The use of biohazardous materials shall comply with the NIH Guidelines, the recommendations in the CDC publication Biosafety in Microbiological and Biomedical Laboratories (BMBL), and the American Biological Safety Association (ABSA) best practices, as well as all federal, state, and local regulations.
Four Biosafety Levels are described in this section, which outline the appropriate laboratory facilities and practices for work with biological agents or material which may carry them. Biological agents are assigned BSL’s based on the risk they pose to human health and the environment. The likelihood to cause disease, severity of disease, and routes of transmission are considered when determining the appropriate BSL. All biological agents or materials used in the laboratory shall be listed on the Biological Material ID Form in the Laboratory Safety Plan. Biosafety Level 1 is the basic level of protection and is appropriate for agents that are not known to cause disease in normal, healthy humans. Examples of BSL-1 materials are non-pathogenic strains of bacteria and fungi. Biosafety Level 2 is appropriate for handling materials potentially containing moderate-risk agents that cause human disease of varying severity by ingestion or through percutaneous or mucous membrane exposure. BSL-2 is appropriate when work is done with:
- ❖ Any infectious pathogenic microorganisms.
- ❖ Any human primary cells or cell lines.
- ❖ Other Potentially Infectious Materials (OPIM), defined as any human-derived blood, body fluids, or tissues, where the presence of an infectious agent may be unknown.
- ❖ Cells or other materials from animals known to have been infected with human pathogens.
Biosafety Level 3 is appropriate for agents of indigenous or exotic origin with a known potential for aerosol transmission and for agents that may cause serious and potentially lethal infections. Research with BSL-3 agents is CURRENTLY NOT APPROVED AT UNCG.
Biosafety Level 4 is appropriate for agents that pose a high individual risk of life threatening disease by infectious aerosols and for which no treatment is available. Research with BSL-4 agents is CURRENTLY NOT APPROVED AT UNCG.
Cell & Tissue Culture
Cells and tissue may contain human pathogens. It is prudent to consider all human cell lines to be potentially infectious. Most cell and tissue cultures can be safely manipulated using BSL-2 practices and containment.
- ❖ All primary and permanent human or other primate cell lines or tissue cultures shall be handled using BSL-2 practices and containment.
- ❖ Personnel handling human cell and tissue cultures shall follow the UNCG Bloodborne Pathogen Exposure Control Plan.
- ❖ If any cell or tissue cultures are known or suspected to contain a specific pathogen or oncogenic virus, appropriate biosafety practices for handling that virus shall be used when working with the cell or tissue culture.
- ❖ BSL-1 practices and containment may be used for cell lines that meet all of the following criteria. Cells shall:
- • Not be of human or other primate origin.
- • Be confirmed not to contain human or other primate pathogens, including viruses, pathogenic bacteria, mycoplasma, or fungi.
- • Be well-characterized in regard to low risk.
Recombinant and Synthetic DNA
All work with rDNA must be registered with the Institutional Biosafety Committee and comply with the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules.
The Occupational Safety and Health Administration (OSHA) Bloodborne Pathogens Standard (BBP, 29 CFR 1020.1030) requires specific training and inoculation for individuals with an occupational exposure to bloodborne pathogens. Bloodborne pathogens are disease-causing microbes present in blood, unfixed tissue, and other potentially infectious material. Pathogens include, but are not limited to Human Immunodeficiency Virus (HIV) and Hepatitis B and C (HBV, HBC), other infectious viruses, bacteria and protozoa. Occupational exposure is reasonably anticipated skin, eye, mucous membrane, or parenteral (subcutaneous) contact with blood or other potentially infectious materials that may result from the performance of an employee’s duties. Laboratory workers with an occupational exposure must complete Laboratory Biosafety Training within 10 days of their work assignment or commencement of activities presenting an occupational exposure, and annually thereafter. Once initial training is completed, workers must complete and submit the Bloodborne Pathogens Exposure Control Plan Enrollment Form and complete or decline the HBV vaccine series, at no cost to them. The vaccination is not required if the individual has completed the vaccine series previously or tests positive, but the declination form must be completed. Gove Student Health Center administers the vaccine to students and employees as a series of three injections, the second and third administered one and six months after the first. The principal investigator or laboratory director is responsible for identifying workers with an occupational exposure and ensuring that required training and vaccinations (ordeclinations) are completed. The laboratory department is responsible for the cost of the HBV vaccination.
All potentially infectious materials shall be treated prior to disposal to the general waste stream. All cultures, stocks, and specimens of microorganisms or other potentially infectious materials and any lab equipment or waste coming in contact with these materials must be treated prior to disposal. Infectious agents (RG2/BSL-2) and some bodily fluids fall under the NCDENR Regulated Medical Waste Rules and require specific treatment procedures, including weekly validation of the autoclave cycle parameters or pre-approval of chemical disinfection method, as described in the UNCG Regulated Medical Waste Policy. Exceptions to this policy are items contaminated with blood or bodily fluids contained on bandages and personal hygiene products, and cleanup materials not related to research activities. These items are not required to be treated prior to disposal. The following table identifies the appropriate treatment method for specific types of biological waste, which includes any other materials or equipment, including PPE, contaminated with the biological material.
|Biological Material||Treatment Method|
|Stocks and cultures of nonpathogenic materials and microorganisms (RG/BSL-1)||Autoclave or chemical treatment|
|Pathogenic microorganisms (RG/BSL > 2)||1Validated autoclave or approved chemical treatment|
|Human blood, blood components and products, and 2OPIM, in individual containers less than 20 mL||Autoclave or chemical treatment|
|Human blood, blood components and products, and 2OPIM, in individual containers greater than 20 mL||1Validated autoclave or chemical treatment|
|Animal or human primary cells, cell lines, and culture media||Autoclave or chemical treatment|
|rDNA waste, transgenic flies and plants||Autoclave or chemical treatment|
|3BSL1 rDNA waste which is not in organisms or viruses||None required|
|Unfixed tissue, cells (and culture media), and fluids from humans or animals known to have been infected with human pathogens.||1Validated autoclave or approved chemical treatment|
|Animal tissues, organs, parts||Autoclave or incineration|
|Animal carcasses||Incineration by commercial vendor|
1See UNCG Regulated Medical Waste Policy for specific treatment requirements.
2Other Potentially Infectious Materials (OPIM) are defined as human semen; vaginal secretions; cerebrospinal, synovial, pleural, pericardial, peritoneal, and amniotic fluids; and body fluids visibly contaminated with blood or in situations where it is difficult to differentiate between body fluids. 3BSL1 rDNA waste which is not in organisms or viruses (e.g., DNA, RNA, oligos waste, PCR waste, Microarrays, etc.). rDNA waste from experiments which use Escherichia coli K-12 host-vector systems that use only non-conjugative plasmids as vectors (e.g., pBR322, pBR313) and does not contain conjugation proficient plasmids or generalized transducing phages. rDNA waste from experiments involving Saccharomyces cerevisiae and Saccharomyces uvarum host-vector systems.