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 Asheville’s EH&S Professional makes recommendations to Faculty for all research and teaching activities involved with potentially hazardous biological materials and recombinant 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 facility safeguards that can prevent laboratory-associated infections (LAI).

Risk Assessment and Biosafety Levels

Laboratories must assess the hazards of working with microorganisms and the need to practice safe handling, containment, and disposal of microorganisms. A risk assessment for each laboratory activity and organism is necessary in order to identify the proper procedures and safety equipment needed. Risk assessment determines the biosafety level of the workspace. A thorough risk assessment takes into account the microorganism being used, the manipulations performed with the organism, and the risks inherent in performing the lab activity. Although microbes are commonly handled at a particular biosafety level, the microbe alone does not determine the biosafety level of the lab. For example, manipulations that generate aerosols, create splash potential, or require large volumes of culture increase the risk associated with a particular microbe.

Summary or Recommended Biosafety Levels for Infectious Agents

Biosafety LevelAgentsPracticesPrimary Barriers and Safety EquipmentFacilities

(Secondary Barriers

  • 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
  • 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
  • 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
  • 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 and Teaching Laboratories.

Guidelines for Teaching Laboratories

The following guidelines from the American Society for Microbiology (ASM) are designed to encourage awareness of the risks, promote uniformity in best teaching practices, and protect the health and wellness of our students. These guidelines are not mandatory, but are designed to promote best practices in the teaching laboratory. Many best practices should be adopted to minimize the risk of laboratory-aquired infections and to train students in the proper handling of micro-organisms.

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.


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.

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

The following potentially hazardous biological agents and by-products are to be registered with the Environmental Health and Safety Offfice:

  1. Pathogenic agents (bacteria, rickettsia, fungi, viruses, protozoa, parasites, prions, and Select Agents.)
  2. Recombinant or synthetically derived nucleic acid, including those that are chemically or otherwise modified analogs of nucleotides or both. The NIH defines synthetically derived nucleic acid molecules as follows:
    • Molecules that (a) are constructed by joining nucleic acid molecules and (b) can replicate in a living cell (i.e., recombinant nucleic acids);
    • Nucleic acid molecules that are chemically or otherwise modified but can pair with naturally occurring nucleic acid molecules (i.e., synthetic nucleic acids);
    • Molecules that result from the replication of those described in (a) or (b) above.
  3. Recombinant DNA molecules, organisms, vectors (e.g., plasmids, viral vectors), and viruses containing recombinant DNA molecules
  4. Human and non-human primate blood, tissue, body fluid, and cell culture (primary and established cell lines)
  5. Plants, animals, or derived waste which contains or may contain pathogenic hazards (including xenotransplantation tissue)

Biosafety Research Protocol

Fill out the applicable Appendix(ces) to accompany you Biosafety Research Protocol.

Complete the following “Request for Changes of Personnel to IBC Protocol” form to update personnel working on your protocol.

Biological Training

Biological training is a vital component of UNC Asheville’s Biological Safety Program and is required under a variety of regulations and guidelines. UNC Asheville uses an independent, web-based program (CITI – Collaborative Institutional Training Initiative) for initial biological training. Additional hands on training may be required for your work.

Mandatory Training for All Biological Research Personnel

At a minimum, all biological researchers must complete the “Initial Biosafety Training” courses. This training is valid for three years, at which point you will be reminded to complete the refresher course. If there is potential to be exposed to occupational or workplace exposure to human blood or other potentially infectious materials (OPIM), then OSHA Bloodborne Pathogens Training is required. This training is required annually.

To sign up for training, visit the CITI Homepage (
Select the “Register” button under “Create an Account”
Search for the “University of North Carolina at Asheville” as your Participating Institution and continue the registration process
When prompted to select a curriculum, select “Initial Biosafety Training” and/or OSHA Bloodborne Pathogens Training.

The Learner Menu will then display the course. Click “Enter” to begin. You do not have to finish all modules at one time; CITI saves your scores each time you submit answers so you can log out and resume later

When an individual selects UNC Asheville as their participating institution during registration, their course completion history is automatically accessible to the Office of Research and Sponsored Programs (ORSP) for verification. You may also save a copy of your completion letter for your own records. If research personnel have completed the CITI modules while working at another institution, the user can add UNC Asheville as an affiliated institution which will give ORSP access to their records.