UCR Biosafety Manual
The UCR Biosafety manual defines the responsibilities, procedures, and guidelines for the safe handling, use, and disposal of biohazardous materials in research and teaching activities performed at UCR. The Biosafety Manual compliments the UCR Exposure Control Plan which contains additional policies and procedures for UCR personnel exposed to blood or other potentially infectious material.
If you have questions, please contact EH&S Biosafety at (951) 827-5528 or email@example.com
- Campus Biosafety Manual
- Document History
- 1. Purpose and Scope
- 2. Roles and Responsibilities
- 3. Biohazardous Research Approval Process
- 4. Laboratory Evaluations
- 5. Risk Groups
- 6. Biosafety Principles
- 7. Animal Biosafety
- 8. Plant Biosafety
- 9. Arthropod Biosafety
- 10. Personal Protective Equipment (PPE)
- 11. Laboratory Equipment
- 12. Sharps Safety
- 13. Transfer of Biological Materials
- 14. Decontamination, Disinfection, and Sterilization
- 15. Select Agents
- 16. Exposures and Injuries
- 17. Spill Procedures
- 18. Waste Management
- 19. Training
- Appendix A. Regulations, and Guidelines
- Appendix B. Links and Documents
- Acknowledgement Form
The UCR Biosafety Manual applies to all laboratories that use, store, or handle all biohazardous materials including recombinant/synthetic nucleic acids, human/animal/plant pathogens, and human and non-human primate materials. The Biosafety Officer will review the Biosafety Manual as least annually and update it as necessary.
|Date||Description of Changes||
Type of Change
(Review or Revision)
|3/18/2020||Revised and posted to EH&S website by Nick Noriea||Revision|
|3/15/2021||Reviewed by Tran Phan - no changes||Review|
|3/15/2022||Revised for change of Institutional Biosafety Committee administration from Office of Research Integrity to Environmental Health & Safety. Verified and updated references. Revised by Tran Phan||Revision|
|3/10/2023||Verified and updated figures and links.
Revised by Rob Miers
I. Purpose and Scope
The purpose of the UC Riverside (UCR) Biosafety Program is to define and implement biological safety policies and practices to safeguard all UCR personnel, the community, and the surrounding environment from exposure to biohazardous materials. The policies and procedures are defined by the UCR Biosafety Program to comply with all federal, state, and UC requirements.
The UCR Biosafety Manual defines the responsibilities, procedures, and guidelines for the safe handling, use, and disposal of biohazardous materials in research and teaching activities performed at UCR. The Biosafety Manual compliments the UCR Exposure Control Plan (ECP) which contains additional policies and procedures for UCR personnel exposed to blood or other potentially infectious material.
All UCR Principal Investigators (PIs), researchers, and UCR personnel who may enter biohazardous research areas should adhere to these biological safety policies and procedures in the conduct of their research and in the management of their laboratories.
II. Roles and Responsibilities
INSTITUTIONAL BIOSAFETY COMMITTEE (IBC)
The National Institutes of Health (NIH) requires the establishment of an Institutional Biosafety Committee (IBC) at any institution that receives NIH funding. At UCR, the IBC is appointed by the Office of Research Integrity (ORI) under the auspices of the Vice Chancellor for Research and Economic Development (VC-RED). Administrative support for the IBC is provided by Environmental Health & Safety (EH&S).
The IBC is a faculty-led review committee responsible for approval and oversight of activities involving the storage and use of biohazardous materials. The IBC consists of at least five individuals so selected that they have experience and expertise in recombinant/synthetic nucleic acid (r/sNA) technology and can assess the safety of biohazardous experiments. The committee must have at least two community members who are not affiliated with UCR, an appropriate recombinant or synthetic DNA expert, a plant or animal containment expert (when applicable), and the institutional Biosafety Officer (BSO). The IBC membership is primarily composed of UCR faculty with expertise and capability to identify potential risks to public health or the environment.
The IBC is responsible for:
- Ensuring research conducted at UCR is compliant with the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules (hereinafter as “NIH Guidelines”)
- Drafting and establishing campus biosafety policies and procedures for proper handling of biohazardous materials
- Reviewing individual research proposals for biosafety concerns
The IBC meets on the final Thursday of each month to review proposals from faculty. For more information, visit https://ehs.ucr.edu/ibc
ENVIRONMENTAL HEALTH AND SAFETY (EH&S)
Environmental Health and Safety oversees the Biosafety Program at UC Riverside.
EH&S is responsible for:
- Provide administrative support for the IBC.
- Monitors compliance with university, CDC, NIH, OSHA, USDA and state policies regarding the use of recombinant/synthetic nucleic acids, potentially infectious materials, pathogenic agents of livestock and agriculture, and exotic/invasive organisms
- Advises researchers and UCR personnel on appropriate safe work practices and procedures, containment controls, and personal protective equipment required for experimental protocols.
- Reviews and approves of transfer of biohazardous materials by PIs.
- Along with the IBC, reviews and approves all use of biohazardous materials.
- Provides training for proper handling, storing, disposing, and transporting of biohazardous materials in pursuant to regulatory requirements.
- Conducts periodic inspections of laboratories to ensure practices and procedures are rigorously followed and facilities are acceptable.
- Develops emergency procedures for occupational exposures and biohazardous spills.
- Investigates all reported accidents which may result in personnel or environmental exposure to biohazardous materials.
- Designate a Biosafety Officer (BSO) as defined by the NIH guidelines with duties including briefing the Institutional Biosafety Committee (IBC) on biohazardous research.
PRINCIPAL INVESTIGATOR (PI)
The Principal Investigator (PI) is directly and primarily responsible for the safe operation of their laboratory and for compliance by all their laboratory personnel with all UCR biosafety policies and procedures. The PI’s knowledge and judgement are critical in assessing risks and appropriately applying campus biosafety guidelines.
Principal Investigators (PIs) who wish to perform research using biological materials should submit a Biological Use Authorization (BUA) application to the IBC for approval prior to beginning work.
The PI is responsible for the following:
- They will not initiate research using biohazardous material and/or recombinant/synthetic nucleic acids until all applicable requirements outlined in this manual are met.
- They will consult and refer to NIH Guidelines and CDC’s Biosafety in Microbiological and Biomedical Laboratories (BMBL) to determine the appropriate Risk Group classification of the microorganisms to be used, and that they will use the prescribed microbiological practices and laboratory techniques required by the biosafety level assigned to their work by the IBC.
- They will report immediately to the BSO all deviations of the policies and procedures and all significant research-related accidents (spills, needle-sticks, exposure, injuries, etc.) which result in overt or potential exposure to infectious materials, or their release into the environment, whether contained within the laboratory or not.
- They are prepared to implement methods for dealing with accidents, spills, and personnel contamination.
- They maintain all appropriate state and federal permits required for certain animal and plant pathogens, exotic organisms, and plant pests prior to beginning work. PIs are strongly encouraged to contact EH&S Biosafety program at firstname.lastname@example.org for help in obtaining local, state, and federal permits.
- They will obtain the appropriate permits and Materials Transfer Agreements (MTA) for all appropriate importation, exportation and interstate shipping requirements for certain biological materials.
Prior to beginning research, the PI shall:
- Submit a BUA application for research and receive appropriate approval for all research projects.
- Submit BUA application amendment for any protocol changes that modify previously approved BUA procedures or alter the list of biohazards upon which approval was originally based.
- Determine the usefulness of serological screening, the requirements of medical surveillance, and the availability of vaccination for Risk Group 2 and 3 agents. Inform the laboratory staff of any precautionary medical practices advised or requested, such as vaccination or antibody titers. Where appropriate, the Hepatitis B vaccination should be offered free of charge to the employee.
- Assure that personnel working with infectious agents or biohazardous materials are appropriately trained, have a complete understanding of the hazards involved, and are proficient in the practices and techniques required for the safe handling of such materials.
During research, the PI shall:
- Appropriately supervise the performance of their staff to ensure that the required safety practices and techniques are employed.
- Investigate and report in writing to the IBC any significant biosafety problems pertaining to the pursuit of the research goals, specifically, new information which was not available at the time of the application.
- Correct any conditions or behaviors that might expose their personnel or release of biohazardous materials into the environment.
- Implement the procedures prescribed for dealing with laboratory incidents/accidents.
All personnel working in UCR research facilities, regardless of employment status, are responsible for the following:
- Follow all appropriate laboratory practices as outlined in this manual.
- Abide by all UC Riverside regulations and policies regarding safe conduct of research.
- Report all incidents, accidents, and possible unsafe work conditions to https://ehs.ucr.edu
- Familiarize and follow all protocols related to organisms and materials used in the laboratory regardless of whether or not they work directly with them.
- Follow all emergency procedures established by EH&S and the Principal Investigator (PI).
- Complete all appropriate training and verify documentation of training as required by EH&S.
III. Biohazardous Research Approval Process
All biohazardous research at UCR requires an approved Biological Use Authorization (BUA) prior to beginning research. Principal Investigators who plan to carry out research involving the use of biohazardous materials need to complete and submit a BUA through the online portal prior to beginning work. The BUA need to be approved prior to initiation of research.
BUA applications should be submitted at least three weeks before the scheduled meeting and have adequately addressed any issues raised during the pre-review. The IBC reviews all research involving infectious agents or for which sections III-A,III-B, III-C, III-D, or III-E of the NIH Guidelines apply. Exempt work (section III-F of the NIH Guidelines without any other biohazards under the purview of the IBC) requires BUA submission and Biosafety pre-review only.
WHAT REQUIRES A BUA?
A BUA submission is required if working with any biohazardous material.
Biohazardous material covered under the IBC Charter includes:
- Recombinant or Synthetic Nucleic Acids (r/sNA)
- Human or Non-Human Primate Material (including blood, tissue, body fluids, and organs)
- Immortalized/Established cell lines from humans (e.g. Caco2, HEK293) or non-human primates
- Infectious agents (including Risk Group 1 agents that are infectious to vertebrates other than humans)
- Toxins of biological origin
- Select Agents and Toxins as defined by CDC and USDA/APHIS, including federally-exempt quantities
- Cellular penetrating peptides and other peptides capable of infecting living cells
- Transgenic animals in which the genome will be altered by stable introduction of recombinant DNA into the germ-line (e.g. making founder lines and breeding existing lines to create new genetic lines, but NOT including purchasing or maintaining already established lines)
- USDA-regulated plant pests
- Noxious weeds or plants that can outcross with noxious weeds
- Human Gene Transfer
- Wild caught animals
New BUA submissions are processed by EH&S Biosafety program (EH&S) in Pre-Review. During Pre-Review, EH&S will review the BUA and may request the PI to modify the BUA. EH&S will make a formal determination of whether the BUA meets exempt status. For more information on the BUA approval process including Pre-Review, see https://ehs.ucr.edu/laboratory/biosafety/bua.
Once the BUA has completed Pre-Review, EH&S places it on the next IBC agenda to be formally reviewed by the Institutional Biosafety Committee (IBC). The IBC will vote whether to request additional information/modifications, decline, approve as is, or approve with conditions/stipulations. Only after the PI has been formally approved can research be initiated.
All research involving r/sNA needs to be submitted in a BUA. Exempt research is research that only contains r/sNA under section III-F of the NIH Guidelines and contain no other biohazards. The PI does not determine exempt status, EH&S will notify the PI in pre-review if the BUA qualifies as exempt.
APPLYING FOR BUA
Preparing for a BUA
Prepare to establish the following when preparing a BUA for submission:
- Describe the scope of work to be conducted with biohazardous materials
- Propose the biosafety level of containment
- Propose the applicable sections of the NIH Guidelines
- Identify the researchers, lab workers and collaborators
- Identify the approved work locations
- Specify the safe work practices and procedures that should be adhered to
Submitting a BUA
The UCR EH&S website has guidance for PIs to submit new BUAs, renew expiring BUAs, or amend current BUAs: https://ehs.ucr.edu/laboratory/biosafety/bua
Federal law requires significant deviations of NIH guidelines to be reported to NIH Office of Science Policy (OSP) within 30 days. Potential violations of NIH guidelines include research without prior IBC approval and failure to adhere to approved procedures and requirements.
Significant BUA deviations require the PI to stop work immediately. EH&S will issue the work-stoppage order to the PI. The PI may not continue work covered under the work-stoppage until formal notice to resume work.
Examples of BUA Deficiencies:
- Failure to obtain IBC approval prior to initiating work involving biohazardous materials
- Failure to amend BUA to include new biohazardous materials not described on the approved BUA
- Failure to follow procedures and policies established by the IBC and approved BUA
- Continuance of work after BUA expiration
- Continuance of work after work-stoppage notification by the IBC
- Significant research-related accidents resulting in overt exposure to personnel
IV. Laboratory Evaluations
EH&S conducts both scheduled and unannounced evaluations of laboratories on a regular basis. Evaluations are not limited to biosafety, but rather encompass additional research-related categories such as chemical, fire and life, radiation, and general safety. A typical evaluation of a BSL1 or BSL2 laboratory will include, but may not be limited to, evaluation of the following items for compliance and/or accuracy:
- Emergency procedures and phone numbers posted
- Active personnel listed on BUA
- Biohazard labels present at the entrance to the laboratory, on equipment, and waste containers
- Biohazard labels on secondary containers for red-bag biohazard waste
- Use of appropriate Personal Protective Equipment (PPE)
- Adherence to applicable medical surveillance program(s)
- Safety practices followed for aerosol-generating procedures
- Safe usage of sharps
- Decontamination of work areas
- Biosafety cabinet annual certification and work practices
- Eyewash present and inspected
- Biosafety Training
- Bloodborne Pathogens Training (if applicable)
- Hazardous Materials and Waste Management
- Laboratory Safety Fundamentals
- Fire Extinguisher Training
- Lab-specific Training
- Use of appropriate biohazard bags used for waste
- Use of appropriate red rigid sharps bins for biohazardous sharps waste
- Proper disposal of dry and liquid biohazardous waste
- Decontamination procedures followed
V. Risk Groups
When evaluating biohazards, it is helpful to categorize the risk present when handling various infectious agents. Categorizing infectious agents into Risk Groups (RG) allow researchers and biosafety professionals to anticipate expected procedures and practices necessary to work safely with that agent.
The principal hazardous characteristics of an infectious agent are:
- Capability to infect and cause disease in susceptible human/animal host
- Virulence as measured by severity of disease
- Availability of preventative measures (e.g. vaccines) and effective treatments
The NIH Guidelines established a classification of infectious agents (viruses, bacteria, fungi, parasites, etc.) into 4 risk groups (RG1 – RG4) using the characteristics listed above. The risk group increases with the risk of the agent
VI. Biosafety Principles
The Biosafety in Microbiological and Biomedical Laboratories (BMBL) manual describes four Biosafety Levels (BSL): combinations of laboratory safety practice and techniques, safety equipment and laboratory facilities. The Biosafety Levels were chosen to represent the conditions under which the agents can be ordinarily handled safely.
Biosafety Level 1
BSL-1 represents a basic level of containment that relies on standard microbiological practices only and a sink for basic hand washing. BSL-1 labs are appropriate for undergraduate/graduate teaching labs and for other labs in which work is done with defined microbes that are not known to cause disease in humans.
Biosafety Level 2
BSL-2 is the minimum biosafety level required to work with human pathogens. BSL-2 laboratories may be appropriate for a spectrum of moderate-risk agents associated with diseases of varying severity.
Biosafety Level 3
BSL-3 is required for work with indigenous or exotic agents with a potential for respiratory transmission and which may cause serious or potentially lethal infection. At this level emphasis is placed on aerosol transmission safeguards.
Biosafety Level 4
BSL-4 applies to work with dangerous and exotic agents that pose a high individual risk of life-threatening disease, may be transmitted via the aerosol route, and for which there is no treatment or intervention.
STANDARD MICROBIOLOGICAL PRACTICES (BSL-1)
All labs which contain biohazardous materials should adhere to standard microbiological practices. These practices and procedures encompass all the requirements of Biosafety Level 1 (BSL1).
- The laboratory supervisor can control access to the laboratory.
- The campus-wide biosafety manual should be available and accessible.
- Laboratory personnel should wash their hands after handling cultures, removing gloves and before leaving the laboratory.
- Eating, drinking, smoking, handling contacts lenses, application of cosmetics, and storing food or beverage for human consumption are prohibited.
- Pipetting by mouth is prohibited.
- Animals and plants not involved in the experiment are not permitted in the laboratory.
- Policies for handling sharps such as needles, blades, pipettes, and broken glass are implemented. Use of sharps is reduced as much as possible.
- Needles should not be bent, cut, removed from disposable syringes, or otherwise manipulated.
- Disposable sharps should be placed in specially designed puncture and leak-proof sharps containers and disposed of appropriately as medical waste.
- Non-disposable sharps should be placed in a hard-walled container for transport to decontamination, preferably by autoclaving.
- Broken glass should never be handled directly. Use tongs, forceps, or dustpan and brush. Use plastic instead of glass whenever possible.
- All laboratory procedures should be performed to minimize the creation of aerosols or splashes.
- Decontaminate all work surfaces after completing work, or after splashes or spills.
- Decontaminate all potentially biohazardous materials before disposal by an effective means.
- If decontamination of materials occurs outside the immediate laboratory, materials to be decontaminated should be packed in accordance with all applicable, local, state, and federal regulations and transported in durable, leak proof containers that are secured prior to transport.
- Signage incorporating the biohazard symbol should be posted when infectious agents are present.
- The laboratory supervisor should post safety procedures and equipment required for entry into animal housing areas containing known or potentially infected animals.
- An effective integrated pest management program is required.
- The laboratory supervisor should ensure that all laboratory personnel receive and annually refresh:
- Training appropriate to their duties
- Training on appropriate laboratory precautions
- Training on exposure evaluation procedures
- Training whenever a new hazardous material or procedure is introduced
Standard Laboratory Design
All UCR laboratories should adhere to the UC Lab Safety Design Manual. All UCR laboratories designated BSL1 or higher should fulfill the following criteria:
- Laboratories should have doors to isolate them from non-laboratory areas. Doors should be kept closed when experimental animals are present.
- Laboratories should be kept neat; good housekeeping procedures should be in place and in regular use.
- Laboratories should have a sink for hand washing. The sink should be located near the laboratory exit. It may be manually, hands-free or automatically operated.
- Laboratory furniture should be secured and appropriate for anticipated loads and uses. Spaces between benches, cabinets, and equipment should be accessible for cleaning and decontamination.
- Bench tops are impervious to water, moderate heat and chemicals.
- Laboratories are designed for ease of decontamination (e.g. no carpets, sealed surfaces, no unreachable areas, etc.)
- Seating in the laboratory should be covered with a non-porous material that can easily be cleaned and decontaminated.
- All laboratory windows to the outside should be fitted with fly screens.
- All laboratory spaces shall exhaust 100% of the air to the outside. Recirculated air is not allowed.
BIOSAFETY LEVEL 2 (BSL-2)
Biosafety level 2 builds upon BSL1 by adding additional special practices and safety equipment.
- All persons entering the laboratory should be advised of the potential hazards and should meet specific entry and exit requirements.
- Laboratory personnel are provided with medical surveillance and offered immunizations for agents handled or potentially present in the laboratory.
- The supervisor should ensure that laboratory personnel demonstrate proficiency in standard and special microbiological practices including disinfection and spill response before working with any biohazards.
- Potentially infectious and/or recombinant materials should be in durable leak proof containers during collection, handling, processing, storage or transport within a facility.
- Laboratory equipment should be routinely decontaminated as well as after any potential contamination.
- Equipment should be decontaminated before repair, maintenance, or removal from the laboratory. Guidance documents can be found at https://ehs.ucr.edu/laboratory
- Potential infectious agent exposure incidents are immediately evaluated and treated per the laboratory biosafety manual. They are reported to the supervisor. Medical evaluation, treatment and surveillance are provided as appropriate.
- All laboratory procedures that may generate aerosols should be performed in a properly certified biological safety cabinet or other physical containment device.
- Centrifugation of potentially infectious material requires the use of safety cups if the centrifuge is outside of a biosafety cabinet.
- Sonication of potentially infectious material requires the use of a biosafety cabinet or sealed cup horn.
- Primary barriers such as biosafety cabinets should be used when handling biohazardous materials. This may include pipetting, grinding, centrifuging, shaking, blending, sonicating, opening containers of infectious material, intranasal animal inoculation, harvesting infected tissues from animals or eggs, or if high concentrations are used.
- Biosafety cabinets should be certified at least annually, and whenever moved or after repair work is done, if used to contain biohazards.
- Biosafety cabinets are installed so that laboratory air movement does not disrupt cabinet operation; away from doors, windows, high-traffic areas, ventilation registers and other disruptions.
- HEPA-filtered cabinet exhaust may be re-circulated to the laboratory if the cabinet is certified at least annually. The exhaust may also be ducted to the outside via either a thimble- or hard-connected exhaust duct.
- PPE should be disposed of with other contaminated waste or decontaminated before reuse. Contact lens wearers should also use eye protection.
- Double gloving may be appropriate. Recommend using darker color inner glove and lighter color outer glove to more easily identify breaks or tears in an outer glove.
- Doors should be self-closing and have locks
- Vacuum lines should be protected by HEPA filters that are cleaned or replaced as needed. Filter traps may be required.
- Autoclave, chemical disinfection, incineration or other approved decontamination method should be available in the facility for waste treatment prior to disposal.
BIOSAFETY LEVEL 3 (BSL-3)
Biosafety Level 3 (BSL3) builds upon BSL2 with several additional requirements. Laboratories must have anterooms, negative airflow, and single-pass filtered exhaust. Work is restricted to biosafety cabinets. The facility design includes several additional standards and rigorous safety and security checks are required for personnel working at BSL3.
For additional information on High Containment (BSL3) Laboratories, see Biosafety in Microbiological and Biomedical Laboratories (BMBL) 6th edition or contact email@example.com.
VII. Animal Biosafety
All research involving animals at UCR must be reviewed and approved by the UCR Institutional Animal Care and Use Committee (IACUC) using the Animal Use Protocol (AUP). Animal research containing biohazards need to also be described in an approved Biological Use Authorization (BUA). When working with biohazards and animals, consider the following as part of the research risk assessment:
- Dosage and route of administration of biohazards to be used in animal model
- Any infectious agents associated with the animal model (e.g. Herpes B virus, C. burnetii)
- Shedding of infectious organisms/biohazards from the animal through blood, feces, urine, etc.
- Potential allergies
ANIMAL CONTAINMENT PRINCIPLES
The Animal Biosafety Level (ABSL) required for given animal research is assigned by the Institutional Biosafety Committee (IBC). Like general laboratory biosafety levels, animal biosafety levels are assigned 1 to 4 and reflect a standard set of practices, equipment, and containment for the safe handling of biohazardous materials in animal research.
Animal Biosafety Level 1 (ABSL-1)
Suitable for work in animals involving well-characterized agents that are not known to cause disease in immunocompetent adult humans and present minimal potential hazard to personnel and the environment.
Animal Biosafety Level 2 (ABSL-2)
Suitable for work involving laboratory animals infected with agents associated with human disease and pose moderate hazards to personnel and the environment.
Animal Biosafety Level 3 (ABSL-3)
Suitable for work with laboratory animals infected with indigenous or exotic agents that present a potential for aerosol transmission and agents causing serious or potentially lethal disease.
LABORATORY OCCUPATIONAL HEALTH PROGRAM
All UCR animal users are offered enrollment in the UCR Animal Occupational Health and Safety Program. In part, enrollment requires annual evaluation by a medical occupational health specialist. Contact IACUCtraining@ucr.edu for further assistance regarding the Occupational health program.
VIII. Plant Biosafety
The research of plants and their associated organisms present interesting and unique containment concerns. While the risk of human infection is lower, environmental risk can increase. The unintentional release of a transgenic arthropod or exotic weed can have significant effects on the surrounding environment.
Plant-related biohazards include:
- Transgenic plants
- Plants associated with transgenic microbes or arthropods
- Noxious weeds or plants that can outcross with noxious weeds
- USDA recognized plant pests
- Plants containing the genome of any non-exotic infectious agent
- Plants containing exotic agents
- Plants encoding the sequence of potent vertebrate toxins (LD50 < 100 ng/kg)
- Plants associated with non-exotic microorganisms that have a recognized potential for serious detrimental impact on managed or natural ecosystems
Research with plant-associated biohazards may also require permits with conditions specified by USDA/APHIS or CDFA. The EH&S biosafety program will assist PIs in application and compliance for plant-related regulatory permits. Approved USDA/APHIS or CDFA permits are not a substitute for BUA approval.
For experiments in which plants are grown in BSL-1 and BSL-2 laboratory settings, containment practices should be in correspondence with standard biosafety levels. This includes plant tissue culture rooms and growth chambers within laboratory facilities. Some additional containment procedures may be required by the IBC prior to research initiation.
For research that requires research plants of a size, number, or growth requirements that preclude the use of laboratory containment conditions, additional precautions need to be taken. UCR employs over a thousand acres of field research with greenhouses, screen-houses, and two field stations. The UCR main campus additionally contains several greenhouses on site. Recommended containment practices in these greenhouses is described by the Plant Biosafety Levels (BSL-P).
IX. Arthropod Biosafety
Laboratory research involving arthropods present challenges distinct from typical microbiological and biomedical laboratories. Escape and environmental protection are significant concerns. Many arthropods may not be infectious but act as a vector for diseases in the environment. Some arthropods are deleterious to livestock or agriculture (e.g. a plant pest). The small size and motile characteristics of many transgenic arthropods require additional administrative, equipment, and design safeguards to prevent release.
What Arthropod research requires a BUA?
Arthropod-related biohazards include:
- Transgenic arthropods
- Exotic arthropods
- Arthropods associated with exotic agents
- Arthropods that transmit pathogens of public health importance
- Arthropods that are known plant pests as defined by the USDA
- Arthropods associated with transgenic microbes, plants, or other associated animals
Arthropods to be considered for risk assessment include
- Insects (Diptera, Hemiptera, Phtiraptera, Siphonaptera)
- Arachnids (Acari)
All life cycle stages are considered under the term arthropod: eggs, larvae, nymphs, and adults.
ARTHROPOD RISK ASSESSMENT
When containment is required for arthropod research, the following should be considered:
- Is the arthropod exotic, and if so, is it likely to establish locally if it escapes?
- Does the arthropod have acquired or characterized insecticide-resistance (through genotype or transgenics)?
- Is the arthropod a vector for any infectious agents found in the environment?
- Are disabled strains available, whose viability after escape would be limited?
- Is the arthropod transgenic? If so, does this alter capability of pathogen transmission or environmental persistence?
X. Personal Protective Equipment (PPE)
The UC Personal Protective Equipment (PPE) policy is designed to prevent workplace injuries and illnesses for academic appointees, staff, students, and visitors. General laboratory PPE is determined by the PI’s Laboratory Hazard Assessment. Risks involving biohazards may require additional PPE beyond what the Laboratory Hazard Assessment describes. All approved Biological Use Authorization (BUA) describe additional PPE requirements for working with the associated biohazards.
Before discussing PPE, one should become familiar with the NIOSH Hierarchy of Controls that are used to protect workers. The top controls are more effective and preventative than controls at the bottom.
All researchers need to wear full length pants (or equivalent) and closed toe/heel shoe attire at all times while entering or occupying laboratory areas. The area of skin between the pants and shoe should not be exposed.
All researchers working in labs with biological hazards need to wear a lab coat. Lab coats are supplied free of charge by UCR Environmental Health and Safety through the Laboratory Hazard Assessment Tool. There are many types of lab coats available; researchers working with biohazards are recommended the traditional white lab coat or the barrier/fluid-resistant white lab coat. Some biohazards may require additional protection such as disposable gowns.
Lab coats should not be taken home. Researchers working with biohazards may contaminate their lab coats during normal usage. Due to the high risk of lab coats acting as fomites, lab coats should be kept at work. EH&S supplies two lab coats to each researcher to ensure that the researcher may continue to wear the appropriate lab coat while the other is laundered.
Gloves should be worn at all times when in contact, or could potentially come in contact, with any biohazardous material in the laboratory. Standard latex or nitrile gloves are sufficient for working with biological hazards, however if chemical hazards are involved these gloves may not be sufficient. Many types of gloves offer biological protection but may not offer equal levels of chemical protection.
Autoclave gloves should always be used for removing hot material from autoclaves. Cryo gloves should be used for handling liquid nitrogen.
Researchers working with biological hazards may consider several types of eye protection including safety glasses, safety goggles, and face shields. Researchers are required, at a minimum, to wear safety glasses while working with biohazards.
Surgical masks are not respirators. Surgical masks are designed to protect the user’s nose and mouth from liquid droplets, they are not used to protect against aerosols. Surgical masks are used to prevent dust/particle inhalation.
Respirators are devices designed to protect the wearer from airborne hazards such as airborne transmissible microorganisms. The selection of respirator depends on the type of hazard potentially present and potential for aerosol production.
N95 respirators are certified to block 95% of particles 0.3 µm or larger (0.3 µm is roughly the size of a single virus). All researchers required to wear N95 need to be fit tested and complete an occupational medical health questionnaire to ensure that the N95 poses no additional safety hazards to the user. To request a medical questionnaire or schedule a fit test, contact EH&S at firstname.lastname@example.org. Some researchers may not be required to wear a respirator but choose to wear a N95. For volunteer use forms for N95, contact email@example.com.
Powered Air Purifying Respirator (PAPR)
The PAPR provides one of the highest levels of protection against biohazards. PAPRs are commonly found in high containment laboratories to protect against aerosolizable infectious material. A PAPR contains a battery-powered blower, facemask, breathing tube, and HEPA filter. The air is blown across the filter and deliver breathable air into the facemask. The filter cartridge may be exchanged to filter chemical vapors and toxic gases. EH&S requires annual training for all PAPR users.
XI. Laboratory Equipment
Laboratory equipment represent the engineering controls used to control researcher exposure to biohazards. Many pieces of equipment function as primary containment when used with infectious material. Used correctly, laboratory equipment can provide high levels of protection for both the user, their research, and the environment.
TYPES OF PROTECTION
There are 3 types of protection that equipment can provide:
- Personnel Protection: To protect people from hazards
- Product Protection: To protect experimental materials from contamination
- Environmental Protection: To protect the surrounding environment from hazards
When considering equipment for the lab, researchers should always examine what kind of containment they require versus what kind of containment the equipment provides.
Biological Safety Cabinets (BSCs) are designed to provide personnel, environmental and product protection when appropriate practices and procedures are followed. There are three classes of BSCs. At UC Riverside, almost all biological safety cabinets approved for use of biohazards and human material are Class II. Please contact the biosafety office at firstname.lastname@example.org if you plan to use a Class I or Class III cabinet in your research. Class II cabinets can be further subdivided into room-exhausted (Class II-A) or ducted exhaust (Class II-B). Class II-A cabinets recirculate 70% of air back into the work zone while Class II-B cabinets recirculate either 0% (Class II-B2) or 30% (Class II-B1) of air into the work zone. Class II biosafety cabinets provide all 3 types of protection.
Figure XI-1. Types of Protection offered
Figure XI-2. Classes of Type II Biosafety Cabinets
Figure XI-2. Class II-A1 Biosafety Cabinet. (A) front opening; (B) Sash; (C) exhaust HEPA filter; (D) return HEPA filter; (E) common plenum; (F) blower. (BMBL 5th ed)
Figure XI-3. Class II-B1 Biosafety Cabinet. (A) front opening; (B) Sash; (C) exhaust HEPA filter; (D) supply HEPA filter; (E) negative pressure dedicated plenum; (F) blower; (G) additional supply HEPA filter. (BMBL 5th ed)
PROCEDURE FOR USING A BIOSAFETY CABINET
- Never put your head inside the cabinet.
- 10% household bleach may be used safely in biosafety cabinets; however bleach should be followed by sterile water or 70% ethanol to prevent pitting to the stainless steel surface.
- The biosafety cabinet is only effective at the sash height it was certified at. If the sash is at an inappropriate level for your height, adjust the seat- never adjust the sash.
- Biosafety cabinets need to be certified annually to ensure correct operation. Any BSC used for biohazardous materials should be current on certification.
- Wear appropriate PPE.
- Check certification sticker located on the exterior surface of BSC to confirm that BSC has been certified within the past 12 months. Notify PI if BSC certification exceeds 12 months. In this case, do not use BSC.
- Turn on cabinet blowers.
- Check the magnehelic gauge to ensure BSC is working properly.
- Raise the sash to the certified operating height. Wait at least 2– 5 minutes prior to beginning work to allow the BSC to purge.
- Do not ignore alarms.
- Adjust seat height so that researcher’s face is above the front opening.
- Using a squeeze bottle, spray the appropriate disinfectant onto interior surfaces of the BSC (work surface, side walls, back walls, front grill, inner glass of sash etc.). Do not put your head inside the BSC during cleaning.
- Wipe down the work surface area by starting at the left front corner at the grill and wiping across the front grill. Continue wiping horizontally across towards the back of the BSC (see Figure 11-4 – yellow arrows).
- Wipe down inner glass of sash by starting on the lower left corner and wiping up and down while moving horizontally (see Figure 11-4 – blue arrows).
Figure 11-4. Cleaning work surface (yellow arrows) and sash (blue arrows)
- Wipe down side walls and back of BSC (see Figure 11-5).
Figure 11-5. Cleaning side walls and back
- Dispose soiled paper towels into biohazard bag.
- Replace gloves. BSC is ready to be set up before commencing work.
- Place all materials towards the back of the work area away from front and rear grills. Leave at least 6 inches between work area and the side, front, and back grills.
- Set up BSC work flow from clean to dirty as shown in figure XI-6.
UV Lamps in Biosafety Cabinets
The CDC, NIH, NSF, and American Biological Safety Association (ABSA) agree that UV lamps are neither recommended nor required in a Biological Safety Cabinet.
All material inside a biosafety cabinet should be decontaminated with an approved disinfectant prior to removal, regardless if the material was treated with UV light.
All vacuum lines used for biohazardous materials should be protected with a vacuum line HEPA filter placed closest to the vacuum line port. Attach an aspiration flask (“trap”). A second inline aspiration flask is recommended. The aspiration flask(s) should be placed into a non-absorbent, rigid secondary container. Styrofoam and cardboard are not appropriate for use as a secondary container.