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Biological Safety

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  • Virginia Tech's Institutional Animal Care and Use Committee (IACUC) is the oversight body that approves research on whole, vertebrate animals, and monitors the humane care and use of research animals. Visit the IACUC website for information on policies, laws and regulations related to research with animals, protocol submission and management, training, news and announcements.
  • Protocols for research projects involving animals must be submitted to the IACUC for review and approval, and for periodic renewal.
  • Individuals must complete initial IACUC training before participating in research work with animals, and they must stay current with IACUC refresher training thereafter.
  • Animal research involving recombinant material, transgenic or genetically modified animals, and/or infectious agents falls within the purview of the Institutional Biosafety Committee (IBC). Protocols for research projects of this type must be submitted to the IBC (in addition to IACUC) for review, approval, and periodic renewal, and people must complete IBC-required training before performing research with infectious agents or recombinant material.

Primary hazards and safety considerations in animal research

Injury and possible infectious exposure from:

  • Animals bites/ scratches;
  • Accidental injury with needle/scalpel used on an animal; and/or
  • Large animals kicking, stomping, head-butting, etc.

Reduce or eliminate the risk to health and safety of the people working with animals by: 

  • Using safe, effective restraint and anesthesia methods on animals when needed;
  • Avoiding inhalation risks and production of aerosols; 
  • Utilizing sharps with safety features and disposing of sharps appropriately; and
  • Using good work practices and appropriate PPE.

Exposure to animal allergens create risk of allergy development, or serious allergic reaction in those with preexisting allergies:

Contain allergens by using filtered cages, animal transfer stations, respiratory protection, PPE, and good animal handling practices.

Escape of infected or transgenic animals from containment:

Give priority to the safety, security and care of the animals you work with, as well as animals living with them or near them.

Animal-generated aerosols (from bedding, dander, saliva, sprayed or flung excreta, etc.) creating exposure hazards to infectious agents, recombinant material or excreted chemicals:

Employ proper: 

  • Containment equipment/practices;
  • Animal handling practices;
  • Waste handling practices; and 
  • Appropriate PPE

to avoid aerosols of pathogens, recombinant material/ organisms, chemically contaminated material.

Animal biosafety levels:

  • See Biosafety Levels in this website's topics list.

Shipping animals:

Guidance and further information:
IACUC
IBC
BMBL, Section V: Vertebrate Animal Biosafety Level Criteria for Vivarium Research Facilities
Office of the University Veterinarian

  • Refers to research projects involving invertebrate animals known as arthropods (having exoskeletons, segmented bodies, and paired, jointed appendages). Insects, spiders/mites, and crustaceans are three of the largest arthropod groups.
  • Some species of arthropods are vectors or transmitters of disease to other species and are often the focus of research efforts.
  • For research involving vector species or non-vector species which are NOT infected or genetically manipulated, and have been collected in the field or purchased commercially, institutional oversight is usually not required.
    • Examples of non-vector species: fruit flies (drosophila), cockroaches, butterflies/moths (lepidoptera), beetles (coleoptera), etc.
  • The Institutional Biosafety Committee (IBC) oversees research involving vector species infected/ potentially infected with biohazardous agents, or vector species whose infection status is unknown. 
    • Protocols for research projects of this type must be submitted to the IBC for review and approval, and for periodic renewal.
    • People must complete IBC-required training before beginning this type of research with arthropods.
    • Examples of vector species:
      Insects:
      • Diptera – mosquitoes, tsetse flies, black flies, sandflies, midges;
      • Hemiptera – reduvids (kissing bugs);
      • Anoplura – lice; and
      • Siphonaptera – fleas.
      Arachnids:
      • Acari – ticks, mites.
    • Biohazardous agents carried by vector species include viruses, rickettsia, bacteria, protozoa, and helminths that cause disease and may present a public health threat.
    • Researchers must take into account all life-cycle stages (eggs, larvae, nymphs, and adults) when providing arthropod containment. The small size, mobility (especially flying and jumping species), reproductive potential, and the relative longevity and resistance of certain life stages of some vector species often require species-specific containment solutions.
    • Follow the risk assessment process and other guidance found in "Arthropod Containment Guidelines" (see link below).
  • The Institutional Biosafety Committee (IBC) also oversees research involving genetic manipulation of arthropod vector and non-vector species and/or use of genetically modified arthropods (even if purchased from another lab/collaborator).
    • Protocols for research projects of this type must be submitted to the IBC for review and approval, and for periodic renewal.
    • Individuals must complete IBC-required training before beginning this type of research with arthropods.
  • Use of blood to feed a vector species:
    • Blood used from a human volunteer – Submit a protocol to the IRB, the entity that oversees research using human subjects/volunteers.
    • Human blood used from a commercial source – No IRB protocol needed.
    • Research animals used for blood-feeding – Submit a protocol to the IACUC, the entity that oversees the use and treatment of animals in research.

Research scope and approval and compliance

Involving vector or non-vector arthropod species; no genetic modifications; no infection with biohazardous or genetically modified agents:

No oversight required.

Involving vector species and research animals—animals exposed to vector species for blood feeding and/or for study:

Engage the IACUC.

Involving vector species that are fed with human blood/blood product:

  • If human volunteers supply blood to feed arthropods, a protocol must be submitted to the Institutional Review Board (IRB) and IBC.
  • If human blood is purchased commercially, an IRB protocol is not needed; register with IBC as a Special Lab.
  • In either case, lab personnel who handle the human blood that is fed to arthropods are required to take Bloodborne Pathogens training.

Involving vector species OR non-vector species:

  • Infected/ potentially infected with biohazardous agents;
  • Infected with genetically modified biological agents;
  • Where the vector species itself has been genetically modified. 

Engage the IBC.

Primary hazards in arthropod research and safety considerations

Exposure to vector-borne disease from arthropod bite:

Reduce or eliminate the risk to health and safety of the people working with this material, as well as others who could be indirectly exposed by: 

  • Using proper containment practices.
  • Using good work practices and appropriate PPE.

Escape of infected or transgenic arthropods from containment:

Contain the arthropods by following Arthropod Containment Level criteria, and by adhering to specific work and containment practices as determined by risk assessment for each situation. Examples of these practices include:

  • Use appropriately-sized screen mesh to provide primary containment.
  • Disinfect containers effectively to prevent arthropod survival and escape.
  • Use an effective trapping method in the lab to monitor for escaped arthropods.
  • Manipulate arthropods in a secure glove box, walk-in incubator, screen cubicle, etc.

Arthropod containment guidelines

See Biosafety Levels in this website's topic list.

Shipping insects and mites

  • A USDA permit is required for the importation, interstate movement and environmental release of most live insects and mites that feed upon or infest plants or plant products, including agricultural crops, trees, shrubs, native plants, etc.
  • The movement of genetically modified organisms requires permits from Biotechnology Regulatory Services.
  • The movement of insects, mites and ticks that affect animals or vector animal diseases requires permits from Veterinary Services.
  • The movement of insects, mites and tick that affect man or vector human diseases requires permits from the Centers for Disease Control.

Biotoxins overview

  • Biotoxins are potent, highly toxic biological substances produced by living organisms; some can be synthesized by modern methods.
    • May cause death or severe incapacitation at relatively low exposure levels.
    • Are handled as chemical hazards.
    • Safety procedures for using toxins are determined by a toxin risk assessment process.
  • Typical research applications include: 
    • Use as a growth factor in cell culture media (e.g., cholera toxin).
    • To produce specific neurologic effects (e.g., tetrodotoxin).
    • To produce localized tissue destruction (e.g., diphtheria toxin).
  • When you are asked (on the IBC Main Form, by Environmental Health & Safety, or by an auditor or inspector) if your lab stores, uses, or will use biotoxins for research purposes, please identify all of the toxins you utilize or store for any purpose in your research efforts (such as an additive to media, etc.), and quantities used/stored.
  • Possession and use of biological toxins or venoms in research requires:
    • Registration with, and approval by the Virginia Tech Institutional Biosafety Committee (IBC).
    • Maintenance of a toxin inventory record to account for current quantities in house, and toxin use and disposition.
    • Storage of toxins in sealed, labeled containers within a secured (locked) storage device that can only be accessed by personnel authorized to work with the toxin.
    • Use of toxins only in designated rooms with posted signage to control/limit access. 

Biotoxins as select agents and the due diligence provision

  • Some biotoxins are classified by the federal government as Select Agents due to their potential to pose a severe threat to public health and safety. Possession, use, and transfer of these toxins are highly regulated. 
  • In small quantities, some of these agents are exempt from Select Agent registration but must still be registered with, and approved for use by the Virginia Tech IBC.
  • Because it would be possible to stockpile toxins in multiples of these amounts which are small enough to be excluded from regulation, the 'Due Diligence' policy places the following responsibilities on Principal Investigators, veterinarians, or medical doctors:

Biotoxins and exempted amount per PI (≤)

Abrin: 1000 mg
Botulinum neurotoxins: 1 mg
Short, paralytic alpha conotoxins: 100 mg
Diacetoxyscirpenol (DAS): 10,000 mg
Ricin: 1000 mg
Saxitoxin: 500 mg
Staphylococcal enterotoxins (subtypes A, B, C, D, E): 100 mg
T-2 toxin 10,000 mg
Tetrodotoxin: 500 mg

If you possess any of these toxins, you must:

  1. Declare your possession and provide inventory documentation to Virginia Tech's Responsible Official (RO) and Alternate Responsible Official (ARO).
  2. Contact the RO and/or ARO to schedule a SHORT informational session regarding requirements related to possession of these toxins.
  3. If no additional toxin is received after the declaration date, but at least annually, you must verify and declare your possession to Virginia Tech's RO and ARO.
  4. Immediately notify Virginia Tech's RO and ARO if your toxin inventory amounts, per toxin, ever exceed specified permissible limits.

If you transfer any of these toxins to another entity, you must:

  1. Apply due diligence in assessing/ensuring that the recipient of these toxins has a legitimate need to handle/use such toxins.
  2. Complete documentation prior to the transfer which records your information and the recipient's intended use of the toxin. The Notification of Toxin Transfer form (template can be found in the Appendices of the University Biosafety Manual and IBC website) must be submitted to Virginia Tech's RO and ARO. 
  3. Immediately report to Virginia Tech's RO and ARO, if a violation of federal law is detected, or if suspicious activity related to the toxin is discerned.

If you receive any of these toxins from another entity, you must:

  • Immediately notify Virginia Tech's RO and ARO of the receipt of the toxin, update your inventory, and submit inventory documentation to the RO (Charlotte Waggoner, 540-231-5864) and ARO (Anna Kroner, 540-231-1122).

Biotoxin risk assessment

  • Risk assessments on biotoxins should include:
    • Biotoxin characteristics (LD50 in solution and dry form; solubility).
    • Risks inherent to experimental procedures and manipulations (e.g., opportunities for accidental needlesticks, the likelihood of dispersal from static build-up when working with powder form, etc.).
    • Total amount of toxin used relative to the estimated human lethal or cytotoxic dose.
    • Volume of material manipulated.
    • Availability of successful treatment, vaccines or antitoxins.
    • Training and experience of the personnel.
  • Safety measures should be determined according to a risk assessment for each manipulation involving the toxin.
  • If toxins/infectious agents/ animals are used in combination, then risks in all of those areas must be considered collectively in the selection of containment equipment and the development of safety procedures.
  • After risk assessment review, selected operations with toxins may require modified BSL-3 procedures; typically, routine operations (e.g., toxin preparation/ animal procedures) with dilute toxin solutions can be conducted under BSL-2 conditions using 1) a certified BSC or chemical fume hood; and 2) appropriate PPE for the hazards involved.
  • Complex operations should be rehearsed without the use of live toxin in supervised practice runs until proficiency is assured.

Primary hazards using biotoxins and hazard mitigation when using biotoxins

Exposure to toxin by direct contact or hand-to-face contact with mouth, eyes, or other mucous membranes.

  • Wrap-around disposable gowns with gathered cuffs are the best choice, and fluid-resistant gloves rated for protection against diluent should be used.
  • Double-gloving is strongly recommended if it does not hinder the worker's dexterity to the point of being unsafe.
  • There should be no unprotected, exposed skin on your body or extremities when undertaking this work. Arms, wrists, and hands must be fully covered; long pants and whole shoes are required.
  • Safety glasses are strongly recommended and must be washed after removal and before storage.
  • Disposable PPE used for this work must be single-use only and disposed of as hazardous lab waste.

Exposure to toxin from inhaling an aerosol.

  • Work in primary containment with directional air flow (biosafety cabinet, chemical fume hood).
  • When working with dried toxin, remove all items that are not necessary for your procedure from the biosafety cabinet or fume hood before handling the toxin to reduce the potential for contamination of item surfaces.
  • Respiratory protection may be needed if you must manipulate a dry toxin in an open vessel, even if in containment. Contact Environmental Health & Safety at 540-231-3600.
  • Primary containers of the toxin should be non-breakable if possible, and vials should be maintained in a closed secondary container that will not allow escape of the product even if dropped. The risk of release is greatly decreased by using a secondary container at all times.

Exposure to toxin through needlesticks or other accidents that compromise intact skin, such as could happen when inoculating animals.

  • Use only syringes with luer-lock or integrated needles.
  • Use vial adapters whenever possible as a substitute for needles when adding diluent through septums on vials.
  • When introducing a needle through a septum, assure that the vial is secured with a device that allows the non-dominant hand to be outside of the 'strike zone' of the needle. Example: secure vial in a rack, or use a clamp to hold vial instead of holding it directly.
  • When inoculating animals, use effective, reliable restraint methods to avoid accidental needlesticks.

For procedures in solubizing and using biotoxins, toxin spill response, and toxin inactivation and disposal, see the University Biosafety Manual, Section 4.23.

Recruitment and use of human volunteers for academic research are regulated by Virginia Tech's Institutional Review Board (IRB). This oversight committee reviews and approves all aspects of research efforts that involve volunteers, such as recruitment practices, survey methods, recordkeeping practices, research procedures, security of volunteer information, etc. The IRB ensures that the rights and safety of human subjects/volunteers are protected.

  • Examples of research areas where IRB oversight occurs:
    • Studies in sociology, psychology, human nutrition, transportation, etc. 
    • Medical or clinical trials for pharmaceutics or other experimental treatments, imaging studies, genetic studies; etc.
    • When human specimen material is collected from volunteers for manipulations in research labs or teaching labs (see below).
  • Protocols for research projects of these types must be submitted to the IRB for review and approval, and for periodic renewal.
  • People must complete IRB-required training before beginning this type of research.

Human Biological Material (Blood/Tissues/Fluids, Human Cell Lines) Used in Research or Teaching

  • The Institutional Review Board oversees research involving human specimen material collected from volunteers for manipulations in research labs/ teaching labs.
    • Teaching labs that use human specimen material (blood from fingersticks, tissue from cheek swabs, saliva, etc.) from lab participants for exercises:
      • An application to the Institutional Review Board may not be needed, if the specimen material is self-collected and self-tested, and if test results are not shared in any way that identifies participants.
      • Contact the IRB to confirm whether your teaching exercises require IRB approval.
  • The Institutional Biosafety Committee (IBC):
    • Reviews research protocols and teaching lab protocols involving culturing and/or manipulation of biological material of human/non-human primate (NHP) origin, including the following:
      • Blood/ blood products/blood components
      • Unfixed tissue or organs from a human or NHP (living or dead)
      • Other potentially infectious materials of human/NHP origin, including sexual fluids, cerebrospinal fluid, organ fluids, joint fluids, amniotic fluid, saliva
      • Human or NHP cell cultures, including primary or established cell lines
    • These materials are considered biohazardous material due to the risk of exposure to bloodborne pathogens; the IBC has determined that any work with these agents must be conducted at BSL2.
    • Clinical diagnostic labs, service labs (such as for electron microscopy or flow cytometry), and some teaching labs using human materials receive a targeted review by the IBC as "Special Labs" rather than the broader review given to academic research labs.
    • OSHA requires that Bloodborne Pathogens training be taken by all lab personnel who have the potential for exposure to human blood or other potentially infectious materials (NHP is also included in this requirement).
Lab Setting Material and Manipulation Oversight Requirements
Research and Teaching Labs Human blood, urine, saliva, etc. cultured and/or manipulated
  • Register with IBC. 
  • Perform risk assessment for BBP.
  • Document training for personnel.
  • Contact IRB.
Clinical Research lab (e.g., VTCRI) Human blood, urine, etc. collected and/or minimally manipulated (ex: tested or shipped in closed containers)
  • Register with IBC as a Special Lab.
  • Perform risk assessment for BBP.
  • Document training for personnel. 
  • Contact IRB.
Service Labs (e.g., Flow Cytometry) Manipulation or testing of tissues/ fluids/ cell suspensions of human, NHP, tumorogenic, transgenic, or infective material
  • Register with IBC as a Special Lab.
  • Perform risk assessment for BBP.
  • Document training for personnel.
  • Contact IRB.
Teaching Labs Human saliva, cheek swabs, blood from fingersticks, etc.
  • No oversight needed IF:
    • Specimens are self-collected & self-tested.
    • Manipulations of specimens are minor, and/or take place in closed containers.
    • Results are not shared or are shared as de-identified information.
    If one or more conditions are not met, contact the IRB.

Primary hazards in research using human/NHP materials and safety considerations:

Exposure to bloodborne pathogens and/or other potentially infectious agents.

Mitigate exposure risk by following OSHA Bloodborne Pathogen Standards and Criteria, and by adhering to specific work and containment practices as determined by risk assessment.

Guidance and further information:

IRB
IBC

  • The term "plant" includes, but is not limited to mosses, liverworts, macroscopic algae, as well as vascular plants (crop, forest, weed, and ornamental species).
  • Non-recombinant plant research, in the greenhouse or in the field, which involves no plant pathogens, pesticides, herbicides, toxins, or releases of non-native/noxious species, is not generally subject to institutional oversight or federal regulation.
  • Three federal agencies regulate genetically engineered plants:
    1. Department of Agriculture (USDA) – provides safety for agriculture and the environment 
    2. Environmental Protection Agency (EPA) – provides safety for the environment
    3. Food and Drug Administration (FDA) – provides safety for food and feed use 
  • The Centers for Disease Control and the USDA – APHIS are involved in regulating work on plant pathogens/toxins that are a biosecurity concern (could be used as biological weapons).
  • Consult with the appropriate federal agencies early in the planning stages of a plant research project to receive guidance.
  • Virginia Tech Institutional Biosafety Committee (IBC) oversight is required for research involving:
    • Transgenic /genetically modified plants
    • Plant toxins 
    • Microorganisms or agents are known to cause/associated with plant disease (viroids, virusoids, viruses, bacteria, fungi, protozoa, arthropods, nematodes, etc.)
  • Protocols for research projects involving these materials must be submitted to the IBC for review and approval, and for periodic renewal.
  • People must complete IBC-required training before beginning research work, and they must stay current with IBC-required training thereafter.

Primary hazards in plant research and safety considerations:

Loss of containment and release of infected or transgenic plant material or plant-associated organisms:

Plant biosafety levels (BSL-P) for greenhouse research:

  • See 'Biosafety Levels' in this website’s topic list.

Guidance and further information:

  • These are biological agents and toxins (often referred to as simply 'select agents') that possess the potential to pose a severe threat to public health and safety, to animal and plant health, or to animal or plant products.
  • list of these agents and toxins is maintained by the Federal Select Agent Program and is regularly updated.
  • Many select agents must be handled using the strict containment and PPE requirements of Biosafety Level 3 or 4, although some can be manipulated at BSL-2. However, ALL select agent research must take place under high-security conditions that are regulated by federal code and overseen by HHS-CDC and/or USDA.

The Centers for Disease Control and Prevention (CDC) is the oversight body that approves and monitors research involving select agents and toxins at Virginia Tech. The Institutional Biosafety Committee (IBC) reviews and approves research involving select agents to ensure that it complies with federal regulations.

  • Principal investigators who are interested in conducting work with select agents must:
    • Contact the Responsible Official.
    • Submit a protocol to the IBC for review and approval, and for periodic renewal.
  • Students who have an interest in participating in research that involves select agents should contact the Responsible Official to learn more.

The Federal Select Agent Program:

  • The program is a joint effort of the Centers for Disease Control and the U.S. Department of Agriculture. Regulatory authority is federally granted by 7 CFR Part 331, 9 CFR Part 121, and 42 CFR Part 73.
  • This program oversees the possession, use, and transfer of biological select agents and toxins by:
    • Developing, implementing, and enforcing the Select Agent Regulations.
    • Inspecting entities that possess, use, or transfer select agents.
    • Ensuring that all individuals who work with these agents undergo a security risk assessment performed by the FBI/ Criminal Justice Information Service.
    • Providing guidance to regulated entities for achieving compliance with the regulations.
    • Investigating any incidents in which non-compliance may have occurred.
  • All entities performing work on select agents in the U.S. must be officially registered with the program and subscribe to its oversight.

The Virginia Tech Select Agent Program:

  • Ensures compliance with CDC and APHIS regulations which apply to:
    • research activities conducted at Virginia Tech involving select agents, including possession and use, transfers within the U.S., and importations.
    • Virginia Tech facilities where select agents are used and stored, including animal research facilities.
    • individuals (Virginia Tech faculty, staff, students, and visitors) requiring access to select agents or access to an area where select agents are present.
  • Aids principal investigators, researchers, and supervisors working with select agents to achieve and maintain compliance.
  • Is administered by Environmental Health & Safety through the Responsible Official (RO) and Alternate Responsible Officials (ARO), who maintain Virginia Tech's registration with the Federal Select Agent Program.
  • The Responsible Official (or ARO in the RO's absence):
    • Acts as a liaison for Virginia Tech with regulatory agencies, state health agencies, and emergency response providers.
    • Investigates all incidents of loss, theft, or release of select agents, and is the reporting authority for these incidents to regulatory agencies and to academic departments.
    • Conducts regular inspections of select agent facilities, including animal research facilities.
    • Reviews and approves risk assessments, and safety, security, and incident response procedures for select agent research.
    • Coordinates with the EHS Occupational Health group as needed regarding appropriate medical surveillance and care for personnel in Virginia Tech's Select Agent Program.
    • Conducts personnel suitability evaluations and investigations when required.
    • Provides administrative processing and select agent training for new personnel.
    • Provides annual training for all personnel in the Virginia Tech Select Agent Program, and for those professionals who would respond to an incident or emergency at Virginia Tech select agent facilities.
  • Lab worker requirements:
    • To be granted access to Virginia Tech select agent facilities (other than an approved, escorted visitor) and to conduct work with select agents therein, an individual must:
    • Undergo a federal Security Risk Assessment approval and maintain security approval.
    • Undergo a personnel suitability assessment and maintain suitability approval.
    • Initially complete and keep up-to-date with all required select agent training and IBC-required training.
    • Initially complete and keep up-to-date all Virginia Tech Occupational Health requirements.
    • Comply with all approved select agent program written plans and procedures (incident response plan, security, biosafety, lab-specific SOPs, etc.).

Biotoxins as select agents and the due diligence provision:

  • Some biotoxins are classified by the federal government as Select Agents due to their potential to pose a severe threat to public health and safety. Possession, use, and transfer of these toxins are highly regulated.
  • In small quantities, some of these agents are exempt from Select Agent registration but must still be registered with, and approved for use by the Virginia Tech IBC.
    Toxin Exempted Amount (≤)
    Abrin 100 mg
    Botulinum neurotoxins 0.5 mg
    Short, paralytic alpha conotoxins (containing the amino acid sequence X1CCX2PACGX3X4X5X6CX7) 100mg
    Diacetoxyscirpenol (DAS) 1000mg
    Ricin 100mg
    Saxitoxin 100mg
    Staphylococcal enterotoxins (Subtypes A, B, C, D, E) 5 mg
    T-2 toxin 1000mg
    Tetrodotoxin 100mg

Because it would be possible to stockpile toxins in multiples of these amounts which are small enough to be excluded from regulation, the 'Due Diligence' policy places the following responsibilities on Principal Investigators, veterinarians, or medical doctors: IF YOU POSSESS ANY OF THESE TOXINS, YOU MUST: IF YOU TRANSFER ANY OF THESE TOXINS TO ANOTHER ENTITY, YOU MUST:IF YOU RECEIVE ANY OF THESE TOXINS FROM ANOTHER ENTITY, YOU MUST:

  • Declare your possession and provide inventory documentation to Virginia Tech's Responsible Official (RO) and Alternate Responsible Official (ARO).
    1. Contact the RO and/or ARO to schedule a SHORT informational session regarding requirements related to possession of these toxins.
    2. If no additional toxin is received after the declaration date, but at least annually, you must verify and declare your possession to Virginia Tech's RO and ARO.
    3. Immediately notify Virginia Tech's RO and ARO if your toxin inventory amounts, per toxin, ever exceed specified permissible limits.
  • Apply due diligence in assessing/ensuring that the recipient of these toxins has a legitimate need to handle/use such toxins.
    1. Complete documentation prior to the transfer which records your information and the recipient's intended use of the toxin. The Notification of Toxin Transfer form (template can be found in the Appendices of this manual) must be submitted to Virginia Tech's RO and ARO.
    2. Immediately report to Virginia Tech's RO and ARO, if a violation of federal law is detected, or if suspicious activity related to the toxin is discerned.
  • Immediately notify Virginia Tech's RO and ARO of the receipt of the toxin, update your inventory, and submit inventory documentation to the RO and ARO.

Guidance and further information:

Non-pathogenic microorganisms or random environmental isolates that have not been/will not be genetically manipulated:

  • Research involving these organisms typically requires no institutional oversight.
  • If an environmental study randomly yields isolates of pathogenic or suspected pathogenic organisms:
    • If you need to identify such isolates, do so in a biosafety cabinet using BSL-2 practices.
    • Handle identified pathogens using the appropriate containment, work practices, safety equipment, and PPE that is prescribed by the organism's biosafety level.
    • If a Risk Group 3 or select agent is isolated and identified, contact the University Biosafety Officer immediately.
    • If your work will extend beyond isolation and identification of Risk Group 2 or 3 microbial flora, (such as further culturing or manipulation of isolates) contact the IBC.

Biohazardous microorganisms/agents/materials:

  • Virginia Tech's Institutional Biosafety Committee (IBC) is the oversight body that approves and monitors research and teaching protocols involving microorganisms that are potentially pathogenic/ pathogenic for humans, animals, and plants (Risk Group 1-3), as well as the following:
    • Genetically engineered organisms/microorganisms, including animal or plant pathogens and products, and veterinary biologics
    • Select agents and toxins
    • Infected animals/ animal tissues
    • Prions
  • Protocols for research projects involving these materials must be submitted to the IBC for initial approval, and thereafter, for annual review and three-year renewal.
  • People must complete IBC-required training before beginning research work, and they must stay current with IBC-required training thereafter.
 
Research Scope Virginia Tech Entity that Gives Approval and  Oversees Compliance
Involving environmental and/or non-pathogenic agents that are not genetically manipulated/ modified. No institutional oversight needed
Involving pathogenic/ potentially pathogenic and/ or genetically modified microorganisms, viral vectors or other potentially infectious materials. IBC
Involving biological products derived from Risk Group 2 or Risk Group 3 microorganisms. IBC
Involving clinical/ medical diagnostic specimens that are used in research and known (or reasonably expected) to contain pathogenic microorganisms/agents. IBC
Involving the culture of more than 10 liters of a biological agent. IBC
Involving animals in a research setting or in the wild (incl. cages/bedding, nests/dens, waste products, etc.) where there is a risk of exposure to infectious agents for humans and/or animals. IBC, IACUC
Involving research animals experimentally infected with infectious agents. IBC, IACUC

Involving arthropod vector OR non-vector species

  • infected with biohazardous agents
  • infected with genetically modified biological agents or genetically manipulated
IBC
Arthropod Containment Guidelines
Teaching labs utilizing pathogenic/ potentially pathogenic and/ or genetically modified microorganisms, viral vectors or other potentially infectious materials. IBC
American Society of Microbiology: Guidelines for Biosafety in Teaching Laboratories

Appendix to the Guidelines for Biosafety in Teaching Laboratories

Primary hazards in research using microorganisms/biological agents and materials and safety considerations:

Exposure to infectious agents from mucous membrane exposure (hand-to-face contact), from inhalation of an aerosol, or from accidental injury or non-intact skin contact with infectious material:

Reduce or eliminate the risk to health and safety of the people working with this material, as well as others who could be indirectly exposed by:

  • Using primary containment (biosafety cabinet).
  • Avoiding inhalation risks and production of aerosols in procedures; use aerosol-producing equipment within containment. 
  • Utilizing sharps with safety features and disposing of sharps appropriately.
  • Using good work practices and appropriate PPE.

 

Loss of containment and release of infectious agents/materials into the environment:

Mitigate the risk of release by following Biosafety in Microbiological and Biomedical Laboratories designated Biosafety Level criteria (containment equipment and practices, engineering controls, PPE, and facilities features).

Biosafety Levels (BSL) for Microbiological Research

  • See 'Biosafety Levels' in this website's topic list.


Shipping of biological material:

  • See 'Shipment of Biological Material' in this website's topics list.

Guidance and further information:

IBC
BMBL
CDC
EHS Biosafety Group

  • Virginia Tech's Institutional Biosafety Committee (IBC) is the oversight body that approves and monitors research and teaching protocols involving:
    • Recombinant and synthetic nucleic acid molecules.
    • Viral vectors.
    • Genetically modified or transgenic animals (i.e., all species in the animal kingdom, including invertebrates), plants, and microorganisms.
  • The IBC also oversees research involving:
    • Artificial gene transfer.
    • Human gene transfer.
    • Synthetic biology.
  • Protocols for research projects of these types must be submitted to the IBC for review and approval, and for periodic renewal.
  • Individuals must complete IBC-required training before beginning research work, and they must stay current with IBC-required training thereafter.
Primary Hazards In Recombinant Research Safety Considerations In Recombinant Research
Exposure to recombinant material from accidental injuries, such as a needlestick. Reduce or eliminate the risk to the health and safety of the people working with this material by:
  • Using primary containment (biosafety cabinet).
  • Utilizing sharps with safety features and disposing of sharps appropriately.
  • Using good work practices and appropriate PPE.
Loss of containment and release of recombinant material/ organisms, risking unknown or potentially deleterious impacts on the genomes of humans, agricultural plants/animals, natural flora/fauna, etc. Mitigate exposure risks by following NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules criteria, and by adhering to specific work and containment practices as determined by risk assessment.

Reporting an exposure to/release of recombinant material/organism(S):

  1. Immediately notify:
    1. The Lab Director and/or Lab Manager
    2. The Animal Facility Manager/ Greenhouse Manager (if applicable)
    3. A Biosafety Officer (BSO) by telephone (preferred) or email:
       
    Biosafety Officers Telephone Email
    Charlotte Waggoner 540-231-5864 ren@vt.edu
    Anna Kroner 540-231-1122 akroner@vt.edu
    Michael Miles 540-231-3361 msmiles@vt.edu 
         
  2. The BSO will acknowledge receipt of notification by communicating to the reporting person via phone or email and will begin notifying other appropriate personnel and/or agencies.
  3. IMPORTANT: In the event of an exposure to/ release of recombinant material (e.g., rDNA/ synthetic nucleic acids/ transgenic or genetically modified organisms), the BSO must inform NIH without delay, thus IMMEDIATE reporting is required.
  4. The reporting person and the supervisor of the facility (e.g., Lab Director/ Lab Manager/ Animal Facility Manager/ Greenhouse Manager) must complete a Lab Incident Report and submit it to the BSO via email (preferred) or campus mail (MS 0423) as soon as possible.
  5. BSO will acknowledge receipt of this report via email.
  6. An Employer's Accident Report must be completed immediately by the supervisor per directions found on the linked webpage.
  7. If the supervisor does not complete the report in a timely manner, injured/exposed individuals are encouraged to complete the Employer's Accident Report themselves. If a paper form is completed rather than the online form, submit the paper form to Teresa Lyons (tlyons@vt.edu or MS 0318).
  8. An NIH Incident Report must also be completed and submitted to the University Biosafety Officer, who reviews the report and submits it to the IBC and NIH.


NIH containment/biosafety levels (BL) for recombinant research:

  • See 'Biosafety Levels' in this website's topics list.


Guidance and further information:

IBC
NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acids Molecules

Information Topics

Visitors:

  • Visitor access will be determined by the PI. (Links to Virginia Tech Policies regarding volunteers and minors are provided on the Biosafety Homepage.)
  • Prior to entry into the laboratory, all visitors must be informed of:
    • Basic emergency evacuation procedures.
    • Health hazards specific to the work occurring in the lab, as well as specific safety practices for avoiding those hazards. They must be told to avoid physical contact with all research equipment, material, and working surfaces unless invited and/or approved to do otherwise by authorized personnel, who will provide appropriate supervision.
  • Infrequent visitors (i.e., those who enter the area less than once/month) must be made aware of the above items upon each visit.
  • Frequent visitors (i.e., those who enter the area at least once/ month) can be informed initially and updated as needed if conditions change in the lab.
    • If visitors' time spent in the lab will exceed a short stay, they will be:
    • Shown the emergency exit route from the laboratory.
    • Given a review of the Emergency Procedures list posted in the lab.
    • Shown the locations and proper use of emergency eyewash and shower. 
    • Shown the locations of the nearest fire alarm and extinguisher. 
  • To minimize possible liability issues, it is highly recommended that documentation be kept which would record dates/times of these visits, and verify visitors' receipt of lab hazard and emergency response information (i.e., a visitor sign-in/sign-out/receipt-of-training log).

Housekeeping:

  • The presence of housekeepers in the laboratory should be kept to a minimum. 
  • PIs must coordinate with housekeeping supervisors to schedule housekeeping services that will be provided for the lab, and the frequency of service. Custodians/ housekeepers can be responsible for:
    • Emptying regular household trash.
    • Regular sweeping and mopping of floors.
  • PIs or their designees are responsible for informing housekeeping supervisors of any safety awareness issues for housekeepers entering their labs and performing their duties.

Service providers (Facilities employees, movers, repair technicians, etc.):

When repairs or other work needs to be done in the lab by service providers:

  • Lab hazards must be secured in the service providers' work area.
  • The area should be cleared, cleaned, and decontaminated (if applicable) by the date that the work is to be done. 
  • The PI or designee will ensure that one or both of the following measures is provided:
    • Post a 'Clearance for Lab Access' form on the main lab entry, indicating that preparations have been made for workers' safety, and documenting any hazards present about which they need to be informed. 
    • Ensure that a responsible person from the lab will serve as an escort for service worker entry into the lab, and convey the pertinent hazard information to workers. 
  • To minimize possible liability issues, it is highly recommended that documentation be kept which would record dates/times of receipt, and a brief description of any lab-specific hazard awareness training provided to housekeeping and service personnel.
  • Steam autoclaves are pressure chambers, ranging from the most basic units to complex machines offering many programmable features. 
  • The purpose of an autoclave is to provide a reliable means for sterilizing equipment and supplies, and for decontaminating biohazardous waste. This is accomplished by the application of steam and pressure to items that have been placed in the autoclave. NOTE: Sterilization systems exist that use means other than steam, such as ethylene oxide. If you need information on an ethylene oxide sterilizer, please contact EHS.
  • The industry standard: an autoclave's steam-filled chamber must be held at a temperature of 121˚ C and at a pressure of 15 PSI for a minimum of 15 minutes to achieve sterilization/ decontamination conditions within the chamber. The actual cycle time used will depend on the characteristics of the load in the chamber. Longer sterilization/decontamination times are needed as load sizes, load densities, or liquid volumes increase. An average time for a typical waste cycle is 45 minutes.
  • Because of the variety of materials and wastes that need to be processed in autoclaves, they are typically equipped to offer the user several different cycles; these can be set to run for different lengths of time. More sophisticated models offer a variety of operating parameters per cycle type to more specifically meet users' needs.
  • For sterilization/decontamination of materials to take place:
    • Steam penetration and actual surface contact with steam are required. 
    • The entrapment of air must be prevented because it blocks contact with steam. 
  • Steam for autoclaves can be supplied as a utility, or be generated by the autoclave unit itself. For larger units, an auxiliary compressor may be needed to achieve steam delivery at the required pressure.
  • Examples of lab materials:
    • Metal items;
    • Borosilicate (Pyrex) glass items;
    • Heat-resistant plastic items (Nalgene, polypropylene, polycarbonate);
    • Pipette tips;
    • Aqueous solutions;
    • Water;
    • Animal food and bedding; and/or
    • Soill.
  • Biohazardous waste: 
    • Solid waste in autoclave bags: gloves, paper towels, empty tubes, agar media in Petri dishes, other solid lab debris contaminated with biohazards.
    • Water-based biological waste: exhausted culture media, supernatants, etc. containing no chemicals and no heat-stable antibiotics.
    • Sharps waste in rigid containers: glass slides, needles, syringes, pipettes, tips, blades, etc.
    • NOTE: Ensure that biowaste is autoclaved in a secondary container such as an 'autoclave safe pan.
  • Chemical solvents;
  • Corrosives;
  • Flammable liquids;
  • Other chemicals or chemical containers;
  • Kit chemical bottles;
  • Antibiotic bottles;
  • Vials or tubes with chemical residue;
  • Radioactive material;
  • Any sealed container; and/or
  • Any material that cannot withstand 121˚C for ≥18 minutes and/or 15-18 PSI, such as polyethylene plastics (PE, recycle #1), or high-density polyethylene plastics (HDPE, recycle #2).
  • Solid/gravity:
    • Supplies steam to the chamber with no mechanical vacuum assistance, i.e., by gravity. When process time is complete, steam is quickly exhausted from the chamber. 
    • Items such as upright containers that trap air within them may not be fully sterilized/ decontaminated using this cycle because the trapped air can prohibit full steam penetration and surface exposure.
    • Use the solid/gravity cycle for: 
      • Pyrex/borosilicate glassware (empty, inverted, no closures);
      • Dry hard items (unwrapped or in porous wrap); and/or
      • Metal items with porous parts, other porous materials.
    • Use the solid/gravity cycle for decontamination of solid biological waste if you have no Pre-vacuum cycle available on your autoclave; compensate for poor steam penetration in the waste by lengthening the processing time as needed.
    • Do not use this cycle for liquids or media that require a slow exhaust.
  • Liquid:
    • Supplies steam to the chamber with no mechanical vacuum assistance. When process time is complete, steam is slowly exhausted from the chamber to prevent boil-over of liquids.
    • NOTE: Because this slow exhaust phase takes extra time to complete, plastic materials that will withstand fast-exhaust cycles can melt using the Liquid cycle due to prolonged exposure to heat.
    • Use the liquid cycle for: 
      • Pyrex/borosilicate glass containers up to 2/3 full of liquid (liquid media, aqueous solutions, liquid biowaste).
      • Do not use this cycle for items that could melt during slow exhaust.
  • Pre-vacuum (available on newer autoclaves):
    • Mechanically removes air from the chamber by pulling a vacuum in a series of pulses, then supplies steam to the chamber. The vacuum allows full steam penetration of dense materials. When process time is complete, steam is quickly exhausted from the chamber.
    • Is the preferred cycle for decontaminating solid biological waste.
    • If available on your autoclave, use the Pre-vacuum cycle for: 
      • Decontamination of solid biowaste and sharps biowaste;
      • Sterilization of glassware that must be processed in an upright position; and/or
      • Sterilization of dry items that can trap air (e.g., pipette tip boxes).
    • Make sure that bags containing agar plates have only a one-inch opening so that the bag can properly contain melted agar during processing.
    • Do not use this cycle for liquids, media, lighter-weight plastic containers, dry items that will collapse in a vacuum.
  • Burns
    • From contact with hot autoclave chamber surfaces;
    • From residual steam when the door is opened at end of the cycle; and/or
    • Hot fluid scalds from boiling or overboiling liquids/spillage.
  • Hand and arm injuries related to door operation.
  • Injury or death from the explosion of the pressurized chamber.
  • Inhalation or other exposure to fumes/vapors from chemicals that should not have been autoclaved.
  • Exposure to biohazardous materials before autoclaving, or after autoclaving if the material was unsuccessfully decontaminated.
  • Successful completion of the EHS online module Safe Autoclave Use.
  • Hands-on training given by an experienced individual in your facility is also required prior to operating an autoclave due to the serious hazards associated with use of this equipment, and because operating procedures differ from building to building, depending on the specific autoclave provided.
  • Hands-on training must cover the topics provided on the EHS Autoclave Training Record.
  • It is highly recommended that departments which provide autoclaves for research should designate trainers from their pool of people who are experienced in autoclave use, and can knowledgeably provide the hands-on training required for new personnel.
  • Wear required personal protective equipment: Buttoned lab coat, closed-toe shoes, heat-resistant gloves (make sure they are dry; if wet, they transfer heat), safety glasses.
  • Use only autoclavable packaging material and containers.
  • Make sure the autoclave drain is clear of debris before each use.
  • Before operating, check to see if the autoclave is functioning properly (no ABORT message, etc.).
  • Include a verification device such as a chemical Integrator (CI) with every waste load.
  • Adjust closures on bags of solid waste so they have at least a one-inch opening.
  • Check to see that closures on liquid-containing vessels are loosened.
  • Arrange items in the load so they are spaced evenly, allowing room between items (no stacking/ crowding/ overpacking).
  • Make sure containers of liquids are no more than 2/3 full.
  • Load items so that they do not touch each other, or sides and top of the autoclave chamber.
  • Properly place bags of waste and liquid items in autoclavable pans or trays: bags must not overflow the pans; vessels of liquid are in a pan deep enough to contain potential boil-over.
  • Process items for sterilization in separate loads from waste for decontamination.
  1. Sign in on autoclave use log and check that drain is clear of debris.
  2. For a waste load, put on disposable gloves and place a verification device into proper position within the waste, (or in an empty, like-sized container if running a liquid waste load). Only one bag/ container per load needs a device placed in it. After placing the device, adjust bag openings to approximately one inch to allow steam penetration.
  3. Wearing appropriate PPE for burn protection, place load into chamber.
  4. Close chamber door and ensure that it closes completely.
  5. Select and start cycle.
  6. Ensure door has sealed and cycle is successfully underway before leaving the autoclave facility.
  7. Set a personal timer to remind you when your cycle will end.
  1. Ensure that cycle is completely finished and chamber pressure is zero before proceeding.
  2. Wearing PPE, open autoclave door and stand aside to avoid contact with any escaping steam.
  3. Allow load to cool somewhat with door open; let liquids cool in autoclave for at least 15 minutes.
  4. Wearing PPE, carefully remove load from chamber. If liquids begin to boil over, stop and allow more cooling time before removal.
  5. Close autoclave chamber door when finished.
  6. If running a waste load, check the verification result. Results of autoclave verification must be documented; record CI results on autoclave user log, and BI results in BI log.
  • If you see an error message, evidence of an aborted cycle or other operational problems, always record your observations on the autoclave user log, and promptly report them to the person responsible for the autoclave.
  • If you discover that the autoclave is dysfunctional, and/or if you think it is unsafe to use, immediately post an 'Out of Service' sign on the unit and report it to the person responsible for the autoclave. Never attempt repairs on a malfunctioning autoclave.
  • If you feel an emergency situation is developing or has developed with an autoclave, immediately remove yourself and others from the area. Secure the area, and post a sign to warn others to stay away. Immediately report the situation to the appropriate authorities/emergency response providers.
  • Autoclaves at Virginia Tech which achieve >175 PSI or have a chamber volume of 60 gallons/8 cubic feet, which includes most floor models and built-in units, are inspected as pressurized vessels every two years by Virginia state inspectors through the Office of Risk Management. Smaller tabletop models that do not meet these criteria are not subject to this inspection.
  • Current inspection certificates are posted on/at the equipment.
  • University departments are responsible for arranging for this certification. 
  • EHS checks autoclave certification when conducting general building inspections, and will notify building managers and/or department heads if autoclave certifications have expired, or are not present.
  • Autoclaves used for decontamination of biohazardous waste must be regularly tested to verify performance in reaching proper decontamination conditions for hazardous biological waste. This is necessary to ensure that infectious agents in waste are inactivated prior to leaving the university for final disposal.
  • NOTE: Performance verification is highly recommended for autoclaves used to sterilize media, reusable glassware, medical devices, etc. This is considered a best practice and improves the quality and reliability of research data and laboratory operations.
  • Verification testing methods:
    • So-called "autoclave tape" reacts to exposure to heat with a light-to-dark color reaction. It can serve as an easy visual means of identifying items that have been through an autoclave cycle from those that have not. However, this tape it is NOT a definitive indicator of sterilization/ decontamination conditions having been met when an item is autoclaved, and must not be used as a verification testing method.
    • Approved testing devices include Biological Indicators (BIs), and Chemical Integrators (CIs).
    • BIs require incubation to determine results; CIs provide immediate results after a cycle is run.
    • CIs are not as accurate as BIs; however, they are reliable. They are also less expensive.
    • BIs must be used when testing autoclaves:
      • For routine performance verification.
      • Following autoclave installation and repair.
      • When new autoclave cycles are added, or when cycle parameters are changed.
      • When a new load configuration or packaging material is introduced.
    • An autoclave verification program must include both testing methods:
      • Verify autoclave performance monthly with BIs and record results in BI Log.
      • Verify decontamination of each waste load with CIs and record results on Autoclave Use Log.
    • Location of the test BI or CI in a waste load is important. Placement of the test device on the outside of an autoclave bag of solid biological waste will not yield the information needed about conditions on the inside of the bag. Instead, the test device must be placed within the bag, preferably in the center of the waste where steam will have the greatest penetration challenge, but not so deeply buried that it cannot be safely and successfully retrieved after decontamination in the autoclave.
    • Depending on the size of your autoclave chamber, a 'load' of waste in need of decontamination may consist of multiple items (e.g., several bags of waste, or a bag and several sharps containers, etc.). You only need to use one test device per load, not in every bag or container in the load.
    • When decontaminating liquid biological waste:
      • Use BIs that are designed for use with liquid wastes.
      • Place BI ampule directly in liquid for the autoclave run, then retrieve the ampule after the liquid has cooled.
      • DO NOT place CIs directly in liquid; they are not designed for direct use in liquids. Place CI in an empty vessel similar or identical in size to the vessel(s) containing the liquid waste (e.g., bottle, flask, etc.) and cap the vessel with the same type of closure as used on the waste container(s). Then place the test vessel in the autoclave with the waste vessel locating it in load position that will pose the greatest challenge to the CI.
  • BI and CI Verification Testing Procedures

Biological Indicators (BI):

  • Label each BI with the date, autoclave identification, and the cycle type you are testing.
    1. Place the BI inside a bag of solid biological waste. Alternatively, the BI can be placed within a bag of mock waste which approximates the density of actual waste; the mock waste can be run as a test load on its own, or along with other bags of actual waste (depending on the size of the autoclave). Retrieval of BIs from mock waste is often easier, and less messy.
    2. Label one extra BI as a positive control for the lot number being used. This BI will not be autoclaved; it will be activated and incubated to confirm that the bacterial spores of the test organism will germinate under favorable conditions.
    3. Run the autoclave cycle.
    4. Fill out the monthly autoclave verification log.
    5. Upon cycle completion and cooling of the load, retrieve the BI.
    6. Wearing appropriate PPE, follow product instructions to release the culture media in all test vials to put it into contact with the bacterial spores.
    7. Make sure the vial incubator is turned on and set at the recommended temperature. Place vials in the incubator.
    8. Observe and record any color changes at 24 and 48 hours. The positive control should grow (and vial liquid should change color); test vials that have been exposed to successful decontamination conditions in the autoclave should not grow (and vial liquid will not change color).

Chemical Integrators (CI):

  • Make sure your CIs have been stored in a manner that protects them from exposure to light, which can affect their performance.
    1. Check the expiration date on a CI before use.
    2. Wearing appropriate PPE, place a CI within a bag of waste, then adjust the closure of the bag to approximately a one-inch opening. If autoclaving liquid waste, place CI in a like-sized container.
    3. Run the load on an appropriate cycle for the waste type.
    4. When the cycle is complete and load has been removed and cooled, retrieve the CI, interpret the reaction according to information supplied with the CI product.
    5. Record results on Autoclave Use log.
       

BI or CI failure to verify:

  • When BIs or CIs indicate decontamination conditions were not met, an investigation of autoclave performance must take place; the machine must be taken out of service until its status can be determined, and malfunction corrected.
  • Waste that has been autoclaved with a verification device that failed to validate kill conditions must not be discarded, but re-autoclaved in a different autoclave, or held until the cause of the problem is identified and it can be determined that the waste was in fact sufficiently decontaminated.
  • Possible causes of BI or CI failure to verify (other than autoclave malfunction):
    • Test devices "Use By" dates have expired.
    • The wrong kind of test device was used (e.g., a test device designed to test dry heat or gas sterilization equipment); the correct test device will specify that it is meant to be used for steam autoclaves.
    • CIs which have been overexposed to light, completely soaked with water, or crimped/bent during use in an autoclave run can result in their performance being compromised.
    • Steam did not fully penetrate the load because 1) the bag was packed too densely with material; 2) the bag was not opened at least one inch before the decontamination cycle was run; 3) the autoclave chamber itself was packed too densely which prohibited steam penetration.

BMBL
EHS Online Training: Safe Autoclave Use

  • Biosafety Cabinets (BSCs) are specialized pieces of lab equipment designed to safely contain biohazardous agents/ materials. Also, all but one class of BSCs protect sterile items and culture materials from contamination when they are manipulated in the BSC workspace. Different classes and types of BSCs meet different, specific needs. Choosing the right BSC for purchase should be based on a thorough risk assessment of all material (biological and chemical) being handled and the procedures involved with the work. 
  • BSCs must not be confused with chemical fume hoods, laminar flow hoods, or tissue culture hoods. In most cases, BSCs and chemical fume hoods have distinctly different functions and cannot be used interchangeably.
    • Chemical fume hoods utilize directional airflow to protect lab workers from exposure to toxic chemical fumes or particulates by venting them to the outside.
    • Material or items inside a chemical fume hood are not protected from room air contamination. 
    • Chemical fume hoods should never be used for the containment of biologicals.
    • Minute amounts of volatile chemicals/radionuclides may be used ONLY with certain types of BSCs which exhaust them to the outside.

HEPA filters:

  • The HEPA filters are important features of the BSC, capturing potentially infectious particles from your work as well as room air contaminants, and contaminants that you may shed. HEPA filters should be replaced by the BSC's service provider every 3-5 years, depending on cumulative hours of operation, the cleanliness of the lab, and the materials being used in the BSC. Changing or cleaning the pre-filter on a regular basis extends the life of a HEPA filter.
  • The HEPA-filtered directional airflow in a BSC: 
    1. Protects the work material from contaminants,
    2. Protects the worker from exposure to aerosols, and
    3. Prevents the release of aerosolized material into the environment. IMPORTANT: BSC HEPA filters do not entrap or filter chemical vapors or gases; they entrap particulates.

Magnehelic gauge:

  • The magnehelic gauge on the BSC shows the air pressure difference across the HEPA filter and can indicate when the airflow system and filter are not operating properly. 
  • BSC users should know their BSC's acceptable gauge readings and limits; ask your service provider, or look on the BSC certification label for this information. 
  • Before each work session, check the gauge and look for changes higher or lower than this acceptable range. A higher resistance reading indicates the filter is loaded or blocked; a lower resistance reading may indicate a hole or tear in the HEPA filter. In either instance, do not use the BSC; contact a service provider.

SIDE-BY-SIDE BSC CLASS COMPARISON

Classes of Biosafety Cabinets Personnel Protection Product Protection Environmental Protection Use
Class I

Yes

Inward airflow through the sash opening

No

Unfiltered room air is dawn across the work surface.

Yes

Exhaust air is HEPA-filtered.

  • Not in use today for bioagents.
  • May be used to enclose equipment or procedures with aerosol potential.

Class II

A1, A2, B1, B2

Yes

Inward airflow through sash opening.

Yes

By HEPA filtered air drawn down onto work surface & room air kept away.

Yes 

Exhaust air is HEPA-filtered.

  • Mosy common class of BSC used today, esp. Type A2.
  • Used to handle specimen material. biological toxins, cell tissue culture, biohazardous agents/

Class III

(Glove Box)

Yes

Complete containment of interior work area.

Yes

HEPA filtered air is supplied to the work surface; total containment keeps room air out.

Yes

Exhaust air is double HEPA-filtered.

  • Provides the highest level of containment for handling the most dangerous microorganisms.

SIDE-BY-SIDE COMPARISON OF BSC CLASS II TYPES

BSC HEPA-Filtered Work Surface Air  Interior Design Air Inflow Rate Chemicals
II, A1 ALL is RECIRCULATED May have contaminated air under POSITIVE pressure, so if plenum leaks, contaminants will escape into lab. 75 linear feet per minute (lfpm) Use with biologicals; Should NOT be used with chemicals.
II, A2 MOST or ALL is RECIRCULATED Contaminated air under/surrounded by NEGATIVE PRESSURE; if outside exhaust is present, uses flexible connection. 100 lfpm Use with biologicals; reccomended for use with MINUTE amounts of volatile chemicals if some air is exhausted outside.
II, B1 MOST is EXHAUSTED OUTSIDE Contaminated air under/surrounded by NEGATIVE PRESSURE; outside exhaust must be hard-ducted. 100 lfpm Use with biologicals & MINUTE amounts of volatile or toxic chemicals.
II,B2 ALL is EXHAUSTED OUTSIDE Contaminated air under/surrounded by NEGATIVE PRESSURE; outside exhaust must be hard-ducted. 100 lfpm Use with biologicals & SMALL anounts of volatile or toxic chemicals.

C1 biosafety cabinets:

  • The Labconco Purifier Axiom Class II, Type C1 biosafety cabinet is designed to be more flexible than other biosafety cabinets. It can be used as a re-circulating Class II Type A2 cabinet for standard microbiological work, or it can be ducted to the outside (or used with a manifold exhaust system) for working with small amounts of volatile chemicals and radionuclides, as with a Type B2 cabinet. 
  • This cabinet does not have the full functionality of a chemical fume hood, and must not be used as a substitute for a chemical fume hood.
  • Special HVAC requirements may be involved with the installation of this type of cabinet and all manufacturer's recommendations must be followed. 
  • Specific EHS training is required for new users of C1 biosafety cabinets to ensure a full understanding of the limitations and proper uses of the C1 cabinet.

Using volatile chemicals/radionuclides in a BSC:

  • In general,
    • Minute amounts can be handled in Class II Type A2 or B1 BSCs vented to outside.
    • Small amounts can be handled in Class II Type B2 BSCs vented to outside.
  • Specifically,
    • Check the Safety Data Sheets for volatile chemicals to learn explosion limits, and avoid approaching those concentrations, as chemicals can volatilize and concentrate to hazardous levels inside a BSC, or in ductwork after being pulled through HEPA filters.
    • Make sure the volatile chemicals you use will not damage HEPA filters.
  • You have exceeded the quantity of a volatile chemical you can safely use in your BSC if you can smell or otherwise detect chemical fumes out in the lab. In these circumstances, discontinue use of that BSC; instead, locate and use a BSC that exhausts most or all air to the outside.

Purchasing:

  • Choosing the right BSC for purchase should be based on a thorough risk assessment of all material (biological and chemical) being handled and the procedures involved with the work.
  • Key questions to consider:
    • What type and quantity of biological and chemical material will be used in the BSC?
    • What biosafety level (and containment level) will be required?
    • Are you working with biologicals only, or with minute/small amounts of volatile chemicals/radionuclides?
    • What type of exhaust (recirculating, or ducted to the outside) will be needed?
  • Contact EHS (540-231-3600) for assistance in evaluating needs for a BSC purchase.

 

Room location:

  • Room location is important to the proper functioning of a BSC:
    • Ideally, BSCs should occupy lab space that is removed from other work areas, especially high traffic areas. 
    • The cabinet should be placed 12-14 inches from the ceiling and walls.
    • The cabinet should be placed away from windows, air supply vents, the lab features creating air movement (chemical fume hoods, centrifuges, vacuum pumps), and entry points into the lab.

 

Required certification:

  • All new BSCs to be used for handling potentially infectious/biohazardous material (BSL-2, BSL-3; ABSL-2, ABSL-3; ACL-2, ACL-3) must be certified before being used; certification involves a standardized check of proper function, performed by a qualified technician.
  • YEARLY CERTIFICATION IS REQUIRED FOR ANY BSC BEING USED WITH POTENTIALLY INFECTIOUS MATERIALS.
  • Cabinets must be re-certified: 
    1. following a major relocation (change of building, campus, state, country); 
    2. when a HEPA filter is replaced; 
    3. following any other repair or service to the unit.  

BSC certification vendors at Virginia Tech:

  • Principal Investigators can choose from these three providers for cabinet certification services.
  • A Purchase Order (PO) must be obtained through the PI's departmental business office for this service; the procedure for this is outlined in the next item.

Decontamination procedures for BSCs:

  • Biosafety cabinets must be properly decontaminated prior to: 
    • certification technician's arrival
    • any service visit for in situ repairs
    • moves within a building.
  • Prepare a BSC for these events by removing all items and disinfecting all work area surfaces, including vertical side and back surfaces, and sash glass.
  • Gas or vapor decontamination of the entire unit (including inaccessible areas--plenums, etc.) by a qualified contractor is required:
    • prior to a major relocation to another building, campus, state, country, etc.
    • prior to going to surplus 
    • after receiving extensive repairs
    • following a high-volume spill 
    • if the unit will be used in a lower containment lab (e.g., from BSL-3 to BSL-2)
  • Disinfecting agents typically used by professionals include formaldehyde gas and vaporized chlorine dioxide.
  • If you need assistance in determining whether whole-unit decontamination is needed, or for information on scheduling decontamination service with a contractor, contact EHS.
  • Whenever you have decontaminated a BSC (by either method listed above), you must attach a completed Lab Equipment Decontamination Form to the unit prior to the date you are expecting the movers, certification vendor, etc. Please follow the directions on the back of the form.
  • Combinations of 1) laboratory practices and techniques, 2) safety equipment including Personal Protective Equipment, and 3) facilities features that define the conditions used for safe manipulation of infectious agents.
  • Levels of containment range from the lowest Biosafety Level, BSL-1, to the highest at BSL-4 where the most stringent measures are taken to prevent exposure to/ release of biohazardous agents.
  • See a summary of Biosafety Levels in Biosafety in Microbiological and Biomedical Laboratories, Section III.
  • Biosafety Levels should not be confused with Risk Groups. Risk Groups are a means of classification of microorganisms based on their association with, and resulting severity of, disease in humans.
  • The Risk Group of a biological agent should be one of a number of factors used in determining the appropriate Biosafety Level to be employed in working with that agent.
  • Often, the Risk Group number assigned to a microorganism used in research will be the same as the Biosafety Level number at which research with that microorganism is conducted, but not always. In some risk-prone circumstances, the Biosafety Level must be heightened for added protection from exposure/release of a biohazardous agent.
  • NOTE: Risk Groups, as defined below, apply to healthy adults; immunocompromised individuals are more vulnerable to infectious disease and thus could be at greater risk if exposed.
  • See a discussion of Risk Groups in the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules, Appendix B.
  • As defined in the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules, Appendix G, these levels define parameters that serve to 1) contain recombinant or synthetic nucleic acid molecules, 2) confine organisms that contain them, and 3) reduce the potential for exposure of lab workers, persons outside the lab, and the environment to that recombinant material.
  • These containment levels are designated as BL1 – BL4, with the highest BL level defining the most stringent containment measures and protective practices.
  • NOTE: The descriptions of NIH’s Biosafety Levels (BLs), are based on existing approaches to the containment of pathogenic organisms, i.e., biosafety levels (BSLs), as described in Biosafety in Microbiological and Biomedical Laboratories. However, because different combinations of measures may be appropriate for specific research activities involving recombinant studies, The NIH Guidelines allow for alternative selections of primary containment in certain circumstances.
  • While BL levels encompass certain lab practices, containment equipment, and facility design features just as the BSL designations do, the emphasis here is placed on the primary means of physical containment, which are provided by lab practices and containment equipment.
  • See the NIH Guidelines, Appendix G, for details.
  • The practices, safety equipment and facilities features used for experiments with animals involved in infectious disease research or other studies that may require containment.
  • The four levels, designated Animal Biosafety Level (ABSL) 1-4, provide increasing levels of protection to personnel and to the environment, and are recommended as minimal standards for activities involving infected laboratory animals.
  • See BMBL, Section V, for details.
  • These levels specify containment and confinement practices for recombinant research involving larger, often loose-housed animals, including:
    • Those in which the animal’s genome has been altered by the stable introduction of recombinant or synthetic nucleic acid molecules (or DNA derived therefrom) into the germ-line (transgenic animals)
    • Experiments involving viable microorganisms modified with recombinant or synthetic nucleic acid molecules which are tested on whole animals.
  • As defined by The NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules, Appendix Q, these levels apply when research animals are of a size or have growth requirements that preclude the use of containment for lab animals as described in the NIH Guidelines, Appendix G. Such larger species include, but are not limited to, cattle, swine, sheep, goats, horses and poultry.
  • These containment levels are designated by NIH as BL1-N – BL3-N, with the highest BL-N level defining the most stringent containment measures and protective practices.
  • Examples of measures specified by BL-N include:
    • Disposal procedures for animals
    • Animal facility access
    • Decontamination and inactivation procedures
    • Signage
    • Protective clothing
    • Recordkeeping
    • Transfer of materials procedures
    • See NIH Guidelines, Appendix Q, for details.
  • This biosafety level was created for special situations in which high containment is required in an agricultural setting to prevent agent escape into the environment, due to the risk the agent poses in causing animal disease, zoonotic disease, and/or serious economic impact.
  • BSL-3Ag describes facility parameters and work practices used for research on high-risk pathogens and large, loose-housed, agricultural animals, or other similar situations, where the facility barriers serve as primary containment.
  • Examples of measures prescribed by BAL-3Ag are:
    • Personnel change and shower rooms.
    • Self-closing and lockable access doors.
    • Double-door autoclaves (all waste is decontaminated before leaving containment).
    • Dedicated, directional air flow with pressure gauges and alarmed monitoring.
    • HEPA filtered supply and exhaust airflow.
    • Liquid effluent is collected and decontaminated in a central liquid waste system before disposal into sanitary sewers
    • Interior surface penetrations are sealed; ductwork is airtight; doors are sealed to airtight requirements
    • Plant research has sometimes been placed under this designation.

For more information, refer to the American Committee of Medical Entomology of the American Society of Tropical Medicine and Hygiene Arthropod Containment Guidelines, Version 3.1.

  • These levels address the unique containment challenges presented by arthropods and provide increasingly stringent measures for standard and special practices, equipment providing primary barriers, and facilities providing secondary barriers.
  • ACL 1-4 are designed to be consistent with the NIH Guidelines for recombinant research, and with BMBL for research involving arthropod-transmitted microbiological pathogens
  • Examples of containment measures prescribed by these levels: 
    • Use of appropriately-sized screen mesh;
    • Effective trapping methods for escaped arthropods;
    • Specific waste handling procedures;
    • Limits on lab access; and
    • Primary containment provided by glove box, walk-in incubator, screen cubicle, etc.
    • See Table 1. Summary of Arthropod Containment Levels in Arthropod Containment Guidelines, Version 3.1, p. 50.

For further information, see:
Arthropod Containment Guidelines, Version 3.1
BMBL, Appendix E.

NOTE: Plant growth for research carried out in the laboratory (in tissue culture rooms, in growth chambers, at the bench, etc.) should be conducted under the guidelines of the appropriate Biosafety LevelPlant Biosafety Levels are specific to greenhouse settings.

  • The principal purpose of plant containment as defined by these levels is to protect the environment from the transmission or release of recombinant or synthetic nucleic acid molecule-containing plant genomes. These levels specify greenhouse practices and greenhouse facility features for physical containment of experiments using recombinant plants, plant-associated microbes, and small animals: 
    • As defined in NIH Guidelines, Appendix P, applicable plants include (but are not limited to) mosses, liverworts, macroscopic algae, and vascular plants including terrestrial crops, forest, and ornamental species.
    • Plant-associated microorganisms include viroids, virusoids, viruses, bacteria, fungi, protozoans, certain small algae, and microbes that have a benign or beneficial association with plants, including those which are being modified to foster an association with plants.
    • Plant-associated small animals include 
      • Arthropods that are in obligate association with plants are plant pests or pollinators, that transmit plant diseases,
      • Other small animals such as nematodes for which testing of their biological properties require the use of plants. 
  • Plant biosafety levels BL- 1P – 4P for greenhouse research, as described in NIH GuidelinesAppendix P, are summarized below:

PLANT BIOSAFETY LEVEL

APPLIES TO:

Biosafety Level 1P

Research plants that have low environmental risk; no evidence of harmful impact, survival or spread.

Biosafety Level 2P

Research plants that could live in the environment with negligible impact, or impact could be readily managed. Includes transgenics capable of interbreeding with weeds/related species, transgenics that use the entire genome of an indigenous infectious agent/pathogen, and plant-associated, indigenous or exotic microbes that are potentially harmful, but harm could be managed.

Biosafety Level 3P

Research plants that have the potential for detrimental impact on the environment (exotic infectious plants, transgenic plants, transgenic microbes).

Biosaftey Level 4P

Plant research that involves readily transmissible infectious agents that are potentially serious pathogens for major US crops.

  • Examples of criteria to be used for Plant Biosafety Levels include:
    • Greenhouse access
    • Recordkeeping requirements
    • Decontamination and Inactivation requirements
    • Facilities requirements/ greenhouse design
    • Signage
    • Transfer of materials requirements

For additional details, see A Practical Guide to Containment – Plant Biosafety in Research Greenhouses.




Click here to learn more about bloodborne pathogens and exposure control plans.

Choosing a disinfectant:

Choose the most effective disinfectant(s) for your lab based on the following criteria:

  • The type of biohazardous materials you are working with (fungal/ bacterial/ vegetative vs. spore formers, etc.) and their risk assessments.
  • The degree of contamination you typically encounter.
  • Whether organic material is/could be present, which reduces the effectiveness of some disinfectants.
  • How the disinfectant works chemically, and in what quantity and concentration.
  • What contact time and temperature are needed for the disinfectant to be effective.
  • How the disinfectant affects materials, such as corrosiveness, or leaving a residue on surfaces.
  • What environmental impact it has, such as toxicity, creating noxious fumes, or being an irritant for the user.
  • What sort of shelf life it has.
  • How expensive it is.

Labs working with different biohazardous materials may find it necessary to stock several disinfectants to supply effective decontamination for all agents. See the following chart, The Antimicrobial Spectrum of Disinfectants, for general information on common types of disinfectants used in labs. (The Center for Food Security and Public Health, Iowa State University)

Bleach:
  • Bleach is the common, product name often used for this disinfectant. Sodium hypochlorite is the active ingredient in bleach that actually disinfects.
  • Bleach is an effective disinfectant but a poor cleaner. When both cleaning and disinfecting are needed, they must be done in a two-step process, where cleaning with a detergent is performed and allowed to completely dry prior to disinfection with bleach. Bleach is a reactive material that can form toxic compounds when mixed with other cleaning chemicals. Therefore do not mix bleach with a cleaner in an attempt to produce a one-step cleaner/disinfectant.
  • Bleach solutions are inactivated by the presence of organic matter, so physically remove organic material before disinfecting surfaces with bleach.
  • CDC recommendations for contact time: Wait ≥ 10 minutes after application of bleach solution before wiping nonporous surfaces; wait 30 minutes if immersing in a bleach solution.
  • Bleach solutions are corrosive to metal surfaces and the skin. When disinfecting metal surfaces with a bleach solution, follow it with a sterile water rinse to avoid corrosion.
  • Contact with bleach can degrade disposable gloves.
  • Never use bleach in the presence of formaldehyde.
  • Never mix bleach with ammonia, as toxic chlorine gas will be released.
  • Bleach disinfectant solutions must be made up fresh weekly, if not daily, as the effectiveness of sodium hypochlorite decreases rapidly with time.
  • Expiration dates of undiluted bleach must always be checked; up-to-date stocks must be maintained in the laboratory.
  • In the U.S., bleach designated for general purpose or household use was previously formulated between 5.25% and 6% (industrial strength bleaches are often formulated at concentrations greater than 20%). Leading manufacturers such as Clorox are now producing household bleach at 8.25%, sometimes called "Ultra".

 

Using bleach as a disinfectant:

Purpose Dilution of Standard Household Bleach (min. 5.25% sodium hypochlorite)  % Sodium Hypochlorite (NaOCl / ppm) Precautions
Spills of material with large amounts or concentrations of organic matter (e.g., blood), liquid media 1:5 dilution

1 part bleach + 4 parts water or contaminated liquid

(e.g., 20 mls bleach + 80 mls water/media/contaminated liquid)
1% NaOCl

(10,000 ppm)
  • Bleach-based disinfectants can cause skin, eye, and lung irritation. Always wear appropriate PPE. 
  • Skin and eye protection must be worn when handling undiluted bleach solution.
  • Make sure you are in a well-ventilated area.
Surfaces with large amounts or concentrations of organic matter 1:10 dilution

1 part bleach + 9 parts water (e.g., 10 mls bleach + 90 mls water)
.5% NaOCl

(5,000 ppm)
  • Bleach-based disinfectants can cause skin, eye, and lung irritation. Always wear appropriate PPE. 
  • Skin and eye protection must be worn when handling undiluted bleach solution.
  • Make sure you are in a well-ventilated area.
Surfaces with low amounts or concentrations of organic material 1:50 dilution

1 part bleach + 49 parts water (e.g., 2 mls bleach + 98 mls water)
.1% NaOCl

(1000 ppm)
  • Bleach-based disinfectants can cause skin, eye, and lung irritation. Always wear appropriate PPE. 
  • Skin and eye protection must be worn when handling undiluted bleach solution.
  • Make sure you are in a well-ventilated area.
Liquid waste decontamination

(e.g., aspiration flasks, etc.)
1:5 dilution

1 part bleach + 4 parts water or liquid containing biohazardous material
1% NaOCl

(10,000 ppm)

 

 

 

Ethanol/isopropanol:

  • The disinfecting ability of ethyl alcohol and isopropyl alcohol drops sharply when either is diluted below 50% or at dilutions higher than 90%. Optimum disinfection occurs at 70% in solution with water. Reason: Alcohol’s mode of action as a disinfectant is protein denaturation, and water supports the denaturing of proteins. Because pure alcohol is very dehydrating to microbial cell walls (which can interfere with protein denaturation) the presence of a certain amount of water in alcohol more readily denatures microbial proteins.
  • A minimum contact time of 10 minutes is necessary for disinfection using alcohols.
  • Air-drying of alcohol on surfaces (vs. wiping surfaces dry) can be practiced, due to the fact that alcohols do not leave residue behind. In many instances, air-drying is the preferred method.
  • Avoid spraying/applying alcohols on surfaces too thinly, resulting in quick evaporation and not enough contact time to achieve disinfection. 
  • Frequently spraying disposable gloves with alcohol can degrade gloves and increase their permeability to biological agents.

Contact time and wiping technique for surface disinfection:

  • After applying a disinfectant to a surface, it is critical that you wait for the specified contact time to allow the active ingredient(s) in the disinfectant to be effective in killing microorganisms on the surface. Simply applying a greater volume of disinfectant to the surface does not lessen the contact time needed.
  • Wipe a surface in a grid pattern, proceeding in one direction that contacts each sector only once, vs. repeated circular wiping that can re-introduce dust, contaminants, etc. on your newly disinfected surface.

 

Dual use research of concern (DURC) overview:

  • If proposed research could potentially provide knowledge, information, products or technologies that, if misapplied, could pose a significant threat with broad consequences to public health and safety, to agricultural crops and other plants, to animals or the environment, or to national security, then it qualifies as DURC.
  • The U.S. government has designated the following agents as potentially involving DURC: 
    • Avian influenza virus
    • Bacillus anthracis
    • Botulinum neurotoxin
    • Burkholderia mallei
    • Burkholderia pseudomallei
    • Ebola virus
    • Foot-and-Mouth Disease virus
    • Francisella tularensis
    • Marburg virus
    • Reconstructed 1918 flu virus
    • Rinderpest virus
    • Toxin-producing C. botulinum
    • Variola major virus 
    • Variola minor virus
    • Yersinia pestis
  • The U.S. government has designated these experiments as potentially involving DURC: 
    • Enhancing the harmful consequences, or altering the host range/ tropism of an agent or toxin
    • Disrupting immunity or effectiveness of immunization against an agent or toxin without justification
    • Conferring resistance to an agent or toxin to interventions, or facilitating its ability to evade detection
    • Increasing stability, transmissability, or ability to disseminate of an agent or toxin
    • Enhancing susceptibility of a host population to an agent or toxin
    • Generating or reconstituting an eradicated or extinct agent or toxin listed above
  • If your research involves the use of one or more of the agents and experiments listed, it may be considered DURC. DURC Researchers must comply with the U.S. Government Policy for Institutional Oversight of Life Sciences Dual Use Research of Concern. The government’s oversight of DURC aims to preserve the benefits of the life sciences while minimizing the risk of misuse of this research.

When your research involves DURC:

  1. You must submit a research protocol to the Virginia Tech IBC for review.
  2. The AVP for the Office of Research Compliance, the IBC, and the Office of Export and Secure Research Compliance will meet with you to discuss the research.
  3. A mitigation plan will be drafted by you and those listed above, in accordance with the U.S. Government Policy for Institutional Oversight of Life Sciences Dual Use Research of Concern. The draft plan will be submitted to NIH and/or your funding agency for review and approval.
  4. You will be provided with information and guidance by those listed above throughout the planning and approval process.
  5. Your research cannot be initiated until your research protocol and risk mitigation plan is approved.

 

  • Field research often takes place in locations where native flora/ fauna/ parasitic and microbial agents can present novel health hazards, especially to non-native people. Medical care can be difficult to obtain, and/ or made more complicated to obtain due to language barriers, remote field locations, etc. You should develop a field research safety plan and assure all participants have been provided this information.  Further guidance can be found on the Field Research webpage.
  • Consult with the EHS Occupational Health Program on appropriate prevention and treatment strategies, vaccinations and so forth AT LEAST SEVERAL MONTHS before you begin your field work. The Occupational Health physician can recommend specific first aid provisions, and supply medication such as anti-malarials. Consultation with your primary care physician is also recommended.
  • Safety training that can be customized for your needs is available through EHS for groups or individuals who will travel to remote areas for field research. Contact EHS at 540-231-3600. 
  • Contact individuals in your field or department who have traveled/ worked in your destination area in order to gain critical knowledge regarding: 
    • local flora/fauna hazard awareness
    • routine and emergency communications, transportation, medical/emergency treatment available locally
    • language and culture of local populations
  • Review the important information provided at Office of Export and Secure Research Compliance. This site includes links to the U. S. State Department.
  • Review the U.S. State Department's travel information about your destination area, and know the location of the nearest U.S. Embassy. Register your travels online using the Smart Traveler Enrollment program on the State Department's website. This registration will provide you with information updates on your destination.
  • Learn about current health risks that could be encountered at your destination by going to the "Destinations" page on the Centers for Disease Control and Prevention website.
  • For students participating in study-abroad programs, register with Virginia Tech's Global Education Office. See their website for valuable travel advice and information, and review Virginia Tech's Global Travel Policy.

Procedures for lab personnel:

  1. COMPLETE THE Lab Equipment Decontamination Form WHEN:
    • Lab equipment needs to be certified or serviced by technicians, or by Facilities staff.
    • Lab equipment requires relocation or transport by VT movers, hired movers or VT Surplus.
  2. ACTIONS TO TAKE:
    • Clean all dirty equipment surfaces that you can access with a detergent solution.
    • Disinfect/ decontaminate accessible inner and outer surfaces according to hazards present.
    • Clean and disinfect/decontaminate the area around the unit, if needed.
  3. WHEN YOU CAN'T MOVE HEAVY EQUIPMENT TO CLEAN AROUND IT:
    • Be present when service providers/movers arrive and ask them to reposition the unit.
    • Clean/decon equipment surfaces and floor area that become accessible after the unit is moved.
  4. COMPLETING THE FORM
    • Fill out all required sections (1st block) and all applicable sections (2nd block) of this form after cleaning/decontamination is complete.
    • Attach the completed form to the equipment, and keep a copy with your equipment's service records.
  5. PROCEDURES FOR SPECIFIC HAZARDS:
    • Chemicals -- Deactivate all hazardous chemicals by appropriate methods, esp. mutagens, corrosives, toxins, mercury, or other metals.
    • Biological Agents -- Use an appropriate disinfectant for the full contact time.
    • Radioactive Materials -- Decontaminate using the appropriate methods. Equipment must be wipe tested by the person performing the decontamination and found free of radioactive contamination.
  6. IMPORTANT:
    • Equipment lacking a completed, attached form will not be serviced or transported.
    • In addition, equipment displaying a Decontamination Form will not be serviced or transported if it is found to be in a questionable state of safety or cleanliness. Service providers/movers can suspend their activity until such time as the unit and/or the area around the unit has been sufficiently cleaned and decontaminated/disinfected.

Procedures for Service Providers/Movers/Surplus Property
(Also supplied on the reverse side of form)

  1. WHEN MOVING EQUIPMENT:
    • Put on PPE and reposition large or heavy units, if necessary, so lab workers can finish cleaning.
    • Check that the Decontamination Form for the equipment is present and completely filled out.
    • Check that equipment, equipment location, & your work area are clean and free of hazards before moving the equipment.
    • Keep the Decon Form and attach it to your work order documents or Surplus Property Report.
    • Wear appropriate PPE when handling/ moving equipment (minimum: disposable gloves).
  2. Report any problems to your supervisor and/or the responsible party listed on the form; do not proceed with the work order until the situation has been corrected.
  3. In the unlikely event that tools or moving equipment come into direct contact with hazardous materials in the lab, ask lab personnel to apply appropriate disinfecting/decontaminating agents.

EHS Biosafety Officers perform biosafety lab inspections; these inspections are conducted using checklists that evaluate biosafety compliance according to federal, state and university requirements. 

  • The results of these inspections are reported to the PI, IBC administrators, and members of the IBC. In some instances they are also reported to department heads and facility coordinators/managers.
  • Biosafety lab inspections are required:
    • As part of the initial IBC approval process for a research project.
    • As part of a 3-year renewal of IBC-approved projects.
    • When a major amendment or change is proposed for a research project.
    • When a lab moves to another physical location on campus.
  • These inspections cover many of the same safety elements as general building inspections, but focus closely on biohazard containment methods, equipment and facilities features, handling procedures of hazardous biological agents/materials, and biosafety training compliance. 
  • The EHS Biosafety Office and the IBC Administrators work collaboratively with PIs and lab personnel to encourage and support the correction of inspection violations in a timely manner, and to see that biosafety compliance is maintained.
  • The IBC requires modifications of research protocols to address violations found upon inspections. 
  • Principal Investigators typically are given 30-45 days from receipt of notification to respond to required modifications for correction of biosafety inspection violations. Violations and deficiencies must be corrected before IBC protocol approval/ renewal is granted, and research can begin or continue.
  • Criteria for EHS inspections are based upon federal regulations, state regulations and university safety standards, and are conducted using comprehensive checklists.
  • EHS personnel conduct general building inspections of all Virginia Tech facilities on a two- or three-year cycle, based on relative safety risks of the activities that take place in the building. In general, these inspections focus on fire safety, electrical safety, physical hazard safety, proper hazardous materials storage, etc.
  • General building inspections include offices, classrooms, laboratories, storage rooms, mechanical rooms, conference and break rooms, classrooms, hallways, stairwells, auditoriums, shops, custodial closets, electrical closets, etc.
  • Reports for general building inspections are submitted to building administrators and/or department heads; violations must be addressed in 30-45 days. Violations which fail to be addressed in that timeframe will be reported to deans and/or other responsible administrators by EHS.

When EHS conducts a general building inspection, a lab safety inspection of each PI laboratory in the building is also performed.

Just as with the building inspection, the criteria for these lab inspections are based upon federal regulations, state regulations and university safety standards, and are conducted using comprehensive checklists.

  • Reports for lab safety inspections are submitted to the PIs who oversee those labs; violations must be addressed in 30-45 days.

Laboratory personnel can use a self-audit checklist to conduct their own inspections as a way to prepare for an EHS inspection, and to maintain regulatory compliance as well as efficient, safe operations. This document is based on the checklists used by EHS inspectors. Comprehensive Self Audit Checklists can be found using this link, or on the IBC website; look under 'IBC Offline Documents.'

  • Reportable lab incidents are unwanted occurrences for which you must:
    1. contact the appropriate response providers FIRST (if there is a serious injury, major toxic spill, etc.), and THEN
    2. report the incident to EHS.
  • Examples of reportable lab incidents:
    • Accident;
    • Injury;
    • Spill – chemical, biological, or other hazardous material;
    • Serious lab equipment failure;
    • Facilities failure (HVAC, plumbing, gas line, etc.);
    • Fire;
    • Security breach or failure;
    • Theft;
    • Threat;
    • Loss of containment/ release into the environment of biohazardous or recombinant material;
    • Exposure to biohazardous agents/materials, or to hazardous chemicals;
    • Non-compliance with regulations; and
    • Any combination of these.

IMPORTANT: EHS is not a first response unit. Responders who provide medical aid, firefighting expertise, policing or security enforcement, or large-scale hazardous spill mitigation must be contacted first in emergencies, according to the nature of the emergency.

The role of EHS is to provide:

  • Subject matter/ safety experts in biological, chemical, radiological, electrical, physical and other hazard areas who can advise and consult with responders;
  • Incident investigations;
  • Required reporting to federal and state agencies; and/or
  • An interface with regulatory agencies during incident response and follow-up.
  • Give priority to addressing injuries or medical emergencies, then direct efforts to containment and cleanup needs. If the situation is life-threatening, treatment must be obtained in the Emergency Department of the nearest hospital; DIAL 911.
  • Elapsed time can be critical after exposure to infectious agents or chemicals. Act quickly on behalf of the victim so that prophylactic medications for pathogens, or proper treatment for chemical exposure can be given as soon as possible to lessen infection risk or toxic effects.
  1. Immediately inform coworkers of the exposure incident.
  2. Remove PPE and provide immediate care to the exposure site:
    • Wash exposed skin/ wound with soap and water for 15 minutes.
    • Flush eyes/mucous membranes with fresh water for 15 minutes.
    • Call 911 if serious injury has occurred; administer first aid as needed.
  3. Tell your supervisor about the exposure incident as soon as possible, even if it was a potential exposure.
  4. Visit a medical provider or Emergency Department for evaluation within 1-2 hours of the incident. Inform the medical provider about the specific material to which you have been (or may have been) exposed so that any appropriate post-exposure treatment can be considered and/or implemented.
  1. As soon as any initial response is complete and the incident is stable, immediately notify:
    1. The Lab Director and/or Lab Manager
    2. The Animal Facility Manager/ Greenhouse Manager (if applicable)
    3. A Biosafety Officer (BSO) by telephone (preferred) or email.
    Biosafety Officers Telephone Email
    Charlotte Waggoner 540-231-5864 ren@vt.edu
    Anna Kroner 540-231-1122 akroner@vt.edu
    Michael Miles 540-231-3361 msmiles@vt.edu
  2. The BSO will acknowledge receipt of notification by communicating to the reporting person via phone or email, and will begin notifying other appropriate personnel and/or agencies.
  3. IMPORTANT: If the incident involves a known exposure to recombinant material (e.g., rDNA/ synthetic nucleic acids/ transgenic or genetically modified organisms/ select agents), the BSO must inform NIH and/or CDC without delay, thus IMMEDIATE reporting is required.
  4. The reporting person and the supervisor of the facility (e.g., Lab Director/ Lab Manager/ Animal Facility Manage/ Greenhouse Manager) must complete a Virginia Tech Lab Incident Report and submit it to the BSO via email (preferred) or campus mail (MS 0423) as soon as possible.
  5. BSO will acknowledge receipt of this report via email.
  6. If an injury or exposure has occurred, an Employer's Accident Report must also be completed immediately by the supervisor per directions found on the link webpage.
  7. If the supervisor does not complete the report in a timely manner, injured/exposed individuals are encouraged to complete the Employer’s Accident Report themselves.

While working with biohazardous materials in this laboratory, personnel must tie back long hair and wear:

  • Closed-toe shoes;
  • Street clothing that fully cover the legs;
  • Disposable or cloth lab coat; and
  • Disposable gloves of appropriate type for work being done.
  • If determined by risk assessment, the following may be required:
    • Eye and face protection (e.g., goggles/safety glasses, face shield or another splatter guard);
    • Hair covers;
    • Disposable sleeves;
    • Shoe covers; and
    • Respirators.
      NOTE: Use of respirators requires enrollment in the Respiratory Protection Program.


PPE use, disposal, and decontamination:

  • Gloves:
    • Consider all the hazards your gloved hands may contact during your work session: chemical, biological, radiological, sharps, animals, cryogenic items, heat, and combinations of these.
    • Do not use gloves with talc powder; these are no longer commercially available but may be encountered in previously purchased stock. The talc used with disposable gloves has been shown to be extremely hazardous to people who react allergically to it.
    • Consult a glove permeation/degradation chart for your preferred vendor (many are online) and select gloves made from the material that has the longest breakthrough time for the hazards you will contact, or are specifically engineered to withstand the hazards you may encounter.
    • Never use regular disposable gloves for heat or cold protection; use insulated gloves.
    • When autoclaving, make sure your insulated gloves are not wet; wet gloves transfer heat and will not protect your hands.
    • Be sure your gloves fit well for maximum safety—always use the right size of the glove.
    • Gloves must be changed when contaminated when integrity has been compromised, or when otherwise necessary.
    • Disposable gloves must not be washed or reused.
    • Be aware that frequent spraying of gloved hands with a disinfectant can increase permeability to biohazards and chemicals with some glove materials.
    • Contaminated disposable gloves must NOT be disposed of in regular trash that housekeepers empty. They must be discarded into lab solid waste containers. 
    • Glove removal video
    • Glove removal written instructions:
      • Pinch one glove at wrist level and peel it off the hand without touching your skin, allowing the glove to turn inside out.
      • Holding the removed glove in the gloved hand, and slide fingers of ungloved hand between the glove and the top of the wrist. Remove the second glove by rolling it down the hand, and fold it into the first glove so that it is captured within, and the outer glove is inside out.
      • Holding an inside surface of the combined gloves with your bare hand, discard them into solid biological waste. Wash hands immediately.
  • Lab coats:
    • Lab coats must have closures fully fastened to protect from contamination; button your lab coat.
    • Ensure that there is no gap between the lab coat cuff and the glove cuff—no exposed skin. Use disposable sleeves if necessary. Tuck the cuff of the lab coat sleeve into the glove cuff.
    • Disposable lab coats/gowns:
      • Best suited for higher biosafety level work, especially back-closing gowns.
      • Normally discarded after a single use; dispose of in the solid biological waste.
      • Limited re-use can be allowed, depending on agents/ procedures used, PI discretion, and if the coat/gown is intact, unsoiled, and not contaminated.
    • Cloth lab coats:
      • Best suited for BSL-1 and general bench work.
      • If the coat is intact, unsoiled, and not contaminated, it can be re-used for a period of time determined by the PI.
        • Cloth Lab Coat Decontamination And Laundering For Re-Use
          • BSL-2 cloth lab coats must be decontaminated by autoclaving or other approved method prior to laundering; decontamination by autoclaving is recommended for BSL-1 lab coats.
          • Collect soiled lab coats in a dedicated, labeled, a lidded container with Biohazard label until they can be decontaminated; line the container with an autoclave bag prior to collection of lab coats.
          • Autoclave lab coats in a clear autoclave bag, or label an opaque bag well so contents will not be mistaken for biowaste.
          • Decontaminate lab coats using an autoclave Solid or pre-vac cycle using a short cycle time; use a Chemical Integrator (CI) for load verification.
          • It is strongly recommended that 1) decontaminated lab coats are laundered by a professional service, and 2) that individuals do not take them home to launder them after autoclaving.
  • Eye/face protection:
    • Persons who wear contact lenses in laboratories must also wear eye protection if their work is not confined to a BSC, chemical fume hood, or behind a bench shield that blocks potential splashes to face. Eye protection can be provided by tight-fitting safety goggles or safety glasses with side shields.
    • Evaluate the splash/droplet potential for exposure to eyes, nose, and lips; wear a face shield over safety glasses/goggles to be fully protected.
    • Protect eyes from UV radiation by using face shields, safety goggles, equipment eye shields, etc. that have the ANSI Z87.1 safety rating imprinted on the browband or earpiece.
    • Pull down the glass sash on the BSC when UV is on to protect the eyes.
    • Non-disposable safety glasses, goggles, face shields, etc. must be decontaminated and cleaned after each use prior to storage.

Further information and guidance:

BMBL
University Biosafety Manual

  • Respiratory protection may be required when personnel:
    • Are working with a highly infectious respiratory pathogen, even in containment;
    • Are working out of containment in circumstances where exposure to an agent capable of causing respiratory infection is possible;
    • Must be fully protected from a potential respiratory pathogen when responding to a biological spill; and
    • Working with potential sources of allergens (animal dander, mold spores, etc.) are at risk of an allergic reaction, or of developing serious allergies.
  • Respiratory protection for biohazards must filter microscopic particulates from breathed air; typically, they do not remove chemical fumes/vapors. Respirators for chemical hazards typically have canisters that scrub air for specific fumes and vapors; only certain categories of canisters filter out inorganic particulates. Thus, respirators worn for chemical protection usually do not offer biohazard protection.
  • A need for respiratory protection to prevent exposure to biohazards is determined through the Occupational Health Services review of your medical questionnaire, through an IBC protocol review for proposed research, or it may also be initiated by a Principal Investigator. In any case, you will be notified by EHS and given guidance on the next steps to follow for provision of your respiratory protection. You must be medically cleared by EHS before you are fit tested unless you are wearing an N95 respirator on a voluntary basis. Voluntary users must comply with the requirements in the Respiratory Protection Program.
  • Respiratory protection may also be used in instances where, although not occupationally required, an individual feels more comfortable working with added protection. In this instance, respiratory protection can be provided at the discretion of the Principal Investigator who employs/supervises the voluntary user.
  • When respiratory protection is required for biohazardous work, the individual must participate in the Respiratory Protection Program, which involves:
    • A pulmonary function test conducted by EHS Medical Services staff.
    • A quantitative fit test, if a filtering facepiece respirator is to be used, to ensure proper fit.
    • Respiratory protection training (provided as an EHS online training module).
    • Annual renewal of fit test and training.

EHS approved respirators used in the presence of biohazards fall into two categories:

  1. Disposable N95, N100 filtering facepiece, two-strap respirators.
    (NOTE: P-series filtering facepiece respirators protect the user from oil droplets as well as particulates; oil is seldom a concern in a biolab, but P series disposable respirators can also be worn in these labs to protect against inhalation of biohazardous material).
  2. Non-disposable loose-fitting powered air-purifying respirators (PAPR) that are equipped with HEPA (P100 or N100) cartridges.
                              Filtering Facepiece Respirators Powered Air-Purifying Respirators
Protection for: Aerosols of infectious agents Aerosols of infectious agents
No protection for: Chemicals, gases, vapors Chemicals, gases, vapors
How it works: Upon inhalation, air filtered through microfiber body of mask; filtering protection is completely dependent on creating & maintaing a tight seal on face. Hood is equipped with battery-powered blowers which forces air through particulate filters & supplies it to user.
Disposable? Yes (Re-use only 1-2x in specific circumstances that have been approved by EHS) No -- Reusable
User-friendly? Takes work (creates negative pressure) to breathe through mask, but is lighter and less bulky to wear; requires fit-testing; cannot be worn with facial hair (beards). Easier to breathe (utilizes positive pressure), but is bulkier & heavier to wear; loose-fitting PAPRs can be worn with beards & require no fit-testing.
Special requirements Must be replaced when discolered, damaged or clogged; must be disposed of as biohazardous waste. Requires a fully charged battery to work properly; must be disinfected & stored properly after each use; must be inspected before each use.
  •  Note that PAPR can be used with combination (stacked) cartridges that protect against chemicals and other agents if needed. Please contact EHS at respirators-g@vt.edu for assistance.
  • Surgical masks, single-strap dust masks, and bandanas are NOT approved to provide respiratory protection against biohazardous agents, aerosols, etc.

 

  • Risk assessment is an important responsibility for directors and Principal Investigators of microbiological and biomedical laboratories [or in other disciplines utilizing biohazardous agents/materials in their research]. Institutional biosafety committees (IBC), animal care and use committees, biological safety professionals and laboratory animal veterinarians share in this responsibility.
  • Risk assessment is a process used to identify:
    • The hazardous characteristics of a known infectious or potentially infectious agent or material,
    • The activities that can result in a person’s exposure to an agent,
    • The likelihood that such exposure will cause an LAI [lab-acquired infection],
    • And the probable consequences of such an infection.
  • The information identified by risk assessment will provide a guide for the selection of appropriate biosafety levels and microbiological practices, safety equipment and facility safeguards that can prevent LAIs.
  • Laboratory directors and Principal Investigators should use risk assessment to alert their staff to the hazards of working with infectious agents and to the need for developing proficiency in the use of selected safe practices and containment equipment. Successful control of hazards in the laboratory also protects persons not directly associated with the laboratory, such as other occupants of the same building, and the public.
  • Risk assessment requires careful judgment. Adverse consequences are more likely to occur if the risks are underestimated. By contrast, imposition of safeguards more rigorous than actually needed may result in additional expense and burden for the laboratory, with little safety enhancement. An unnecessary burden may result in the circumvention of required safeguards. However, when there is insufficient information to make a clear determination of risk, it is prudent to consider the need for additional safeguards until more data are available.
  • The primary factors to consider in risk assessment and selection of precautions:
    • Agent hazards, and
    • Laboratory procedure hazards.
  • In addition, the capability of the laboratory staff to control hazards must be considered. This capacity will depend on the training, technical proficiency, and good habits of all members of the laboratory, and the operational integrity of containment equipment and facility safeguards.

For Principal Investigators at Virginia Tech, the process of completing an IBC protocol for review and approval is designed to serve as a detailed risk assessment.

Further information and guidance:

BMBL
IBC
EHS Biosafety Group

  • Risk Groups are a means of classification of microorganisms based on their association with, and resulting severity of, infectious disease in humans.
  • The Risk Group of a biological agent should be one of a number of factors used in determining the appropriate Biosafety Level to be employed in working with that agent.
  • Risk Groups are not to be confused with Biosafety Levels. Often, the Risk Group of a microorganism used in research and the Biosafety Level at which that research is conducted have the same number, but not always. In some risk-prone circumstances, the Biosafety Level must be heightened for added protection from exposure/release.
  • NOTE: Risk Groups, as defined below, apply to healthy adults; immunocompromised individuals are more vulnerable to infectious disease and thus could be at greater risk if exposed.
  • See more details on Risk Groups, as described in the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules, Appendix B.
  • As more research groups work in open or shared laboratory spaces and use common equipment, biohazard information must be shared across the groups for the safety of all.
  • What you can do if you work in shared lab spaces:
    • Do not assume that the biosafety measures you employ for the agents/materials you work with will be appropriate for the agents used by other lab groups. Biosafety measures are agent-specific.
    • Always thoroughly clean and decontaminate shared equipment and surfaces that you use.
    • Ensure that you receive awareness training on the specific agents/materials used by others in shared spaces. Training must include pertinent information about the agents, procedures, equipment used, disinfectant needed, spill response, etc.
    • Principal Investigators or their designees are required to provide awareness training to all others who share workspaces and equipment with their group. If you begin work in a facility that has shared work spaces and you do not receive initial or periodic awareness training as needed, contact your PI, EHS or the IBC.

Further information and guidance:

IBC
EHS Biosafety Group

  • Do not attempt to ship biohazardous/ recombinant materials or agents, chemicals, or radiological materials. Contact EHS when you have these shipping needs so that certified, trained shippers can ensure all packaging and documentation regulations are followed. Significant financial penalties can result from non-compliance with shipping regulations.
  • Contact an EHS Biosafety Officer if you need to ship biohazardous materials to domestic or international destinations:
    BIOSAFETY OFFICERS 8 am - 5 pm  
    Michael Miles 540-231-3361 msmiles@vt.edu
    Anna Kroner 540-231-1122 acastigl@vt.edu
         
  • Certified shippers in the Biosafety group will package and ship biohazardous materials in accordance with international/ federal/ state regulations. They perform this task for the university community and supply the outer package materials.
  • Make contact with the EHS Biosafety Group at least 3 working days before your preferred shipment date; allow longer for international shipments.
  • Shipments should be mailed on Monday, Tuesday or Wednesday to expedite reaching their destination within that business week, which is particularly important for shipments on dry ice that could warm if delivery is delayed over a weekend.
  • A certified shipper of hazardous materials at EHS will work with you to prepare the shipment:
    1. The sender completes a Biohazardous Shipping Request Form and submits it to the EHS certified shipper, who 1) determines how the shipment must be classified according to biosafety level, presence of chemicals/preservatives, etc., and 2) schedules a shipment date with the sender.
    2. EHS certified shipper discusses specifics with the sender: 1) sender must obtain needed permits prior to shipment; 2) sender must arrange for payment of the vendor; 3) sender must provide cold packs or dry ice in appropriate quantity, if needed, at time of packaging for shipment. Your EHS certified shipper will let you know the quantity of dry ice you will need to provide.
    3. EHS certified shipper will determine and provide the appropriate packaging system, attach required labels/ markings to outer package, and prepare airway bill and other needed documents.
    4. EHS certified shipper arrives at sender’s research laboratory on agreed-upon date/time, and works with sender to package the shipment. The EHS certified shipper will take the sealed package to EHS for pickup by the carrier.

Moving biohazardous materials from one work area to another:

  • Transport outside of the laboratory but within the building -- Place within a durable, leak-proof container, such as a plastic sterilite box plus lid; close lid and disinfect the outside container before removing from the room. Either the primary or the secondary container must be labeled with the biohazard symbol.
  • Transport outside the building -- Seal the material in a primary tube/flask or other leak-proof container; place biohazard symbol on this container. Disinfect the container before placing in a durable, leak-proof, lidded secondary transport box or container. The secondary container will be disinfected and securely closed for transport.
  • For added safety, containers should be transported on a cart to further minimize spill hazards.
  • If material must be transported in a vehicle from one building to another on campus, use of a state vehicle is recommended.


Transporting biohazardous waste to autoclave facilities:

  • Waste must be securely closed and sprayed thoroughly with 70% ethanol.
  • Waste must be placed within a secondary container (e.g., Nalgene or stainless steel pan, plastic lidded tub, etc.) which is dedicated for this function and labeled with a Biohazard label.
  • Place in secondary containers on a cart to autoclave facilities.

Contact the EHS Biosafety team.

These are laboratories that collect and/or perform minor manipulations with human/NHP blood, tissues or other potentially infectious materials for research or teaching purposes. The scope of activities of such labs may require 1) registration with the IBC and 2) meeting EHS biosafety requirements below, but may not require full protocol submission and review with the IBC. See table below for details.

LAB TYPE MATERIAL AND MANIPULATION (Manipulation = removing material from closed container & doing something with it) REQUIREMENTS for the IBC REQUIREMENTS for BIOSAFETY
Research Lab Blood draw only – no or minor manipulation
  • Contact the IBC.
  • Document training for personnel.
  • Contact IRB.
  • Confirm that lab has contacted the IBC.
  • Lab/area inspection could take place.
  • Complete written procedures + Exposure Control Plan & supply copies in work area.
  • Training: BBP
  • Have in place:
    • EHS phlebotomy guidelines
    • Biohazard door signage
    • Appropriate holding/storage area for labeled specimens
    • Proper lab waste collection/disposal; spill kit
Research Lab Extensive manipulation of human blood, tissues, cell cultures, etc.
  • Register with IBC (full review and approval).
  • Document training for personnel.
  • Contact IRB.
  • Confirm registration with IBC as BSL-2 lab.
  • Biosafety lab inspection will take place.
  • Complete Lab-Specific Biosafety Manual (includes Exposure Control Plan) & supply copy of LSBM in work area.
  • Training: BBP; site-specific training as needed
  • Have in place:
    • BSL-2 door signage
    • Biohazard stickers on equipment
    • Appropriate holding/storage area for labeled specimens
    • Proper lab waste collection/disposal; spill kit
Clinical Diagnostic Labs (e.g., Schiffert, Vet. Med.) Manipulation of tissues/fluids/ cell suspensions that are potentially infective If not involved in generating research data, then lab is not subject to IBC oversight.
  • Follow CDC "Guidelines for Safe Work Practices in Human and Animal Medical Diagnostic Laboratories."
  • If the lab is not accredited or inspected by an outside agency then EHS will inspect it.
Clinical Research Labs (e.g., VTCRI) Urine collection
Saliva collection
Blood collection
  • Contact the IBC.
  • Document training for personnel.
  • Contact IRB.
  • Confirm that lab has contacted the IBC.
  • Lab/area inspection could take place.
  • Complete written procedures + Exposure Control Plan & supply copies in work area.
  • Training: BBP; site-specific training as needed
  • Have in place:
    • Biohazard door signage
    • Appropriate holding/storage area for labeled specimen
    • Proper lab waste collection/disposal; spill kit
Service Labs (e.g., Flow Cytometry, Microscopy) Manipulations of fixed or live tissues, fluids or cell suspensions of:
  • human/NHP material
  • tumorogenic material
  • recombinant material
  • infected or possibly infective material
  • Contact the IBC.
  • Document training for personnel.
  • Confirm that lab has contacted the IBC.
  • Lab/area inspection could take place.
  • Complete written procedures + Exposure Control Plan & supply copies in work area.
  • Training: BBP; site-specific training as needed
  • Have in place:
    • Biohazard door signage
    • Biohazard labels on equipment
    • Proper lab waste collection and disposal; spill kit

Detailed list of biosafety requirements for special labs:

Signage/labeling:

  • Post biohazard sign (or biosafety level sign) on all access doors.
  • Post any other warnings or requirements to enter, such as 'No eating/drinking'.
  • Place Biohazard labels on 1) equipment used with biohazardous material and 2) storage units (freezers/refrigerators) containing biohazardous material.

Waste handling:

  • Have in place well-labeled lab waste containers with lids and visible biohazard labels.
  • Use the appropriate bag in the waste container, according to the type of waste being generated.
  • Autoclave biowaste, then dispose of in Regulated Medical Waste.

Disinfection:

  • Ensure that appropriate disinfectant at proper concentration is adequately supplied.
  • Ensure that spill cleanup procedure is posted or readily available, and spill supplies are available.

PPE:

  • Ensure that lab coats and gloves (minimum requirement) are supplied.
  • Based on risk assessment, ensure other necessary PPE is supplied: safety goggles, face shields; respiratory protection may be required to protect against inhalation of aerosols.

Universal precautions:

  • Ensure that the Exposure Control Plan or Lab-Specific Biosafety Manual (inclusive of ECP) is in place.

Training:

  • Online EHS Bloodborne Pathogens training for lab workers: See 'Training' section.
  • Site-specific training is dependent on the equipment, work procedures, material, and hazards involved; Principal Investigators, lab managers, or area supervisors or their designees are responsible for delivering site-specific training.
  • Records – Documentation of all training must be kept current, and maintained for 3 years post-employment.

Documentation:

  • Exposure Control Plan – complete, and make a copy available for reference and training.
  • Written procedures involving biohazardous material – complete and include containment practices and safe handling/sharps/storage/disposal measures; make copies available for work area.

Further information and guidance:

IBC
EHS Biosafety Group

Minor biological spills:

A spill of lesser volume and/or with agents of lesser pathogenicity, for which cleanup can usually be handled by lab personnel using absorbent materials/ disinfectants routinely kept on the bench or in lab spill kits.

General response procedure:

  1. Immediately inform coworkers that a spill has occurred.
  2. Remove any contaminated PPE or clothing.
  3. If possible without risking exposure, quickly place absorbent material (such as paper towels) on the spill to prevent spread and seepage in the next 30 minutes.
  4. Restrict personnel access to spill area for 30 minutes for aerosols to settle.
  5. Report the spill to a supervisor as soon as possible.
  6. If you are trained, able and equipped to handle the spill, and you have determined that it is not a major spill (beyond your scope to handle), proceed with the cleanup.

General cleanup procedure:

  1. Use supplies from biological spill kit, or from the bench.
  2. Don clean PPE (one or two pairs of gloves, apron or gown, shoe covers if needed). If splash protection is needed, don goggles and/or faceshield. If respiratory protection is needed and you have been trained and fit-tested to use it, don the appropriate respirator.
  3. Cover spill with absorbent paper towels. Start at edge of spill and work inward.
  4. Apply a disinfectant to the towels and let sit for 10 minutes (or for the specific contact time needed for the disinfectant you are using). Use full strength disinfectant on larger spills.
  5. Remove any sharps from the spill with tongs, forceps, cardboard, dustpan, etc. – not your hands – and discard into biosharps container.
  6. You can remove soaked towels with tongs, especially if small bits of glass are still in spill area; discard soaked towels as chemical waste.
  7. Apply disinfectant again to the spill area. Wipe dry or allow to air dry.
  8. Remove PPE carefully in this order, and avoiding direct touch to contaminated PPE surfaces: shoe covers, gloves, lab coat, face/eye protection. Discard in autoclave bag, and wash your hands.
  9. Inform coworkers that spill cleanup is complete.
  10. IMPORTANT:
    • Chemical disinfectants require contact time with the spill to effectively decontaminate it. Be aware of the specific contact time of the disinfectant you use and allow that time to elapse before clean-up.
    • For metal surfaces, follow all bleach disinfectant treatments with a water rinse.
    • If a chemical disinfectant is not used (or cannot be used) with contaminated items, decontaminate by autoclaving or other method approved by EHS if items can withstand the process (e.g., contaminated lab coats).
       

Major biological spills:

  • A major spill is one which, in your judgment, could represent a significant environmental risk or serious human health risk as a result of release or exposure,
  • and/or is a larger-volume spill of biohazardous agents or recombinant/synthetic nucleic acids which is beyond the capacity and training of lab personnel to safely execute cleanup, and will require cleanup by haz-mat professionals.

How do I decide if a spill is major or minor?

Consider:

  • Pathogenicity,
  • Concentration, and
  • Aerosol hazard and/or environmental hazard of the agent(s) in the material, and
  • The volume of the spilled material.

When making this decision. Consult the risk assessment(s) for the agent(s) if necessary, but make the decision as quickly as possible.

What to do in the event of a major spill:

  1. Immediately notify everyone in the lab /area and clear the area of all personnel.
  2. Secure the area as needed by locking doors, standing guard to keep people out, posting signs, etc.
  3. If possible, quickly place whatever absorbent material is at hand to lessen spread or seepage, but only if this can be done without increasing your exposure risk significantly beyond its current level.
  4. Call 911 or 540-231-6411 for Virginia Tech Police; ask for EHS to be informed immediately.
  5. Inform your PI or lab manager/supervisor as soon as you possibly can.
  6. You are not responsible for trying to clean up a major spill, or arranging for the cleanup of a major spill. In some cases, EHS evaluates major spill situations and schedules the appropriate hazardous material response crews or contractors; in other instances, hazmat teams are dispatched via a 911 call.
     

Biological spill kits:

  • Contents should be contained within a handled, lidded bucket and include:
    • Disposable lab coat or coveralls,
    • Disposable gloves,
    • Face shield/mask,
    • Disposable shoe protectors,
    • Spray disinfectant,
    • Clean-up supplies (forceps, dustpan, autoclave bags, spill pillows and socks)
    • Sign that reads "Biohazard Spill DO NOT ENTER."
  • Ensure that sufficient quantities of materials (on bench, and/or in spill kit) are always available to clean up the maximum spill volume that could be anticipated in your lab.
  • If a respiratory hazard has been identified through risk assessment of the BSL-2 agents used in the area, respirators must be provided separately by the laboratory for use in spill situations. N-95 respirators must be fit-tested by EHS. Contact EHS to arrange.
  • The Biological Spill Kit should be well-labeled, and a wall sign should indicate the location of the Biological Spill Kit in the lab so it can be located quickly, particularly if it is kept under the bench.
  • All personnel working with BSL-2 materials must receive training by the PI or designee for Biohazard Spill Kit use.

Further information and guidance:

BMBL
University Biosafety Manual
EHS Biosafety Group

  • Faculty in charge of teaching laboratory courses (or their designees) must submit a protocol to the Institutional Biosafety Committee (IBC) when one or more of the following materials are used:
    • Risk Group 2 agents;
    • Recombinant plant/animal/microbial material (Risk Group 1 or 2);
    • Human/ non-human primate specimen material or cell lines; and/or
    • Animals inoculated with infectious/recombinant material.
  • Likewise:
    • Faculty in charge of teaching labs that utilize live animals must submit a protocol to the Institutional Animal Care and Use Committee (IACUC).
    • Faculty in charge of teaching labs that collect and/or utilize human specimen material obtained from class participants or other volunteers, and/or derive data from that material, must contact the Institutional Review Board (IRB) to learn if their projects require IRB protocol submissions.
  • Teaching protocols are evaluated for approval by the IBC following initial submission, after which they are reviewed annually. Teaching protocols must be renewed every three years, just the same as research protocols.
  • Teaching labs that utilize Risk Group 1 non-recombinant agents perform a self-inspection which is submitted to the IBC; teaching labs utilizing Risk Group 2 agents and/or recombinant material are subject to a biosafety inspection by a Biosafety Officer.
  • Virginia Tech IBC and EHS apply the standards and practices outlined in the ASM Guidelines for Biosafety In Teaching Labs when reviewing teaching protocols, setting requirements to be met in teaching labs, and inspecting teaching lab facilities.
     

Further information and guidance:

IBC
EHS Biosafety Group

New lab personnel biosafety training needs: General and specific:

General training:

  • Initial training is required for performing work in a laboratory using microbiological/ biomedical/ recombinant techniques and equipment. This training is available online, accessed through the EHS website. The following EHS training modules must be completed PRIOR to beginning work at the bench:
    • General Laboratory Safety (EHS online module);
    • Biosafety For Research Labs (EHS online module; can include Bloodborne Pathogens training — see below);
    • Introduction to Biological Safety Cabinets (EHS online module);
    • Safe Autoclave Use (EHS online module).
    • Laboratory Hazardous Waste (EHS online module)
    • These trainings have a 3-year expiration, at which time they must be renewed. Certain other trainings, such as Bloodborne Pathogens and Respiratory Protection, have an OSHA-based annual renewal requirement
  • Persons who work with rDNA molecules or synthetic nucleic acids must complete training on NIH Guidelines for Research Involving Recombinant DNA molecules. This is accessed through the IBC website.
  • Additional training may be required or recommended, according to the person’s occupational tasks/risks in the lab, such as Compressed Gas Cylinder Safety and Portable Fire Extinguisher training. Further determinations may be made by EHS, the Institutional Biosafety Committee (IBC), or Occupational Health in consultation with the PI.
  • Successfully completed EHS online training is recorded electronically. Trainees can access their online training profiles at any time through the EHS Safety Management System (SMS); it will display a list of completed trainings and their expiration dates, registrations for training not yet taken, and expired training. A record of completed EHS trainings can be printed from the SMS if needed. The SMS also allows PIs/lab managers to view/monitor the training records of their lab groups for tracking completions, expired training, etc.

Specific training:

  • Initial orientation and training that is specific to the lab’s practices, equipment, procedures, and research --- this is what is meant by "lab-specific" training, and must be provided by the Principal Investigator or designee. Lab-specific training also can be tracked and managed in the SMS by PIs/lab managers.
  • The form provided by EHS, Biosafety Training for New Lab Personnel, outlines the required lab-specific training that should be provided to new lab workers. This form must be completed, with signatures of the trainee and PI (or designee) to document that lab-specific training has occurred; it also includes a statement that the PI (or designee) has found the trainee to be proficient in standard and special microbiological practices before being allowed to work with biohazardous agents. One of these forms must be completed and signed for every individual working in the lab, documenting that 1) lab-specific training has taken place and 2) the individual has demonstrated the needed proficiency. These forms should be kept in your Lab-Specific Biosafety Manual (or other file location in lab) for ready reference.

"Biosafety For Research Labs" (BSRL) online training provides Bloodborne Pathogens (BBP) training for lab workers if needed:

  • BSRL  is customized to the needs and work situations of research lab workers.
  • BSRL exists in two versions, one that includes BBP training components, and one that does not. Therefore it can accommodate lab workers who need biosafety training AND Bloodborne Pathogens (BBP) training for working with human cell lines, specimen materials, etc., in their lab work, as well as those who need biosafety training only. NOTE: All lab personnel who work with human/non-human primate materials must receive BBP training through the online BSRL module.
  • To receive the BBP-inclusive version of this training, simply answer YES to the question that will be asked of you when you register for the training: "Do you work with human/NHP materials (including human cell lines?" Those who answer NO will receive biosafety training without BBP required material, so take care to answer the question correctly for your situation.
  • Answering YES at registration for BSRL will launch the full biosafety + BBP version when you take the training initially, and every three years thereafter. When you successfully complete the BBP-inclusive version of BSRL initially, you will get credit for having taken both BBP training and BSRL in your EHS training profile, and they will both show a 1-year expiration.
  • To keep your training in compliance, you will need to register for BSRL annually; you DO NOT need to register for BBP training separately. In interim years when you register for BSRL, the system will launch a concise, BBP-only online refresher for you to take, NOT the full biosafety training module. The database will track your training profile, and will launch whichever module you need to keep both your biosafety training (3-year expiration) and your BBP training (1-year expiration) in compliance.

How do I register for EHS online trainings?

  • Go to ehss.vt.edu.
  • Click on ‘Training’ in the left gray bar.
  • Click on ‘Class Schedule and Registration.’
  • Click on the topic area you need. For example, clicking on ‘Biological Safety’ will open a list containing "Safe Autoclave Use," "Introduction to Biosafety Cabinets," etc. Click on the ‘Register’ button beside the training you want to take. The registration process will require your two-factor authentication.
  • At the bottom of the Training Class Registration Form, click on the SUBMIT button.
  • When you are ready, click on LAUNCH to begin the training.

NOTE: When trainings are viewed using older computers, older versions of software, or on older devices such as tablets, Ipads, etc., the likelihood of the training not loading or advancing properly is slightly increased. Before you click LAUNCH, refresh your browser. If you experience difficulty, try launching the training using a different browser, and/or a different computer, if possible.

Besides new personnel in the lab, who else needs biosafety training?

  • Existing lab personnel could need:
    • Training in new lab topics/ techniques, using new equipment, etc.;
    • Refresher training to increase proficiency;
    • Remedial training following mistakes, exposures, accidents, etc.; and/or
    • Higher-level training if preparing to manage the lab.
  • Other lab personnel (with whom you share lab spaces or common-use equipment) will need:
    • Awareness training – to provide information about the hazardous biological materials manipulated in your research efforts using shared spaces or common equipment, such as disinfectants needed, etc.
  • Personnel who work in or around the lab but do NOT work with biohazards will need:
    • Awareness training on how to avoid exposure to biohazards in the lab.

Requirements for documenting biosafety-related training:

  • Q: Why do we have to document this training, and how must we do it?
  • A: For liability and regulatory compliance purposes, all training (i.e., initial, refresher, lab topics, awareness, remedial) must be documented via a written record which includes, at a minimum, the trainee’s name, type of training received, and date of receipt.
  • Q: Under what circumstances are these training records examined?
  • A: Training records are subject to examination during inspections/ audits by oversight and regulatory agencies, and especially during investigations following an incident or exposure. Being able to prove that appropriate training took place can greatly impact the outcome of an investigation in the wake of a lab accident (the size of fines imposed, criminal prosecution, loss of funding, etc.).
  • Q: How must these training records be stored?
  • A: Training records can be stored as paper copies or scanned documents:
    • Printed copies of training records can be kept in the Lab-Specific Biosafety Manual; records for separated personnel can be filed elsewhere in an accessible location.
    • Electronic copies of training records can be stored in the lab’s SMS records, or another local electronic storage site that is easily accessible to PI, Lab Manager, and lab personnel.
  • Q: How long do these training records have to be kept?
  • A: Completed training records (electronic or printed) must be retained for the duration of each person’s employment or activity period in the lab, and for at least 3 years after a lab worker’s separation date.
  • Q: Who can view a person’s EHS electronic training record, and how is it viewed?
  • A: Any individual who is a Virginia Tech employee or student can view their "Training Profile" of successfully completed EHS online trainings, as follows:
    • Go to ehss.vt.edu.
    • Click on ‘Training’ in the left gray bar.
    • Click on "View Training Profile."

    You can also see expiration dates for each training, as well as those for which you have registered but not yet completed.

    PIs or other supervisory personnel in the lab who have been designated as administrators for the lab in the SMS can access and track EHS training records for all individuals in their lab group.

BIOSAFETY TRAINING SUMMARY TABLE

People To Train Type of Biosafety Training Needed Training Document To Use What the Document Certifies Who Provides Training
New lab personnel who will be working with biohazards EHS online training Training is documented electronically in EHS training database and can be accessed on an individual’s EHS Training Profile or through the Safety Management System.
  • The trainee is credited with successful completion of the module.
  • Trainee name, date of completion of training, and when training expires
EHS -- online training modules
New lab personnel who will be working with biohazards
  • Biosafety training specific to your lab’s procedures, equipment & agents.
  • Safety orientation (e.g., evacuation routes, safety equipment, reporting procedures, etc.)
Biosafety Training Record for New Lab Personnel

(Serves as a lab-specific training template for topics to cover, as well as a record of training, and certification of proficiency by the trainer.)
  • Trainee understands topic areas covered.
  • The trainee is generally proficient in lab procedures covered.
  • Trainee/trainer names, signatures & date of training
Principal Investigator or designee
Existing lab personnel working with biohazards
  • Biosafety refresher or review
  • Specific training on new procedures, equipment, etc.
Lab Topics Training Record

(Trainer fills in the lab topic covered; trainees/ participants sign on a signature page.)
  • Topics covered
  • Trainee/trainer names & date of training
Principal Investigator or designee
Existing lab personnel working with biohazards
  • Higher-level, comprehensive lab training
  • Remedial training in one or more areas
Biosafety Proficiency Checklist

(Serves as a template for topics to cover and proficiency evaluation; can be customized according to need.)
  • Trainee understands all topic areas covered.
  • The trainee has demonstrated proficiency in specific lab skills & methods covered.
  • Trainee/trainer names, signatures & date of training
Principal Investigator or designee
Lab personnel from other groups with whom you share lab space or equipment
OR
Personnel working in/around lab but NOT with biohazards
Lab-specific training to provide awareness of biohazards present in the lab, & appropriate response to exposures, signs of disease, lab incidents Bioawareness Training Record

(Serves as a template for topics to cover and training record sheet; can be customized according to the situation.)
  • Trainee understands topic areas covered.
  • Trainee/trainer names, signatures & date of training
Principal Investigator or designee

Biological Waste Management

Click here for more information on handling and disposing of biological, biohazardous, and regulated medical waste.

Contaminated waste items generated in a biosafety cabinet

Items may be removed from the BSC after they are decontaminated with an appropriate disinfectant, such as being sprayed with 70 percent ethanol, or they can be placed within a bag or sealed container in the biosafety cabinet, which is sprayed with disinfectant before removal from the biosafety cabinet.

Liquid biohazardous waste

Preferred method for liquid biowaste containing no disinfectant or other chemical components:

  • Collect the waste in a well-labeled autoclavable container (containing no bleach), then autoclave the waste on a liquid cycle at 121˚ C, with the sterilization time determined by the liquid volume.  After cooling, the waste may be poured down the lab drain
  • Alternative method:
    • If using the above method is not possible, liquid biowaste can be discarded into a container containing a sufficient quantity of bleach (e.g., pure bleach to yield a 1:5 dilution. Example:  100 mls. household bleach added to 400 mls. tissue culture media.
    • After the required exposure time in the biosafety cabinet, the liquid waste plus bleach may be disposed of down the lab drain, followed by a water flush.  If a different disinfectant is used, it must be disposed of as chemical waste.
    • Decontamination by bleach a less reliable decontamination method because of the opportunities for a failed result if the bleach used is 1) expired; 2) present in the wrong proportion to liquid waste volume; and 3) not given sufficient contact time before disposal. 
  • Liquid biowaste containing bleach, another chemical disinfectant, or other chemical constituents such as sodium azide: 
    • Dispose of as liquid chemical waste.  DO NOT AUTOCLAVE THIS WASTE.
  • Liquid biowaste containing antibiotics:
    • Media wastes containing the following antibiotics can be autoclaved, cooled, and flushed down the lab sink drain because the antibiotics are broken down by heat, and are then environmentally safe to go into the domestic sewer.
    • Examples of heat-sensitive antibiotics:
Heat-sensitive antibiotics    
Ampicillin Kanamycin Geneticin
Amphotericin Neomycin Tetracycline
Carbenicillin Puromycin  
Penicillin Streptomycin  
  • Media wastes containing the following heat-stable antibiotics cannot be autoclaved and discarded into the domestic sewer because of their relatively long half-life which persists in the environment; add bleach in a 1:5 vol/vol concentration to deactivate biologicals, then dispose of as liquid chemical waste.
    • Examples of heat-stable antibiotics:
Heat-stable antibiotics     
Hygromycin B Vancomycin Gentamicin
Chloramphenicol Nalidixic acid  
Ciprofloxacin Zeomycin  
  • If antibiotics are used that are not on these lists, contact Environmental Health & Safety or disposal consultation, or simply submit for disposal as liquid chemical waste.
  • Media with additives such as growth factors, metals, or other chemicals must be disposed of as liquid chemical waste.
  • Liquid decontamination method(s) should be verified and documented.
  • Liquid biowaste containers must be appropriately labeled as “WASTE”, with biohazard signage; liquid chemical waste containers must be appropriately labeled and well-identified as a different waste stream. 
  • Liquid Wastes Containing Extracted DNA/ Cellular Components/ Lysates, etc.
    • Biological material treated with extraction kit chemicals or other chemical treatments that lyse cell membranes renders bioagents non-viable and no longer hazardous. This material must be disposed of as chemical waste.

Solid biohazardous wastes

  • BSL-1:  Collect in a non-colored autoclave bag with no Biohazard symbol.
  • BSL-2:  Collect in an ORANGE autoclave bag with a Biohazard symbol.
  • Autoclave bags must be kept inside appropriately labeled biowaste containers, equipped with a closeable lid. 
  • Solid biowaste containers must remain closed except when in use.
  • Proceed with autoclaving the waste according to standard procedures Bags of solid biowaste must be autoclaved on a gravity or pre-vacuum cycle at 121 degrees celsius; the length of cycle used must be determined by the size and density of the autoclave load.  Each load must be checked with a verification device such as a chemical integrator strip to confirm that kill conditions were met.
  • Bags must be autoclaved daily when possible, or as soon as an autoclave is available.  No full bags should be left in the laboratory or the autoclave/glassware room OVER WEEKENDS OR BREAKS.
  • Bags must remain closed until they are ready to be autoclaved, at which time their closures should be loosened to at least a 1-inch opening to allow steam penetration.
  • All BSL-2 solid waste is to be decontaminated by autoclaving and disposed of as Regulated Medical Waste.
  • Decontaminated BSL-1 waste can be discarded into regular trash, or disposed of in Regulate Medical Waste.
  • Agar Plates With Antibiotics In The Medium:
    • BSL-1 -- It is acceptable to autoclave BSL-1 waste containing heat-stable or heat-labile antibiotics, and once decontaminated, dispose of waste in dumpster trash bound for landfill.  Bagged waste is well contained, allowing adequate time for breakdown of antibiotics when disposed of in a landfill.
    • BSL-2 – It is acceptable to autoclave BSL-2 waste containing heat-stable or heat-labile antibiotics, and once decontaminated, this waste must be discarded in Regulated Medical Waste---the same disposal method as BSL-2 plates containing no antibiotic.
  • Solid biowaste/lab debris mixed with chemical waste:
    • Contact Environmental Health & Safety to determine the best disposal method.

Sharps biohazardous wastes

  • Sharps contaminated with biohazardous materials include anything that could puncture an autoclave bag. Examples include:
    • Pipette tips
    • Wood applicator sticks/ swabs
    • Syringe + needles
    • Blades
    • Glass slides/cover slips
    • Serological pipettes
    • Glass Pasteur pipettes
    • Disposable plastic pipettes
    • Broken glass
    • Blood tubes/capillary tubes
  • Discard all sharps used with biologicals into lidded, rigid, labeled Bio-sharps containers. Environmental Health & Safety supplies containers that meet the requirements of being closeable, puncture-resistant and leakproof; they can be requested through the Environmental Health & Safety online Safety Management System.
  • Sharps waste containers:
    • Must be located for ease of accessibility, i.e., at or near point-of-use.
    • Must be replaced when full.
    • Must be securely closed before removal from point-of-use.
    • Must have NO chemical or liquid waste placed within (with the exception of specimen tubes containing blood).
  • Never re-cap needles or scalpel blades before disposal into a sharps container.
  • Avoid forcing sharps waste into a full container; this can cause puncture/ cut injuries.
  • All sharps containers are to be autoclaved using the same cycle as used for biological solid waste and then disposed of in the same manner as BSL-2 waste, i.e., in regulated medical waste.
  • In situations when collection of Sharps waste is intermittent or of smaller volume, avoid collecting them in large sharps containers.  Use smaller Sharps. containers whenever possible, as these will fill more quickly and thus can be decontaminated more expeditiously.
  • Rigid plastic laundry detergent bottles with lids can be used but must be well labeled.

Regulated medical waste (RMW)

  • RMW collection boxes are used for end-point disposal of DECONTAMINATED (i.e., autoclaved) biological Sharps and ALL BSL-2 solid waste.
  • Decontaminated BSL-1 waste can be disposed of in regular trash cans lined with black bags, or in RMW.
  • RMW must not contain any liquids in containers, or free liquids.
  • RMW box assembly and use:
    • Tape bottom center seams 2-3X, and tape open side seams one time with wide sellotape.
    • Line each RMW box with 1-2 red RMW plastic liner bags.  If using 2 bags, place one inside the other. Remember, these liners cannot substitute as autoclave bags—they disintegrate in high heat.
    • Do not overfill RMW boxes, or pack them so that they are too heavy; limit is 50 lb.
  • RMW final packaging:
    • When a RMW box is full, twist the top of the inner bag enough times so that you can loop it over on itself in a “gooseneck."
    • Seal the twisted top by wrapping with tape.  Seal a second, outer bag by the same method.
    • Close the box’s top and seal X3 on center seam with wide sellotape, then tape the open side seams 1X.
    • Fill out the white label for the box that identifies the generators of the waste, and the date the box is closed and sealed.  Be sure to list all labs that contributed waste to the box.  Apply the label in the designated area on the box.
    • Be sure you write the date of closure on the box, not the date the box was assembled and began to be used.
    • Notify EHS when you have RMW waste for pickup using the Environmental Health & Safety online request system.

Animal and animal-related wastes

  • Lab workers who produce a minimal volume of biological material with no chemical present (e.g., tiny pieces of dissected unfixed tissue) can dispose of that material in solid biowaste which will be autoclaved.
  • Lab workers who produce larger volumes of this waste or other types of animal/animal-related wastes should refer to Virginia Tech’s Animal and Animal-Related Waste Procedures charts to determine proper waste handling and disposal.  Categories of wastes included in the charts are:
    • Animal tissue with/ without fixative or other chemicals present
    • Animal tissue or carcasses with/ without hazardous biological agents, rDNA, etc.
    • Companion animal carcasses
    • Related wastes:  bedding, disposable containers, fecal material, dressings, etc.
    • Associated sharps and liquids
    • Blood collection tubes
    • Radioactive wastes

Plant and plant-related wastes

  • BSL- 1P, BSL2-P Greenhouse Plant Waste 
    • Examples include transgenic, exotic, or infected plant material; soil; pots; etc.
    • Collect in clear autoclave bags or other bags/containers appropriate for the method of biological deactivation to be used.
    • Heat inactivation can be accomplished by:
      • Autoclaving (for smaller volumes) at 121˚ C, 15-30 psi, for 15-180 minutes, depending on type and state of material.
      • Treatment in greenhouse steam box/soil sterilization box (for larger volumes, for fungal, viral or nematode plant pathogens under permit) at ≥ 104˚ C for 3 hours.   NOTE:  Permission to use greenhouse steam box must be obtained from the greenhouse manager.
    • BSL-1P material can be composed or desiccated, according to IBC approved protocols.
    • Deactivation of plant/seed material must be confirmed before disposal.  Acceptable methods of confirmation include:
      • Recording time/temperature criteria of heat treatment used.
      • If autoclaving, use verification devices.
    • Disposal:
      • Following successful deactivation, material can be discarded in regular trash cans with black liners.
  • Plain plant waste
    • Non-transgenic, non-exotic, non-infective laboratory plant waste is still laboratory-generated waste, and can appear identical to other more hazardous plant wastes.
    • According to your lab-specific situation, this type of waste can be handled in several ways, at the PI’s discretion:
      • Collect, deactivate and discard this waste as if it is BSL-1P or BSL2-P waste.
      • Collect material in opaque trash bags; when full, close securely and discard bags in dumpster.

 

Biosafety Definitions

Sometimes referred to as good microbiological technique.

Work methods designed to reduce or prevent contamination by microorganisms in the environment when working with cultures of microorganisms, sterile materials or sterile areas; elements of aseptic technique are 1) use and maintenance of sterile work areas and containment equipment (BSC), 2) good personal hygiene, 3) use of sterile reagents and media, and 4) sterile handling methods for equipment, instruments, supplies and other materials.

A biological agent or condition that constitutes a hazard to human beings or the environment, including:

  • Infectious agents (bacterial, viruses, protozoans, fungi, etc.);
  • Biotoxins;
  • Human or non-human primate blood, fluids, cells or tissue culture;
  • Recombinant DNA and synthetic nucleic acid molecules;
  • Transgenic microbes, animals, invertebrates and/or plants;
  • Select agents;
  • Prions;
  • Opportunistic pathogens;
  • Poorly characterized or uncharacterized cell lines, tumors or other tissues;
  • Cell lines and research materials from other labs that have not been verified;
  • Replication defective pathogen vectors; and
  • Attenuated strains and fixed materials that have not been tested to verify the attenuation or inactivation of the material.

The application of knowledge, techniques and equipment to prevent personal, laboratory and environmental exposure to infectious/biohazardous agents and materials.

Sometimes called a biological safety cabinet.

An enclosed laboratory work surface equipped with filtered directional air flow. BSCs provide the best place to work with biohazardous materials because they protect workers and the environment from exposure to agents, and protect cultured agents and sterile items from airborne contaminants.

Combinations of laboratory practices, safety equipment and facilities features that define the conditions under which infectious agents can be safely manipulated. Levels of containment range from the lowest Biosafety Level, BSL-1, to the highest at BSL-4, where the most aggressive measures are taken to prevent exposure and release of biohazardous agents. At Virginia Tech, most research laboratories operate at BSL-1 and BSL-2. Two BSL-3 facilities are maintained by highly trained laboratory personnel. Virginia Tech has no BSL-4 facility.

Also includes associate biosafety officer.

Individuals responsible for establishing and monitoring workplace safety procedures designed to minimize or prevent exposures and releases of biohazards.

Also known as biological toxin.

Toxic substances of biological origin that may cause death or severe incapacitation at relatively low exposure levels. To utilize or produce biotoxins in research, PIs must have IBC approval and adhere to explicit safety and ‘Due Diligence' requirements. Certain biotoxins have been designated as select agents. 

A process or treatment that removes or neutralizes biologic, chemical or radiologic contamination from a person, object or area. In the biosafety realm, this term can be correctly applied to the autoclaving of biological lab waste prior to disposal, or chemically treating a lab surface with a disinfectant. The term can refer to achieving an acceptable reduction of microbial contaminants, whereas sterilization refers to a complete elimination of microbes.

The Virginia Tech form that injured employees are expected to complete in order to report an accident or illness that has arisen out of (or during the course of) their employment; completed forms are submitted to Virginia Tech Human Resources. This form must be completed as soon as possible for the injured employee to be eligible for workers' compensation.

When a biohazardous material has made contact or may have made contact with:

  • A puncture injury site on your body (e.g., needlestick or cut with sharp object);
  • The mucous membranes of your eyes/ nasal passages/ mouth; and/or
  • Non-intact skin surface (e.g., exposed skin that is chapped, abraded, has dermatitis, etc.).

The oversight body tasked with ensuring that instructional and research activities involving biohazardous materials at Virginia Tech are in compliance with federally-mandated responsibilities and obligations for proper containment and safe handling of biohazards.

An event that results in an undesired circumstance. For a lab conducting research using biohazardous materials, examples of incidents are:

  • Spills, in or out of containment;
  • Potential or known exposures;
  • Loss of containment;
  • Release of biohazardous agent into the environment;
  • Injuries/illness caused by lab activities;
  • Breaches of biosecurity; and/or
  • Any combination of these.

The Virginia Tech form to be completed by Principal Investigators, Lab Managers and/or lab personnel after the occurrence of an incident/accident in the laboratory; completed forms are submitted to EHS.

Protective clothing, headgear, eyewear or other garments or equipment designed to protect the wearer's body from physical injury or exposure to hazardous substances/agents. Hazards addressed by PPE include physical, electrical, temperature extremes, chemical, biohazards, and airborne particulates. Typical PPE for biosafety in laboratory research: disposable gloves, lab coat or disposable gown, safety glasses/goggles, face shield, insulated gloves for autoclaving or working with liquid nitrogen storage, respirator for working with respiratory pathogens.

The principal investigator (PI) is the primary individual responsible for the preparation, conduct and administration of a research grant, for achieving technical success of the research project, and for oversight of laboratory operations and personnel involved in working on the research project.

Safety devices which are physical barriers which are used to prevent direct contact between a hazardous agent/material and the worker. These include biosafety cabinets, personal protective equipment, etc. which are intended to protect laboratory workers.

Also includes alternate responsible official.

The individuals who are accountable for Virginia Tech's Select Agent Program and its compliance with federal select agent regulations. The RO must be familiar with the regulations, have authority to act on behalf of Virginia Tech regarding the Select Agent Program, maintain required records, and conduct annual inspections. The ARO serves as RO when the RO is not available.

The process of researching, evaluating and identifying hazards/potential hazards in procedures, experiments, materials handling, etc. Risk must be assessed in every step of a procedure. Risk assessments also must determine the actions and controls required to eliminate or reduce risks to workers. 

Consists of a variety of safety features and devices (air filtration systems, decontamination equipment, facilities design features, etc.) meant to prevent exposure to/release of hazardous agents/materials if primary barriers fail.

Also known as select agents and toxins.

Bioagents that have been declared by the U.S. Department of Health and Human Services or by the U.S. Department of Agriculture to have the "potential to pose a severe threat to public health and safety." At Virginia Tech, select agents can only be utilized in research with IBC approval and within the parameters of the Select Agent and Toxins Program.

An object that can pierce or cut skin, causing injury and possible hazardous exposure. Higher risk sharps include needles, scalpel blades, razor blades, glass Pastuer pipettes, broken glass, glass slides, etc.; lower risk sharps capable of piercing non-rigid containment and creating an exposure risk or minor injury include plastic pipettes, tips, wood applicator sticks, etc. 

Materials assembled in a leak-proof bucket or lidded box for use in cleaning up a spill of a biohazardous material in the laboratory. The spill kit should be well-labeled, and a wall sign should indicate the location of the biological spill kit in the lab, particularly if it is kept under the bench, so it can be located quickly.

Contents of the biological spill kit:

  • Disposable PPE (lab coat, gown or apron, gloves, shoe covers, face shield);
  • Absorbent paper towels; 
  • Dustpan; 
  • Tongs/forceps; 
  • Autoclave bags; 
  • Disinfectant; 
  • Copy of spill cleanup procedure; and
  • Spill warning sign to post. 
  • Respiratory protection should be provided in labs where agents are used that would present a respiratory hazard when spilled.

Basic hygiene practices or guidelines for working safely with microorganisms, including:

  • Wash hands after working with potentially hazardous materials and before leaving lab.
  • Do not consume food/drink, smoking, applying cosmetics or contact lenses, and no storing food in the lab.
  • Do not mouth-pipette.
  • Safely handle and dispose of all sharps.
  • Minimize splashes and aerosols.
  • Decontaminate biological material before disposing of it.
  • Post biohazard signage on lab entrance when working with infectious agents.
  • Maintain an effective pest management program.
  • Ensure that lab personnel receive appropriate training and information that may be relevant to their particular health status.

The complete elimination of microorganisms to achieve a sterile, microbe-free condition; this can be done using a physical procedure (e.g., autoclaving) or chemical procedure (e.g., treatment by a strong disinfectant).


Biosafety Policies and Procedures

  • Virginia Tech Policy 1005: Health and Safety: Policy link.
  • 3.1.5 Directors/Department Heads: Policy link.
  • 3.1.6 Faculty, Principal Investigators, Supervisors, and Safety Representatives: Policy link.
  • 3.1.7 Employees and Students: Policy link.
  • 3.1.8 Contractors: Policy link.
  • 3.1.9 Human Resources: Policy link.
  • 3.1.10 Environmental Health and Safety: Policy link.
  • 3.1.11 Office of Research Compliance: Policy link.
  • Virginia Tech Policy 433: Guidelines for the Use of Volunteers: Policy link.
  • Virginia Tech Policy 4815: Minors on Campus or Participating in University-Related Programs: Policy link.
  • Employee Accident Reporting Procedures: Procedures link.

Contact Information

Charlotte Waggoner
Assistant Director
ren@vt.edu | 540-231-5864

Michael Miles
Associate Biosafety Officer
msmiles@vt.edu | 540-231-3361

Anna Kroner
Associate Biosafety Officer
acastigl@vt.edu | 540-231-1122

Sarah Owen
Industrial Hygienist
sowen@vt.edu | 540-231-4034

Contact Biological Safety

Environmental Health & Safety mainline phone: 540-231-3600 (Monday-Friday, 8 a.m. - 5 p.m.)

Emergencies: 911 (after 5 p.m.; weekends)

Email: ehs-bsg-g@vt.edu