Bobby Clark, CSPBiological Safety Officer

Clemson University Office of Research Safety

How to Manage Biohazardous Waste in Laboratories

2. This is a regulatory issue.

Compliance with the regulations protects Clemson University from violations and fines.

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.

Why is this important?

Completing this training will increase understanding of these concepts and help achieve

proper management of this waste in the labs.

Types of Biological Waste

We have a new permit

1Non-infectious Waste

2Items that must

be treated prior to disposal

3Infectious Waste

Non-infectious Waste

Cannot cause human or animal disease, have not touched recombinant DNA and cannot be mistaken for medical waste

1

• Internal protocols or permits may require you to treat or containerize your waste even though it is non-infectious (APHIS permit, IBC protocol, etc.)

• 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!)

Non-infectious waste

• Solid media and media containing liquefied agar should NOT be poured down the drain.

Colle

ction

is a

lway

s an

optio

n 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.

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)

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!

Special consideration is needed for these items!!!

Petri Dishes, Biohazard Bags and Syringes; oh my!

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

All of these appear to medical wasteUse this instead!

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…

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.

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

Cultures and Stocks of Etiological Agents

Human Pathological Waste, Bodily Fluids and Infected Body Substances

Non-human Primate Waste

Animal Waste

Recombinant DNA from risk group 2 or above

Lab Waste that Has Come into Contact with a Biohazard

Additional Wastes

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

» 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

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

Infectious Waste-Liquids

There are three different disposal methods for liquids. They are:

These are liquids that fall under any of the definitions given for infectious waste and contain no solids

Chemical disinfection Steam sterilization Collection for incineration

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:GlovesLab coatSafety 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:o No infectious waste may be left in the open

air without first being disinfected chemically or thermally (closed containers).

o Waste must be properly packagedo The weight of the container must not

exceed 40 pounds.o Liquids must be in sealable containers and

then placed in sealable plastic bags.o 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

1) If an empty plastic container is what is provided, put two, red biohazard bags in the container to line it.

2) Place the materials in the innermost bag. Twist the top of the bag and tape or tie it shut (Remember PPE!).

3) Place a label on each bag.4) Seal the outer bag in the same

way.5) Place the lid on the container

and place a label on the outside.6) Request pickup of the waste.

Infectious Waste-Liquids and Solids (Collection)

• Overfill the containers (making them too heavy)

• Underfill the containers• Put putrescent waste into boxes. Putrescent

waste must be frozen until pickup.

Here are a few things you should NOT do:

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.

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 Clark120C Long Hall(864) 710-2933rclark3@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