For current Biosafety standards, please visit the University of Utah’s IBC website by clicking here or visiting www.ibc.utah.edu

 

I am a PI of a research laboratory, do I need to register in BioRAFT?

Yes. PIs of all research laboratories are required to register in BioRAFT.

I conduct biological research, do I need to register my work with the Institutional BioSafety Committee?

Yes. PIs of work with biological materials will need to register their work with the IBC through BioRAFT.

If your work involves any of the following, some level of review and oversight by the IBC or the Biosafety Office is necessary:

• Non-exempt recombinant or synthetic nucleic acid molecules research.
• Studies using human or animals pathogens, including materials known to harbor pathogens (for example, blood from HBV-positive patients).
• Generation of de novo transgenic animals. The breeding of transgenic animals to generate additional transgenic offspring does not require IBC approval. Those transgenic animals that already exist or which have been purchased also do not require IBC approval.
• Work with Acute Biological Toxins.
• Human Subjects research involving the introduction of recombinant molecules or biohazards into human subjects: these studies must be approved by the IBC and by the IRB.
• Animal Subjects: All research involving the use of recombinant molecules or human or animal pathogens in whole animals requires both IBC and IACUC approval.
• Materials potentially containing human pathogens (for example, unfixed human specimens, human blood).
• Work with human cell lines that are not well-characterized or require BSL 2 containment. This includes all cell and organ cultures of human origin (except well-established cell lines that have had comprehensive pathogen testing), human embryonic stem cells, and pluripotent cells and their derivatives.
• The administration of human or non-human primate cells (primary cultures and established cell lines) or tumors into whole animals requires both IBC and IACUC approval.

What are my responsibilities as the PI of a biological research laboratory?

• Ensuring compliance with all pertinent policies and requirements as indicated by the Biosafety Program within your laboratories
• Performing risk assessments (link to a risk assessment form?) of all research projects and lab activities involving biological materials and addressing specific hazards associated with each project
• Drafting, maintaining and ensuring laboratory staff training on a laboratory-specific Biosafety Manual
• Establishing appropriate safety practices within your laboratory
• Training laboratory staff and any visitors on laboratory-specific safety practices, potential hazards and emergency responses (e.g., spill, potential exposure) as it relates to their research activities.
• Ensuring that laboratory staff have the necessary resources to perform their work safely
• Maintaining your laboratory facility and equipment
• Submitting research protocols for review and approval to the Biosafety Program, IBC , IACUC and the IRB, depending on the type of research being performed
• Ensuring that staff who handle animals, animal tissue or have vivarium access participate in the Occupational Medicine Surveillance Program
• Reporting emergencies to OEHS within the required timeframe
• Notifying the Biosafety Program (biosafety@ehs.utah.edu) of plans to commission laboratory space for use with biohazardous material
• Notifying the Biosafety Program (biosafety@ehs.utah.edu)of plans to clear or renovate laboratory space that previously was used for biohazardous material

NIH has produced a useful brochure that describes the Investigator Responsibilities under the NIH Guidelines.  This can be downloaded at: “Investigator Responsibilities under the NIH Guidelines for Research Involving Recombinant DNA Molecules” (National Institutes of Health Office of Biotechnology Activities)

Additional information regarding investigator responsibilities can be found at University of Utah’s Research and Integrity guidelines.

How do I get started?

Principal Investigators (PIs) must:

1. Complete the General Laboratory Setup and Biological Registration Wizards in BioRAFT at least 2 months prior to the planned start date.
2. Create a laboratory Biosafety Manual, as described in the CDC Biosafety in Microbiological and Biomedical Laboratories (BMBL) 5th Edition.
3. Ensure all lab personnel are trained and proficient prior to beginning experiments.

What should my Biosafety Manual include and are there templates?

Include laboratory-specific Standard Operating Procedures (SOPs) for all experiments involving biological agents, risk assessments for the agent, descriptions of personal protective equipment, procedures for using and decontaminating equipment, spill procedures and post-exposure plans, waste disposal and emergency equipment and practices. The PI should develop SOPs describing experimental procedures for working with r-sNA agents and human or animal pathogens.

The University of Utah maintains a Biosafety manual which can be used as a source of information and can be downloaded here.

• The IBC has developed Biosafety Manual/SOP templates for BSL2 and BSL2-enhanced laboratories that may be used by PIs as the basis for laboratory-specific SOPs. These address most areas of review by the IBC but should be customized to reflect the work being performed in the laboratory and the facilities available. BSL1 laboratories can also use these as templates, modified as appropriate.

How do I register with the Institutional BioSafety Committee?

Complete the General Laboratory Setup and Biological Registration Wizards in BioRAFT at least 2 months prior to the planned start date or expiration of approved protocols.

1. For work with recombinant or synthetic nucleic acids, human or animal pathogens, materials that may harbor human pathogens, or involve the generation of genetically modified animals, complete the relevant surveys in BioRAFT.
2. For work with biological toxins, submit a signed registration form for review: links to these forms can be found below. The completed forms must be attached to the Laboratory registration in BioRAFT.

Registration Forms:

• Biological Toxins
• Human Gene Transfer Registration
• Human Gene Transfer Closure Notification
• Human Gene Transfer Amendment Form
• Recombinant DNA and Synthetic Nucleic Acid and Biological Materials Incident Reporting Template
• IBC Closure Notification Form

If you have any questions, contact OEHS Biosafety at 801-585-6950 or biosafety@OEHS.utah.edu.

What is the Institutional BioSafety Committee?

An Institutional Biosafety Committee (IBC) is required at institutions that receive funding from the National Institutes of Health (NIH) for research involving recombinant or synthetic nucleic acid molecules (r-sNA). All non-exempt recombinant DNA and synthetic nucleic acid research at the University of Utah, regardless of funding source, must be conducted in accordance with the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules and must be registered with the IBC. At the University of Utah, as at many other institutions, the IBC also has the responsibility of reviewing a variety of experimentation that involves biological materials, such as risk group (RG) 2 or higher pathogens, hazardous biological materials, and other potentially hazardous agents, such as acute biological toxins.

The IBC works in conjunction with the University of Utah Occupational and Environmental Health and Safety (OEHS) Department to advise the Vice President for Research on policies concerning the safe conduct of research involving recombinant or synthetic nucleic acid molecules in living cells, human gene transfer, microbial pathogens, acute biological toxins, select microbial agents and biological toxins produced by microorganisms that are listed as potential bioterrorism agents, and restricted microbial pathogens of domestic animals and plant crops, and to ensure compliance with current University, local, state, and federal regulations.

The committee consists of a Chair, Administrator (the University of Utah Biosafety Officer), and a minimum of 5 additional members. In order to ensure the competence necessary to review and approve recombinant or synthetic nucleic acid molecule activities, the IBC includes:

1. At least one practicing scientist with experience in microbiology, molecular biology, or virology.
2. At least one practicing scientist with expertise in recombinant or synthetic nucleic acid molecule technology, biological safety, and physical containment.
3. At least one practicing scientist with expertise in human gene transfer experiments
4. At least one member representing laboratory technical staff.
5. At least two community members, not affiliated with the University of Utah in any way other than as a member of the IBC, and who does not have an immediate family member who is affiliated with the University of Utah. At least one community member has primary experience in a nonscientific area (for example, an ethicist, lawyer, or member of the clergy).

Committee members serve at least one 3-year term. Members each have a personal commitment to laboratory safety in general and biosafety in particular.

What are the NIH Guidelines for the conduct of research with Recombinant or Synthetic Nucleic Acids?

The NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules (NIH Guidelines) detail safety practices and containment procedures for basic and clinical research involving recombinant or synthetic nucleic acid molecules, including the creation and use of organisms and viruses containing recombinant or synthetic nucleic acid molecules.

In the context of the NIH Guidelines, recombinant and synthetic nucleic acids are defined as:

1. molecules that a) are constructed by joining nucleic acid molecules and b) that can replicate in a living cell, i.e., recombinant nucleic acids;
2. 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
3. molecules that result from the replication of those described in (i) or (ii) above.

All projects (independent of whether they are funded by NIH) involving recombinant or synthetic nucleic acid molecule conducted at or sponsored by an institution that receives NIH funds for projects involving such techniques must comply with the NIH Guidelines. Noncompliance may result in: (i) suspension, limitation, or termination of NIH funds for recombinant or synthetic nucleic acid molecule research at the institution, or (ii) a requirement for prior NIH approval of any or all recombinant or synthetic nucleic acid molecule projects at the institution.

I work with recombinant or synthetic Nucleic Acids. Do I need to register with the Institutional Biosafety Committee?

Likely yes.

According the NIH Guidelines, recombinant or synthetic DNA molecules are defined as either: (i) molecules that are constructed outside living cells by joining natural or synthetic DNA segments to DNA molecules that can replicate in a living cell, or (ii) molecules that result from the replication of those described in (i). It is the responsibility of each investigator to make sure that his/her laboratory is in compliance with the NIH guidelines. Work with nucleic acids that fall under these guidelines must be reviewed by the Institutional BioSafety Committee (IBC), as described below. Work with recombinant nucleic acids that do not meet these criteria, such as the use of plasmids to express genes in mammalian cells, may be exempt from IBC review. If your experiments require registration with the IBC, check the Guidelines for the appropriate biosafety level and relevant section. For additional information, copies of the Guidelines or registration forms, or if you are unsure into which category your experiments fall, please contact the University of Utah Biosafety Officer at (801) 581-6950 or biosafety@ehs.utah.edu.

Experiments which must be registered and approved by the IBC prior to initiation

• Deliberate transfer of a drug trait to a microorganism not known to acquire it naturally (if it could compromise the use of the drug to control disease agents in humans, animal or agriculture).
• Human gene transfer experiments.
• Cloning of DNA encoding molecules lethal to vertebrates at an LD 50 of <100ug/kg body weight.
• Cloning using human or animal pathogens as host-vector systems.
• Cloning of DNA from all Risk Group 3, 4 or restricted human or animal pathogens (including HIV and related viruses, and human tumor viruses).
• Experiments using more than 2/3 of the genome of infectious animal or plant viruses or defective viruses grown in the presence of helper virus.
• Recombinant DNA experiments involving whole animals or plants.
• Large scale DNA project (>= 10 liters of culture combined).

Experiments that require IBC registration simultaneous with initiation

• Experiments using as vectors <= 2/3 of the genome of a eukaryotic virus, free of helper virus
• Low risk plant rDNA experiments
• BL1 transgenic or knockout rodent experiments. (Note: the purchase of transgenic rodents for BL1 experiments is exempt from registration).

Exempt experiments that require registration but not IBC review

• rDNA containing less than 1/2 of an eukaryotic viral genome propagated in cell culture. The following categories are not exempt from the NIH Guidelines: (i) experiments described in Section III-B which require NIH/OSP and Institutional Biosafety Committee approval before initiation, (ii) experiments involving DNA from Risk Groups 3, 4, or restricted organisms (see Appendix B, Classification of Human Etiologic Agents on the Basis of Hazard, and Sections V-G and V-L, Footnotes and References of Sections I through IV) or cells known to be infected with these agents, (iii) experiments involving the deliberate introduction of genes coding for the biosynthesis of molecules that are toxic for vertebrates (see Appendix F, Containment Conditions for Cloning of Genes Coding for the Biosynthesis of Molecules Toxic for Vertebrates), and (iv) whole plants regenerated from plant cells and tissue cultures are covered by the exemption provided they remain axenic cultures even though they differentiate into embryonic tissue and regenerate into plantlets.
• rDNA work involving E.coli K12, S. cerevisiae, and B. subtilis host-vector systems. The following categories are not exempt from the NIH Guidelines: (i) experiments described in Section III-B which require NIH/OSP and Institutional Biosafety Committee approval before initiation, (ii) experiments involving DNA from Risk Groups 3, 4, or restricted organisms (see Appendix B, Classification of Human Etiologic Agents on the Basis of Hazard, and Sections V-G and V-L, Footnotes and References of Sections I through IV) or cells known to be infected with these agents may be conducted under containment conditions specified in Section III-D-2 with prior Institutional Biosafety Committee review and approval, (iii) large-scale experiments (e.g., more than 10 liters of culture), and (iv) experiments involving the deliberate cloning of genes coding for the biosynthesis of molecules toxic for vertebrates (see Appendix F, Containment Conditions for Cloning of Genes Coding for the Biosynthesis of Molecules Toxic for Vertebrates).
• Those synthetic nucleic acids that: (1) can neither replicate nor generate nucleic acids that can replicate in any living cell (e.g., oligonucleotides or other synthetic nucleic acids that do not contain an origin of replication or contain elements known to interact with either DNA or RNA polymerase), and (2) are not designed to integrate into DNA, and (3) do not produce a toxin that is lethal for vertebrates at an LD50 of less than 100 nanograms per kilogram body weight. If a synthetic nucleic acid is deliberately transferred into one or more human research participants and meets the criteria of Section III-C of the NIH Guidelines, it is not exempt under this Section.
• Other types of experiment that fall under Section-III-F of the NIH Guidelines
• See the FAQs on the NIH OSP website for more details

I am conducting human gene transfer experiments, how do I register with the IBC?

Experiments involving deliberate transfer of rDNA into human subjects must also be reviewed by the NIH Office of Science Policy (Appendix M of the NIH Guidelines) and the University of Utah IRB. Details can be found here. The IRB will not approve gene transfer protocols until the IBC has approved the protocol. OEHS staff will advise the IRB staff on the status of IBC review and the BSO or Associate BSO will participate in IRB reviews. Human gene transfer studies require the submission of the following.

1. A completed and signed copy of the Human Gene Transfer registration form.
2. A complete packet of documentation to be submitted to NIH OSP as described in Appendix M-I-A of the NIH Guidelines. If the University of Utah is not the lead/initial site please include copies of documentation from the lead site on the determination for the requirement for RAC review.
3. Copies of correspondence from the NIH OSP and/or RAC.
4. Training records for personnel involved in the study
5. If the University of Utah is the lead/initial site, detailed responses to the questions; (i) Does the protocol use a new vector, genetic material, or delivery methodology that represents a first-in-human experience, thus presenting an unknown risk? (ii) Does the protocol rely on preclinical safety data that were obtained using a new preclinical model system of unknown and unconfirmed value? and (iii) Is the proposed vector, gene construct, or method of delivery associated with possible toxicities that are not widely known and that may render it difficult for oversight bodies to evaluate the protocol rigorously?
6. Vector Maps.
7. Copies of the Informed Consent Documents. The IBC is tasked with reviewing the description of the risks to ensure that they are adequately communicated to the participants.
8. For BSL2 or higher protocols “Caregiver Information Sheets” should be created. These should be used to communicate the risks of the agent to patient caregivers (such as family members) and should include descriptions of how to treat wounds, dispose of dressings, clean up spills of bodily fluids, signs and symptoms of exposure, and information on who to contact if an exposure may have occurred.
9. One copy of any portions of FDA, IND correspondence that discussed safety issues and/or agent description information attached.
10. Investigator’s brochure.

At the end of the human gene transfer study, submit the Human Gene Transfer Closure Notification Form to the Biosafety Office.

I work with Biological Toxins. Do I need to register with the Institutional Biosafety Committee?

Likely yes.

Biological toxins are toxic substances produced by bacteria, fungi, protozoa, insects, animals, or plants that have the capability of causing harmful effects when inhaled, ingested, injected or absorbed. They may be transmitted via surface contact with contaminated object(s) and subsequently spread to mucus membranes (eyes, nose, and mouth) and/or to open sores on skin. Some biological toxins can be absorbed through intact skin, especially if solubilized in substances such as dimethyl sulfoxide (DMSO). Accidental needle-stick is a mode of transmission within research laboratories. Accidental ingestion of contaminated materials and inhalation are other routes of transmission. While they cannot replicate and are not infectious they can be extremely hazardous, even in minute quantities. The health effects of exposure can vary greatly depending on the toxin, the amount, and the route of exposure, ranging from minor (skin or eye irritation, headache, nausea) to severe (respiratory distress, muscle weakness, seizures, death).

“The University of Utah Institutional Biosafety Committee (IBC) reviews registrations for work with, possession of, use of, and transfer of acute biological toxins (mammalian LD50 <100 µg/kg body weight) or toxins that fall under the Federal Select Agent Guidelines, as well as the organisms, both natural and recombinant, which produce these toxins

The following is a list of toxins that are required to be registered with the IBC.  However, the list is not comprehensive and principal investigators can confirm that toxins they propose to work with do not require IBC registration (LD50 >100 µg/kg body weight and not on Select Agent list) by contacting the Biosafety Office (biosafety@ehs.utah.edu or 801-581-6590).”

Abrin
Aflatoxin
Bacillus anthracis edema factor
Bacillus anthracis lethal toxin
Botulinum neurotoxins
Brevetoxin
Cholera toxin
Clostridium difficile toxin
Clostridium perfringens toxins
Conotoxins
Dendrotoxin (DTX)
Diacetoxyscirpenol (DAS)
Diphtheria toxin
Domoic acid
Pertussis toxin
Pfeisteria spp. toxin(s)
Ricin
Saxitoxin
Shiga-like ribosome inactivating proteins
Shigatoxin
Staphylococcal enterotoxins
T-2 toxin
Tetanus toxin
Tetrodotoxin (TTX)

PIs or their delegates working with acute toxins must complete the General Laboratory Setup and Biological Registration Wizards. Currently there is not a Toxin registration form. Please complete the Toxin Registration form that can be downloaded below and add as a “Document” in BioRAFT.

Click here to access the University of Utah BioRAFT page.

Biological Toxins Registration

The laboratory will be expected to maintain detailed standard operating procedures for work with toxins: the IBC has developed a template that can be adapted for this purpose. Alternatively labs can adapt their existing chemical hygiene plan (University of Utah template can be found here)

I work with human pathogens. How do I find information to help conduct a risk assessment?

Laboratories that use Risk Group (RG) 2 or higher biological agents (see ABSA Guidelines for risk group designations) are required to register with the Institutional Biosafety Committee (IBC) prior to commencing work with the agent. Note agents that are RG2 or higher in animals but RG1 in humans need to registered only if the work involves exposure of susceptible animals to the agent. For example, work with murine cytomegalovirus in mice requires IBC registration.

The Public Health Agency of Canada has generated a series of Pathogen Safety Data Sheets (PSDSs), which are technical documents that describe the hazardous properties of human pathogens and recommendations for work involving these agents in a laboratory setting, including appropriate disinfection protocols and personal protective equipment. These PSDSs can be found here.

The University of Utah IBC has developed a Fact Sheet for work with Zika virus, ZIKV.  This includes information on the risks associated with working with this virus and precautions that should be taken while working in a laboratory setting.  This can be found here.

What are the requirements for laboratories working with blood, human cell lines or other potentially infectious materials?

Occupational Safety & Health Administration (OSHA) Bloodborne Pathogen Standard, 29 CFR 1910.1030), specifies practices to limit occupational exposure to blood and other potentially infectious materials, including human and non-human primate cell lines since exposure could result in transmission of bloodborne pathogens, which could lead to disease and/or death. This protocol is mandatory for all University of Utah employees who could reasonably be anticipated, as the result of performing their job duties, to contact blood or other potentially infectious materials. Where exposure to human blood may occur, adherence to the controls, decontamination and disposal sections of this policy is mandatory for students and visitors.

PIs and Supervisors must ensure that the procedures of this policy are followed. This includes maintaining an Exposure Control Plan for their lab or work area, making it available to the workers, enforcing compliance with the plan, ensuring new employees are trained and vaccinations offered, performing follow-ups on incident exposures and providing personal protective equipment as needed. Training must be repeated annually.

PIs or their delegates working with blood, human cell lines or other potentially infectious material must complete the General Laboratory Setup and Biological Registration Wizards.

Click here to access the University of Utah BioRAFT page.

I generate animal models. Do I need to register with the Institutional Biosafety Committee?

Possibly Yes.

The creation, generation, breeding and propagation of transgenic animals are covered under Section III-D-4 of the NIH Guidelines. These activities are not exempt from the NIH Guidelines and must be reviewed by the IBC. This includes models that involve the introduction of DNA into the germline. Knock-out (gene silencing, gene ablation, etc.) models may be exempt from IBC review if the method used to generate the model does not leave any “new” genetic material or any markers behind in the genome after the procedure. If the recombinant or synthetic nucleic acid molecules that are used to create the knock-out are permanently inserted into the genome the experiment must be reviewed by the IBC. Models that have mutations or genetic modifications that are the result of natural variation, chemical mutagenesis or radiation exposure, and that have not had any molecular manipulation, are not typically reviewed by the IBC.

Follow these instructions only for experiments involving the generation of transgenic animals that require BSL-1 containment. For experiments that require BSL-2 containment or higher, please register the work as a Recombinant and Synthetic DNA protocol. If you are unsure what biosafety level your work falls under, please contact the Biosafety Office (biosafety@ehs.utah.edu or 801-581-6590).

Transgenic and knockout animals made in the mouse core facility will be registered on an annual basis by the core facility. Thus, if your transgenic animals are made by the mouse core facility, you do not need to register separately.

PIs or their delegates generating transgenic or recombinant animals must complete the General Laboratory Setup and Biological Registration Wizards.

Click here to access the University of Utah BioRAFT page.

I inject animals with human or non-human primate cells or tumors. Do I need to register with the Institutional Biosafety Committee?

Yes. There is potential risk of exposure while working with primary and commercially available cell lines. Cell lines may carry unknown agents that are potentially infectious to humans and animals. Human cell lines may carry agents directly infectious to humans. Animal cells may carry zoonotic agents. Work with human cell lines must adhere to the OSHA Bloodborne Pathogen Standard and must be conducted at Biosafety Level 2 and Animal Biosafety Level 2. Human and animal cell lines that are not well characterized or are obtained from secondary sources may introduce an infectious hazard to the laboratory. For example, the handling of nude mice inoculated with a tumor cell line unknowingly infected with lymphocytic choriomeningitis virus resulted in multiple laboratory-acquired infections.

Animals injected with well-characterized established cell lines that are from a commercial vendor (or other source) with documentation of being free of pathogens (ATCC tests for HBV, HCV, HIV, CMV, and EBV for cell lines accessioned into their inventory since 2010), may be handled at ABSL-1 after IBC approval of the risk assessment. Cell lines can also be tested: IDEX Bioresearch and Charles River offer a comprehensive pathogen service. This is still at the discretion of both the IBC and IACUC and each protocol will be risk assessed for any factors that may warrant a higher level of containment. PIs should provide documentation to support the determination that cells are pathogen-free.

PIs or their delegates injecting animals with cells or tumors must complete the General Laboratory Setup and Biological Registration Wizards.

Click here to access the University of Utah BioRAFT page.

I work with recombinant lentiviral vectors, do I need to test for Replication Competent virus?

Possibly.

Lentivirus vector stocks generated with 2nd generation packaging systems AND transfer plasmids with wild type LTRs (i.e., not self-inactivating transfer plasmid constructs) must be tested for replication-competent virus (RCV) by serial transfer in a cell line documented to be capable of propagating wild type HIV if:

1. The PI proposes to reduce biocontainment after target cell transduction,
2. The PI proposes to transduce mammalian cells capable of supporting lentiviral replication, introducing the cells into animals (e.g., xenografts) and maintain the animals at ABSL1, or
3. The PI proposes to introduce lentiviral vectors or transduced mammalian cells capable of supporting lentiviral replication into animals that could support viral replication.

Testing should be conducted on the initial preparations of viral stocks.

Lentivirus vector stocks generated with 3rd generation (4 plasmid, including self-inactivating transfer vectors) or with 2nd generation packaging systems and 3rd generation transfer vectors (SIN) are exempt from these requirements. However, if the PI proposes to introduce lentiviral vectors, or transduced mammalian cells capable of supporting lentiviral replication, into animals that could support viral replication, then the PI should provide a complete risk assessment to the IBC.

If none of these criteria apply RCV testing will not typically be required. However, the Institutional Biosafety Committee (IBC) may determine that, following the risk assessment, RCV testing is still required.

The vector stock should be tested for RCV at a limit of sensitivity of 1 infectious unit per milliliter (mL). Several sensitive RCV assays have been described, including an ELISA-based p24 Gag antigen assay, a product enhanced reverse transcriptase (PERT) assay that involves the vector’s reverse transcriptase, a PCR-based assay that detects Psi-Gag sequences from a recombination event between vector and packaging constructs, and PCR-based assays to detect the VSV-G Env used for pseudotyping. All represent acceptable methods for RCV testing. The protocol must be provided to the IBC for approval.

I will be conducting Gene Editing experiments.  What information do I need to provide the IBC?

To assist Principal Investigators with their responsibilities for ensuring that recombinant or synthetic nucleic acid research utilizing new gene-editing technologies is appropriately registered, reviewed and approved, the University of Utah Institutional Biosafety Committee (IBC) requires PIs to address risks associated with gene-editing technology, such as CRISPR/Cas9.  To assist in this review the IBC has a questionnaire, which can be accessed here, to be completed and attached to the IBC registration in BioRAFT. Note, for gene editing research involving viral vectors, potential off-target effects by the guide RNA should be identified.  This is helpful in assessing the risk of potential exposure in the event of an incident. The IBC recommends GT Scan (http://bioinformatics.csiro.au/gt-scan/), but other utilities may be used. Please identify which utility you have used and provide a list of off-target genes (with up to 3 mismatches).

What do I do if someone in my lab was exposed to recombinant or synthetic Nucleic Acids or a hazardous biological material?

Immediately wash affected areas with soap and water, or if exposure to eyes or mucous membranes occurred, immediately flush affected area with water for 10-15 minutes. 2. If the injury is life threatening call 911. If the injury is not life threatening, go directly to the Occupational Medical Clinic for medical evaluation and follow-up. 3. Complete the Employers First Report of Injury E1 Form. This form can be downloaded from the human resources website under “Forms.”

All significant research-related accidents and illnesses must be reported to the Institutional Biosafety Committee (IBC) immediately, using the template below:

Minor spills of low-risk agents not involving a breach of containment that were properly cleaned and decontaminated generally do not need to be reported. If you are uncertain whether the nature or severity of the incident warrants reporting, contact the University of Utah Biosafety Officer (BSO), at (801) 581-6950 or biosafety@ehs.utah.edu,

Adverse events in human gene transfer trials are subject to a separate reporting requirements. These are found in Appendices M-1-C-3 and M-1-C-4 of the NIH Guidelines. Serious adverse events that are unexpected and possibly associated with the gene transfer product should be reported to the NIH Office of Science Policy (OSP) within 15 calendar days of sponsor notification, unless they are fatal or life threatening, in which case they should be reported within 7 calendar days. Other serious adverse events should be reported to OSP as part of the Principal Investigator’s annual report to OSP.

Downloads:
In the event of an incident please complete an Incident Report here

What Procedures should I follow to work with Biological Toxins?

Laboratory Practices
Guidelines for working with biological toxins can be found in Appendix I of the Biosafety in Microbiological and Biomedical Laboratories. These are summarized below.

Routine operations with dilute toxin solutions are conducted using Biosafety Level 2 (BSL2) practices and these must be detailed in the IBC protocol and will be verified during the inspection by OEHS staff prior to IBC approval. BSL2 Inspection checklists can be found here. All labs working with toxins must have detailed standard operating procedures (SOPs) for work with the toxins; the IBC has developed a template that can be adapted for this purpose. All personnel working with biological toxins or accessing a toxin laboratory must be familiar with the SOPs and training in the theory and practice of the toxins to be used, with special emphasis on the nature of the hazards associated with laboratory operations and should be familiar with the signs and symptoms of toxin exposure.  This includes how to handle transfers of liquids containing toxin, where to place waste solutions and contaminated materials or equipment, and how to decontaminate work areas after routine operations, as well as after accidental spills. The worker must be reliable and sufficiently adept at all required manipulations before being provided with toxin. Laboratory work with toxins should be done only in designated rooms with controlled access and at pre-determined bench areas. When toxins are in use, the room should be clearly posted: “Toxins in Use—Authorized Personnel Only.”

Researchers working with a toxin should be vaccinated if a vaccine is available (e.g. diphtheria toxin, tetanus toxin). Routine operations with dilute toxin solutions are conducted using BSL2 practices and facilities shall be used for activities involving biological toxins. These include:

• Biohazard signs and labels must be displayed in areas and on equipment where biological toxins are used and stored. This includes, but is not limited to, laboratory entrance doors, biological safety cabinets, chemical fume hoods, refrigerators, and freezers.
• Use a biological safety cabinet (BSC) or a chemical fume hood for resuspension of biological toxins or manipulations of stock solutions of toxins that can generate aerosols, such as pipetting, harvesting, infecting cells, filling tubes/containers, and opening sealed centrifuge canisters. When using an open-fronted fume hood or BSC, workers should wear suitable laboratory PPE to protect the hands and arms, such as laboratory coats, smocks, or coveralls and disposable gloves.
• Whenever possible, use needle-free techniques to resuspend biological toxins.
• If a quantity of powder-form toxin must be weighed, then the scale must be located in a certified chemical fume hood.
• When conducting liquid transfers and other operations that pose a potential splash or droplet hazard in an open-fronted hood or BSC, workers should wear safety glasses and disposable facemask, or a face shield.
• Toxin should be removed from the hood or BSC only after the exterior of the closed primary container has been decontaminated and placed in a clean secondary container. Toxin solutions, especially concentrated stock solutions, should be transported in leak/spill-proof secondary containers.

Emphasis must be placed on evaluating and modifying experimental procedures to eliminate the possibility of inadvertent generation of toxin aerosols. Pressurized tubes or other containers holding toxins should be opened in a BSC, chemical fume hood, or other ventilated enclosure. Operations that expose toxin solutions to vacuum or pressure, for example, sterilization of toxin solutions by membrane filtration, should always be handled in this manner, and the operator should also use appropriate respiratory protection. If vacuum lines are used with a toxin, they should be protected with a HEPA filter to prevent the entry of toxins into the line.

Centrifugation of cultures or materials potentially containing toxins should only be performed using sealed, thick-walled tubes in safety centrifuge cups or sealed rotors. The outside surfaces of containers and rotors should be routinely cleaned before each use to prevent contamination that may generate an aerosol. After centrifugation, the entire rotor assembly is taken from the centrifuge to a BSC to open it and remove its tubes.

Experiments should be planned to eliminate or minimize work with dry toxin (e.g., freeze-dried preparations). Unavoidable operations with dry toxin should only be undertaken with appropriate respiratory protection and engineering controls.

Personal Protective Equipment (PPE)
Work with biological toxins shall be conducted using BSL2 PPE:

• Disposable gloves – consider the use of double gloves for enhanced protection. Ensure your gloves are compatible with any solvent your toxin may be dissolved in.
• Lab coat or back-closing disposable gown
• Eye protection (safety glasses or goggles) is recommended, but not required when working within a biosafety cabinet or a chemical fume hood.

An inventory control system should be in place to account for toxin use and disposition. If toxins are stored in the laboratory, containers should be sealed, labeled, and secured to ensure restricted access; refrigerators and other storage containers should be clearly labeled and provide contact information for trained, responsible laboratory staff.

Decontamination and Spills
Toxin stability varies considerably outside of physiological conditions depending upon the temperature, pH, ionic strength, availability of co-factors and other characteristics of the surrounding matrix. Literature values for dry heat inactivation of toxins can be misleading due to variations in experimental conditions, matrix composition, and experimental criteria for assessing toxin activity. Moreover, inactivation is not always a linear function of heating time; some protein toxins possess a capacity to re-fold and partially reverse inactivation caused by heating. In addition, the conditions for denaturizing toxins in aqueous solutions are not necessarily applicable for inactivating dry, powdered toxin preparations. Inactivation procedures should not be assumed to be 100% effective without validation using specific toxin bioassays.

Many toxins are susceptible to inactivation with dilute sodium hydroxide (NaOH) at concentrations of 0.1-0.25N, and/or sodium hypochlorite (NaOCl) bleach solutions at concentrations of 0.1-0.5% (w/v). Use freshly prepared bleach solutions for decontamination; undiluted, commercially available bleach solutions typically contain 3-6% (w/v) NaOCl: a 1:10 dilution of bleach gives an appropriate concentration.

Exposures
Antitoxins are available for some biological toxins and immediate medical “first-aid” interventions may help prevent or lessen the severity of the reaction. If you know or suspect a biological toxin exposure.

• Irrigate the site of exposure
  • If exposure was by a needle stick or other route which breaks the skin, wash with soap and water for 5-15 minutes and cover with a bandage.
  • If exposure was by splash to eyes or mucous membranes, irrigate thoroughly for 15 minutes at an appropriate eyewash station.
• Report to your Laboratory Supervisor and the Biosafety Officer IMMEDIATELY. Seek medical attention as outlined in your laboratory-specific safety manual.

Export Controlled Toxins

Several biological toxins are restricted for export by the U.S. Department of Commerce and require an export license prior to any shipment out of the U.S. Many of the toxins on the export control list do not meet the definition of an acute toxin and do not require registration with the IBC but the Principal Investigator is responsible for ensuring that they are in compliance with the export control laws and regulations.  If you have questions about export controls, please visit the Office of Sponsored Projects webpage or contact OSP’s Export Control Officer, Todd Nilsen.

What are Select Agents?

“Biological Select Agents or Toxins” (BSATs) — or simply Select Agents for short — are bio-agents which since 1997[1] have been declared by the U.S. Department of Health and Human Services (HHS) or by the U.S. Department of Agriculture (USDA) to have the “potential to pose a severe threat to public health and safety”. The agents are divided into (1) HHS select agents and toxins affecting humans; (2) USDA select agents and toxins affecting agriculture; and (3) Overlap select agents and toxins affecting both.

The Public Health Security and Bioterrorism Preparedness and Response Act of 2002, Subtitle A of Public Law 107–188 requires the Department of Health and Human Services (HHS) to establish and regulate a list of biological agents and toxins that have the potential to pose a severe threat to public health and safety. The Agricultural Bioterrorism Protection Act of 2002 requires the United States Department of Agriculture (USDA) to establish and regulate a list of biological agents that have the potential to pose a severe threat to animal health and safety, plant health and safety, or to the safety of animal or plant products (Select Agents). CDC and APHIS share responsibility for some agents because they potentially threaten both humans and animals (overlap agents). The laws require HHS and USDA to review and republish the lists of Select Agents and toxins on at least a biennial basis. Please visit the CDC/USDA website on select agents for more information.

Links to the list of Select Agents and toxins and information about additions or deletions that have been made to the list based on recommendations made from the biennial review or advances in research can be found below.
Select Agents and Toxins List
Select Agents and Toxins Exclusions
Select Agents and Toxins Restricted Experiments
Permissible Toxin Amounts

What research needs to be registered as Dual Use Research of Concern (DURC)?

Research that uses one or more of the agents or toxins listed in section (i) below, and produces, aims to produce, or can be reasonably anticipated to produce one or more of the effects listed in section (ii) below, must be evaluated for DURC potential.

1. Agents:
   1. Avian influenza virus (highly pathogenic)
   2. Bacillus anthracis
   3. Botulinum neurotoxin: For the purposes of this Policy, there are no exempt quantities of botulinum neurotoxin. Research involving any quantity of botulinum neurotoxin should be evaluated for DURC potential.
   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
2. Categories of experiments:
   1. Enhances the harmful consequences of the agent or toxin
   2. Disrupts immunity or the effectiveness of an immunization against the agent or toxin without clinical and/or agricultural justification
   3. Confers to the 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 agent or toxin
   5. Alters the host range or tropism of the agent or toxin
   6. Enhances the susceptibility of a host population to the agent or toxin
   7. Generates or reconstitutes an eradicated or extinct agent or toxin listed above

Experiments that fall under the potential Dual Use Research of Concern (DURC) guidelines must be registered with the IBC, and be reviewed by the University of Utah Institutional Review Entity (IRE) for risk assessment and mitigation, as necessary. The PI should contact the BSO or Associate BSO immediately if experiments covered by these guidelines are considered.

What training should my laboratory staff complete?

For laboratories where staff have occupational exposure to blood and other potentially infectious materials training in bloodborne pathogens is required. This is required at the time of hire and must be repeated annually.

For laboratories working with animals at Biosafety Level 2 (ABSL2), attendance at an ABSL2 training session is required annually. These are organized through the University of Utah Research Administration Training Series (RATS). Details can be found here: https://education.research.utah.edu/

All research personnel working with recombinant vectors where the parental virus is classified as Risk Group 2 or higher, including lentiviral, retroviral, and adenoviral vectors, as well as recombinant adeno-associated virus (AAV) vectors requiring BSL2 containment, are required to take the following:

1. Either:
   1. Bloodborne Pathogen/BSL2/ABSL2 training offered by the University of Utah Research Administration Training Series: information and registration can be found at https://education.research.utah.edu/, OR
   2. Bloodborne Pathogen training offered by the University of Utah Research Administration Training Series: information and registration can be found at https://education.research.utah.edu/, AND the vector specific online training kindly available from the University of Cincinnati Office of Research Integrity: http://researchcompliance.uc.edu/Biosafety/Training/ViralVectorWebtraining.aspx (Module 1 plus relevant vector modules), OR
   3. Bloodborne Pathogen offered by the University of Utah Research Administration Training Series: information and registration can be found at https://education.research.utah.edu/, AND the Recombinant Viral Vector training offered at the Huntsman Cancer Institute: contact the Biosafety Office for details on registration.AND
2. Laboratory-Specific Training

Research personnel working with recombinant AAV vectors requiring BSL1 containment are requested to complete the AAV specific online training kindly available from the University of Cincinnati Office of Research Integrity: http://researchcompliance.uc.edu/Biosafety/Training/ViralVectorWebtraining.aspx (Modules 1 and 5). Please note that if you are working with human or non-human primate cell lines you are required to take Bloodborne pathogen training, as described above.

For individuals taking the University of Cincinnati Office of Research Integrity training, please complete the Online Training Assurance form and send to the Biosafety Office at biosafety@ehs.utah.edu or attach in the documents section of your laboratory BioRAFT registration. Copies of laboratory-specific training records should be submitted in the same manner.

Research personnel working with non-recombinant Risk Group 2 pathogens are required to take the Bloodborne Pathogen/BSL2/ABSL2 training offered by the University of Utah Research Administration Training Series, as described above, and Laboratory-specific training.

All training should be repeated annually.

I share laboratory space with other PIs. What are our responsibilities for cross-training staff?

All BSL2 (or higher) lab spaces that are shared by 2 or more PIs using different biohazardous agents, who are not co-PIs or co-investigators on an approved IBC protocol(s), must conduct a joint “Biohazards Awareness and Training” session for all the personnel working in the space.

The training must be presented by each PI or delegate, outlining the agents, and risks associated with the agents used for their specific research, covering at a minimum the topics listed on the following page. If you need assistance with these training sessions please contact the Biosafety Office at 801-581-9325 or ehs@biosafety.utah.edu. Training should be provided for all new employees and repeated at least annually for all staff.

All attendees at the training session(s) must sign a sign-in sheet that identifies the trainer(s) and agent(s). All attendees at the training session(s) must sign a sign-in sheet that identifies the trainer(s) and agent(s) discussed: a template is provided here. Please edit the template as applicable. Submit the sign-in sheet to ehs@biosafety.utah.edu.

Do I need to have a Laboratory Safety Manual or Exposure Control Plan?

Yes. If you are working at BSL2 or higher, a Laboratory Biosafety Manual, as described in the CDC Biosafety in Microbiological and Biomedical Laboratories (BMBL) 5th Edition, is required. This should include laboratory-specific Standard Operating Procedures (SOPs) for all experiments involving biological agents, risk assessments for the agent, descriptions of personal protective equipment, procedures for using and decontaminating equipment, spill procedures and post-exposure plans, waste disposal and emergency equipment and practices. In addition, the PI should develop SOPs describing experimental procedures for working with r-sNA agents and human or animal pathogens.

• The manual can be used for laboratory-specific training.
• The University of Utah maintains a Biosafety manual which can be used as a source of information and can be downloaded here.
• The IBC has developed Biosafety Manual/SOP templates for BSL2 and BSL2-enhanced laboratories that may be used by PIs as the basis for laboratory-specific SOPs. These address most areas of review by the IBC but should be customized to reflect the work being performed in the laboratory and the facilities available. BSL1 laboratories can also use these as templates, modified as appropriate.
• The manual must be reviewed annually by all lab personnel and updated as appropriate.
• The manual must be readily available to all laboratory personnel and EHS personnel during laboratory inspections/audits.

Laboratories working with bloodborne pathogens must develop an Exposure Control Plan: a template is available here , as required under OSHA Standard 1910.1030.  The plan must include descriptions of:

1. Universal precautions shall be observed to prevent contact with blood or other potentially infectious materials. Under circumstances in which differentiation between body fluid types is difficult or impossible, all body fluids shall be considered potentially infectious materials.

2. Engineering and work practice controls that will be used to eliminate or minimize employee exposure.

3. The type and use of personal protective equipment. When there is occupational exposure, the employer shall provide, at no cost to the employee, appropriate personal protective equipment such as, but not limited to, gloves, gowns, laboratory coats, face shields or masks and eye protection, and mouthpieces, resuscitation bags, pocket masks, or other ventilation devices. Personal protective equipment will be considered appropriate only if it does not permit blood or other potentially infectious materials to pass through to or reach the employee’s work clothes, street clothes, undergarments, skin, eyes, mouth, or other mucous membranes under normal conditions of use and for the duration of time which the protective equipment will be used.

4. Spill cleanup procedures: Contaminated work surfaces shall be decontaminated with an appropriate disinfectant after completion of procedures; immediately or as soon as feasible when surfaces are overtly contaminated or after any spill of blood or other potentially infectious materials; and at the end of the work shift if the surface may have become contaminated since the last cleaning.

5. Disposal of sharps. Contaminated needles and other contaminated sharps shall not be bent, recapped, or removed. Shearing or breaking of contaminated needles is prohibited. Sharps must be disposed of in a hard sided, leak-proof, sealable, and puncture resistant container.

6. Eating, drinking, smoking, applying cosmetics or lip balm, and handling contact lenses are prohibited in work areas where there is a reasonable likelihood of occupational exposure.

7. Food and drink shall not be kept in refrigerators, freezers, shelves, cabinets or on countertops or benchtops where blood or other potentially infectious materials are present. Food and drinks are not permitted in University of Utah laboratories.

8. Mouth pipetting/suctioning of blood or other potentially infectious materials is prohibited.

9. Specimens of blood or other potentially infectious materials shall be placed in a container which prevents leakage during collection, handling, processing, storage, transport, or shipping. The container must be labeled with the Universal Biohazard Symbol.

10. The laboratory must be maintained in a clean and sanitary condition. The employer shall determine and implement an appropriate written schedule for cleaning and method of decontamination based upon the location within the facility, type of surface to be cleaned, type of soil present, and tasks or procedures being performed in the area.

11. Post exposure procedures. Following a report of an exposure incident, the employer shall make immediately available to the exposed employee a confidential medical evaluation and follow-up.

12. Injury Log: The employer shall establish and maintain a sharps injury log for the recording of percutaneous injuries from contaminated sharps. The information in the sharps injury log shall be recorded and maintained in such manner as to protect the confidentiality of the injured employee.

The Exposure Control Plan must be reviewed and updated at least annually and whenever necessary to reflect new or modified tasks and procedures which affect occupational exposure and to reflect new or revised employee positions with occupational exposure. The review and update of such plans shall include consideration and implementation of appropriate commercially available and effective safer medical devices designed to eliminate or minimize occupational exposure

What is an Exposure Control Plan and Do I need one?

Designed to eliminate employee exposure to bloodborne pathogens, laboratories working with such as blood, tissue, potentially infected materials and human and non-human primate cell lines, must develop and implement an Exposure Control Plan. . All human blood and other potentially infectious materials (OPIM) are considered to be infectious for Human Immunodeficiency Virus (HIV), Hepatitis B virus (HBV), and other BBP, and will be treated as if infectious, i.e. with universal precautions. Since animal blood is not readily distinguished from human blood by appearance, this document’s guidance for handling and disposal of human blood and sharps is recommended for all blood. A template for a Laboratory Exposure Plan can be downloaded here: Laboratory Exposure Control Plan (DOC, 112KB)

I work with recombinant viral vectors. How do I find information to help conduct a risk assessment?

The IBC has generated guidelines to help investigators with their risk assessment when developing projects involving recombinant viral vectors. This provides information on many of the recombinant viral vectors used in laboratories to express foreign genes in cultured cells and animal models. Investigators should use these guidelines as part of their risk assessment when planning experiments with these vectors and preparing applications to the Institutional Biosafety Committee (IBC). Note the listed containment levels are the minimum that should be employed with these vectors: some experiments, such as the expression of toxins or oncogenes, may require higher levels of containment. The appropriateness of the containment should be considered as part of the investigator’s risk assessment and will be reviewed by the IBC.

What is the deadline for submitting protocols for IBC review?

Registrations will be reviewed following the schedule outlined below. The IBC recommends that the PI or a representative attend the IBC meeting. For meeting time and location, contact the Occupational and Environmental Health and Safety at 801-581-6590.

Registrations received by the submission date will be reviewed in that review cycle. All registration forms submitted after the due date will be reviewed during the following review cycle. Approval/disapproval or requests for additional information may be sent in writing to the Principal Investigator during the review cycle.

Review Month	Submit By	Review Date
January	January 2, 2019	January 17, 2019
February	January 31, 2019	February 21, 2019
March	February 28, 2019	March 18, 2019
April	March 28, 2019	April 15, 2019
May	May 2, 2019	May 20, 2019
June	May 30, 2019	June 17, 2019
July	June 27, 2019	July 15, 2019
August	August 1, 2019	August 19, 2019
September	August 29, 2019	September 16, 2019
October	October 3, 2019	October 21, 2019
November	November 4, 2019	November 25, 2019
December	November 28, 2019	December 16, 2019

 

What are the different types of Biological Safety Cabinets?

Biological Safety Cabinets or BSCs are designed to provide personnel, environmental or product protection when appropriate practices and procedures are followed handling biological materials. Three kinds of biological safety cabinets (designated as Class I, II and III) have been developed to meet varying research and clinical needs, and provide various levels of protection. A complete description of the BSC can be found in the 5th edition BMBL: http://www.cdc.gov/biosafety/publications/bmbl5/. Below is a summary description of the different classes of BSC.

Class I Biological Safety Cabinets

The Class I BSC provides protection for personnel and the environment, but NO product protection. It is similar in air movement to a chemical fume hood, but has a HEPA filter in the exhaust system to protect the environment.

In the Class I BSC, unfiltered room air is drawn across the work surface. Personnel protection is provided by this inward airflow as long as a minimum velocity is maintained through the front opening. The use of the Class I BSC has declined due to lack of product protection but, in many cases, Class I BSCs are used specifically to enclose equipment (e.g., centrifuges, harvesting equipment or small fermenters), or procedures with potential to generate aerosols (e.g. cage dumping, culture aeration or tissue homogenization). Some Class I models used for animal cage changing are designed to allow recirculation of air into the room after HEPA filtration and may require more frequent filter replacement due to filter loading and odor from organic materials captured on the filter. The re-circulating Class I BSC should be annually certified for sufficient airflow and filter integrity.

Class II Biological Safety Cabinets

The Class II BSC provides protection for personnel, the environment, AND the product. Class II BSCs are partial barrier systems that rely on a continuous unidirectional air movement at a fixed velocity along parallel lines (“laminar flow”) to provide containment. If the air curtain is disrupted (e.g., movement of materials in and out of a cabinet, rapid or sweeping movement of the arms) the potential for contaminant release into the laboratory work environment is increased as is the risk of product contamination. All Class II cabinets are designed for work involving microorganisms assigned to Biosafety Levels 1, 2 and 3. Class II BSCs provide the microbe-free work environment necessary for cell culture propagation and also may be used for the formulation of nonvolatile anti-neoplastic or chemotherapeutic drugs.

There are 4 types of Class II BSC (Types A1, A2, B1 and B2) each with unique characteristics, however, in all four, the airflow is drawn into the front grille of the cabinet, providing personnel protection. In addition, the downward laminar flow of HEPA-filtered air provides product protection by minimizing the chance of cross-contamination across the work surface of the cabinet. Because cabinet exhaust air is passed through a certified HEPA filter, it is particulate-free (environmental protection), and may be recirculated to the laboratory or discharged from the building via a canopy connection (Type A1 and A2). Exhaust air from Types B1 and B2 BSC must be discharged to the outdoors via a fixed or canopy connection. HEPA filters are effective at trapping particulates and thus infectious agents but do not capture volatile chemicals or gases. Only Types A2, B1 and B2 BSCs exhausting to the outside should be used when working with volatile, toxic chemicals, but amounts must be limited. (See below for more information on Working with Chemicals in BSC).

Class III Biological Safety Cabinets

The Class III BSC is designed for work with highly infectious microbiological agents and for the conduct of hazardous operations. It provides maximum protection from the environment and the worker. The Class III BSC is a gas tight enclosure with a non-opening view window. Long, heavy-duty rubber gloves are attached in a gas-tight manner to ports in the cabinet to allow direct manipulation of the materials isolated inside. Access for passage of materials into the cabinet is through a dunk tank, that is accessible through the cabinet floor, or double-door pass-through box (e.g., an autoclave) that can be decontaminated between uses. Both supply and exhaust are HEPA filtered on a Class III BSC. Exhaust air must pass through two HEPA filters, or a HEPA filter and an air incinerator, before discharge directly to the outdoors. Class III BSC are not exhausted through the general laboratory exhaust system.

Whom do I contact if I have a problem with my BSC or it is due for certification?

Contact OEHS to discuss options for repairs or certifications of BSCs (801-581-6590).  All repairs and certifications must be performed by an NSF-certified technician and the University of Utah has a contract with ENV Services.

No work on the BSC may be performed by University of Utah Facilities staff, with the exception of work/servicing on the exterior of the cabinet, such as connection of the cabinet to the vacuum system.

Can I relocate my BSC within the lab or move it to another lab space?

Cabinets must be fully decontaminated before they are moved, and then re-certified after they are in place. Please contact OEHS (801-581-6590) to assist with scheduling the decontamination and re-certification by our contracted vendor ENV Services. Labs are also responsible for arranging the movement of cabinets with University Facilities Management. If the cabinet is destined for surplus, please remove biohazard stickers after the cabinet has been fully decontaminated by ENV Services and contact surplus for pick-up.

What is the difference between the surface decontamination and the decontamination service by ENV Services?

Upon completion of work, lab staff should decontaminate the interior surfaces of the cabinet by wiping down all accessible surfaces with an appropriate disinfectant. Additionally, monthly or as needed, lab staff should remove the cabinet work surface to access and surface disinfect the lower plenum (a.k.a. drain pan). These surface decontaminations are needed for proper maintenance of your cabinet and are the responsibility of lab staff.

The decontamination service provided by ENV involves decontaminating the entire interior of the cabinet (including the blower/motor and HEPA filters) by fumigation with vaporized hydrogen peroxide. This decontamination service is required prior to repairs in which service personnel must access potentially contaminated areas of the cabinet (i.e. HEPA filter or motor/blower replacement). Additionally, this type of decontamination is required prior to cabinet relocation to ensure biohazardous material is not released during transportation. This service does not include cleaning of the cabinet interior surfaces. As indicated above, lab staff are responsible for routine surface cleaning/disinfection.

How can I hook up gas/vacuum lines to my cabinet?

Facilities Management should be contacted to hook up gas or vacuum lines to your cabinet. Remember—Bunsen burners are not allowed in BSCs.  If heating is required one of the options described below.

Safe alternatives to Bunsen burners:

Early microbiologists had to rely on open flames to ensure sterility. However, with the advancement of modern technology, including the introduction of the biosafety cabinet, the use of an open flame is almost always no longer necessary. Alternative options include:

• Use disposable sterile loops and sterile lab supplies. This eliminates the need to use open flames for sterilizing.
• Autoclave utensils and equipment prior to use. Place loops, spreaders, needles, forceps, scalpels and other tools in autoclavable plastic or wrap in autoclavable foil.
• Use a Bacti-Cinerator to sterilize loops and needles safely and conveniently while preventing infectious spatter and cross-contamination.
• The Electrical Bunsen Burner combines the efficiency of a gas burner with the safety and control of an electric heater. It is ideal for sterilizing inoculating needles and loops, and for heating small flasks, test tubes, and beakers.
• The Glass Bead Sterilizer provides a safe, effective, and convenient method for sterilizing small instruments without using flames, gases, or chemicals.
• If it is deemed absolutely necessary for the experiment being done, use a pilotless burner or touch-plate microburner (Touch-O-Matic) to provide a flame on demand.

The use of Bunsen burners inside of a biological safety cabinet is not recommended because it:

• Disrupts airflow, compromising the protection of the worker and the product. The Class II BSC maintains product protection through delivery of laminar flow (air volumes traveling in a single direction at a constant speed – without turbulence) down over the work area of the cabinet. The heating of air from the Bunsen burner causes up‐flow of air that mixes with the down flowing airstreams to produce turbulence and recirculation within the working area. The notion of laminar flow may be completely destroyed and any aerosols generated beneath the burner may be carried to other parts of the cabinet, jeopardizing the product and personnel working within the cabinet.
• Causes excessive heat build‐up within the cabinet. As most Class II BSCs recirculate the majority of the air within the cabinet, heat from the Bunsen burner builds up over time. The excessive heat can inactivate and degrade components in media such as vitamins, amino acids and growth factors, possibly below the threshold for finicky cell lines. In addition, the excessive heat may make it an uncomfortable environment for the worker, leading to errors and mistakes.
• May damage the HEPA filter or melt the adhesive holding the filter together, compromising the cabinet’s integrity.  An open flame has the capacity for melting the bonding agent that holds the HEPA filter media to its frame. This destroys the HEPA filters effectiveness, leading to loss of containment in the positive pressure plenum. ENV will charge $250 for decontamination of the cabinet, $250‐$1000 for the filter, and $145 for recertification each time the HEPA filter needs to be replaced.
• Presents a potential fire or explosion within the cabinet. The cabinets are not constructed to be explosion proof. If the flame was to go out, there was a leak, or the valve was not shut off completely, flammable gas would be introduced to the cabinet at a steady rate. In the case of a Class II A2, where 70% of the air in the BSC is recirculated, concentrations of the flammable gas could reach explosive potential and pose a serious risk to not only the cabinet, but to the user and the laboratory it is occupied in. Electrical components like the fan motor, lights, or electrical outlets could ignite a flash fire with a spark in this volatile environment. Manufacturers often post their cabinets with warning labels stating that flammable materials should not be used in the cabinet.
• Inactivates manufacturer’s warranties on the cabinet. Biological safety cabinet manufactures are opposed to the practice and will assume no liability in the event of fire, explosion or worker exposure due to the use of a flammable gas in their cabinet.
• Automatically voids UL approval. Underwriters Laboratories Inc. (UL) is an OSHA approved independent product safety certification organization that develops standards and test procedures for products, materials, components, assemblies, tools and equipment, chiefly dealing with product safety.  The use of a Bunsen burner in the cabinet will void UL approval of that piece of equipment.
• Requires hook‐up of central gas source. Some laboratories may not be fitted with gas lines and will require costly room renovations for retrofitting. Facilities will also need to install plumbing from the house lines to the cabinet. If the cabinet needs to be moved, this will incur additional costs from Facilities. In addition, gas connectors are generally not supplied with new biosafety cabinets without customer insistence and at an additional cost.

 

Quotes on Open Flames in BSCs:

NIH/CDC: National Institutes of Health and the Centers for Disease Control and Prevention (Appendix A of the BMBL): “Open flames are not required in the near microbe‐free environment of a biological safety cabinet. On an open bench, flaming the neck of a culture vessel will create an upward air current which prevents microorganisms from falling into the tube or flask. An open flame in a BSC, however, creates turbulence which disrupts the pattern of HEPA‐filtered air supplied to the work surface. When deemed absolutely necessary, touchplate microburners equipped with a pilot light to provide a flame on demand may be used. Internal cabinet air disturbance and heat buildup will be minimized. The burner must be turned off when work is completed. Small electric “furnaces” are available for decontaminating bacteriological loops and needles and are preferable to an open flame inside the BSC. Disposable or recyclable sterile loops can also be used.”

WHO: World Health Organization’s Laboratory Biosafety Manual: “Open flames should be avoided in the near microbe‐free environment created inside the BSC. They disrupt the airflow patterns and can be dangerous when volatile, flammable substances are also used. To sterilize bacteriological loops, micro‐burners or electric “furnaces” are available and are preferable to open flames.”

Public Health Agency of Canada; The Laboratory Biosafety Guidelines: “The provision of natural gas to BSC’s is not recommended. Open flames in the BSC create turbulence, disrupt airflow patterns and can damage the HEPA filter. When suitable alternatives (e.g., disposable sterile loops, micro‐incinerators) are not possible, touch‐plate micro‐burners that have a pilot light to provide a flame on demand may be used.”

NSF/ANSI Standard 49 – 2009 published by NSF International, Annex G; Section G.3.3.1: “Service valves allow inert gases, air, or vacuum lines to be plumbed into the BSC. Although many users connect gas to a service valve in the cabinet, this practice should be avoided if possible, because open flames in a Class II BSC disrupts the airflow, and there is the possibility of a buildup of flammable gas in BSC’s that recirculate their air.”

Are Laminar Flow Cabinets or Hoods (Clean Bench) the same as Biological Safety Cabinet?

No.

Horizontal Laminar Flow: Horizontal laminar flow “clean benches” are NOT Biological Safety Cabinets (BSCs). These pieces of equipment discharge HEPA-filtered air from the back of the cabinet across the work surface and toward the user. These devices only provide product protection. They can be used for certain clean activities, such as the dust-free assembly of sterile equipment or electronic devices. Clean benches should never be used when handling cell culture materials or drug formulations, or when manipulating potentially infectious materials. The worker will be exposed to the materials being manipulated on the clean bench potentially resulting in hypersensitivity, toxicity or infection depending on the materials being handled. Horizontal airflow “clean benches” must never be used as a substitute for a biological safety cabinet.

Vertical Laminar Flow: Vertical laminar flow clean benches also are NOT BSCs. They may be useful, for example, in hospital pharmacies when a clean area is needed for preparation of intravenous solutions. While these units generally have a sash, the air is usually discharged into the room under the sash, resulting in the same potential problems presented by the horizontal laminar flow clean benches. These benches should never be used for the manipulation of potentially infectious or toxic materials.

How do I Work Safely in a Biological Safety Cabinet?

Adapted from the University of Vermont EHS website

Biosafety cabinets (BSC) afford the best protection and are more effective when maintained and used properly. To accomplish this goal, it is necessary that:

• The BSC must be certified upon installation, after it is moved, after repairs and annually thereafter. Please contact OEHS at (801) 581-6950 to arrange certification or repairs to the BSC.
• Researchers learn must how to work in and take care of a biological safety cabinet, as summarized below. OEHS provides training as part of their Bloodborne Pathogen and BSL2/ABSL2 training series. The Yale University Department of Environmental Health and Safety has prepared a useful instructional video that can be accessed here.

General Considerations

1. Minimize the storage of materials in and around the BSC. Avoid storing items on top of the cabinet since it could damage the HEPA filters.
2. Try to avoid blocking the grill at the back of the cabinet as much as possible to avoid disruptions to the airflow. Keep materials to the side and/or with a gap to the vents.  Never place materials on the front grill.
3. Never operate the blower with the sash closed – an excessive air inflow could damage the HEPA filters.
4. The use of open flames/Bunsen burners in the BSC is not permitted at the University of Utah. Open flames cause a disruption in the airflow, can cause damage to the HEPA filter, and create a fire hazard.
5. When using a vacuum system to aspirate liquids, attach a flask containing a suitable disinfectant, such as bleach or Wescodyne, to collect and disinfect the aspirated materials, and an in-line HEPA filter, placed between the flask and vacuum valve, to protect the vacuum system from aerosolized microorganisms.
6. Turn the BSC off after use – do not leave it running overnight.
7. If an UV light is present, use only as needed – never have the UV on when somebody is in the room. However, please note that there are considerable data showing that these UV lights have minimal efficacy. Click here for an interesting article talking about this.
8. Clean any spill immediately to prevent staining of the surfaces. If you use bleach to clean up the spill wipe down with 70% ethanol or water afterwards. To avoid damage to the stainless steel surface.

To operate a BSC safely and effectively, follow the instructions in your laboratory Exposure Control Plan/Biosafety Manual.  A brief summary of important considerations follows:

PRIOR TO USE:

1. Wash your hands with soap and water and put on the appropriate PPE, as described in your laboratory Exposure Control Plan/Biosafety Manual.
2. Turn off the UV light if used.
3. Raise the sash to the proper height.
4. Turn on the blower and light and allow to run for 10 to 15 minutes to purge the air.
5. Wipe down the surface, walls, grills, and viewscreen of the BSC with an appropriate disinfectant.
6. Load the cabinet with the necessary materials and reagents. Make sure that you disinfect the outer surface of media bottles and pipettes before putting them in the BSC and, if possible, place a container with disinfectant and/or sharps containers to discard serological/pasteur pipettes and tips.
7. Let the air purge for another 5-10 minutes.

WHILE WORKING IN THE BSC:

1. Do not block the front and rear air intake grills.
2. Do not disrupt the protective airflow pattern. Avoid rapidly moving your arms in and out of the cabinet, people walking rapidly behind you, and open laboratory doors, which may reduce the effectiveness of the BSC. Try to move arms into the cabinet at a perpendicular angle.
3. Establish working areas and always work from the clean to the dirty area.
4. Clean spills as soon as they occur. Cover spills with a paper towel and then soak with disinfectant. Leave for the appropriate contact time (>20 minutes) and then clean up. Remove contaminated gloves in the Biohazardous waste. Wash hands and put clean gloves on.

AFTER USE:

1. Disinfect the materials that have been inside the cabinet before taking them out.
2. Disinfect the surfaces of the BSC. Do not place your head inside the BSC.  If you cannot reach all surfaces use a device to extend the cleaning materials, such as a handle.
3. Purge the air inside the BSC for a 10-15 minutes.
4. Turn off the blower and light and close the sash.
5. If the BSC is equipped with a UV light, turn it on for 15 minutes if nobody is present and abandon the room. Post a sign on the door notifying others that the UV light is on. Return later to turn the UV light off. Do not expose yourself unnecessarily to the UV light, since this can result in skin burns and eye lesions.
6. Remove PPE and place disposable materials in the Biohazardous waste. Thoroughly wash hands.

If you have any questions about the correct and safe use of the BSC, please contact EHS at (801) 581-6950.

Can I work with chemicals in a Biological Safety Cabinet?

Evaluation of the inherent hazards of the chemicals must be part of the risk assessment when selecting a Biological Safety Cabinet (BSC) . Flammable chemicals should not be used in Class II, Type A1 or A2 cabinets since vapor buildup inside the cabinet presents a fire hazard. For more information regarding the risks associated with using chemicals in Biological Safety Cabinets, or chemical fume hoods, contact OEHS (801-581-6590).

Additionally, chemical fume hoods are NOT BSC. They are designed for worker protection only and do not provide any environmental protection against microbiological agents since the air in a chemical hood is not filtered through a HEPA filter. Chemical hoods do not provide any protection from room contaminants for the work done in the hood. Chemical fume hoods operate by pulling large quantities of air into the front or face of the hood, through the hood box and out through a series of slots in the rear of the hood box using large fans (normally located on the facility roof). One hundred percent of the air that enters the hood is exhausted from the building. Thus, chemical fume hoods do not provide environmental protection against infectious agents because the exhausted air is not filtered through a HEPA filter.

When EHS inspects my lab what are they looking for?

A lab inspection/audit is required as part of the review process for all work at biosafety level 2 (BSL-2) or above. After the registration has been submitted, the BSO or Associate BSO will contact the Principal Investigator to arrange a time for the lab audit. Copies of the lab audit checklist can be found here (https://OEHS.utah.edu/research-safety/biosafety/biosafety-laboratory-audits). The inspection will ensure that safe practices are in place, that physical containment is appropriate, training is up to date and that laboratory staff have access to, and are familiar with, Biosafety manuals and/or standard operating procedures. If an incident occurs resulting in an exposure to a hazardous agent, the BSO or Associate BSO will conduct additional audits. Subsequent lab inspections will occur annually and may include unscheduled inspections.

I ship hazardous materials, including infectious or recombinant agents. Do I need training for this?

Yes!

Strict government regulations must be followed when transporting hazardous materials, which include dry ice. Shipments must arrive at their destination in good condition and present no hazard during shipment. All individuals who ship hazardous materials must undertake training: training for the shipment of Category B substances is available through the University of Utah RATS program. Category A shipments must be made by OEHS.

To read the regulations concerning the transport of a diagnostic or infectious specimen, see:

• U.S. Department of Transportation, Office of Hazardous Materials Safety, 49 CFR Regulation 173.134.
• International Air Transportation Association (IATA) Dangerous Goods information Online.

For more information, please visit Shipping and Receiving Hazardous Materials page.

Prior to shipping any hazardous materials, please contact the Biosafety Officer at 801-581-6590.

If I am working with Biological Materials do I need to undergo medical surveillance or vaccination?

All employees in research laboratories working with, or who may be exposed to, potentially infectious agents, including recombinant viral vectors, must be aware of signs or symptoms consistent with diseases caused by these agents and their parental strains. For example personnel working with recombinant lentiviral vectors should be aware of the signs and symptoms of human immunodeficiency virus (HIV) infection. Personnel exposed to these agents may or may not become sick; however, they may have the potential to transmit them to others outside the laboratory if proper biosafety practices have not been followed. Therefore, laboratory-specific training must include hazard communication related to the risks of these agents, anticipated signs/symptoms associated with these agents to facilitate recognition of potential occupational illnesses, and procedures to follow if a potential exposure has occurred.

For certain activities, medical surveillance should be undertaken prior to working with biological agents. This may be a requirement for approval of work that is reviewed and approved by the University of Utah Institutional Biosafety Committee (IBC). Examples include laboratories working with human pathogens, such as HIV or Zika virus, or with agents for which vaccination may offer protection, such as pertussis toxin (PT). In addition, all personnel should be made aware by their employers/supervisors that certain medical conditions increase their risk of potential health problems when working with pathogenic microorganisms and/or animals. These conditions include pregnancy, immunosuppression, animal related allergies, and chronic skin conditions. All personnel should discuss their work with an Occupational Health physician or their personal physician/health care professional if any of these conditions apply.

Certain types of work may require the use of a respirator to protect against aerosol exposures. In such cases, personnel must get medical clearance from the Department of Occupational Medicine: for most personnel this can be achieved by completing and submitting an OSHA Respirator Medical Evaluation form that can be downloaded here. Once Occupational Medicine has provided clearance, call the Biosafety Officer (801-581-6590) to arrange an appointment for Respirator Fit Testing. A Respiratory Protection Plan must also be submitted to the Biosafety Officer: a template can be found here. Fit testing must be repeated on an annual basis.

Vaccinations

Personnel may be required by the IBC to be offered vaccinations to protect them from workplace hazards. Examples include the Hepatitis B vaccine for all workers with reasonable expectation of exposure to human blood or other potentially infectious materials (OPIM), which includes human and non-human primate cell lines, including those acquired from commercial sources. Tdap vaccination, which is highly effective for the prevention of diphtheria, tetanus and pertussis, should be offered to personnel working with PT or handling animals dosed with PT. Vaccine recommendations can be found at http://www.cdc.gov/vaccines/hcp/acip-recs/vacc-specific/index.html and http://www.cdc.gov/vaccines/hcp/vis/index.html?s_cid=cs_748.

Protective vaccines, if available and appropriate based on workplace hazards, will be provided by the University of Utah at no cost to the employee. In most cases, if there is limited public health concern, employees may choose to decline the recommended vaccinations after understanding their risks. In these circumstances, the University of Utah is obligated to document the offer and obtain a signed declination by the employee that they understand the risks, yet chose to decline the vaccination. If the employee changes his/her mind, the vaccination will be made available to them upon request.

Post Exposure Procedures

Exposures or potential exposures should be reported to the supervisor and the Biosafety Officer (801-581-6590), and the affected individuals should report to the Occupational Medicine Clinic at the Redwood Health Center or to the RedMed Clinic at the Student Union building. In the event of a life-threatening event call 911 immediately. Information about the University of Utah Health Care Occupational Medicine Clinics can be found here.

Employees must also follow the Incident Reporting Policy described in their laboratory Exposure Control Plan or Biosafety Manual. The medical professionals at Occupational Medicine will determine the need for post-exposure prophylaxis, treatment, and continued medical surveillance at that time. Employees must notify the medical professionals if the agent involved is modified in any way to allow the medical professionals to treat the agent appropriately. The Principal Investigator (PI) or Biosafety Officer may be required to provide additional information about agent modifications and their potential effects on treatment. PI’s should make available to all personnel post-exposure procedures for all agents used in the laboratory: a template can be found here .

The University of Utah IBC requires plans to address how a biological exposure incident be developed by the PI: details must be incorporated into the laboratory IBC registration and should be part of the laboratory-specific exposure control plan and/or Biosafety Manual. The IBC has developed a template for a post-exposure SOP that can be downloaded here. This should include identification of any post-exposure prophylaxis options and/or medical monitoring plans for those who may have been exposed to the agents, documentation of important aspects of the experimental design and procedures, such as changes in drug sensitivity and/or genetic modifications, which may modify the risks of exposure of these agents.

In the event of exposure it is recommended that laboratory personnel reporting to the Occupational Medicine clinic after an exposure should bring completed post-exposure SOPs with them to the health care provider to ensure proper communication to those who may be providing care, particularly for agents which are genetically modified agents