Biological Safety

Research laboratories working with biohazardous material, microorganisms and/or recombinant or synthetic DNA technologies are special, often unique, work environments. The materials being used may pose special risks to persons working in or near the laboratory or to the environment should the material escape containment procedures established for the laboratory.  UNC Asheville's Biosafety Manual has been established to ensure the safe handling of biohazardous agents, ensure the appropriate assessment of potential risks, and reduce the opportunity of personnel exposure or accidental environmental release.

UNC Aheville's EH&S Professional makes recommendations to Faculty for all research and teaching activities involved with potentailly hazardous biological materials and recombinanat or synthetic DNA in a way to minimize or eliminate risks associated with their research or teaching activities.  

Biological Hazard

Biological hazards are those materials of biological origin that could potentially cause harm to humans, animals, or plants.  Examples include infectious agents, toxins, pathogenic microorganisms, human or primate sourced materials (primary and established cell lines) and/or recombinant or synthetic DNA.

Principles of Biosafety

Biosafety is the maintenance of safe conditions in biological research to prevent harm from infectious agents or hazardous biological materials that present a risk or potential risk to the health of humans, animals, or the environment. 

The Center for Disease Control (CDC) guidelines, Biosafety in Microbiological and Biomedical Laboratories (BMBL) has become the code of practice for biosafety - the discipline addressing the safe handling and containment of infectious microorganisms and hazardous biological materials.

The principles of biosafety are containment and risk assessment. The fundamentals of containment include the microbiological practices, safety equipment, and facility safeguards that protect laboratory workers, the environment, and the public from exposure to infectious microorganisms that are handled and stored in the laboratory. Risk assessment is the process that enables the appropriate selection of microbiological practices, safety equipment, and faciltiy safeguards that can prevent laboratory-associated infectioins (LAI).

Summary or Recommended Biosafety Levels for Infectious Agents

Biosafety Level Agents Practices Primary Barriers and Safety Equipment

Facilities

(Secondary Barriers

BSL1

  • Not known to consistently cause diseases in healthy adults
  • Standard microbiological practices
  • No primary barriers required
  • PPE: laboratory coats and gloves; eye, face protection, as needed
  • Laboratory bench and sink required
BSL2
  • Agents associated with human disease.
  • Routes of transmission include percutaneous injury, ingestion, mucous membrane exposure

BSL1 practice plus:

  • Limited access
  • Biohazard warning signs
  • "Sharps" precautions
  • Biosafety manual defining any needed waste decontamination or medical surveillance policies

Primary barriers:

  • BSCs or other physical containment devices used for all manipulations of agents that cause splashes or aerosols of infectious materials
  • PPE: Laboratory coats, gloves, face and eye protection, as needed

BSL1 plus:

  • Autoclave available
BSL3
  • Indigenous or exotic agents that may cause serious or potentially lethal disease through the inhalation route of exposure

BSL2 practice plus:

  • Controlled access
  • Decontamination of all waste
  • Decontamination of laboratory clothing before laundering

Primary barriers:

  • BSCs or other physical containment devices used for all open manipulations of agents
  • PPE: Protective laboratory clothing, gloves, face, eye and respiratory protection, as needed

BSL2 plus:

  • Physical separation from access corridors
  • Self-closing, double door access
  • Exhausted air not reciirculated
  • Negative airflow into laboratory
  • Entry through airlock or anteroom
  • Hand washing sink near laboratory exit
BSL4
  • Dangerous/exotic agents which post high individual risk of aerosol-transmitted laboratory infections that are frequently fatal, for which there are no vaccines or treatments.
  • Agents with a close or identical antigenic relationship to an agent requiring BSL4 until data are available to redesignate the level
  • Related agents with unknown risk of transmission

BSL3 practices plus:

  • Clothing change before entering
  • Shower on exit
  • All material decontaminated on exit from facility

Primary barriers:

  • All procedures conducted in Class III BSCs or Class I or II BSCs in combination with full-body, air-supplied, positive pressure suit

BSL3 plus:

  • Separate building or isolated zone
  • Dedicated supply and exhaust, vacuum, and decontamination systems
  • Other requirements outlined in the text

ONLY BSL1 and BSL2 agents are used in UNC Asheville's Biological Research Laboratories.

ONLY BSL1 agents are used in UNC Asheville's Biological Teaching Activities.

Laboratory Biosafety Level Criteria for BSL1 Laboratories

Biosafety Level 1

Biosafety Level 1 is suitable for work involving well-characterized agents not known to consistently cause disease in immunocompetent adult humans, and present minimal potential hazard to laboratory personnel and the environment. BSL-1 laboratories are not necessarily separated from the general traffic patterns in the building. Work is typically conducted on open bench tops using standard microbiological practices. Special containment equipment or facility design is not required, but may be used as determined by appropriate risk assessment. Laboratory personnel must have specific training in the procedures conducted in the laboratory and must be supervised by a scientist with training in microbiology or a related science.

The following standard practices, safety equipment, and facility requirements apply to BSL-1.

Standard Microbiological Practices

  1. The laboratory supervisor must enforce the institutional policies that control access to the laboratory.
  2. Persons must wash their hands after working with potentially hazardous materials and before leaving the laboratory.
  3. Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human consumption must not be permitted in laboratory areas. Food must be stored outside the laboratory area in cabinets or refrigerators designated and used for this purpose.
  4. Mouth pipetting is prohibited; mechanical pipetting devices must be used.
  5. Policies for the safe handling of sharps, such as needles, scalpels, pipettes, and broken glassware must be developed and implemented. Whenever practical, laboratory supervisors should adopt improved engineering and work practice controls that reduce risk of sharps injuries. Precautions, including those listed below, must always be taken with sharp items. These include:
  • Careful management of needles and other sharps are of primary importance. Needles must not be bent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated by hand before disposal.
  • Used disposable needles and syringes must be carefully placed in conveniently located puncture-resistant containers used for sharps disposal.
  • Non-disposable sharps must be placed in a hard walled container for transport to a processing area for decontamination, preferably by autoclaving.
  • Broken glassware must not be handled directly. Instead, it must be removed using a brush and dustpan, tongs, or forceps. Plastic ware should be substituted for glassware whenever possible.
  1. Perform all procedures to minimize the creation of splashes and/or aerosols.
  2. Decontaminate work surfaces after completion of work and after any spill or splash of potentially infectious material with appropriate disinfectant.
  3. Decontaminate all cultures, stocks, and other potentially infectious materials before disposal using an effective method. Depending on where the decontamination will be performed, the following methods should be used prior to transport.
  • Materials to be decontaminated outside of the immediate laboratory must be placed in a durable, leak proof container and secured for transport.
  • Materials to be removed from the facility for decontamination must be packed in accordance with applicable local, state, and federal regulations.
  1. A sign incorporating the universal biohazard symbol must be posted at the entrance to the laboratory when infectious agents are present. The sign may include the name of the agent(s) in use, and the name and phone number of the laboratory supervisor or other responsible personnel. Agent information should be posted in accordance with the institutional policy.
  2. An effective integrated pest management program is required.
  3. The laboratory supervisor must ensure that laboratory personnel receive appropriate training regarding their duties, the necessary precautions to prevent exposures, and exposure evaluation procedures. Personnel must receive annual updates or additional training when procedural or policy changes occur. Personal health status may impact an individual’s susceptibility to infection, ability to receive immunizations or prophylactic interventions. Therefore, all laboratory personnel and particularly women of childbearing age should be provided with information regarding immune competence and conditions that may predispose them to infection. Individuals having these conditions should be encouraged to self-identify to the institution’s healthcare provider for appropriate counseling and guidance.

Special Practices

None required.

Safety Equipment (Primary Barriers and Personal Protective Equipment)

  1. Special containment devices or equipment, such as BSCs, are not generally required.
  2. Protective laboratory coats, gowns, or uniforms are recommended to prevent contamination of personal clothing.
  3. Wear protective eyewear when conducting procedures that have the potential to create splashes of microorganisms or other hazardous materials. Persons who wear contact lenses in laboratories should also wear eye protection.
  4. Gloves must be worn to protect hands from exposure to hazardous materials. Glove selection should be based on an appropriate risk assessment. Alternatives to latex gloves should be available. Wash hands prior to leaving the laboratory. In addition, BSL-1 workers should:
  • Change gloves when contaminated, glove integrity is compromised, or when otherwise necessary.
  • Remove gloves and wash hands when work with hazardous materials has been completed and before leaving the laboratory.
  • Do not wash or reuse disposable gloves. Dispose of used gloves with other contaminated laboratory waste. Hand washing protocols must be rigorously followed.

Laboratory Facilities (Secondary Barriers)

  1. Laboratories should have doors for access control.
  2. Laboratories must have a sink for hand washing.
  3. The laboratory should be designed so that it can be easily cleaned. Carpets and rugs in laboratories are not appropriate.
  4. Laboratory furniture must be capable of supporting anticipated loads and uses. Spaces between benches, cabinets, and equipment should be accessible for cleaning.
  • Bench tops must be impervious to water and resistant to heat, organic solvents, acids, alkalis, and other chemicals.
  • Chairs used in laboratory work must be covered with a non-porous material that can be easily cleaned and decontaminated with appropriate disinfectant.
  1. Laboratories windows that open to the exterior should be fitted with screens

 

Laboratory Biosafety Level Criteria for BSL2 Laboratories

Biosafety Level 2

Biosafety Level 2 builds upon BSL-1. BSL-2 is suitable for work involving agents that pose moderate hazards to personnel and the environment. It differs from BSL-1 in that: 1) laboratory personnel have specific training in handling pathogenic agents and are supervised by scientists competent in handling infectious agents and associated procedures; 2) access to the laboratory is restricted when work is being conducted; and 3) all procedures in which infectious aerosols or splashes may be created are conducted in BSCs or other physical containment equipment.

The following standard and special practices, safety equipment, and facility requirements apply to BSL-2.

Standard Microbiological Practices

  1. The laboratory supervisor must enforce the institutional policies that control access to the laboratory.
  2. Persons must wash their hands after working with potentially hazardous materials and before leaving the laboratory.
  3. Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human consumption must not be permitted in laboratory areas. Food must be stored outside the laboratory area in cabinets or refrigerators designated and used for this purpose.
  4. Mouth pipetting is prohibited; mechanical pipetting devices must be used.
  5. Policies for the safe handling of sharps, such as needles, scalpels, pipettes, and broken glassware must be developed and implemented. Whenever practical, laboratory supervisors should adopt improved engineering and work practice controls that reduce risk of sharps injuries. Precautions, including those listed below, must always be taken with sharp items. These include:
  • Careful management of needles and other sharps are of primary importance. Needles must not be bent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated by hand before disposal.
  • Used disposable needles and syringes must be carefully placed in conveniently located puncture-resistant containers used for sharps disposal.
  • Non-disposable sharps must be placed in a hard walled container for transport to a processing area for decontamination, preferably by autoclaving.
  • Broken glassware must not be handled directly. Instead, it must be removed using a brush and dustpan, tongs, or forceps. Plastic ware should be substituted for glassware whenever possible.
  1. Perform all procedures to minimize the creation of splashes and/or aerosols.
  2. Decontaminate work surfaces after completion of work and after any spill or splash of potentially infectious material with appropriate disinfectant.
  3. Decontaminate all cultures, stocks, and other potentially infectious materials before disposal using an effective method. Depending on where the decontamination will be performed, the following methods should be used prior to transport:
  • Materials to be decontaminated outside of the immediate laboratory must be placed in a durable, leak proof container and secured for transport.
  • Materials to be removed from the facility for decontamination must be packed in accordance with applicable local, state, and federal regulations.
  1. A sign incorporating the universal biohazard symbol must be posted at the entrance to the laboratory when infectious agents are present. Posted information must include: the laboratory’s biosafety level, the supervisor’s name (or other responsible personnel), telephone number, and required procedures for entering and exiting the laboratory. Agent information should be posted in accordance with the institutional policy.
  2. An effective integrated pest management program is required.
  3. The laboratory supervisor must ensure that laboratory personnel receive appropriate training regarding their duties, the necessary precautions to prevent exposures, and exposure evaluation procedures. Personnel must receive annual updates or additional training when procedural or policy changes occur. Personal health status may impact an individual’s susceptibility to infection, ability to receive immunizations or prophylactic interventions. Therefore, all laboratory personnel and particularly women of childbearing age should be provided with information regarding immune competence and conditions that may predispose them to infection. Individuals having these conditions should be encouraged to self-identify to the institution’s healthcare provider for appropriate counseling and guidance.

Special Practices

  1. All persons entering the laboratory must be advised of the potential hazards and meet specific entry/exit requirements.
  2. Laboratory personnel must be provided medical surveillance, as appropriate, and offered available immunizations for agents handled or potentially present in the laboratory.
  3. Each institution should consider the need for collection and storage of serum samples from at-risk personnel.
  4. A laboratory-specific biosafety manual must be prepared and adopted as policy. The biosafety manual must be available and accessible.
  5. The laboratory supervisor must ensure that laboratory personnel demonstrate proficiency in standard and special microbiological practices before working with BSL-2 agents.
  6. Potentially infectious materials must be placed in a durable, leak proof container during collection, handling, processing, storage, or transport within a facility.
  7. Laboratory equipment should be routinely decontaminated, as well as, after spills, splashes, or other potential contamination.
  • Spills involving infectious materials must be contained, decontaminated, and cleaned up by staff properly trained and equipped to work with infectious material.
  • Equipment must be decontaminated before repair, maintenance, or removal from the laboratory.
  1. Incidents that may result in exposure to infectious materials must be immediately evaluated and treated according to procedures described in the laboratory biosafety manual. All such incidents must be reported to the laboratory supervisor. Medical evaluation, surveillance, and treatment should be provided and appropriate records maintained.
  2. Animal and plants not associated with the work being performed must not be permitted in the laboratory.
  3. All procedures involving the manipulation of infectious materials that may generate an aerosol should be conducted within a BSC or otherphysical containment devices.

Safety Equipment (Primary Barriers and Personal Protective Equipment)

  1. Properly maintained BSCs, other appropriate personal protective equipment, or other physical containment devices must be used whenever:
  • Procedures with a potential for creating infectious aerosols or splashes are conducted. These may include pipetting, centrifuging, grinding, blending, shaking, mixing, sonicating, opening containers of infectious materials, inoculating animals intranasally, and harvesting infected tissues from animals or eggs.
  • High concentrations or large volumes of infectious agents are used. Such materials may be centrifuged in the open laboratory using sealed rotor heads or centrifuge safety cups.
  1. Protective laboratory coats, gowns, smocks, or uniforms designated for laboratory use must be worn while working with hazardous materials. Remove protective clothing before leaving for non-laboratory areas, e.g., cafeteria, library, and administrative offices). Dispose of protective clothing appropriately, or deposit it for laundering by the institution. It is recommended that laboratory clothing not be taken home.
  2. Eye and face protection (goggles, mask, face shield or other splatter guard) is used for anticipated splashes or sprays of infectious or other hazardous materials when the microorganisms must be handled outside the BSC or containment device. Eye and face protection must be disposed of with other contaminated laboratory waste or decontaminated before reuse. Persons who wear contact lenses inlaboratories should also wear eye protection.
  3. Gloves must be worn to protect hands from exposure to hazardous materials. Glove selection should be based on an appropriate risk assessment. Alternatives to latex gloves should be available. Gloves must not be worn outside the laboratory. In addition, BSL-2 laboratory workers should:
  • Change gloves when contaminated, glove integrity is compromised, or when otherwise necessary.
  • Remove gloves and wash hands when work with hazardous materials has been completed and before leaving the laboratory.
  • Do not wash or reuse disposable gloves. Dispose of used gloveswith other contaminated laboratory waste. Hand washing protocols must be rigorously followed.
  1. Eye, face and respiratory protection should be used in rooms containing infected animals as determined by the risk assessment.


Laboratory Facilities (Secondary Barriers)

  1. Laboratory doors should be self-closing and have locks in accordance with the institutional policies.
  2. Laboratories must have a sink for hand washing. The sink may be manually, hands-free, or automatically operated. It should be located near the exit door.
  3. The laboratory should be designed so that it can be easily cleaned and decontaminated. Carpets and rugs in laboratories are not permitted.
  4. Laboratory furniture must be capable of supporting anticipated loads and uses. Spaces between benches, cabinets, and equipment should be accessible for cleaning.
  • Bench tops must be impervious to water and resistant to heat, organic solvents, acids, alkalis, and other chemicals.
  • Chairs used in laboratory work must be covered with a non-porous material that can be easily cleaned and decontaminated with appropriate disinfectant.
  1. Laboratory windows that open to the exterior are not recommended. However, if a laboratory does have windows that open to the exterior, they must be fitted with screens.
  2. BSCs must be installed so that fluctuations of the room air supply and exhaust do not interfere with proper operations. BSCs should be located away from doors, windows that can be opened, heavily traveled laboratory areas, and other possible airflow disruptions.
  3. Vacuum lines should be protected with liquid disinfectant traps.
  4. An eyewash station must be readily available.
  5. There are no specific requirements for ventilation systems. However, planning of new facilities should consider mechanical ventilation systems that provide an inward flow of air without recirculation to spaces outside of the laboratory.
  6. HEPA filtered exhaust air from a Class II BSC can be safely recirculation back into the laboratory environment if the cabinet is tested and certified at least annually and operated according to manufacturer’s recommendations. BSCs can also be connected to the laboratory exhaust system by either a thimble (canopy) connection or directly exhausted to the outside through a hard connection. Provisions to assure proper safety cabinet performance and air system operation must be verified.
  7. A method for decontaminating all laboratory wastes should be available in the facility (e.g., autoclave, chemical disinfection, incineration, or other validated decontamination method).

Safe Use of Equipment and Tools in Biological Research

Bio-hazardous Waste Management

Examples of items that are considered to be biohazardous waste include cultures of infectious microorganisms; human blood, body fluids, or tissues; carcasses, body parts, or fluids from infected laboratory animals; or materials that contain or may have been contaminated with any of the above. All potentially infectious waste generated in laboratories must be segregated from other refuse and placed into waste containers that are impervious to moisture, of sufficient strength and thickness to prevent expulsion, secured to prevent leakage or expulsion, and labeled with the biohazard symbol.

Infectious waste is regulated by state and local regulations. The key requirements with regard to infectious waste are proper labeling with subsequent disposal in a safe manner.

Disposal

Biological Waste Chart

All potentially contaminated sharps must be placed in containers that are leak proof, rigid, and puncture resistant (e.g., plastic "sharps containers"), in addition to meeting the above criteria for infectious waste containers.

Free flowing liquid waste may be chemically decontaminated (e.g. with a freshly prepared 1:10 dilution of household bleach with a 20 minute contact time) and then provided they contain no other hazardous chemicals or materials may be discharged into the sanitary sewer system.

Solids biohazardous waste is collected in white polypropylene containers lined with red bags or puncture resistant (cardboard boxes) lied with red bags for pipettes. The outer container is labeled with the biohazard symbol. The containers should be closed when not in use.  Bags should be autoclaved before disposal.

Autoclave Procedure

Autoclave Testing Log for Biohazard Waste

When waste contains a combination of a biohazard and another hazard, such as radiation or chemical waste, the usual practice is to decontaminate the biohazard and then treat the waste as only a chemical or radioactive waste. Contact UNC Asheville’s EH&S Professional for guidance with mixed chemical or radioactive wastes.

 

Biological Project Registration

coming soon