Working Safely with Biological Materials Basics of Biosafety Working Safely with Biological Materials

What is Biosafety? Principles and practices employed to protect laboratory personnel (& others in the area) 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

Agents and Risks There is always risk! The risk must be identified The “agent” is the what creates risk Risks to the worker or environment are often unknown Determining “acceptable risk”? There is always risk! The risk must be identified The risk is evaluated The risk must be measured Plan to minimize the risk

Who Determines Acceptable 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 We are working on this.

Identifying 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

Evaluating Risk Acceptability Worst case scenario -What might happen? Likelihood of an event Seriousness of the incident Actions needed to resolve the problems

WHO-World Health Organization 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

Biosafety Levels (BSL) 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 Level 1 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 2 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 3 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 4 FORGET ABOUT IT!!! Hemorrhagic fever, deadly viruses, etc. Total containment, airtight labs, “submarine” doors, air pumps, water treatment, HEPA filtration, etc. Positive pressure “moonsuits”

Laboratory Acquired Infections (LAI) Bacterial: 76% from clinical labs 8% 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

Biohazardous/Medical Waste Waste that is potentially infectious to humans, animals or plants. It includes: Medical Waste according to MMWRA Regulated Waste by MIOSHA Regulated 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 “Medical waste” means any of the following that are not generated from a household, a farm operation or other agricultural business, a home for the aged, or a home health care agency: (a) Cultures and stocks of infectious agents and associated biologicals, including laboratory waste, biological production wastes, discarded live and attenuated vaccines, culture dishes, and related devices. (b) Liquid human and animal waste, including blood and blood products and body fluids, but not including urine or materials stained with blood or body fluids. (c) Pathological waste. (d) Sharps. (e) Contaminated wastes from animals that have been exposed to agents infectious to humans, these being primarily research animals.

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 a) Cultures and stocks of infectious agents and associated biologicals laboratory waste biological production waste discarded live and attenuated vaccines culture dishes and related materials contaminated PPE

Biohazardous Waste Categories b) Liquid human and animal waste liquid or semi-liquid blood and blood products and body fluids contaminated 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 c) Pathological waste tissues body parts other than teeth products 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: Is it contaminated with viable biological material or recombinant DNA? Can blood or other regulated body or biological fluids be released? Is it a sharps hazard? If the answer to any of these questions is yes, then you have a biowaste.

Is it contaminated with viable biological material or recombinant DNA? #1 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…)

Can blood or other (regulated) body fluids or viable biological materials be released? #2 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 Autoclave Disposal: THEN Flush to sewer Use proper PPE!

Disinfection 10% bleach solution Ethanol 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 *Researchers should investigate and know effectiveness and contact time for the best disinfectant against your agent!

Managing Non-Sharp Biohazardous Solid Waste labeled container lined with a biohazardous waste bag equipped with a lid.

The Autoclave or Steam Sterilizer

Moist Heat Sterilization Proper autoclaving is important All air must be flushed out of the chamber The chamber should not be overpacked with solids Large volume of liquids may take longer to autoclave due to the high specific heat of water. To determine if the proper conditions have been met, we use indicators Tape – indirect indicator Biological indicator – paper strip covered with Geobacillus spores

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!

Place sharps in approved sharps container for disposal! Is it a sharps hazard? #3 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 containers must be used for both clinic and field 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 a one-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 mercury thermometers

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Sharps Container Disposal Containers must be permanently closed and disposed of via our hazardous waste company Within 90 days of first use When ¾ full Disposal methods: Landfill Incineration We 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.

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. Treat as chemical waste.

Any Questions?