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Biohazardous Waste

Biohazardous waste may include human blood and bodily fluids, human tissues, pathological specimens, pathogenic or recombinant microbiological materials, infected or transgenic animal tissues and carcasses, infected or transgenic plant materials, and any disposable items contaminated with these materials.

This page outlines the types of biohazardous waste most often generated in UK research laboratories and details acceptable decontamination and disposal methods for each. If you will be generating biohazardous waste not described here, please email biosafety@uky.edu for guidance.

Please see the UK Research Safety Laboratory Waste Guidelines available online HERE for additional instructions and guidance, including hazardous chemical and radiological waste disposal.  

Liquid Biohazard Waste



Erlenmeyer flask of liquid in laboratory

Liquid biohazardous waste may consist of blood, blood products, and body fluids from human or animal research and culture media contaminated with pathogenic or recombinant microbiological materials which may be hazardous to humans, animals, plants, or the environment. Liquid biohazardous waste can come in many forms in the laboratory. These include, but are not limited to liquid bacterial cultures, non-adherent human tissue cultures, supernatants of tissue cultures infected with viral vectors, human blood samples and blood products (serum, plasma, and other blood components), and human tissue culture media that is aspirated into a vacuum line trap. 

To disinfect liquid biohazardous waste prior to disposal:

  1. Add unexpired household bleach to achieve a final concentration of 10% bleach.
  2. Allow a contact time of at least 20 minutes. 
  3. Once the appropriate concentration and contact time have been met, test the pH of the liquid using a pH meter or pH test strip (available free upon request - biosafety@uky.edu) to ensure it is within acceptable limits for drain disposal (5.5-11.5 SU). 
  4. Once the mixture has met all requirements (10% bleach, 20 minutes, pH 5.5-11.5), it may be poured carefully and slowly down the laboratory sink drain with copious amounts of water. 

Liquid biohazardous waste must not be left to sit overnight and should be promptly decontaminated as described and disposed of at the end of each workday or when the container is 2/3 full, whichever comes first

Please note that once bleach has been added to liquid biohazardous waste, it cannot be autoclaved. 

If the use of bleach is not a suitable option, please contact the Office of Biological Safety for alternative options and further guidance. 

 

The transport of liquid biohazardous waste should be avoided when possible as this introduces the heightened risk of leakage and spills. Transporting liquid biohazardous waste to an autoclave for processing, while effective for sterilization, is not advised and should only be utilized when disinfection within the lab is not possible. If transporting liquid biohazardous waste for autoclaving is unavoidable, the primary container must be securely closed and placed in an appropriate secondary container that is closed, leak-proof, and shatter-proof. The primary container must be placed in a leak-proof autoclavable container with the lid/cap loosened for autoclaving. Ensure contents are totally cooled prior to removal and dispose of autoclaved liquid down the sink with copious amounts of water.  

Solid Biohazard Waste (non-sharp)



Person throwing away waste into biohazard bin

Solid biohazardous waste consists of any non-sharp items that may be contaminated with human or animal research material (ex. bodily fluids, tissue, etc.), pathogenic organisms, or recombinant and/or synthetic nucleic acid materials. Solid biohazardous waste may consist of used gloves and other disposable PPE (Personal Protective Equipment), culture plates, specimen vials, lab bench paper, etc., and must be collected in orange or clear/opaque autoclave bags clearly marked with a biohazard symbol. Red bags are NEVER to be used for collection and autoclaving of solid biohazardous waste. Bags containing solid biohazardous waste must be held in a non-porous, leak-proof container, preferably with a foot operated lid. 

Once a bag of solid biohazardous waste has become 2/3 full, it must be loosely closed and placed in a leak-proof autoclavable secondary container for processing. Bags should not be overfilled. This waste is then processed in an autoclave designated for biohazardous waste. Look for the signage below.



Autoclave Designated for Biohazardous Waste Sign

Biohazardous waste must always be accompanied by an individual familiar with its contents once it is transported from its collection point. Unprocessed biohazardous waste should never be left unattended waiting to be autoclaved. If an autoclave is not available, waste must be brought back to the lab space that generated it and secured until the autoclave is available. Biohazardous waste should also not be stockpiled in lab spaces and must be processed as soon as reasonably possible once collected. Once bags of solid biohazardous waste have been successfully autoclaved, they must be placed into regular opaque trash bags before disposal with normal trash. 

Most solid biohazardous waste can be autoclaved using gravity steam sterilization cycles with a target temperature of 121°C and sterilization phase of ≥ 40 minutes. However, the length of time required for sterilization can be higher depending on the biohazardous organism or materials to be autoclaved. The Office of Biological Safety can assist you in determining an appropriate sterilization time based on your lab’s work and agent(s). 

Sharps Waste



Sharps container with needles

Sharps Waste can be divided into four categories: biohazardous sharps waste, biohazardous plastic sharps waste, non-biohazardous sharps waste, and non-biohazardous plastic sharps waste.  

  • Biohazardous sharps waste consists of objects that have the high potential to produce punctures or lacerations and have been used in conjunction with biohazards. This includes needles/syringes, lancets, unfixed glass slides, scalpel blades, glass vials, glass Pasteur pipets, razor blades, and disposable surgical instruments. Biohazardous sharps waste must be collected in a hard plastic medical sharps container that is marked with the universal biohazardous symbol or red in color. Once the container is 2/3 full, close and seal it and decontaminate its exterior with a suitable disinfectant. If in the Medical Center (Medical Sciences, HSRB, Roach, Combs, Whitney-Hendrickson, and Gill Heart buildings), housekeeping will pick up these containers for final disposal. If located outside of the Medical Center, please submit an E-trax ticket for pickup. Do NOT autoclave these types of sharps containers. 
  • Biohazardous plastic sharps waste typically consists of plastic serological pipettes, micropipette tips, plastic Pasteur pipets, plastic inoculating loops, plastic plate spreaders, and any other pointy plastic items that have been utilized in conjunction with biohazardous materials that may be likely to puncture an autoclave bag. While these items may not be considered as sharp as needles or glass, they do still pose a risk of puncture or laceration. Loose biohazardous plastic sharps, such as pipettes or tips, are prohibited in autoclave bags. 
  • Small biohazardous plastic sharps waste (micropipette tips, etc.) may be collected in small benchtop biohazardous waste bags. Small benchtop biohazardous waste bags must be loosely closed, placed into a larger biohazardous waste bag with other solid biohazardous waste, and autoclaved before final disposal. Alternatively, small biohazardous plastic sharps waste may be collected as described directly below.  
  • Large biohazardous plastic sharps waste (ex. plastic serological pipettes and plastic Pasteur pipets) must be collected in a plastic lined box (ex. a carboard box lined with a biohazardous waste bag on the interior). Once full, close the cardboard box loosely with tape, and put the closed box into a biohazardous waste bag. Boxes should not be overfilled such that they cannot be safely closed. The bagged box of biohazardous plastic sharps may then be autoclaved as solid biohazardous waste. 
  • Used needles, syringes, lancets, razor blades, and other sharp objects which have NOT been used in conjunction with biohazardous or hazardous chemicals shall be placed in a hard plastic or metal container with a screw-on lid. When full, reinforce the lid with heavy-duty tape, label the container “Not recyclable trash,” and place with regular trash for collection by housekeeping. If unsure as to whether these items have been used in conjunction with biohazardous materials, it is advisable to dispose of them in a medical sharps container as outlined above in Biohazardous Sharps Waste.  
  • Non-biohazardous plastic sharps waste often consists of plastic micropipette tips, plastic transfer pipettes, plastic Pasteur pipets, and plastic serological pipettes not used with biohazardous materials. Pipettes and pipette tips which have not been utilized in conjunction with biohazardous materials or hazardous chemicals shall be disposed of in a hard plastic or metal container with a screw-on lid. Once full, the container’s lid shall be taped securely shut, labeled with “Not recyclable trash,” and placed with the regular trash for removal by housekeeping. Larger plastic sharps (ex. Serological pipets and plastic Pasteur pipets) not used with biohazardous materials or hazardous chemicals shall be disposed of in a plastic lined cardboard box. Once the box is filled to a capacity where it can be safely closed (not filled above the top flap seam), its opening must be secured with tape and “Trash” written on the outside. It can then be placed with regular trash for disposal. Containers and boxes should not be overfilled such that they cannot be safely closed.  

Small Animal Carcasses



Lab mice

Small Animal carcasses (intact or partial) must be disposed of according to how the animal/carcass was treated and where it was originally obtained. If the animal/carcass was acquired from a DLAR (Division of Laboratory Animal Resources) facility, and the animal/carcass was not exposed to chemical or biohazardous agents, it must be collected in a leak proof opaque bag and returned to the appropriate location in DLAR. If the animal/carcass was acquired from a DLAR facility, and the animal/carcass was exposed to biohazardous materials, it must be collected in a red bag and placed in designated location in DLAR. Do not place carcasses in with other biohazardous waste to be autoclaved. For small animal carcasses not originally obtained in DLAR, contact the Office of Biological Safety to determine a suitable disposal method. Infected animal carcasses or tissue that are also contaminated with hazardous chemicals or radioactive materials is a type of mixed waste, please contact UK Biosafety at biosafety@uky.edu for guidance regarding mixed waste.

Large Animal



Horse and technician

Large animals that are used in conjunction with biohazardous materials pose an added challenge for final disposal and many factors must be considered. Please contact the Office of Biological Safety for guidance during planning of projects that may generate large animal carcasses and associated bedding and/or manure contaminated with biohazardous materials. 

Plants



Arasystem for Arabidopsis thaliana containment

Plants, plant parts, and other plant materials from transgenic or infected plants (ex. plants infected with a pathogen or pest) must be devitalized/sterilized before leaving the greenhouse or laboratory housing them. Devitalization of plant material may be accomplished by steaming. Sterilization of plant material may be accomplished by autoclaving. The method utilized is dependent on the specific plant material, associated hazards, and regulatory requirements. Plant materials requiring steam devitalization, autoclaving, or other special disposal procedures can include but are not limited to transgenic/infected plant clippings, seeds, plant pests, stems, soil containing roots, flowers, plant reproductive structures, and associated plant microorganism contaminated items. For materials requiring USDA-APHIS permitting, the approved permit will often specify the required treatment. The Office of Biological Safety can assist you in determining an appropriate devitalization/sterilization procedure for your project’s specific plant material disposal requirements in accordance with all required permitting. 

Regulated Medical Waste



red biohazard bag

Regulated Medical (Pathological) Waste consists of large volumes (greater than 500 ml) of human blood, recognizable human organs, large amounts of unfixed human tissues or blood-soaked materials. This includes blood-soaked bandages, discarded surgical gloves after surgery, used sharps, cultures, stocks, and swabs used to inoculate cultures, and removed bodily organs. At the University of Kentucky, medical waste is NOT autoclaved and must instead be incinerated by a licensed vendor; therefore, laboratory disposal of this waste requires special designation using a red bag and specific containers. Red bags should never be used for regular or autoclaved waste. Please contact the Office of Biological Safety and the Department of Environmental Quality Management (EQM) if you will be generating Regulated Medical Waste outside of the UK Healthcare clinical environment.  

Mixed Waste

Mixed Waste is waste material contaminated with a combination of chemical, biological, and/or radiological hazards. Special consideration must be given to the disposal of this type of waste. Autoclaving is often NOT appropriate for mixed waste. Please reach out to the Department of Research Safety office (Biological Safety, Chemical &Laboratory Safety, Radiation Safety) and the Department of Environmental Quality Management for guidance.  

Prion Waste



Microscopy of prions

Biohazardous waste known or suspected to contain prion materials require special consideration and disposal methods. Prions are composed of abnormal, misfolded, pathogenic isoforms of normal prion protein. Prions are highly resistant to conventional inactivation by heat and chemicals and require special biosafety precautions and disposal methods. Please contact the Office of Biological Safety for guidance when planning projects that may generate known or suspected prion contaminated waste.