Laboratory Safety Manual - Chapter 15: Safe Handling Of Biological Hazards


Laboratory Safety Manual - Chapter 15: Safe Handling Of Biological Hazards


This chapter is an overview of the requirements for working with biological hazards. You can find more detailed information about working with biological hazards in the UNC Exposure Control Plan (Bloodborne Pathogens), the UNC Biological Safety Manual and on our website.

Table of Contents

  1. Introduction
  2. Registering Biohazards for Use at UNC
    1. Recombinant DNA
    2. Use of Transgenic Animals or Plants
    3. Schedule F
    4. BSL-2 Checklist
    5. Standard Operating Procedures (SOPs)
  3. Human/Non-Human Primate Blood, Blood Products, Body Fluids, Tissues, and Cells
    1. Regulation
    2. Cultured Cells and Tissue
  4. Training
  5. Signage for Laboratories Using Biological Hazards
  6. Medical Surveillance
  7. Exposure Reporting
  8. Biohazard Waste Disposal Policy
  9. Packaging and Shipping

I. Introduction

Work with biological hazards at UNC is documented on Schedule F: Biological Hazards Registration of the online Laboratory Safety Plan. Because laboratory practices, facilities, and safety equipment for work at or above Biosafety Level 2 is regulated at federal, state, and local levels, this chapter will cover some basic information about biological hazards, including the use of established human cell lines and those found in human blood and other potentially infectious materials (OPIM). For more information on this topic, consult the UNC Biological Safety Manual.

II. Registering Biohazards for Use at UNC

Recombinant DNA.

University policy requires Principal Investigators comply with the National Institutes of Health “Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules”, regardless of the funding source supporting that research. The Principal Investigator is responsible for registering all research studies involving Recombinant DNA with the Institutional Biosafety Committee. Use Schedule G of the online Laboratory Safety Plan to submit applications to use Recombinant or Synthetic Nucleic Acid Molecules. The types of experiments that require reporting to the IBC or obtaining prior approval from the IBC include: those using human or animal pathogens as host-vector systems; cloning DNA from human or animal pathogens into a non pathogen host-vector system; and, experiments involving whole animals and plants, including transgenic species.

Use of Transgenic Animals or Plants.

Use Schedule H of the online Laboratory Safety Plan to submit applications to the IBC for the use of transgenic animals or plants that are imported, bought or transferred from other labs, institutions or companies. For transgenic animals that your laboratory prepares, use Schedule G. For Schedules G and H, you must fully describe the genetic alterations and/or foreign gene expression.

Schedule F.

Lab reagents requiring BSL-2 containment or above must be registered on the PI’s Laboratory Safety Plan; examples include human pathogens, pathogens of concern in UNC vivariums, viral vectors used in recombinant or synthetic nucleic acid molecule research, human source material such as blood (including well established cell lines), non-human primate material, and biological toxins. Refer to Chapter 2 for further requirements when submitting a new or updated Laboratory Safety Plan.

BSL-2 Checklist.

To ensure laboratories meet basic requirements at the federal, state, and local levels for BSL-2 practices and containment, the BSL-2 checklist must be completed for each space designated at BSL-2. In 2007, the CDC enhanced requirements for all work at BSL-2.

Standard Operating Procedures (SOPs).

Certain work with biological hazards requires written standard operating procedures to define facilities, practices, and responses. An example is work conducted at enhanced containment where all workers are required to conform to a specific set of procedures. Submit SOPs to EHS for review by the Institutional Biosafety Committee.

III. Human/Non-Human Primate Blood, Blood Products, Body Fluids, Tissues, and Cells


Biosafety level 2 practices and containment must be followed when handling human or non-human primate materials. The OSHA Bloodborne Pathogens (BBP) Standard (29 CFR 1910.1030) applies to all occupational exposure to blood or other potentially infectious materials. Under the OSHA BBP Standard, employers are required to develop a written Exposure Control Plan, offer employees the hepatitis B vaccination, and provide annual training. Since the mid-1990s OSHA’s position has been that workers handling human cell cultures (primary or established) fall under the purview of the Bloodborne Pathogen (BBP) Standard. For more information on the impact of the OSHA BBP standard on the laboratory setting at UNC, see the Lab Worker Exposure Control Plan.

Cultured Cells and Tissue

Cultured cells which are known to contain or be contaminated with a biohazardous agent (e.g. bacteria or viral) are classified in the same biosafety level as the agent. Cell lines that are not human or non-human primate cells and which do not contain known human or animal pathogens are designated biosafety level 1.

The following cells and tissue must be registered with EHS and handled at BSL2.

  • Human and non-human primate primary cells, established cell lines, and unfixed tissue
  • Cell lines exposed to or transformed by a human or primate oncogenic virus
  • Cells, cell lines or tissue infected with pathogens requiring BSL2 containment.

IV. Training

Training on the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules is required of all Principal Investigators, regardless of work with recombinant or synthetic nucleic acid molecules. The training is available online.

Workers at BSL-2 are required to complete the online BSL-2 training prior to beginning work at BSL-2.

Lab workers expected to handle human or non-human primate material (including well established human cell lines) in the lab or by treating lab waste are also required to complete the online Bloodborne Pathogens training annually.

Lab workers at enhanced BSL-2 or at BSL-3 receive classroom instruction and emergency drills by EHS at least annually. Lab access will not be granted to laboratory employees until all of the required training is completed.

V. Signage for Laboratories Using Biological Hazards

Laboratories in which employees handle biosafety level 2 (including laboratories handling human blood, OPIM and human cell lines) or biosafety level 3 agents, must be posted with the Universal Biohazard Sign and the Principal Investigator’s name and contact information. The lab entrance signage system can be used or a separate BSL2 door placard may be generated.

Figure 15.1

Figure 15.1. Example of the Universal Biohazard Symbol. This version is available on the EHS Safety Labels webpage.

VI. Medical Surveillance

A medical surveillance program is provided through the University Employee Occupational Health Clinic (UEOHC) for personnel who are occupationally at risk of exposure to bloodborne pathogens (BBP), have direct contact with research animals, or receive vaccines for various infectious agents (e.g. vaccinia, rabies, measles), used in the laboratory. The program includes free hepatitis B vaccine and post-exposure evaluation and follow-up. Contact the Clinic for more information at 919-966-9119.

In addition to being offered recommended vaccines, lab workers may have baseline serum samples collected as appropriate and tests for agents handled in the lab (e.g. TB skin test).

VII. Exposure Reporting

Employees that have had an incident that results in occupational exposure to biohazardous agents (e.g., needlestick or cut with contaminated object, splash to mucous membranes, contact with non-intact skin, large spill, etc) must follow appropriate exposure procedures. Wash exposed skin and sites of parenteral exposure thoroughly with soap and water. Flush eyes with water at an eyewash station for 15 minutes. If exposed, flush the mouth with clean water.

Employees are to report all incidents to the UEOHC. To ensure prompt attention, University employees are to call:

  • During daytime hours (8:30 a.m.-4:30 p.m., M-F): Call the UEOHC, 919-966-9119, for consultation and assessment.
  • For exposures to human blood or needle sticks after-hours call Healthlink, 919-966-7890, for consultation and assessment.

The employee’s supervisor must complete a Form 19, “Employer’s Report of Injury to Employee” directly following the incident. EHS will investigate the circumstances of the exposure incident, and make a report with recommendations to avoid further exposure incidents. As with all injuries, the employee must complete an Employee Accident Report Form, and the supervisor must complete a Supervisor Accident Report Form.

VIII. Biohazard Waste Disposal Policy.

The procedures of the UNC Biological Waste Disposal Policy are consistent with the North Carolina medical waste rules (15A NCAC 13 B .1200) and the applicable sections of the OSHA Bloodborne Pathogens Standard 29 CFR 1910.1030.

All biohazard waste generated in UNC-CH research laboratories will be properly treated and tagged prior to its transfer and final burial in the landfill. This biohazard waste includes:

  • Materials contaminated or potentially contaminated during the manipulation or clean-up of material generated during research and/or teaching activities requiring biosafety level 1, 2, or 3 or animal or plant biosafety level 1, 2, or 3. Refer to your laboratory’s Biological Hazards Registration section of the Laboratory Safety Plan (Schedule F) to identify these materials in your lab.
  • Human liquid blood and body fluids.
  • Human tissue and anatomical remains.
  • Materials contaminated with human tissue or tissue cultures (primary and established) because these are handled at BSL-2.
  • Animal carcasses, body parts, blood, fluids and bedding from animals infected with BSL2 and BSL3 agents.

Weekly testing with indicators that contain an average bacterial spore population of 104 to 106 (e.g. Bacillus stearothermophilus) meet the requirements of the North Carolina Medical Waste Rules. An example of a proper protocol for handling and treating biohazard waste at UNC follows:

  1. Safety First: Ensure BSL-2 Training, BBP training, and Hepatitis B vaccination are completed.
  2. Minimum protective apparel for operating the autoclave is a lab coat, safety glasses, gloves, and closed-toed shoes.
  3. Ensure biohazard waste is disposed of in an orange autoclavable bag.
  4. Place autoclave tape “X” over biohazard symbol.
  5. Always leave the bag open in the autoclave (to allow steam penetration).
  6. Always use a leak-proof secondary container in the autoclave.
  7. Secure the indicator to the end of a string, paper, or serological pipette to aid retrieval.
  8. Carefully place the indicator in the densest portion of the biohazard waste load.
  9. Run the autoclave cycle as usual and log the cycle parameters and indicator lot number.
  10. Carefully remove the indicator from the waste, and incubate with a control indicator.
  11. Document the results on the weekly log form.
  12. A positive result on the indicator requires the autoclave be posted as “NO USE” and EHS contacted and/or repairs initiated.

Waste does not need to be held until the indicator grows out. However, once the result is positive, the autoclave may not be used and must be reported.

For more information on proper collection and treatment of biohazard waste, refer to the UNC Biological Waste Disposal Policy.

IX. Packaging and Shipping.

Biological materials require specific packaging and labeling according to their potential hazards. Biological materials must be shipped according to DOT and IATA regulations. These regulations state that you must receive specific training to legally ship a hazardous biological material. Contact EHS to receive this training and to identify the specific requirements for your materials.

Back to Chapter Fourteen

Proceed to Chapter Sixteen

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