Basics of Biosafety

Working Safely with Biological Materials

Central Michigan UniversityCollege of Science and Technology

Principles and practices employed to protect laboratory personnel and the environment from exposure or infection while working with living organisms, biological materials, or agents.

Included are any materials that may be potentially infectious.

Includes recombinant DNA research

What is Biosafety?

The “agent” is the what creates risk

Risks to the worker or environment are often unknown

Determining “acceptable risk”?

Agents and Risks

There is always risk!The risk must be identified The risk is evaluated The risk must be measured Plan to minimize the risk

Assessing Risk

Assessment is conducted by a Biosafety Professional in partnership with and based on information provided by the Principal Investigator

The assessment is presented to the Institutional Biosafety Committee (IBC) for approval

Who Determines Acceptable Risk?

Understand the biology of the agent Susceptibility and transmission within the

host Hazards associated with equipment and

procedures Goal:

Provide the highest practical protection and the lowest practical exposure

Identifying Risk

Worst case scenario -What might happen?

Likelihood of an event

Seriousness of the incident

Actions needed to resolve the problems

Evaluating Risk Acceptability

Since there is no such thing as “no risk” “Safe” means risk has been judged acceptable Judging risk is a subjective- humans make

decisions Measuring risk is objective- use available

guidelines, data, and documentation Keep records of how determinations were made

due to subjective nature of the process

What is Acceptable Risk?

Agents Assigned Risk Groups

RG-1 Unlikely to cause disease in humans or animals low individual or community risk

RG-2 May cause disease but typically not serious individual risk, low community risk, treatable

RG-3 May cause serious disease, usually treatable High individual but low community risk, serious respiratory agents

RG-4 Serious or fatal, often not treatable, Easy transmission, high individual and community risk

WHO-World Health Organization

Different than the Risk Groups!! Risk groups used in risk assessment BSL are used in risk management

BSL are ways to control the agent facilities, safety equipment, practices, PPE, etc.

Once risk is assessed then the appropriate BSL is determined

Biosafety Levels (BSL)

Well characterized, non-pathogenic organisms or agents

Open bench- no containment

Use good laboratory practices, waste disposal, and aseptic techniques

Example: E. coli K-12 strains

BioSafety Level 1

Agents of moderate hazard to personnel or environment

Basic lab, but restricted access, containment during certain processes (i.e. aerosols, large volumes, etc.)

Autoclave and Biological Safety Cabinet desired

Use good laboratory practices, waste disposal, and aseptic techniques

Example: most non-respiratory, non lethal, agents

BioSafety Level 2

Agents of high hazard to personnel or environment

Respiratory exotic or indigenous agents which are easily transmissible causing serious or lethal disease

All work is contained, engineering controls and controlled environments we currently do not have the facilities to handle.

Example: Mycobacterium tuberculosis, SARS, etc.

BioSafety Level 3

FORGET ABOUT IT!!!

Hemorrhagic fever, deadly viruses, etc.

Total containment, airtight labs, “submarine” doors, air pumps, water treatment, HEPA filtration, etc.

Positive pressure “moonsuits”

BioSafety Level 4

Bacterial:76% from clinical labs8% from research labs

Exposure:60% acquired from inhalation

Other exposures include: digestion, sharps, splashes, direct and indirect contact

Laboratory Acquired Infections (LAI)

Viral 16% from clinical labs 70% from research labs

32% from animal related activities

Laboratory Acquired Infections (LAI)

Biohazardous/Medical Waste

Waste that is potentially infectious to humans, animals or plants. It includes:

Medical Waste according to MMWRARegulated Waste by MIOSHARegulated Waste by CDC/NIH

Michigan Medical Waste Regulatory Act (MMWRA)

Defines “medical waste” Requirements for waste handling and

disposal Requires generators to register with

DEQ and implement a Medical Waste Management Plan

Michigan Medical Waste Regulatory Act (MMWRA)

Defines “medical waste” Requirements for waste handling and

disposal Requires generators to register with

DEQ and implement a Medical Waste Management Plan

Biohazardous Waste Management Plan

Must outline how generating facility complies with the MMWRA: Types of wastes generated Storage and disposal of wastes Contingency plans Training

Biohazardous Waste Categories

Cultures and stocks of infectious agents and associated biologicals

laboratory wastebiological production wastediscarded live and attenuated vaccinesculture dishes and related materialscontaminated PPE

Biohazardous Waste Categories

Liquid human and animal waste liquid or semi-liquid blood and blood

products and body fluidscontaminated items that would release

blood or items that are caked with blood or other potentially infectious materials; NOT including urine or materials stained with blood or body fluids

infectious animal waste (research)

Biohazardous Waste Categories

Pathological waste tissuesbody parts other than teethproducts of conception fluids removed by trauma or during

surgery or autopsy/necropsy or other medical procedure and not chemically fixed.

…And More Biohazardous Waste Categories

Animal and plant pathogen waste Recombinant DNA waste Sharps

Biowaste vs. Trash

3 basic questions to differentiate:1. Is it contaminated with viable

biological material?

2. Can blood or other regulated body or biological fluids be released?

3. Is it a sharps hazard?

#1Is it contaminated with viable biological material?

Examples:• Contaminated lab waste

• Personal protective equipment used for handling potentially infectious materials (including handling infected animals or their products)

• Wastes from infectious disease research (carcasses, body fluids…)

#2Can blood or other (regulated) body fluids or viable biological materials be released?

Some Examples…

Tubes of blood

Vacuum flasks containing body fluids or cell line waste

Managing Liquid Biohazardous Waste

Storage: Label and secure bulk

vessels if not disposed of immediately

Treatment: Chemical disinfection OR AutoclaveDisposal: THEN Flush to sewer Use proper PPE!

10% bleach solution good for general disinfection High organics use 20% Needs to be made weekly Test contact time

Ethanol Use 70% solution (most effective) Longer contact time and flammable

*Should research and know effectiveness and contact time for the best disinfectant against your agent!

Disinfection

WRAPPERS/NON-ABSORBENT MATERIALS CONTAMINATED WITH BLOOD

BANDAGES/OTHER ABSORBENTS SATURATED OR CRUSTED WITH BLOOD

STAINED?….

or SATURATED?

Managing Non-Sharp Biohazardous Waste

labeled container

lined with a biohazardous waste bag

equipped with a lid.

Managing Non-Sharp Biohazardous Waste

Securely tie bags for transport to treatment/collection site.

When moving wastes, use secondary containment; avoid using public halls and elevators.

“Breakable” Non-sharps Biowaste

Store in labeled containersthat are puncture-resistant, closable and will capture leakage, BUT….

…Do NOT use SHARPS containers!

Effective Waste Autoclaving

Leave bag open during autoclaving or loosely closed

Add water to bag prior to autoclaving if primarily dry materials

Steam must contact materials

Place bag in autoclavable tray with sides

Treated Waste Bag Disposal

Allow waste bag to cool Use fume hood to reduce

odors Securely tie bag shut Place bag in a non-

transparent black bag for regular disposal

Remember: NO ORANGE BAGS IN DUMPSTER!

#3Is it a sharps hazard?

Examples:– needles– syringes– scalpels– all biologically contaminated objects that

can easily penetrate skin (Pasteur pipettes,razor blades, etc.)

Place sharps in approved sharps container for disposal!

…Syringes in research settings should be disposed of as a sharp to avoid public relations concerns!

Sharps Containers Containers must be leak-proof,

puncture-resistant, closable & labeled with the biohazard symbol.

Proper sharps containersmust be used forboth clinic andfield work.

Proper Use of Sharps Containers

Place tops on containers before use on lab bench

Don’t forget to date the container when first put into use

Remember: sharps containers are aone-way disposal system

Proper Use of Sharps Containers

Use sharps containers for sharps ONLY!

• No solid biohazardous waste (i.e. gauze, un-broken pipettes, gloves)

• No mercurythermometers

What’s wrong with this picture?

Sharps Container Disposal Containers must be permanently closed

and disposed of through the animal facility manager:Within 90 days

of first useWhen ¾ full

Disposal methods:Landfill IncinerationWe use waste hauler

Safety Notes on Sharps Use Do not re-cap sharps

Keep sharps container in close proximity to point of use (i.e. limit handling) for easy disposal

Do not leave needles in pockets of coveralls or smocks

Carcasses and Body Parts Human tissues

Unfixed tissues are medical waste Make waste unrecognizable!

Animal tissues, carcasses When generated in infectious disease or recombinant DNA research, these are

medical waste

These items must be stored in biolabeled, leakproof containers for incineration.

Waste service- see Audrey Brown

Managing All That Other Waste…

Drain bottles of non-hazardous materials before disposal in trash

<3% of volume is considered empty Higher volumes must not be thrown

in the trash

Managing All That Other Waste…

Do NOT discard medications in the trash.

Return to source for disposal or seek assistance from your campus waste group.

See Jaime Stock!

Any Questions?

Thank You for your attention!

Thanks to Carol Stevens at CMU & Robin Mecklem at MSU for their assistance with this presentation