardous Waste

Following proper decontamination, most liquid biological waste may be disposed of by pouring it into a sink drain and flushing it with a ten-fold excess of water. Proper decontamination with bleach is the most common method. Dilute bleach with liquid 1:10 (one part bleach to nine parts liquid), swirl, and let stand twenty minutes. When working, take care to avoid generation of aerosols. Remember, if the fluid is contaminated with infectious agents or biological toxins, it must be decontaminated by chemical disinfection before being released down the drain.

3. Chemical Disinfection

Chemical disinfection kills potentially infectious materials, reducing viability. The disinfectant should be freshly prepared at a concentration effective against the biohazardous agent. 

Remember, a chemical disinfectant does not kill everything but reduces the viability and potential for infectivity.

An effective disinfectant kills/inactivates the agent at the lowest concentration and with minimal risk to the user. Other considerations, such as the economy and shelf life, are also important. Allow sufficient contact time to ensure complete inactivation. It is essential to be aware that common laboratory disinfectants can cause harm. For example, ethanol and quaternary ammonium compounds may cause contact dermatitis. Bleach solutions may irritate the skin and the upper respiratory tract when inhaled.

a. Halogenated Chemicals (e.g., hypochlorites)

Halogenated chemicals such as hypochlorite (household bleach) are the least expensive and are also highly effective in decontaminating large spills. Their drawbacks include short shelf life, easy binding to non-target organic substances, and corrosiveness. Hypochlorite typically is diluted between 1:10 and 1:100 such that the available halogen is 0.01-5.0%. A 1:10 dilution of household bleach (one part bleach to nine parts liquid) will achieve this concentration. To ensure disinfection, the recommended contact time is twenty minutes. The more proteinaceous the material, the more bleach solution should be used to ensure disinfection.

Bleach solutions need to be made fresh each day. Be aware that using chlorine compounds to disinfect some substances may inadvertently release toxic compounds.  For example, using chlorine compounds to disinfect substances co-contaminated with radioiodine may cause gaseous release of the isotope. Household bleach is an inexpensive and effective disinfectant against many microorganisms, including bacterial spores (considered the most difficult to kill). It is the preferred disinfectant at Northwestern. Bleach solutions are corrosive. If used on stainless steel, follow with a sterile water rinse.

b. Alcohols

Alcohols, such as ethanol and isopropanol, are effective against vegetative forms of bacteria and fungi and enveloped viruses. They will not destroy spores or non-enveloped viruses (e.g., adenovirus and adeno-associated virus).

Alcohols (ethanol or isopropyl alcohol) are usually most effective at 70% concentration (a 7:10 dilution of alcohol in water). At greater concentrations (say 95%), alcohol is much less effective as a disinfectant. Some intrinsic characteristics of alcohols, such as flammability, poor penetration, presence of protein-rich materials, and rapid evaporation, limit their usefulness. Alcohol is not recommended for larger spills.

c. Phenolics

Two phenol derivatives commonly found as constituents in hospital disinfectants are ortho-phenylphenol and ortho-benzyl-para-chlorophenol.  The antimicrobial properties of these compounds and many other phenol derivatives are much improved over those of the parent chemical.  Porous materials absorb phenolics and the residual disinfectant can become an irritant.  In high concentrations, phenol acts as a gross protoplasmic poison: penetrating and disrupting the cell wall and precipitating cellular proteins.  Phenolics are effective against most microbes including bacteria (including M. tuberculosis), fungi, and viruses. Phenolic germicides are Environmental Protection Agency (EPA)-registered as disinfectants for use on environmental surfaces (e.g., laboratory surfaces).

d. Quaternary Ammonium Compounds

Quaternary ammonium compounds (also called quaternaries or quats), are a diverse class of disinfectants with varied chemical structure. The microbial properties are unique to each compound. Quaternaries are good cleaning agents but their microbicidal properties can be diminished by high water hardness and absorption or inactivation of the active ingredient by some cleaning materials. Quaternaries have been shown to be effective against fungi, gram-positive bacteria, and enveloped viruses. Generally, quaternaries are not effective in disinfection of spores, many gram-positive bacteria, mycobacteria, or enveloped viruses. Quaternaries play an essential role in sanitation but are not the first choice as a disinfectant.

e. Formaldehyde (and Other Aldehydes)

Formaldehyde is used as a disinfectant in both its liquid and gaseous states.  Liquid formaldehyde is used most commonly whereas gaseous formaldehyde is used only in specific circumstances such as high containment labs or when the entire atmosphere within a large space must be decontaminated.  Formaldehyde is most commonly sold and used in a water-based solution called formalin.  This aqueous solution is effective at decontaminating bacteria, including mycobacteria, fungi, viruses, and spores.  Depending on the target agent and other factors such as media, effective concentrations can range from 2% to 8% formalin in water.  It is important to also note that the contact time required varies widely.  Formaldehyde poses a number of health risks to laboratory staff that are not seen with other disinfectants.  Consult with Research Safety if you are considering using formaldehyde as a disinfectant at researchsafety@northwestern.edu, or call (312) 503-8300 (Chicago) or (847) 491-5581 (Evanston). Formaldehyde is one method used to decontaminate biological safety cabinets, and it is not to be used as a routine disinfectant in any laboratory at Northwestern.

f. Peroxides

Hydrogen peroxide is commonly used for decontamination in healthcare and sometimes biomedical laboratories. Hydrogen peroxide is active against bacteria, spores, viruses, and fungi by producing destructive hydroxyl free radicals that attack membrane lipids, nucleic acids, and other essential cellular components. Cellular catalase, produced by aerobic and facultative anaerobic microbes to protect them from metabolically produced hydrogen peroxide, is overwhelmed by concentrations used for disinfection. 

Effective concentrations for decontamination range from 6% to 25%, depending on the target agent. The stability of the peroxide during storage is a consideration; prepare fresh solutions of hydrogen peroxide each day. Note: concentrations of peroxide as low as 3% have been shown to cause dermal irritation when direct exposure occurs. Never use concentrations of hydrogen peroxide great than 30%, as they pose an unacceptable risk in the laboratory.

g. Iodophors

An iodophor is a combination of iodine and a solubilizing agent. The resulting complex provides a sustained release reservoir of iodine and releases small amounts of free iodine in an aqueous solution. Iodophors have been used as both an antiseptic and a disinfectant. The best-known and most used iodophor is povidone-iodine (often known as Betadine®), sometimes used in first aid cleaning wounds. 

Iodophors are thought to function by allowing the iodine to penetrate the cell wall and disrupt protein and nucleic acid structure and synthesis. Iodophors are effective in decontaminating bacteria, mycobacteria, and viruses but may require prolonged contact to kill certain fungi and bacterial spores.

Wescodyne® is a commercial product that combines povidone-iodine and a detergent. Used undiluted in aspiration traps, it provides a practical and stable disinfectant. As the trap fills, the Wescodyne is diluted and remains effective until the aspiration trap is emptied and replaced with fresh disinfectant. Research Safety provides a proper sticker to help note when an aspiration trap was last cleaned.

A similar disinfectant is Wex-cide 128. This EPA-registered disinfectant is suitable for agents handled at BSL-2. It has excellent detergent properties and is well-suited for use in aspiration traps, similar to Wescodyne®.

Table 6: Chemical Disinfectants and Their Efficacy

Efficacy of disinfectants with recommended dilutions or concentrations

DISINFECTANT CLASS	EFFECTIVE AGAINST						RECOMMENDED DILUTION OR CONCENTRATION
	Fungi	Bacteria (Gram-positive and Gram-negative)	Mycobacteria	Spores	Enveloped Viruses	Non-enveloped Viruses
Phenolics	Good	Good	Good	Ineffective	Mild	Varies	1%-5%
Hypochlorites	Mild	Good	Fair	Fair	Mild	Mild	0.005-0.5% free chlorine 1-10% household bleach
Alcohols	Ineffective	Good	Good	Ineffective	Mild	Varies	70-85%
Formaldehyde	Good	Good	Good	Good1	Good	Mild	2-8%
Glutaraldehyde	Good	Good	Good	Good2	Mild	Mild	2%
Iodophors	Good	Good	Good	Mild	Mild	Mild	0.5%
1. above 40°C 2. above 20°C

Some laboratory mishaps require an emergency response. The laboratory supervisor must follow institutional requirements and prepare emergency instructions specific to the biological hazards of the laboratory. These instructions should be displayed prominently in the laboratory and periodically reviewed with laboratory personnel. A best practice is to review these every three months during a lab meeting and ensure that action items are addressed.

Your Research Safety laboratory safety specialist can assist you.

A. Medical Surveillance and Occupational Health

Laboratory personnel are provided with medical surveillance, as appropriate, and offered available immunizations for agents handled or potentially present in the lab.

All Northwestern employees with potential exposure to human or non-human primate blood, bodily fluids, or other substances covered by the Bloodborne Pathogens program are eligible to receive the Hepatitis B (HepB) vaccination.

The HepB vaccination schedule includes a series of intramuscular injections, with a defined number of doses at defined intervals. The number of doses and timing may depend upon the country in which they are given. Additional vaccinations may be required depending on the agents in use and will be identified during the IBC review process.

B. Laboratory Animal Medical Surveillance

Animal care staff are provided information on signs and symptoms of disease, receive occupational medical services including medical evaluation, surveillance, and treatment, as appropriate, and are offered available immunizations for agents handled or potentially present in the facility based upon a risk assessment. An animal allergy prevention program is included in medical surveillance for those with potential exposure.

C. Spill Protocols

For spills outside of a primary containment device (BSC):

1. In the event of an extensive or explosive spill of virulent pathogen, everyone should leave the affected area immediately. Contaminated clothing should be removed and properly disinfected prior to laundering at home. Exposed skin should be washed thoroughly with plenty of soap and water.
2. Leave the lab, close the laboratory door, and post a “No Entry” sign indicating the hazard. Notify the laboratory supervisor, PI, and the Office for Research Safety at (312) 503-8300 (Chicago) or (847) 491-5581 (Evanston).
3. Determine the necessity and extent of medical treatment for individuals exposed to infectious microorganisms. In the event of life-threatening injuries, contact emergency personnel; call 911. For non-life-threatening injuries, personnel accidentally exposed via ingestion, skin puncture, or apparent inhalation of infectious agents should be given appropriate first aid and, if necessary, transported to the nearest emergency room.
4. Contact Risk Management at 1-5582 from a campus phone. Risk Management will make an appointment for individuals with injuries through Corporate Health (Chicago Campus) or Omega (Evanston Campus).
5. Only reenter the room once large droplets have settled and aerosols have been cleared by the building ventilation system (a minimum of 30 minutes). The extent of the hazard and its dissemination have been determined.
6. Each person who enters the laboratory for cleanup should wear proper PPE.
7. Use an appropriately concentrated disinfectant to clean up and decontaminate the area. A supply of disinfectant should always be available. 
8. Please dispose of all contaminated, non-reusable materials in biohazard bags and discard them as biohazard waste.
9. Decontaminate all reusable materials used in cleanup procedures.

 Small spills within a BSC:

1. Do not turn off the BSC.
2. Cover the spill with absorbent paper towels
3. Carefully pour an appropriate disinfectant onto the towel-covered spill; a 1:10 bleach solution is recommended to reduce the amount of disinfectant needed due to dilution in addition to the spill.
4. Remove the decontaminated towels.
5. Place contaminated material into the biohazard bag
6. Splatter onto items within the cabinet, and the cabinet interior, should be immediately wiped with a disposable towel dampened with fresh disinfectant.
7. Remember to remove your gloves, wash your hands, and wear clean gloves when finished. Hands should be washed whenever you change your gloves.

 Large spills within a BSC:

1. Do not turn off the BSC.
2. Ensure the drain valve is closed.
3. Decontaminate and remove all items from within the BSC.
4. Pour an appropriate disinfectant directly onto the work surface and through the grilles into the drain pan; undiluted bleach is recommended.
5. Wait at least 20-30 minutes.
6. Empty the drain pan into a collection vessel containing more disinfectant.
7. Pour the disinfectant solution down the drain, followed by 10-fold excess water.
8. If the spill contains radioactive material, Radiation Safety staff should be alerted before proceeding with the clean-up.

In an emergency situation in a BSL-2 lab, attention to immediate personal danger overrides containment considerations. No biological agents managed at BSL-2 would prohibit emergency personnel wearing appropriate PPE from entering any BSL-2 laboratory in an emergency.

D. Exposure Response Protocols

Determine the necessity and extent of medical treatment for persons exposed to infectious microorganisms. Personnel accidentally exposed via ingestion, skin puncture, or apparent inhalation of an infectious agent should be given appropriate first aid and, if necessary, taken to the nearest emergency room. Immediately go to the nearest sink and begin washing the area with plenty of soap and water for at least 15 minutes. If available (free in the Stockrooms) use generic povidone-iodine. Avoid damaging the skin.

For exposures to the eyes or mucous membranes, the exposed area should be flushed with running water for 15-20 minutes. 

Seek immediate medical attention for all exposures.

E. Reporting

The importance of reporting accidents, spills, or other exposure events immediately to your laboratory or animal facility supervisor is not to identify fault or failure within the laboratory or on the part of the laboratory personnel. The goal of reporting such incidents is to identify opportunities to refine standard operating procedures for the laboratory and improve laboratory safety. Importance is always placed on personal health and the health and safety of coworkers, the research community, and the public. All incidents will be evaluated at the discretion of the medical provider. Appropriate records of the incident, response, and evaluation must be maintained.

The secure and responsible conduct of life sciences research depends, in part, on observation and reporting by peers, supervisors, and subordinates. Individuals working with potentially infectious material and/or rDNA constructs with either direct or indirect, acute or latent disease potential (e.g., insertional mutagenesis due to exposure to a viral vector) must understand and acknowledge their responsibility to report activities that are inconsistent with a culture of responsibility or are otherwise troubling. Likewise, institutional and laboratory leadership must acknowledge their responsibility to respond to reports of concerning behavior and undertake actions to prevent retaliation stemming from such reports.

There are numerous methods for reporting concerning behavior, as described above in the Code of Conduct and Culture of Responsibility section. Where appropriate, an individual should report to his/her PI, supervisor, Department Chair, or Dean. In instances where confidentiality is of importance, reports may be made to the Northwestern Research Integrity office (https://researchintegrity.northwestern.edu/), Ethics Point (a third-party vendor that will allow you to report your concerns anonymously at https://secure.ethicspoint.com/domain/media/en/gui/7325/index.html or call (866) 294-3545), or Research Safety (researchsafety@northwestern.edu or call (312) 503-8300 (Chicago) or (847) 491-5581 (Evanston)).

Persons with health conditions, whether chronic or acute, that have the potential to place themselves or others at risk in the laboratory should consider self-reporting to their personal physician and/or Corporate Health (Chicago) or Omega (Evanston). A decision about continued involvement with research involving infectious agents must be an informed decision that includes appropriate medical expertise.

A. Transportation Within Northwestern University

When transporting biohazardous materials within the Northwestern University campus or between laboratory or animal facility locations, they must be contained to protect against spill or exposure to the individual hand-carrying them or others in public spaces. 

Biohazardous materials must be secured in a closed primary container designed to hold the type of material inside. This container must be placed in a secondary container that is hard-walled, puncture-proof, and effectively cleaned and disinfected. 

If wet or dry ice is needed, additional requirements for this secondary container must be considered to ensure the handler’s safety. This secondary container must be secured so it would not come open if dropped and be labeled with the biohazard symbol and lab contact information. 

Avoid traveling through high traffic, public areas or places where food is prepared or consumed, whenever possible. If several items are being transported, consider using a cart. 

Although PPE should be removed before leaving the laboratory or animal facility, one clean glove may handle the secondary container leaving the other hand ungloved to open doors, push elevator buttons, etc. If a spill occurs during transportation, contact Research Safety at (312) 503-8300 (Chicago) or (847) 491-5581 (Evanston) immediately.

Individuals may not personally transport materials between campuses or any other Northwestern facility. To transfer materials between campuses, a professional courier service or commercial carrier must be used. IATA DGR requirements (see “Packaging and Shipping Outside of Northwestern University” section below) must be met for packaging all samples transported via commercial carrier. Professional courier services may allow packing materials in a manner different from IATA DGR requirements. This should be discussed with the courier service.

B. Transportation from Northwestern Memorial Hospital to Northwestern University

When human materials are transported from Northwestern Memorial Hospital to a Northwestern University lab they must be well contained and protected against spill or exposure to the individual hand-carrying them or others in public spaces.

See above for the details about packaging and transportation. In addition, before transportation, a transport protocol must be reviewed and approved by the IBC at a convened meeting. For the transport protocol and instructions, you should send a request to ibc@northwestern.edu.

C. Tranportation of Human Samples in Personal Vehicles

Transportation of human samples for research purposes in personal vehicles is subject to Northwestern policy and significant oversight by Research Safety and the IBC. Such transportation is forbidden without prior approval by the IBC. The IBC will only grant approval with appropriate scientific justification. Financial justifications will not be considered. For more information contact the Biosafety Officer or IBC staff at ibc@northwestern.edu.

D. Packaging and Shipping Outside of Northwestern University

For transportation, dangerous goods are those substances or articles that have the potential to cause harm to individuals, property, or the environment in the event of an accident or incident. Since infectious substances pose a risk to health (in the form of the disease), if an individual is exposed to them during transport, they are considered dangerous goods when transported. This is important to note, as the hazard level of a particular substance may be different in transport than when being manipulated in a laboratory. National and international regulations govern how shipments of infectious substances are prepared and transported. 

National and international regulations dictate that any individual transporting hazardous materials must be trained, tested, certified, and retain a record of their training. This includes any individuals responsible for preparing and packaging a shipment, marking and labeling packages, preparing shipping documents, loading and unloading transport vehicles, and supervising any of the activities above.

Various national and international regulatory bodies have provided guidance and requirements for the shipment of dangerous goods. The United States Department of Transportation (DOT) regulates the transport of dangerous goods via roadways and railways. Dangerous goods shipped internationally via air are subject to the International Air Transport Association (IATA) Dangerous Goods Regulations (DGR). Compliance with IATA DGR meets or exceeds the requirements of the US DOT. 

Northwestern University provides access to training through Research Safety. The Safe Shipping of Biological Materials and Dry Ice course provides shipping training following IATA DGR. Alternative training may be obtained through other agencies; however, Northwestern University recommends that all personnel involved in the shipping of dangerous goods be trained in compliance with IATA DGR. IATA DGR training must be renewed every two years.

For more information regarding the Safe Shipping of Biological Materials and Dry Ice course, or any other training offered by Research Safety, please visit https://researchsafety.northwestern.edu/training/what-training-do-i-need.html.