Biorisk Management

Download the Biosafety and Biosecurity Manual Program Document.

This document outlines the minimum standards and requirements for the safe handling, use, storage, transfer, and disposal of biological materials in research and instructional laboratories at Penn State, focusing primarily on Biosafety Levels 1 and 2. It integrates federal, state, and local regulations and establishes a comprehensive biorisk management program to protect human, animal, and environmental health.

Coming Soon.

Biohazardous waste includes materials potentially contaminated with pathogens or recombinant/synthetic nucleic acids (rDNA). Examples:

• Cultures/stocks of infectious agents (BSL2)
• Human/primate blood, body fluids, tissues, and contaminated items
• Animal waste from research involving infectious agents
• Used sharps exposed to infectious materials

Key Notes:

• Non-biohazardous rDNA waste may go in clear autoclavable bags (no biohazard symbol).
• All biohazardous waste must be decontaminated before disposal.

Disposal Procedures

Solid Waste

• Use red, orange, or clear autoclavable bags with a biohazard symbol.
• Double-bag petri dishes with agar.
• Collect serological pipettes separately to avoid punctures.
• Place bags in leak-proof containers; loosely tie bags before autoclaving.

Sharps

• Use rigid, puncture-resistant red/orange sharps containers.
• Close when ¾ full; never force items in.
• Avoid cardboard or food containers.

Liquid Waste

• Use non-breakable containers.
• Add bleach (10% final concentration).
• Label with biohazard symbol and disinfectant name.
• Let sit 30 minutes before sewer disposal; rinse sink thoroughly.
• Never autoclave pretreated liquids.
• Consult EHS for non-bleach disinfectants.

Ethidium Bromide and Acrylamide Waste

• Treat all electrophoresis gels and related materials as chemical waste.
• Green-tag waste for chemical waste disposal.
• Do not use red biohazard bags.
• Dispose of ethidium bromide-contaminated buffer and sharps as chemical waste.

Biologically-Derived Toxins

• Autoclave or chemically deactivate waste.
• Use at least 60-minute contact time for disinfectants.
• Refer to Table 7 for toxin-specific methods.
• Consult EHS for conotoxin and other special cases.

Plant Waste

• Composting allowed after inactivation (if rDNA involved) or per APHIS permit.
• Use clear bags for autoclaved compostables.
• Red/orange bags reserved for biohazardous RMW.
• Refer to Plant Containment Guidelines.

Mixed Waste

• For biological + chemical or radioactive waste, consult EHS before generation.
• Requires special handling for compliance and safety.

For Biosafety Level 1 labs, follow these basic precautions:

• Wash hands after handling biological materials and before leaving the lab.
• Limit access to trained personnel during experiments.
• Prevent exposure to eyes, face, and mouth in hazardous areas.
• Follow and enforce restrictions that minimize eye, face, or mouth exposure in areas with hazardous materials. Prohibit eating, drinking, chewing (tobacco, gum, etc.), applying cosmetics or lip balm, and pipetting or siphoning by mouth.
• Keep food and drinks out of the lab; store them only in designated areas outside.
• Minimize splashes and aerosols during procedures.
• Use mechanical pipettes—never pipette by mouth.
• Handle sharps carefully; avoid use unless necessary. Follow Section 15.4.4 for disposal.
• Work on open benches or in biosafety cabinets (BSCs) when needed (e.g., tissue culture).
• Decontaminate work surfaces daily and after spills.
• Decontaminate all biological waste before disposal (see Chapters 13 & 14 of the Biosafety and Biosecurity Manual).

For Biosafety Level 2 labs, in addition to precautions for BSL1, enhanced precautions are required:

• Minimize aerosols; use BSCs, sharps safety devices, and PPE (e.g., face shields, eye protection, double gloves).
• Apply Universal Precautions for human/primate materials (see Exposure Control Plan).
• Decontaminate equipment before removal or maintenance.
• Keep spill kits tailored to your lab’s materials and volumes.
• Use safety features on aerosol-generating equipment (e.g., sealed centrifuge rotors, enclosed sonicators).

Proper lab design supports research, instruction, and safety.

BSL-1 Facility Requirements

• Door: Closable entrance.
• Surfaces: No carpets or porous materials; use easy-to-clean, chemical-resistant work surfaces.
• Sink: Handwashing sink inside the lab.
• Windows: Exterior windows must have screens.
• Furniture: Non-porous and easy to decontaminate.
• Eyewash: Install per Penn State safety standards.

BSL-2 Additional Requirements includes all BSL-1 features, plus:

• Lockable doors to control access.
• In-lab decontamination for infectious waste.
• HEPA filters on vacuum lines used with biological materials.
• Biosafety cabinets placed away from doors and traffic.
• Eyewash station must be easily accessible.
• Ventilation: ideally, HVAC should provide 10–12 air changes per hour.
• Windows: Must be sealed (non-operable) to maintain airflow control.

Enhanced Containment (BSL-1+ / BSL-2+): When risk assessments indicate the need for added precautions—without requiring full BSL-2 or BSL-3 protocols—labs must adopt selected higher-level practices. These may include:

• Use of negative air pressure rooms to contain airborne contaminants.
• Vaccinations for personnel handling specific agents.
• Occupational health monitoring for exposure tracking.
• Restricted access to authorized personnel only.
• Respiratory protection during aerosol-generating or high-risk procedures.
• Centrifuge loading/unloading inside a biosafety cabinet.
• Double gloves to reduce skin exposure.
• Disposable sleeve covers for arm protection.
• Solid-front gowns for full torso coverage.

Explore the Bloodborne Pathogens Program. Access the Resources and documentation. 

Decontamination reduces microbial contamination to safe levels. It includes sterilization, disinfection, and antisepsis. Cleaning is essential before these processes to remove organic matter and ensure effectiveness.

Sterilization kills all microorganisms. Methods include steam (autoclaving), dry heat, gas/vapor, plasma, and radiation.

• Steam Sterilization: Autoclaving uses steam at 121°C and 15 psi to sterilize materials. Loosely close bags to allow steam penetration. Use Type 5 chemical integrators in every load to verify time, temperature, and moisture. Conduct annual validation with biological indicators (e.g., Bacillus spore strips).

• Dry Heat Sterilization: Used for materials like oils and powders. Requires 160–170°C for 2–4 hours. Less efficient than steam.

Disinfection eliminates most pathogens (not spores). Effectiveness depends on microbe type, organic matter, surface, and temperature. Use appropriate PPE and ensure good ventilation. Choose disinfectants based on activity level:

• High-level: Kills most microbes and viruses (e.g., concentrated bleach). Not for surfaces.
• Intermediate: Kills TB, fungi, most viruses (e.g., hospital disinfectants).
• Low-level: Kills some bacteria and viruses. Used for general cleaning.

Selected Disinfectants Examples:

• Hydrogen Peroxide (0.5–3%): Fast-acting.
• Bleach (500–6000 mg/L): Broadly effective. Use 1:10 dilution. Avoid mixing with other chemicals.
• Phenolics: Effective but odorous and toxic.
• Quats: Low toxicity, not compatible with bleach.

UV Radiation is not reliable for disinfection. Ineffective on porous/dirty surfaces, in shadows, or high humidity. Bulb intensity declines with age—check annually. Not recommended as a primary method. Alcohols are not recommended as a primary disinfectant because rapid evaporation makes achieving the required 10-minute contact time difficult.

Equipment Decontamination: Before removing equipment from the lab, clean with 10% bleach and allow contact time. Attach a completed Decontamination Certificate. Contact EHS if unsure how to decontaminate.

Toxin Inactivation: Toxins used in research must be inactivated before disposal. Contact EHS for proper procedures.

Bunsen Burners and Induction Sterilizers: Open flames were once standard for sterilization, but modern labs—especially those using biosafety cabinets (BSCs)—should avoid them due to safety risks.

Open Flames Are Prohibited in BSCs

• Disrupt airflow: Flames disturb laminar flow, spreading aerosols.
• Overheat the cabinet: Can damage sensitive materials.
• Melt HEPA filters: Compromises containment.
• Void warranties: Manufacturers ban flammable gases in BSCs.
• Fire/explosion risk: Gas buildup and electrical sparks can ignite fires.

Safer Alternatives

• Disposable sterile tools: Eliminate the need for flame.
• Bacti-Cinerators: Flame-free loop/needle sterilization.
• Glass bead sterilizers: For small instruments.
• Electric Bunsen burners: Controlled heating without open flame.

For help choosing the right alternative, contact EHS.

Explore the Personal Protective Equipment program. Access the Resources and documentation. 

Download the Proper Use of Autoclaves.

Download the Proper Use of Biosafety Cabinets procedure document.

Sharps are items that can puncture or cut skin, posing a risk of exposure to biohazards.

Examples:

• Needles (hypodermic, surgical)
• Syringes
• Scalpels, razor blades
• Pasteur pipettes
• Contaminated glass slides, cover slips, broken glass

Note: Pipette tips aren’t classified as sharps but can puncture waste bags. Dispose of them in sturdy containers (e.g., empty media bottles).

Safe Handling and Disposal

• Use plasticware instead of glass when possible.
• Only use sharps when no safer alternative exists.
• Prefer safety-engineered sharps (e.g., retractable needles).
• Use Luer lock or integrated needle syringes to prevent disconnection.
• Never place sharps in bags—use red sharps containers only.

Best Practices

• Keep sharps containers within reach, including inside BSCs.
• Dispose of sharps immediately—no handling or recapping.
• Never bend, shear, or remove needles/blades by hand.
• Never recap needles unless absolutely necessary.

If recapping is unavoidable, use:

• One-handed scoop method: Slide the needle into a cap placed on a flat surface.
• Conical tube or rack method: Place the cap in a tube or rack and insert the needle safely.
• Commercial recapping devices are also available.

Transport sharps in secondary containers, especially in public areas.

Clean-up: Use tools (forceps, tongs, cardboard, broom/dustpan) for broken glass—never use hands.

Ampule Safety: Opening glass ampules can cause cuts and aerosol exposure. Always open ampules inside a BSC at BSL-2.

To open safely:

1. Score the neck with a file.
2. Wrap in a disinfectant-soaked towel.
3. Snap open while upright.

To reconstitute contents:

• Add liquid slowly to avoid aerosols.
• Mix gently and transfer contents to a clean container.
• Dispose of ampule parts and towel as biohazardous waste.

Avoid using glass ampules in liquid nitrogen. Use polypropylene tubes instead—they’re safer and explosion-resistant.