How to Manage Biohazardous Waste in Laboratories Bobby Clark, CSP Biological Safety Officer Clemson University Office of Research Safety How to Manage Biohazardous Waste in Laboratories

Why is this important? 1. This is a safety issue. Managing biologically hazardous waste properly will protect workers and the environment. Remembering the big picture will cause us to handle this waste properly. 2. This is a regulatory issue. Compliance with the regulations protects Clemson University from violations and fines. Completing this training will increase understanding of these concepts and help achieve proper management of this waste in the labs.

Items that must be treated prior to disposal Types of Biological Waste 1 Non-infectious Waste 2 Items that must be treated prior to disposal 3 Infectious Waste We have a new permit

1 Non-infectious Waste Cannot cause human or animal disease, have not touched recombinant DNA and cannot be mistaken for medical waste

Non-infectious waste This is waste that requires no treatment or processing. Many times these wastes can go directly into the trash or sewer. Examples: Certain plants, items that have touched biologicals (not recombinant DNA!) Internal protocols or permits may require you to treat or containerize your waste even though it is non-infectious (APHIS permit, IBC protocol, etc.) Solid media and media containing liquefied agar should NOT be poured down the drain.

Collection is always an option Not sure if your waste is infectious? This training should help you answer that question, but if you are in doubt, collection is always an option. Collection is always an option These containers are not supplied by our office, but collection of waste is a free-of-charge service. Contact emails: Biological Safety Officer (rclark3@clemson.edu) Hazardous Waste Officer (juneb@clemson.edu)

Items That Must be Treated Prior to Disposal 2 Items That Must be Treated Prior to Disposal These items cannot go in the general trash!

Two categories in this section Items that appear to be medical waste Cells, bacteria, viruses and anything that touches recombinant DNA Items in this section DO NOT have to be recorded on the treatment log!

Petri Dishes, Biohazard Bags and Syringes; oh my! Special consideration is needed for these items!!!

Items that appear to be medical waste: Petri Dishes Petri Dishes that look like this: May go here:

Items that appear to be medical waste: Biohazard Bags Use this instead! All of these appear to medical waste

Items that appear to be medical waste: Syringes The guidance for these is common sense: If you are using a large number of these, containerize them or autoclave them. Otherwise dispose in normal trash. Note that these syringes do NOT have needles!!!

If it appears to be regulated waste it is. Take home message on this section… If it appears to be regulated waste it is. Don’t make me have to dig it out of the landfill or the dumpster!!!

Anything touching recombinant DNA According to NIH guidelines, anything that touches recombinant DNA must be decontaminated. This can be done chemically with bleach, thermally by autoclave or by collection.

Infectious (Regulated) Waste 3 Infectious (Regulated) Waste Must be handled by permit

Infectious Waste- What is it? All of the following wastes must be handled as infectious waste by Clemson University: Sharps Recombinant DNA from risk group 2 or above Cultures and Stocks of Etiological Agents Lab Waste that Has Come into Contact with a Biohazard Human Pathological Waste, Bodily Fluids and Infected Body Substances Non-human Primate Waste Additional Wastes Animal Waste Definitions are taken from the Clemson University Biosafety Manual

Infectious Waste: What’s New? Infectious waste can now be treated by the generator! All treated infectious waste must be logged Chemically treated: date, amount treated (volume) Autoclave: Date, amount treated (volume or weight), temperature, pressure and monthly challenge results Collection: If waste is collected by Research Safety, logging is unnecessary All collected infectious waste must be sealed before pickup

Infectious Waste- Sharps All sharps are infectious waste. This includes: Needles Razor blades Contaminated broken glass This does not include: Pipette tips Pasteur pipettes Serological pipettes Sharps containers are provided by Research Safety and must be requested. Chemically contaminated sharps must be handled as chemical waste and must comply with the hazardous waste regulations.

Infectious Waste- Sharps (continued) Only sharps should be placed in sharps containers. Re-sheathing or bending needles is prohibited. Special considerations for chemically contaminated sharps. Sharps containers must not be autoclaved. X Can you show an example of the safe type of retractable type needles?

Infectious Waste-Liquids These are liquids that fall under any of the definitions given for infectious waste and contain no solids There are three different disposal methods for liquids. They are: Chemical disinfection Steam sterilization Collection for incineration Not sure of first sentence “not solids”

Infectious Waste-Liquids (Chemical Disinfection) Currently, Clorox bleach is the only approved disinfectant for chemical treatment of biohazardous (infectious) waste. A 0.5% concentration of sodium hypochlorite (the active ingredient of household bleach) is required with a contact time of at least 20 minutes before disposal to the sanitary sewer. This concentration is achieved by adding 1 part bleach to 9 parts liquid waste. Solids cannot be treated chemically.

Infectious Waste-Liquids (Chemical Disinfection) Example: You have 100 mL of liquid culture that must be disinfected. How much bleach must be added to achieve the appropriate concentration? 100 mL {volume of liquid waste} = 90% volume of total solution 100 mL / 0.9 = volume of total solution = 111 mL 111 mL {volume of total solution} – 100 mL {volume of liquid waste} = 11 mL {volume of bleach to add} Remember to check the expiration date on your bleach!

Infectious Waste-Liquids (Chemical Disinfection) While chemically disinfecting infectious waste, make sure to wear the appropriate PPE: Gloves Lab coat Safety glasses or splash goggles Remember to use the appropriate contact time; at least 20 minutes Do not pour solid or semi-solid substances down the drain!

Infectious Waste-Liquids (Steam Sterilization) There may be times where chemical disinfection is impossible or impractical. Steam sterilization is allowed, but there are a number of rules.

Infectious Waste-Liquids (Steam Sterilization) If an autoclave is to be used for steam sterilization, the user must: Use the biological indicator Geobacillus stearothermophillus at the center of a load processed under standard operating conditions to confirm the attainment of adequate sterilization conditions. Indicator organisms must be used monthly at a generator facility in each steam sterilizer. Record the temperature and time during each complete cycle to ensure the attainment of a temperature of 121 degrees Centigrade (250 degrees Fahrenheit) for 45 minutes or longer at fifteen (15) pounds pressure, depending on quantity and density of the load, in order to achieve sterilization of the entire load; (Thermometers shall be checked for calibration at least annually.) Ensure the autoclave has a gauge that indicates the pressure of each cycle

Infectious Waste-Liquids (Steam Sterilization) If an autoclave is to be used for steam sterilization, the user must: Use heat sensitive tape or other device for each container that is processed to indicate that the steam sterilization temperature has been reached. The waste will not be considered appropriately treated if the indicator fails. Note: Use of autoclave tape containing lead is not permitted! Maintain records on pressure and temperature for each treated load for a period of three (3) years. Assure that treatment residues are disposed of in accordance with applicable State and Federal Requirements. X

Infectious Waste-Liquids (Steam Sterilization) In addition to the rules regarding the verification of the autoclave, there are requirements for transport of infectious materials to the autoclave: Items must be stored in leak-proof containers bearing the biohazard symbol if the autoclave is outside of the lab. Containers are subject to testing by the BSO. Heat-resistant gloves must be used when removing items from the autoclave Do not chemically decontaminate and then autoclave!!!

Infectious Waste-Solids (Steam Sterilization) The same rules apply for steam sterilization of infectious waste solids (Petri dishes, PPE, paper towels, etc.) Remember to render items that can be mistaken as medical waste as unrecognizable even if that means additional autoclave time. With all thermal treatment, the guidelines in the BSM for autoclave use should be followed. X

Infectious Waste-Liquids and Solids (Collection) So what about items that cannot be chemically or thermally treated? Solid items when no autoclave is available Carcasses of infected animals Other large materials Collection of waste for incineration is an option.

Infectious Waste-Liquids and Solids (Collection) There are rules for collection of infectious waste. Some will be explained in more detail in subsequent slides, but the general rules are: No infectious waste may be left in the open air without first being disinfected chemically or thermally (closed containers). Waste must be properly packaged The weight of the container must not exceed 40 pounds. Liquids must be in sealable containers and then placed in sealable plastic bags. All infectious waste collected from the lab must be sealed before pickup.

Infectious Waste-Collection: Proper Packaging If collection of waste is the option you choose, the waste will have to be packaged properly. You will need to request materials for packaging from Research Safety.

Infectious Waste-Collection: Proper Packaging If an empty plastic container is what is provided, put two, red biohazard bags in the container to line it. Place the materials in the innermost bag. Twist the top of the bag and tape or tie it shut (Remember PPE!). Place a label on each bag. Seal the outer bag in the same way. Place the lid on the container and place a label on the outside. Request pickup of the waste.

Infectious Waste-Liquids and Solids (Collection) Here are a few things you should NOT do: Overfill the containers (making them too heavy) Underfill the containers Put putrescent waste into boxes. Putrescent waste must be frozen until pickup.

What about mixed wastes? Mixed wastes contain both infectious materials and hazardous chemicals. The BSO and the Hazardous Waste Officer need to be consulted about these wastes. They will ultimately be treated as chemical waste so all the regulations regarding Hazardous Waste will apply (see Hazardous Waste training for details).

What about spills? How a spill is cleaned up depends on the size of the spill and where the spill occurs. All spills of infectious materials shall be cleaned up with a 1:9 dilution of household bleach that has been made in the last 8 hours. The spill shall be saturated with the diluted bleach and left for 10 minutes. The materials shall be collected for pickup by research safety. See the Biosafety Manual for more specific details.

Biological waste falls into three categories: Summary Biological waste falls into three categories: Non-infectious waste (this may go in the general trash or down the drain) Waste that is not infectious but requires further processing before regular disposal Infectious waste (this must be treated according to permit rules) Infectious waste can be treated chemically (with bleach), thermally or collected. When in doubt, contact our office!

Questions??? Contact Information: Bobby Clark 120C Long Hall (864) 710-2933 rclark3@clemson.edu Or use the AskRS function: http://www.clemson.edu/research/safety/askRS.html Please go to the quiz: http://www.clemson.edu/research/safety/training/biotraining/biowasteQuiz.html THE END