MGI

BSL-2

Non-aireborne moderate risk agent containment (Bloodborne, foodborne, and waterborne pathogens) Suitable for work involving agents that: Are associated with human disease. There are preventative or therapeutic interventions often available. They are a moderate potential hazard to personnel and the environment. At BSL-2 work is limited to infectious materials which are likely to be transmitted through ingestion of contaminated items, through broken skin contact (like a hangnail or razor burn), and contact through the mucous membranes. Controlling transmission in a BSL-2 laboratory can be accomplished by following the same procedures for BSL-1, and through utilizing additional engineering controls, PPE, and hazard communication.

Non-Emergency Situations

Non-emergency situations involve releases that do not pose significant safety or health hazards to persons in the immediate vicinity or to the person cleaning, and do not have the potential to become emergencies within a short time frame.

Lethal Dose 50

One of the most commonly used measures of toxicity is the LD50. The LD50 is one way to measure the short-term poisoning potential (acute toxicity) of a material. The LD50 is the single dose that is lethal to 50% of the animals tested. A chemical with a low LD50, like osmium tetroxide is highly toxic. The LD50 however, says nothing about the non-lethal toxic effects of chemicals. Some chemicals may have a high LD50, but may produce toxic effects at very small exposure levels, Sulfuric acid has an oral (rat) LD50 of 2,140 mg/kg, but may be corrosive at concentrations as low as 15%, causing severe burns at very low quantities.

Administrative Controls

One way to control worker exposure to workplace hazards is through policies and procedures. Some examples of the administrative controls used in labs are: The MSU CHP Lab-Specific Training Security Training Standard Operating Procedures Chemical Labeling (Material) Safety Data Sheets

Heat-Sensitive Indicators

Operators should use heat-sensitive, lead-free sterilization indicator tape for each load to indicate minimum steam temperature. This tape only indicates that the temperature has reached 1210c, but does not indicate it was heated for the proper length of time.

Permissible Exposure Limit (PEL)

PELs are regulatory limits on the amount or concentration of a substance in the air. PEL's are based on an 8-hour adult time weighted average (TWA) exposure. PEL's refer to airborne concentrations of substances that may be inhaled. When working with materials that have a low PEL (< 50ppm), use administrative and engineering controls to minimize the generation of a vapors or dust in the first place. If these controls are not sufficient, use appropriate personal protective equipment

Personal Protective Equipment

PPE is a device worn on the body to protect the wearer from a hazard. When used correctly, PPE will create a barrier between the hazard and the route of transmission. When selecting PPE consider the following items: Is it designed for the hazards encountered? Is it designed so as not to hinder your ability to work? Is it disposable or reusable equipment such as gloves, lab coats, shoe booties, etc.? Is it available in a range of sizes?

To interrupt or prevent the infection process, the goal is to prevent ________ transmission from the reservoirs to a susceptible host.

Pathogen

The Chemical Hygiene Plan (CHP) contains basic SOPs for:

Peroxide Forming Chemicals Labeling of Chemicals Small Lab Spills However, labs must create SOPs for procedures/chemicals not in CHP: Acid Baths Highly Toxic Chemicals

Peroxidizable Chemicals

Peroxidizable chemicals can undergo auto oxidation to form organic peroxides that can become explosive with impact, heat or friction. These chemicals may become more hazardous as they age; peroxides may form even when the container has not been opened. Good management of peroxidizable materials starts with: -Dating all peroxide formers upon receipt and again after opening -Dispose of, or test for peroxide formation at the expiration date or 18 months if the container has not been opened Never open a bottle that has solid formation around the lid. (Friction caused by unscrewing the cap can lead to explosion.)

Glove Use

Prior to use, inspect for defects, such as discoloration or holes in the gloves. Must be worn when hands may come into contact with potentially infectious materials, contaminated surfaces or equipment. Change when overtly contaminated to prevent the spread of contamination. Must not be reused if they are disposable. When removing, limit contact with outer surfaces, minimizing contact with the skin. WASH your hands after removing gloves. The US Food and Drug Administration has issued a ban on all powdered gloves. Exposure to starch powder from gloves can cause undesirable reactions, which vary from well-known allergy symptoms and upper respiratory-tract disorders to surgical adhesions and infections. The presence of glove powder can also result in many other undesirable effects, such as interference in laboratory testing causing false results (i.e. PCR - Polymerase Chain Reaction, enzyme immunoassay or some HIV tests).

Selecting Appropriate Eye and Face Protection Equipment

Safety glasses are required when: An impact hazard exists When working with low hazard chemicals When a low probability of splash exists Examples: Pipeting Handling closed bottle of injurious chemical Mixing solutions Opening centrifuge tubes Chemical splash goggles are required when: Working with small amounts of corrosive or injurious chemicals Reasonable probability of splash exists Examples: Pouring acid out of a 1 pint or 500 mL bottle Pouring methylene chloride from a 1 liter bottle Working with liquids under pressure Face shield and splash goggles required when: Working with larger quantities of corrosive chemicals A high probability of eye and face injury exists Examples: Working with an acid bath Pouring 4 liters of acid into a container Handling highly reactive chemicals that may spatter

Safety Glasses

Safety glasses do not protect an individual's eye from exposure from the sides, top, and bottom.

Which of the following materials can be safely disposed of into the sink/sanitary sewer system (select all that apply)? 3% Hydrochloric acid solution Buffer solution (pH unknown) Leftover saline in IV bags Soapy water from glassware dishwashing 70% ethanol solution Expired, unused Oxvir cleaning product Cell culture waste containing 10% bleach solution

Saline solutions, bleach solutions of 10% or less, and soapy water solutions are acceptable for sink disposal. Buffer solution may be acceptable for drain disposal; however in this case, the pH of the material is unknown and must be tested prior to sink disposal. Only dilute solutions of cleaning products are allowed down the drain - expired, unused cleaning products must be submitted as hazardous waste.

Biosafety Level 3: Facility

Separate building or isolated zone Double-door entry Directional inward airflow Single-pass air

Corrosives

Skin corrosion means the production of irreversible damage to the skin following the application of a test substance for up to 4 hours. If corrosive chemicals come in contact with skin or eyes, they cause irreversible and visible damage at the site of contact. The damage will be acute. Chemicals with a low or high pH (<2.0 or >12.5) are considered corrosive. Therefore, acids and bases are corrosive. Examples: Hydrochloric acid Ammonium hydroxide Acetic acid

Autoclaving - Don't endanger yourself by accident!

Some plastics are not stable when autoclaved and may release cancer-causing gases (styrene and ethyl benzene), or may melt and ruin other materials. Corrosives will significantly damage the autoclave chamber. Radioactive materials can further break down and release potentially toxic chemicals or radiation. Dangerous chemicals can release toxic gases.

Non-RCRA, Non-Hazardous Waste*

Some waste materials are not considered hazardous waste and can be disposed of in the regular trash. Those materials include: Uncontaminated lab debris Uncontaminated PPE

Standard Operating Procedures (SOPs)

Standard Operating Procedures (SOPs) are required for any lab work that falls outside of the hygiene plans general operating procedures. An SOP is a set of written procedures explaining how to safely work with hazardous chemicals.

_______ sterilization is a time-proven method of killing microorganisms.

Steam

Heat damages the cell's ________, making it no longer infectious.

Structure

Minimizing Contact with Chemicals

The risk of exposure to particularly hazardous substances can be reduced by: -Substituting whenever possible for less toxic/hazardous materials -Using as little material as possible -Handling inside a fume hood -Minimizing contact by the use of personal protective equipment *It is necessary to develop a written SOP for operations involving highly toxic materials Never intentionally smell, inhale or taste a chemical. Wash thoroughly with soap and water after handling any chemical and removing your PPE.

Chemical Storage Guidelines

The storage area and cabinets should be labeled to identify the hazardous nature of the products stored within. Chemicals should be stored according to compatibility. (E.g., oxidizers such as sulfuric and nitric acid away from organics such as acetic acid and acetone). Corrosive and flammable materials should be stored below eye level. Chemicals should not be stored on the floor except in approved shipping containers. Dry chemicals can be stored on shelves. Flammable liquids should be stored in flammable liquid storage cabinets or under fume hoods. Do not store flammable liquids in household refrigerators or walk-in cold rooms; use only flammable-safe refrigerators. Flammable liquids stored outside of flammable liquid storage cabinets in quantities greater than 1 liter need to be in a safety can. Examples: acetone, hexane, acetic acid Concentrated perchloric acid should be stored alone in a cabinet by itself

Potentially Infectious Materials

These may include: Pathological or microbiological wastes containing potentially infectious materials, including body fluids, tissues, other materials known or suspected to be infectious, and items contaminated with these materials. Cultures and stocks of infectious agents and associated biologicals. rDNA Waste items that are contaminated with viable genetically-modified materials.

4 Routes of Transmission

Through the eye: splashes of infectious material or, transfer by contaminated fingers Through the mouth: eating, drinking, mouth pipetting; or transfer by contaminated fingers Through the lungs: inhalation of airborne contaminants Through the skin: accidental inoculation with a sharp object (needle, broken glass) or, entry through broken skin

Safe Operation of Chemical Fume Hoods

To ensure that airflow is not interrupted, and that the fume hood offers as much protection as possible: -Keep sources of emission at least 6 inches inside the fume hood (behind sash). -Minimize storage of chemicals in the hood. -Avoid blocking off baffle exhausts. -Ensure that blower is working (some hoods even have an on/off switch). -Work with sash at proper operating level (between your face and materials in the fume hood). EHS inspects fume hood performance regularly, and at times may put "Match arrows for proper ventilation" stickers on the hood. Use the hood sash at or lower than the sticker to assure that air speed is adequate.

Autoclave Check Exhaust Setting

To prevent spills and accidents, be sure that the exhaust setting is appropriate for the type of material you are autoclaving. Use SLOW exhaust or Liquid Cycle for liquids, liquid wastes (including petri plates with agar), and most solid waste items (including instruments). Use FAST exhaust or a Gravity cycle as directed for specific solid items.

Flammable liquids safety

To safely work with flammables observe the following guidelines: Keep flammables/combustible materials away from sources of ignition, open flames, hot surfaces, electrical equipment and static electricity. Store flammable liquids in NFPA approved containers or inside cabinets or storage rooms designed for flammable materials. Small squirt bottles containing a volume of less that one-liter can be stored on the bench top. Larger volumes (>1 liter) need to be stored appropriately. Ensure fire extinguishers are in the area.

Shipping

To ship diagnostic specimens, infectious substances, genetically modified organisms/micro-organisms or to ship any of your biological materials on dry ice, you must have the proper training. If you intend to ship biological materials, you must: -Take the required biological shippers training (available online), as required by federal agencies. -Contact EHS biosafety team for assistance. -Prepare required specific packaging, labeling, and documentation. Due to an increased level of surveillance on the part of federal and international authorities, a shipper must ensure that materials are properly classified and that all applicable regulatory provisions for the shipment are met.

Toxic Chemicals

Toxic chemicals are substances that have an adverse effect on human health. These adverse effects can be acute (causing immediate harm), or chronic (long term exposure resulting in harm). Toxic chemicals can impart a systemic effect (multiple organ systems affected) or target a specific organ or body system. Five categories of acute toxicity are used by the Global Harmonization System of classifying chemicals. Substances are assigned to one of the five toxicity categories on the basis of LD50 (oral, dermal) or LC50 (inhalation). Chemicals that fall into a Category 1 are more toxic than chemicals in Category 4. The lower the category, the greater risk.

Transportation of Biological Materials

Transporting biological materials for research or diagnostic purposes, from one lab to another located in the same building, or across campus, occurs frequently at MSU. Although these materials may or may not contain pathogenic organisms it is still important to properly package, label and transport them to prevent product loss and to inform emergency responders of the contents in the event of an accident or other delays.

Emergency Situations

Uncontrolled releases of hazardous substances that pose a significant threat to health and safety, or any occurrence of a health or safety hazard are considered emergencies. The situation is unclear to the person causing or discovering the spill. The release requires evacuation of persons. The release involves or poses a threat of: Fire, suspected fire, explosion or other imminent danger. Conditions that are Immediately dangerous to life and health. High levels of exposure to toxic substances. The person(s) in the work area is uncertain they can handle the severity of the hazard with the personal protective equipment and response equipment that has been provided and/or the exposure limit could easily be exceeded.

Vehicular Transport of Research Materials

Use a University-owned vehicle whenever possible. A spill within the vehicle must be decontaminated. Contact EHS (355-0153). Store and secure the transport container in a location in the vehicle. For example, if transporting materials by car or van, secure the container via a seatbelt or other mechanism, keeping it upright and stable in the back seat or cargo bay.

Waste Minimization

Use fewer hazardous chemicals when ever possible. Purchase only needed amounts of hazardous chemical. Consult with your PI before purchasing. Determine amount needed from all lab workers (no double orders). Check inventory to dispose of any hazardous waste not being used.

Biohazardous Waste Management

Use the following prudent practices for disposal of waste items that are marked or labeled with the biohazard symbol: If the item is NOT contaminated with any biological materials or fluids, the biohazard label MUST be defaced before disposal in the regular lab trash. If the item is contaminated with biological materials or fluids, the item must be managed as biohazardous waste. Vacuum flasks of cell line material and waste body fluids are also regulated for disposal and cannot be placed in regular trash. We can reduce the amount of biowaste and help to prevent accidental exposures by the custodial staff and others by answering these three basic questions: -Is it contaminated with recombinant DNA and/or material that is potentially infectious to humans, animals or plants? -Can blood or other regulated body fluids be released? -Is it a "sharps" hazard? *If you can answer "Yes" to any one of these three questions, the item must be treated as biowaste for disposal purposes*

Disposable gloves such as nitrile may be used (select all that apply): When prolonged contact with chemicals is unlikely. When working with non-corrosive solutions or dry chemicals. When laboratory work requires a fine dexterity glove.

When prolonged contact with chemicals is unlikely. When working with non-corrosive solutions or dry chemicals. When laboratory work requires a fine dexterity glove. Disposable gloves may be used when immersion or prolonged contact is not common for chemical work in the laboratory. They are also good for work with non-corrosive solutions and dry chemicals.

How does a biosafety cabinet work?

When used properly, biosafety cabinets protect the person working in the cabinet, the materials being used, and the environment outside of the cabinet.

Gloves in Common Areas - Rules to Remember:

Whenever possible, gloves must not be worn outside the laboratory, even if they are clean! When transporting samples to another part of the building, use a secondary container. If gloves must be worn, use the one glove method shown in the bottom picture. Gloves must be worn in all designated BSL-2 spaces when contact with potentially infectious materials, contaminated surfaces, or equipment is possible. Do not reuse or wash disposable gloves or use them to touch clean surfaces (ie. telephone, keyboard, or doorknobs). Note: If you need to carry materials to another floor, wear only one glove and use the ungloved hand to touch common use surfaces such as elevator buttons and door knobs.

Materials ACCEPTABLE for Sink/Sewer Disposal

While generally all chemical wastes must be submitted to EHS for proper disposal, the following solutions may be sent to the sink: Saline solutions Buffer solutions (pH range 5.5 - 10.0) Soapy water solutions Blood with deactivated cultures Rinse water solutions (from non-hazardous lab glassware) Glucose solutions Food additive solutions Bleach solutions (10% or less) Watercolor dye solutions Disinfected liquid biohazardous waste, blood, or body fluids (disinfected with bleach, TB-Cide, quats, etc.)

Disinfectant Guidelines

You must disinfect any work surface and items within arm's reach of the work surface where specimen materials were manipulated. If bench paper was in use where biologically-active materials are manipulated, it must be removed and the bench underneath disinfected at the conclusion of the procedures or end of the work shift. Disinfection must include any equipment that was part of the material manipulation. If you must have equipment repaired or certified, an equipment release form (a document that assures disinfection of equipment) must accompany each item.

Loading the Autoclave

You must properly load the autoclave to ensure effective processing. Do not overload the chamber with bags for the capacity of your autoclave. Allow the steam to circulate freely throughout the chamber. Keep the drain clear. Do not mix together supplies that must be used for sterile surgery along with biological waste for disposal in the same cycle.

Selecting the "Right" Disinfectant

You want to make sure the product can effectively destroy the agent of concern. Dilution ratio is important. Some agents may be destroyed with 70% Ethanol, or may use a 1% - 10% bleach dilution. Contact time is a key component of disinfection. Apply the disinfectant to the contaminated surfaces and allow the surfaces to remain wet with the product for the manufacturer's prescribed contact time. Most agents require a contact time of 10 minutes. Bleach concentrate according to the EPA has a shelf life of 6 months; once diluted the shelf life becomes 24 hours. All disinfectants contain hazardous ingredients; therefore, you must review the Material Safety Data Sheet (MSDS) for the product before you use it and follow the manufacturer's recommendations for safe use.

When loading and unloading the _________, you may encounter: biohazards, hot containers, steam and spills that can cause severe burns.

autoclave

Substances which on contact with water emit flammable gases

chemicals that, in contact with water, emit flammable gases are solids or liquids which, by interaction with water, are liable to become spontaneously flammable or to give off flammable gases in dangerous quantities.

Pyrophoric substances

compounds which, even in small quantities, are liable to ignite within five minutes after coming into contact with air.

Biosafety defines the ______________ ________________ under which infectious agents or biohazards can be safely manipulated. Containment conditions are also known as biosafety levels (BSLs).

containment conditions

As the biosafety level _______, so do the procedural and facility requirements and the risk to humans, animals, and/or the environment

increases

Self-Heating substances

solids or liquids, other than pyrophoric substances, which, by reaction with air and without energy supply, is liable to self-heat. This differs from a pyrophoric substance in that it will ignite only when in large amounts (kilograms) and after long periods of time (hours or days).

Self-Reactive substances

thermally unstable liquids or solids liable to undergo a strongly exothermic thermal decomposition even without participation of oxygen (air).

Chemical Spill Emergency

(Spills greater than 1 liter and/or potential high-risk chemicals like flammable liquids, corrosive materials or highly toxic) Call 911 (MSU dispatch will contact EHS). Evacuate personnel from the spill area and alert neighbors to the spill. Isolate the spill area and close doors to the room where the spill occurred. Remove ignition sources and shut down equipment. Turn fume hood ON in HIGH or in emergency mode.

Leaving the Laboratory

*After completing a day of work in the laboratory, or anytime you leave the laboratory take some time and make sure that you:* Stabilize your experiment, equipment or process. Label all chemical containers and store them properly. Clean all contaminated instruments or containers including glassware. Dispose of hazardous waste in properly labeled containers. Remove PPE and wash your hands thoroughly. *Housekeeping* Keep work areas clean and uncluttered with chemicals and equipment. Clean up work areas upon completion of an operation or at the end of each workday, including floor. Do not block exits, emergency equipment or controls. Designate waste container for non-contaminated glass.

Managing Liquid Biohazardous Waste There are two different methods of decontamination that can be used:

1. Chemically disinfect the liquid waste using an appropriate disinfectant and following the manufacturer's specifications for use. The waste can then be disposed of down the sanitary sewer by way of the lab sink. 2. Autoclave the contaminated liquid waste and then dispose down the sanitary sewer by way of the lab sink. Safety Notice: Do not chemically disinfect AND autoclave the liquid waste. Autoclaving chemicals is hazardous and should not be done (i.e. Never put solvents, volatile or corrosive chemicals such as phenol, chloroform, bleach, etc., or radioactive materials in an autoclave).

Hazard Identification Goal:

1. Identify the hazard 2. Select appropriate safety measures -Use appropriate engineering controls -Personal Protective Equipment -Develop and follow SOPs 3. Reduce the risk of exposure or injury

After the Cycle Finishes

1. use proper PPE to avoid burns 2. Wait until the chamber pressure gauge reads zero before attempting to open. Waiting at least 10 minutes will significantly reduce your risk of being burned 3. Remove the secondary container carefully. It may have collected liquids or moles agar during the cycle.

The storage limit for hazardous waste in the laboratory is:

90 days

Material Safety Data Sheets/Safety Data Sheets

A Material Safety Data Sheet (MSDS) or a Safety Data Sheet (SDS) is a valuable reference. Prepared by its manufacturer, an MSDS/SDS provides information to help you understand the intrinsic hazards of the chemical, including: Physical and chemical properties Reactivity and stability information Physical and health Hazards Acute and chronic toxicity information Permissible exposure limits Waste disposal Handling and storage information

Germ Cell Mutagenicity

A mutagen is an agent that increases occurrence of mutations in populations of cells and/or organisms. Substances and mixtures with mutagenic effects are assigned to one of two hazard categories, "known or presumed" and "suspected." Examples: Ethidium bromide Formaldehyde, nicotine Other common mutagens

Health Hazards

A person must be exposed (inhalation, skin absorption, ingestion) to a health hazard chemical for it to cause harm. The level and duration of exposure determines the severity of the health effect. The effects of exposure to hazardous chemicals vary with the amount of exposure or "dose."

Decontamination is achieved through ...?

A proper combination of time, temperature, and pressure.

Safer Sharps Devices

A safer sharps device has a built-in safety feature or mechanism that effectively reduces the risk of an exposure incident by preventing the sharps injury before, during, and after use of the device. Safer sharps devices are highly recommended when working with BSL-2 materials and required when working with human materials. For information on safer sharps devices and manufacturers, please contact the Biosafety Staff at EHS.

What is the most complete source of information about a chemical's hazards and properties?

A safety data sheet (SDS) or material safety data sheet (MSDS)

Substances Corrosive to Metal

A substance or a mixture that by chemical action will materially damage, or even destroy, metals is termed "corrosive to metal." These substances can cause damage to equipment, storage cabinets and structural elements. Care must be taken to ensure these chemicals cannot cause damage to surrounding materials; consider the use of secondary, corrosion-proof containment during storage.

Substances Corrosive to Metal Examples include:

Acids Bases Caustics or alkalis Alkali metals Dehydrating agents Strong oxidizers such as concentrated hydrogen peroxide Elemental fluorine, chlorine, bromine and iodine Acid anhydrides

To reduce infection and contamination risks laboratories are instructed to utilize:

Administrative controls such as required training. Engineering controls and facility design. Work practices and procedures to minimize risk.

Unloading - Solid Waste

After carefully removing solid waste, prepare it for landfill: 1. Close the treated biohazard bag tightly. 2. Place treated autoclave bag into a new non-transparent black bag. Do not put multiple orange biohazard bags in a single black bag. 3. Close black outer bag securely. 4. Place in approved dumpster. AUTOCLAVED

Use and Care of Safety Glasses

After repeated use, safety glasses require different types of care, including cleaning, maintenance, and replacement. Cleaning: -It is very important to clean your safety glasses on a regular basis: -Rinse with tepid water to remove grit. -Wipe with soft dry cloth or tissue taking care not to scratch lens. Maintenance: -Safety glasses should be checked before every use for damage or flaws such as cracks, scratches, or dents: -Run your finger over the face of the lenses to find imperfections or scratches that may have occurred during previous use or storage. -Put the glasses on to observe the clarity the lenses. Replacement: -Safety glasses should be replaced if the glasses have significant damage or if they feel uncomfortable.

BSL-3

Airborne Hazard Containment (Mycobacterium tuberculosis complex species) Suitable for work with moderate risk airborne pathogens, such as: Coxiella burnetii Mycobacterium complex species In addition to BSL-2 practices: -Unique facility requirements -Respiratory protection -Further training and restricted access

Handling Contaminated Materials

All procedures are performed carefully to minimize the creation of aerosols. Work surfaces are decontaminated once a day and after any spill of viable material. All contaminated liquid or solid wastes are decontaminated before disposal. Experimental organisms shall be rendered biologically inactive by appropriate methods before disposal outside of the laboratory. Contaminated materials that are to be decontaminated at a site away from the laboratory are placed in a durable leak-proof container which is closed before being removed from the laboratory. Waste is properly disposed of through pick-up or by autoclaving (this area will be covered in depth later in the training).

Tips for using a biosafety cabinet:

Allow the BSC to purge for 3-5 minutes before use. Be careful not to overly disrupt the air currents in the cabinet. This will prevent the proper flow of air through the HEPA filter and in the cabinet, which could possibly cause harmful aerosols to reach the user and cause an exposure. You can help prevent airflow disruption by following these guidelines: -Do not block the inlet grilles in the front or back of the BSC. -Keep a waste collection container in the cabinet for pipettes and tips to prevent excessive movement in and out of the cabinet. -It is strongly recommended to avoid using open flames inside the cabinet. Wear gloves and a lab coat when using a BSC. Work from "clean to dirty." Use disinfectant-filled vacuum traps for aspiration of infectious fluids. -Lentiviral and retroviral work requires an inline filter between vacuum source and flask. Stabilize and contain flasks to prevent spillage. Disinfect the BSC at the end of each use

Materials Pick Up Tag

An MSU Waste Materials Pick Up Tag must be attached to each waste storage container. Complete immediately: location and contact information start date (date first drop of waste is added to container) Complete as waste is accumulated: UNABBREVIATED chemical names of each material poured into container Actual amounts of material added to container (amount of waste in container must match listed materials at all times) Check balance water as necessary Potential hazards (ignitable, corrosive, toxic, reactive) by checking applicable boxes Complete when container is full (or within 90 days): Color Consistency

BSL-2 engineering controls include:

An eyewash facility must be available Sharps containers available (if applicable) Biosafety Cabinet installed, certified, and used Autoclave available Centrifuges with safety features

Organic Peroxides

An organic peroxide is an organic liquid or solid which contains the bivalent -0-0- structure, where one or both of the hydrogen atoms have been replaced by organic radicals. The term also includes organic peroxide formulations (mixtures). Such substances and mixtures may: be liable to explosive decomposition; burn rapidly; be sensitive to impact or friction; react dangerously with other substances Examples include: Benzoyl peroxide Methyl ethyl ketone peroxide (MEKP)

Oxidizing Materials

An oxidizing liquid or solid is a substance which, while in itself not necessarily combustible, may, generally by yielding oxygen, cause or contribute to the combustion of other material. These reactions can result in a fire or an explosion. Oxidizers are incompatible with organics. Store oxidizers separate from organics. Use oxidizers with extreme care and consult the (M)SDS for specific guidelines.

When transferring chemicals to a different container, the container must be labeled with (select all that apply): Any hazards that the substance may have. The unabbreviated name of the chemical. The chemical formula of the substance.

Any hazards that the substance may have. The unabbreviated name of the chemical. The chemical formula of the substance. All chemical containers must be labeled with the unabbreviated chemical name and the hazard warning.

Exposure Incidents

Any situation that results in an exposure incident requires that immediate action be taken. If blood, body fluids or other viable biological materials: -Are accidentally ingested, Splashed into your eyes, nose or mouth, -Enter your body through breaks in the skin, or -Enter your body by way of a biologically-contaminated sharp device (including a bite),

The LD50 can be used:

As an aid in developing emergency procedures in case of a major spill or accident. To help develop guidelines for the use of appropriate safety clothing and equipment. For example, if the dermal LD50 value for a chemical is rated as extremely toxic, it is important to protect the skin with clothing and gloves made of the right chemical-resistant material. As a part of the information in Material Safety Data Sheets. Remember, the LD50 is only an approximate figure so that lethal toxicity can be compared. It says nothing about levels at which other acute toxic, but non-lethal effects might occur.

Autoclave Safe Operator Practices

Autoclave rooms often contain contaminated materials. No Food or Drinks or Food/Drink Containers No Cosmetics No Medicine

A _________ is an agent of biological origin that has the capacity to produce deleterious effects on humans, animals, plants, and/or other living organisms.

BIOHAZARD

_____________ is the application of knowledge, techniques and equipment to prevent personal, laboratory, and environmental exposure to biohazards or other potentially infectious agents.

BIOSAFETY

There are four biosafety levels; with BSL-1 being the (least/most) restrictive and BSL-4 being the (least/most) restrictive

BSL-1 being the least restrictive BSL-4 being the most restrictive

Autoclave Safety Use the Posted Guidelines

Basic Procedure: Follow the guidelines set by the posted autoclave parameter signs when setting cycle time. -Use the Posted Guidelines -121°C is only minimum -Parameters vary -Set by EHS DO NOT override autoclave's built-in safety control features under any circumstance.

Preparing to Ship Biological Materials

Before you package and ship materials to an off campus destination YOU MUST EITHER BE TRAINED TO SHIP BIOLOGICAL MATERIALS or contact the EHS-Biological Safety Officer: Dr. Jamie Sue Willard-Smith (353-1877). There are several items that should be taken care of when preparing to ship biological materials. These paperwork requirements can take several weeks to complete, therefore you should prepare well in advance for them: -Material Transfer Agreements -Export Controls and Trade Sanctions -Permits

Autoclave Burn Injuries

Begin First Aid Immediately: Cool the burn with cool running water or cool wet wash cloth for 5-10 minutes. Remove rings, belts, and loosen tight clothing from burn area before the area swells. For minor burns, cover with gauze or a clean cloth. Seek medical attention for non-urgent events at Lansing Urgent Care. Take the required form on the EHS website ehs.msu.edu/incident-injury.html Report any workplace injuries to EHS. Major Burns - Call 911 -If the burn is deep, includes blisters, or covers large or sensitive areas, is swelling quickly, or is accompanied by other dangerous substance, call 911 immediately after starting first aid. -Until emergency help arrives, lift the wound above heart level, if possible. -Watch for signs of shock - cool, clammy skin, weak pulse and shallow breathing. -If necessary, get transport to professional medical care.

Managing Non-Sharps Biohazardous Waste

Biowaste includes solid materials with minimal liquid content, and do not include sharps. Examples: Disposable labware Contaminated PPE Grown Petri dishes Pipettes

Carcinogens

Carcinogens are chemical substances or a mixture of chemical substances which induce cancer or increase its incidence. Carcinogens are assigned to one of three hazard categories: 1A - Known Human Carcinogen 1B - Presumed Human Carcinogen 2- Suspected Carcinogen Carcinogens are a chronic hazard. That is, they do not cause harm upon initial exposure, but rather take repeated exposures over many years. Common lab carcinogens and their uses are: Chloroform - laboratory solvent Formaldehyde - tissue preservation Carbon tetrachloride - laboratory solvent

Emergency Eye and Body Washes

Chemical burns require urgent first aid. Emergency eyewash stations must be located near the hazard, have unobstructed access, and be maintained for use in case of emergency. Eyewash Stations / Drench Hose: Required in labs where injurious or corrosive chemicals are present. Know the location in your work area. Check once a week with a log for documentation. Six Inches of clearance is required around the area of the eyewash. Emergency Showers / Eyewash Stations: Located in hallway. Should be used for large areas and/or amounts of contaminants.

Alternate Labeling Methods for Multiple Small Containers Key Method

Chemical names can be abbreviated, BUT a key must be conspicuously located and updated frequently. The key must include complete chemical name. The hazard warning(s) can be included on the chemical container or on the abbreviation key. Document that employees are trained on the labeling system

Chemical Splash Goggles

Chemical splash goggles do not protect the individual's face from exposure to chemicals.

Physical Hazards

Chemicals that release energy in a violent fashion either by their natural chemical composition or by the manner they are handled can become a physical hazard. These materials can be safely used if the specific hazard(s) are understood.

Clean Bench (Laminar Flow Hood)

Clean benches protect the product only, Not the worker! A clean bench is not a BSC. The picture below demonstrates the airflow for the hood.

Hazard Communication

Communicate the location of biohazards by using proper labeling. The following items should have a biohazard label: Containers of regulated waste Refrigerators, freezers and other units used for storing potentially infectious materials Containers used to store or transport blood or potentially infectious materials Equipment that is contaminated with blood or potentially infectious materials Benchtop biohazardous waste containers need to have a biohazard label on the lid and on the side of the container.

Compressed Gases

Compressed gas containers must be handled with care and should be stored taking into account the property of the gases contained. Employees should know the hazards of the gases they work with. This information can be found in the MSDS of the material. Flammable gases (i.e. propane, hydrogen) must be stored in areas away from sources from ignition and kept separate from oxidizing gases (i.e. oxygen). Use an appropriate regulator, flame arrestor and excess flow valve when using flammable gases. Toxic gases (NFPA H=3 or 4) like chlorine, carbon monoxide, phosgene or ammonia can pose serious potential hazards to personnel and therefore special storage and handling measures are required. EHS must preapprove the purchase of toxic gases; a list of toxic gases requiring preapproval can be found on the EHS website: Toxic Gas Cylinder Preapproval List (XLSX). Toxic gases will also require a Standard Operating Procedure; a template can be found on the EHS website: Toxic Gas SOP (DOCX)

A biosafety cabinet should be used whenever:

Conducting aerosol generating procedures such as grinding, blending, pipetting, shaking, mixing, etc. Using high concentrations of BSL-2 materials Using large volumes of BSL-2 materials

Small size cylinders (fit inside hood)

Control procedures: Use and store in hood

Large size cylinders

Control procedures: Use and store in ventilated cabinets with air monitoring and alarm system

Autoclave these categories of biohazardous waste

Cultures and stocks of infectious agents Liquid human and animal waste Pathological waste Sharps (high containment laboratories only) Waste from animals exposed to infectious agents Waste generated in recombinant DNA research Transgenic plants and the soil Fabrics and clean-up materials Equipment

Disinfectant - Product Hazard Considerations

Disinfectant should be compatible with the surfaces to be treated. The product shouldn't be corrosive to the equipment. For example, bleach is corrosive and should be rinsed thoroughly after use on stainless steel equipment. Refer to Material Safety Data Sheet (MSDS) for exposure hazards.

Decontamination Purpose

Disinfection is a reduction of the concentration of microorganisms to a level that is no longer considered to be an infectious disease risk.

Limitations of Disposable Gloves

Disposable gloves do not offer adequate protection against solvents, corrosive solutions, or against high or low temperatures.

Non-Sharps Biohazardous Waste - Management Practices

Dispose of solid biohazardous waste in a labeled container with a labeled lid lined with a biohazardous waste bag. Waste containers should be closed with a lid when not in use. Securely tie the bags for transport to be autoclaved or to fiber barrels for disposal through a licensed medical waste hauler. When handling bags of biohazardous waste you should always wear disposable gloves to reduce your exposure risk. When transporting the bags within the building they should be placed in a secondary container on a cart to minimize any potential for leaking while en route.

Laminar flow hoods and biosafety cabinets (do/do not) provide protection from chemicals?

Do NOT

Autoclave Maintenance - Testing & Repair

EHS tests at least annually. If you suspect a problem or see damage: -Post a warning sign for others. -Contact your autoclave repair representative. -Contact EHS once the repair is made. EHS will remove the "do not use" warning sign once the autoclave has been recertified. EHS will replace the autoclave parameters sticker.

Chemical Hygiene Engineering Controls

Engineering Controls include devices such as fume hoods, glove boxes or other enclosures that reduce exposure or remove contaminants from the work environment. Fume hoods work by moving air from the lab, into the hood, and exhausting to the outside atmosphere. Use a chemical fume hood anytime your work involves: -Toxic volatile materials (chloroform, formaldehyde) with a PEL <50 mg/kg. -Carcinogens or particularly hazardous substances. -A procedure that may create an aerosol of a toxic substance. -Reactive or explosive materials or chemicals that may spatter. -Toxic gases (NH3, CO, F).

Engineering Controls

Engineering controls are equipment designed to isolate or eliminate the hazard.

Examples of common peroxide formers in MSU labs include:

Ether Dioxanes Isopropyl ether Tetrahydrofuran Never open a bottle that has solid formation around the lid. (Friction caused by unscrewing the cap can lead to explosion.)

Specific Labeling & Packaging Procedure

Ethidium Bromide Solutions Collect ethidium bromide solutions for disposal. Ethidium bromide is mutagenic at higher concentrations. Corrosive Liquids The following corrosive liquids shall not be mixed with any other hazardous waste under any circumstances. These liquids must be packaged in their own separate shipping container. Nitric acid exceeding 40 percent concentration Perchloric acid Hydrogen peroxide exceeding 52 percent strength by weight Nitrihydrochloric or Nitrohydrochloric acid diluted Acids, Bases, and Aqueous Solutions DO NOT mix strong inorganic acids or oxidizers with organic compounds. Keep acids, bases or aqueous solutions containing heavy metals separate from other wastes. Avoid mixing concentrated acids and bases together in the same container. Intermediate storage containers like those connected to laboratory instruments (e.g. High Performance Liquid Chromatograph or HPLC) are required to be: Closed at all times when being stored. Labeled with complete chemical names and hazard warnings. Labeled with the words Hazardous Waste. Emptied at the end of the shift / day into the main hazardous waste container in the laboratory.

Sharps Precautions

Evaluate and use safer needle and sharps devices when feasible Limit unnecessary handling of sharps Contaminated needles and other contaminated sharps must not be bent, recapped, or removed. Do not handle sharps with two hands or pass a sharp to someone else's hand. Always dispose of needles, syringes and scalpels in approved sharps containers regardless if they are contaminated or not. Contaminated sharps (e.g., broken glass, slides) must also be disposed of in approved sharps containers immediately after use. Place reusable sharps in puncture-resistant, closeable containers immediately after use for safe transport and processing. Never pick up contaminated sharps with your bare hands; always use some type of engineering control such as tongs, forceps or a broom and dustpan.

Materials NOT ACCEPTABLE for Sink/Sewer Disposal

Examples of materials forbidden from sink/sewer disposal include, but are not limited to: Metal-containing wastes Solvents (halogenated and non-halogenated) Oxidizers (perchloric acid, chromic acid, etc.) Biohazardous wastes (including animal wastes) Formaldehyde PCBs Phenols Reactive wastes (cyanides, sulfide, etc.) Corrosive wastes Radioactive wastes Liquid industrial by-products (antifreeze, pesticides, etc.) Ammonia Unused or outdated chemicals (including cleaning products) Petroleum oil or grease Sludges or viscous materials Solids or insoluble substances Known and probable carcinogens Wastes or unwanted materials must NOT be intentionally diluted to meet sink/sewer disposal requirements.

Autoclave Loading - Liquids

Explosion & Boil-Over Risk! -Use only borosilicate glass (PyrexTM or KimaxTM). -Fill your containers no more than 2/3 full to help prevent boil-overs. Use approved secondary tray with a solid bottom and walls to contain the contents. Loosely cover with caps or aluminum foil on glass beakers to allow the liquids to allow safe expansion of liquid. Never autoclave liquids in a closed container. Add water to secondary container to maintain uniform heat distribution.

Do NOT Autoclave:

Explosives Flammables Chlorine-Based Products Reactive, Corrosive, Solvents, or Toxic Materials... Radioactive Material Unauthorized Containers

Eye and Face Protection

Eye and face protection equipment must be made available to all employees and visitors where chemicals are used and stored. Appropriate eye and face protection equipment must be worn at all times in those labs (including teaching labs) where eye hazards exist. Eye and face protection equipment must be ANSI Z87 approved.

Emergency eyewash stations (select all that apply): Are required in labs where hazardous chemicals are being handled. Are required to be tested once a week. Need to be clear from obstruction (at least 6" around eyewash).

Eyewash stations are required to be tested once a week, need to be clear from obstruction (at least 6" around the eyewash), and are required in labs where hazardous materials are being handled.

All non-RCRA waste can be disposed of in the regular trash. True False

False Only non-RCRA, non-hazardous waste like uncontaminated filter paper or pipette tips can be disposed of in the regular trash.

The main physical hazards at MSU are:

Flammable liquids Oxidizers Explosives Peroxidizable chemicals Compressed gases Reactive materials Pyrophoric

Flammable Liquids

Flammable liquids are some of the most commonly used hazardous chemicals on campus. Flammable liquid means a liquid having a flash point of not more than 93°C. Substances and mixtures of this hazard class are assigned to one of four hazard categories on the basis of the flash point and boiling point. Flammable liquids include alcohols, ketones, xylenes and carboxilic acids. Most organic chemicals are also flammable or combustible.

Protective Glove Recommendations

For limited chemical contact or when laboratory tasks require fine motor skills use disposable gloves. Vinyl and Nitrile (N-dex) - Appropriate for use with dry chemicals, non-corrosive aqueous solutions. For prolonged chemical contact or immersion use reusable gloves recommended below: Neoprene Recommended for corrosives (pH <2.0or >12.5), solvents and alcohols. Nitrile (Blue or green) Recommended for non-halogenated solvents such as hexane, methanol and puncture or abrasion hazards. Nomex or Zetex Recommended for temperature extremes such as handling cryogenic fluids like liquid nitrogen. Viton Recommended for chlorinated and aromatic solvents such as carbon tetrachloride and benzene. Butyl Recommended for aldehydes, ketones and esters such as acetaldehyde, formaldehyde and propanone.

Packaging requirements

For liquid waste: use EHS-supplied 5-gallon container or empty plastic or glass bottles with closable screw lids. For liquids, fill containers to about 90% of container volume. DO NOT fill containers to the top. Leave at least 2 inches of space in 5-gallon liquid waste containers to allow for liquid expansion. Make sure the caps on all cans and bottles have gaskets and are tightly secured before the pick up. For solid waste (contaminated solid debris, pipette tips, gloves and paper): use a covered box or bucket lined with a bag to retain liquids. Ex: ethidium bromide or acrylamide gel waste

Gases Under Pressure: Use, storage and transport

Gas cylinders with regulators must be individually secured; only cylinders with safety caps can be group chained. When moving a cylinder don't forget to: -Use appropriate cart -Secure cylinder in an upright position -Make sure safety cap is in place

Gases Under Pressure

Gases under pressure are gases that are contained in a receptacle at a pressure not less than 280 Pa at 20°C, or as a refrigerated liquid. This includes gases or gaseous mixtures that can experience a sudden release of pressure or freezing compounds which may lead to serious damage to people, property, or the environment independent of other hazards the gases may pose. Compressed gas cylinders may not be purchased with P-Cards. Cylinders purchased with P-Cards may be subject to immediate return to the supplier, or disposal.

Submitting a Pick-Up Request

Gather the waste containers destined for disposal. Log in to EHS Safety Portal and complete "Hazardous Waste Request."Enter the names of the chemicals from the Hazardous Materials Table, the container size, and the number of containers.Indicate if any replacement containers are needed. Use the comments section of the Pick-Up Request Form to denote any scheduling conflicts e.g., "Lab open in afternoons only" or "will not be here Friday," etc. An EHS technician will visit your lab within 10 working days to remove the material.

Mutagens and Teratogens

Germ Cell Mutagenicity and Reproductive Toxicity

Work Practices Personal Protective Equipment (PPE)

Gloves are required Lab coats are recommended Eye protection is required if there is a potential for splash of hazardous materials While in the same room where research is conducted, restrict activities that involve contact with your face; such as: eating, drinking, smoking, and applying cosmetics. Mouth pipetting is prohibited. Persons should wash their hands regularly. In normal laboratory hygiene, hands should be washed: After removing gloves After cleaning up biological spills When visible contamination is present After work is completed and before leaving the laboratory Before activities that entail hand contact with mucous membranes or breaks in the skin (e.g., eating, applying cosmetics, etc.) applying cosmetics, etc.)

Hazard Control Methods

Hazard control methods must be implemented by the lab supervisor to reduce employee exposure to hazardous chemicals in the laboratory. This section covers the three types of hazard control methods to reduce employee exposure to workplace hazards: Administrative controls Engineering controls Personal protective equipment

Hazardous Waste

Hazardous materials are substances that have characteristics such as flammable, corrosive, reactive, toxic, ignitable, carcinogen, poisonous, radioactive or infectious that make them a potential risk to human health and the environment. RCRA hazardous waste is defined as waste regulated under the Resource Conservation and Recovery Act (RCRA). The EPA developed a regulatory definition of hazardous waste by using two different mechanisms: by listing certain specific wastes as hazardous and by identifying characteristics that would make the waste hazardous. RCRA Hazardous Waste is considered to be a liquid, solid or gas that is a: Listed RCRA Hazardous Waste by EPA: -Discarded commercial product (e.g. an expired bottle of ether) -Process waste (e.g. waste generated by HPLC) or waste mixtures Characteristic Hazardous Waste: -Corrosive (pH < 2 or pH > 12.5) -Ignitable (Flash Point < 140° F) -Toxic -Reactive --Reacts violently with water or forms potentially explosive mixtures with water --Is a cyanide or sulfide bearing waste that when exposed to conditions between pH 2 and 12.5 gives off toxic vapors --Is explosive

What are the correct requirements for hazardous waste containers? Select all that apply. The words "Hazardous Materials" must be written on the container for pick up. Hazardous waste containers must have closed lids while being stored The full chemical name of all chemicals in the hazardous waste container must be written on the pick up tag

Hazardous waste tags must have the full chemical name of the materials being disposed in the hazardous waste container. Hazardous waste containers must also have closed lids while being stored. The words "Hazardous Waste," not "Hazardous Material," must be written on the container or a tag attached to the container. For example; it CANNOT say "Waste Acetone" or "Hazardous Acid Waste."

Biosafety cabinets use __________ filters to capture the particles within the cabinet. The filters can remove particles 0.3 µm in diameter with an efficiency of 99.97%. __________ filters are made of borosilicate fibers and are pleated to increase the overall surface area. This material is very sensitive and should not be handled. The filters require installation by a professional.

High Efficiency Particulate Air (HEPA)

Examples of biohazards include:

Human, animal and plant pathogens; Blood, tissues, and body fluids, or other materials that may potentially contain infectious agents; Animal waste and carcasses; Toxins of biological origin such as Tetrodotoxin, T-2 toxin, and Conotoxin; Recombinant DNA; Any unknown substance should be treated as potentially infectious.

BBP Exposure Incident Response

If an exposure incident occurs: -Flush/wash the exposed area for 15 MINUTES. -Contact your supervisor if he or she is available. -Report to emergency care facility for medical evaluation/follow-up immediately!

Responding to Improperly Disposed Biohazardous Waste

If biohazardous waste such as sharps containers, regulated sharps items, a biohazard bag, or carcasses/animal parts are discovered in or around a dumpster, notify your supervisor who will contact EHS for disposal and follow-up. Do NOT put biohazard bags in the dumpster without proper treatment and containment in a black, non-transparent bag!

When conducting research, some items to consider include but are not limited to:

If infectious material is splashed in the eye, or if your fingers are contaminated and you rub your eyes or adjust contact lenses, there is a potential risk for exposure. Eating, drinking, smoking, applying cosmetics, and mouth pipetting are potential risk of contamination by accidental ingestion. Transmission to the lung can occur by inhaling airborne contaminants trapped within aerosols. Transmission through the skin can occur through accidental inoculation with a contaminated sharp object (needle, broken glass) or entry through broken skin (paper cut, razor burn, hangnail)

Managing Spills during Transport

If the spill occurs outside of a building (i.e. sidewalk), or is too large for personnel to clean up, isolate the area and contact EHS (355-0153). Stay at the spill site until EHS arrives. If the spill occurs in a vehicle and the material is known or suspected to contain pathogens, leave the vehicle with closed windows and locked doors. Contact EHS (355-0153).

Requirements for Hazardous Waste Management

If you are involved in generating hazardous waste or packaging it for pick up you are required by law to: Complete the Initial Chemical Hygiene/Hazardous Waste Training through EHS, and complete the Hazardous Waste Refresher on-line course each year. Follow the General Labeling & Packaging Procedures: -Keep container closed at all times, unless adding to the container. -Label "Hazardous Waste" on container. -Fill out an MSU Materials Pick Up Tag, including appropriate GHS symbols at the bottom, and attach it to each container. -Submit a pickup request before the waste is 90 days old.

Impact Goggles

Impact goggles do not protect an individual's eye from exposures to splashes from the sides, top, and bottom. Impact goggles have vents on the sides for ventilation, these vents prevent the entrance of large solid particles, however does not prevent the entrance of liquids, dusts, and mists.

Safe Work Practices for Carcinogens

In general, keep exposure as low as possible by: Using engineering controls, such as fume hoods or glove boxes Using PPE such as eye protection, lab coat and gloves Developing an SOP Keeping quantities to a minimum Labeling bottle and storage area with the words carcinogen or cancer hazard

Hazard Identification

In order to incorporate safety aspects of handling and storage into your experiment you must understand and recognize the hazards associated with the materials, equipment used and the procedures that are followed. The following section will describe physical hazards as well as special precautions and examples.

Examples of items that are not sharps

Intact blood tubes Glass vials Wrappers, needle covers, paper towel, band aids, gloves, and other paper waste Pipette tips Broken or discarded glass that is not biologically or chemically contaminated should be discarded in a broken glass container.

Irritants

Irritants will cause reversible, but still painful, inflammation when in contact with the skin, eyes, nose or respiratory tract. Skin irritants cause reversible damage to the skin following the application of a test substance for up to 4 hours. Eye irritants cause changes in the eye following the application of a test substance to the front surface of the eye, which are fully reversible within 21 days of application. Examples: Chlorine and ammonia containing products Diluted acids Halogens Nitrogen dioxide Alkaline dusts and mists Ozone Phosphorus chlorides Hydrogen chloride Phosgene Arsenic trichloride Hydrogen fluoride Diethyl/dimethyl sulfate

Alternate Labeling Methods for Multiple Small Containers Box or Tray Method

Label box or tray with chemical name and hazard warning. If containers are removed from the box or tray then they must be properly labeled. Hazard labels on storage areas exempt labeling of individual containers.

Chemical Labeling

Label chemicals in the lab with: Full Chemical Name (no abbreviations, acronyms, or formulas unless a key is clearly posted in the lab) Example: Ethanol, hydrochloric acid Hazard Warning (can either be sticker, written, or NFPA diamond) Example: Flammable, corrosive, cancer hazard

Research Hand Hygiene

Lather up your hands with soap. Be sure to rub your hands together for at least 15-20 seconds. Don't forget your wrists and forearms, between your fingers and around your nail beds. Rinse your hands and dry. Use the paper towel to turn off the faucet.

Managing Liquid Biohazardous Waste

Liquid biohazardous waste must also be stored in a leak-proof container labeled as a biohazard. Appropriate PPE should be worn when handling and disposing of liquid biohazardous waste. This includes: Splash goggles, gloves, and a lab coat.

Before Entering the Lab

Make sure that you minimize exposed skin at all times by wearing the appropriate clothing (long pants/skirts, close toed shoes). The appropriate personal protective equipment must be used at all times in the lab when chemical, radioactive, or biohazardous materials are being handled. It is advised to wear PPE even if not working with hazardous material, because you can be injured by someone else's accident!

Classes of Fire and Fire Extinguishers

Many types of fire extinguishers could emerge in laboratories. Employees must choose the right extinguisher, given the type of fire, to effectively control a fire. The four basic classes of fires are: A B C D

BSL-4

Maximum Containment (Ebola virus, Marburg virus) Suitable for work with agents that are: -Dangerous and exotic -Cause life-threatening disease -No available vaccine or therapy -Transmitted via aerosols

Sink/Sewer Disposal of Chemicals

Michigan State University is required to comply with sewer disposal restrictions established by local wastewater treatment plants; Michigan Department of Environment, Great Lakes, and Energy (EGLE); and the Environmental Protection Agency (EPA). The regulatory governance of certain categories of chemical/hazardous waste is extremely complicated. In general, most chemicals, commercial products, and solutions (including rinseate when cleaning an empty chemical bottle) from a laboratory are forbidden from sink/sewer disposal and should be properly disposed by EHS.

Splash goggles

Must be worn if splashes or splatters of infectious materials is anticipated Must be worn during spill clean up

Laboratory coats

Must be worn while conducting procedures with potentially infectious materials to prevent the spread of contamination outside of the laboratory. Must not be worn outside of the laboratory areas (e.g., into break rooms, administrative areas, etc.). Must be cleaned or disposed of routinely and when visibly contaminated. Note: Lab coats cannot be taken home to be cleaned. They must be cleaned through an approved facility.

Autoclave Spill Response

NEXT If a spill occurs in an autoclave or on the floor in the autoclave room, use the biological spill kit located in your lab to clean up the materials. If the spill is too big for the spill kit in your lab, call the EHS immediately.

Which of the following materials should not be stored together: Acetone and methanol Nitric acid and acetic acid Sulfuric acid and hydrochloric acid Sulfuric acid and phosphoric acid

Nitric acid and acetic acid Acetic acid is an organic compound that is corrosive and combustible. Acetic acid should be stored away from oxidizers such as nitric acid. Acetic acid is better stored in a flammable proof cabinet with other organics. Nitric acid should be stored with other corrosives in a corrosive storage area.

Examples of common oxidizers are:

Nitrous oxide Hydrogen peroxide Nitric acid Perchloric acid Nitrates Use oxidizers with extreme care and consult the (M)SDS for specific guidelines.

Do not work alone in laboratories.

No one should work alone in laboratories when using equipment, processes and/or chemicals with the potential for explosions, fire, electrocution or serious physical harm. This can include: toxic and highly reactive chemicals poison gases high energy lasers power tools dangerous equipment After hours work or work performed alone must meet approval of the Principal Investigator before beginning work. Have a written SOP completed for all high hazard activities.

Personal Electronic Device Use in the BSL-2 Laboratory

Personal Electronic Devices (PED) can include but are not limited to head phones, ear buds, cell phones, smart phones, MP3 players, personal computers, tablet computers, electronic book readers. Items may become contaminated, increasing the potential to transfer the contamination outside of the laboratory, potentially causing illness. These devices may also cause a significant distraction when doing critical work. Ear buds and headphones are prohibited: MP3 players must be placed in a sealed plastic bag (i.e. Ziploc) and disinfected prior to leaving the lab. Utilize a benchtop Bluetooth speaker that remains in the facility and is disinfected regularly instead of any listening devices that are worn on the person. Smart Phones and Tablet Computers: The device is kept in an area far from experiments or hazardous materials, such as a laboratory desk designated for documentation. must be inside of a sealed bag (i.e. Ziploc) and disinfected before leaving the laboratory. Laptops and Computers: Personal computers are discouraged from being brought into the laboratory from home unless: the device will remain within the facility as a laboratory only computer and is disinfected regularly the device is kept in an area far from experiments or hazardous materials, such as a laboratory desk designated for documentation

Personal Protective Equipment (PPE)

Personal protective equipment includes any devices or clothing worn by the worker to protect against the hazards in the environment. Examples are: lab coats aprons protective gloves safety glasses goggles face shields

Autoclave

Piece of equipment used to sterilize articles by way of steam under pressure and/or dry heat -The addition of water in a pressured space greatly speeds the process -The higher the concentration of organisms, the longer it will take to kill all of those cells -At lower temperature, it takes more time to sterilize than at a high temperature -liquids require longer cycle time, but lower temperatures to minimize evaporation

Sharps Containers

Place disposable sharps into a manufactured sharps container. Sharps containers used on campus must also be labeled with the EHS "SHARPS" label to facilitate proper treatment and disposal of containers or if no label available use a permanent marker to identify the date of first use, initials of user, room number, building and date of expiration. A sharps container must be permanently closed and disposed of through the EHS within 90 days of the date that it was first put into service (when you added the first sharp item), or when it is ¾ full- whichever comes first. Keep in mind that the following items are NOT approved sharps containers: -Coffee cans -Reagent bottles -Sharps Keepers boxes

Loading - Solid Waste

Place solid items to be processed in autoclave bags and an appropriate rigid autoclavable secondary container. Examples of acceptable secondary containers are: polypropylene, polycarbonate, and stainless steel. -Loose opening to allow steam release

Autoclave Treatment

Please observe the following when treating waste: -Wear the appropriate PPE. Always wear heat-resistant autoclave gloves when using an autoclave. You should wear a laboratory coat to further protect yourself. -Only autoclaves that have been tested by EHS within the last year can be used for decontaminating biohazardous waste. These autoclaves have a sign and sticker on or near the autoclave indicating the minimum parameters for decontamination and the last test date. -To autoclave the waste, place the bag in a secondary container that is capable of withstanding the autoclave conditions. --Start the cycle for the indicated amount of time posted by EHS. --When the cycle is finished, do not open the autoclave door until the temperature has cooled down, and the pressure is 0 PSI. --Allow the waste to cool, and then place it in a non-transparent black trash bag for disposal as soon as possible. Do not let waste pile-up around the autoclave. Some departments have autoclaves available for biohazardous waste decontamination. These devices may be used for treatment of solid, non-sharps biowaste except for tissues and body parts. Do not let waste pile-up around the autoclave.

Autoclave Unloading - Liquids

Practices for Autoclaving Liquids: Let liquids stand for a full hour before touching with ungloved hands. Be sure others in the area know a heat hazard is present.

Biowaste Spill Response

Prepare for cleanup: -isolate the area -wait a few minutes for aerosols to settle -retroeve a spill kit and review the procedure Biohazard Spill (NO sharps): -cover the spill with paper towels -spray the spill with freshly prepared bleach solution: working from the outside in -wait for the appropriate contact time and then wipe up the spill -disinfect the area again Biohazard Spill (WITH sharps) -Using a mechanical device remove the sharps -cover the spill with Superfine -Sweep the absorbent up and discard in a biowaste bag -spray the spill with freshly prepared bleach solution: working from the outside in -wait for the appropriate contact time and then wipe up the spill -disinfect the area again Spill Response Reminders: -Use proper PPE -Use mechanical tools for handling sharps -Dispose of all spill waste as biohazardous waste -Notify the safety Manager after cleanup

Procedures for Preparing and Transporting Biological Materials

The process used to prepare and transport biological materials is referred to as the "PCAL" system. Package Contain Absorb Label PACKAGE: Leak-proof primary containers CONTAIN: Leak-proof secondary container with secure lid ABSORB: Add enough material to absorb all contents LABEL: Biohazard symbol and Emergency contact information

Glove Selection and Use

Protective gloves are an important aspect of protection against hazardous materials. The use of protective gloves can reduce the risk of a hand injury by 40%. It is critical that users select the correct glove material based on the chemicals used and the permeation data. Consult the glove manufacturer's glove guide for recommendations on how to select the best glove for your application. Ensure protective gloves fit properly and comfortably. To determine the right size for you, measure the circumference of the hand and compare it to the manufacturer's size chart. The natural rubber proteins found in latex gloves may produce an allergic reaction with sensitive individuals. Some people may develop an allergy from latex after repeated use of latex products. The US Food and Drug Administration has issued a ban on all powdered gloves. Exposure to starch powder from gloves can cause undesirable reactions, which vary from well-known allergy symptoms and upper respiratory-tract disorders to surgical adhesions and infections. The presence of glove powder can also result in many other undesirable effects, such as interference in laboratory testing causing false results (i.e. PCR - Polymerase Chain Reaction, enzyme immunoassay or some HIV tests). To reduce the risk of latex allergy: -Substitute non-latex gloves when ever possible or use powder free, reduced protein content.

Chemical Hygiene Glove Use

Protective gloves must be worn when there is a potential for a hand injury or skin contact with chemicals, extreme temperatures or abrasives. All protective gloves have limitations. Chemicals will eventually permeate gloves, however they can be safely used for specific time periods when the conditions and use of the chemicals are known.

Two immediate steps to take if a fire starts in the laboratory are:

Pull fire alarm and call 911 from safe location

Emergency Situations: Fire

Pull fire alarm. Call 911 from a safe location. Notify the unit emergency coordinator. Evacuate.

Microorganisms require _________ in which they can acquire nutrients, reproduce, and spread. Depending on the pathogen, suitable reservoirs may include humans, animals, water, soil, plants, dust, liquids or food. Within the lab, microorganisms may reside within the materials or your research; however, the lab personnel and the surrounding environment may also be an inviting reservoir to with the organisms can spread.

RESERVOIRS

Spill Kits

Ready access to a chemical spill kit is required in laboratories that work with hazardous chemicals. Minimally such a kit should contain: Splash resistant goggles Chemical resistant gloves Plastic bags Multi-chemical sorbent (enough for 2 gallon spill) Scoop Only use a spill kit in a non-emergency situation (spills less than 1 liter of low toxicity and/or flammability hazard material like methanol or diluted acids).

Jacket pressure gauge

Remains static

Reproductive Toxicity

Reproductive toxicity includes adverse effects on sexual function and fertility in adult males and females, as well as developmental toxicity in offspring. Substances and mixtures with reproductive and/or developmental effects, called teratogens, are assigned to one of two hazard categories, "known or presumed" and "suspected." Examples: Ethyl alcohol Mercury compounds Other common teratogens

Respirators and Biosafety

Respiratory protection is the last line of defense against airborne infectious agents or particulate allergens. Respirators are required for working with Mycobacterium species (TB) and other similar work.

Sensitizers

Respiratory sensitizer means a substance that induces hypersensitivity of the airways following inhalation of the substance. Skin sensitizer means a substance that will induce an allergic response following skin contact. The definition for "skin sensitizer" is equivalent to "contact sensitizer." These groups of chemicals that will cause, through an immune response, abnormally severe allergic responses upon repeated exposure to the chemical. A common lab example is formaldehyde. Exposure to formaldehyde/formalin can cause a sore throat and other respiratory problems if inhaled, and eczema-like symptoms upon repeated skin contact. Note that a person who has allergic reactions to a chemical upon the first exposure is considered a sensitive individual. Examples of sensitizers: Formaldehyde/Formalin Latex

Limitations of Reusable Gloves

Reusable gloves may limit fine motor skills for specific tasks.

Use and Care of Protective Gloves

Reusable: Inspect gloves before wearing and replace whenever they become discolored or show signs of damage. Before gloves are removed, thoroughly rinse them off with water and/or a mild detergent and allow drying. Disposable: If a disposable glove becomes contaminated, remove immediately and replace with a new glove. Never re-use disposable gloves.

______________ _____________ are those at risk of acquiring an infection. Certain situations or conditions may put a person at a higher risk for disease. People who are included in the high-risk group include, but are not limited to: immunocompromised, the elderly, children, neonates, chronically ill patients and pregnant women. Vaccination status is also important in terms of susceptibility. It is important to take greater precaution when working with biohazardous materials when high-risk individuals are present.

SUSCEPTIBLE HOSTS

Sarah is pipeting acetone solution. What is the minimum eye protection for this situation?

Safety glasses Safety glasses are appropriate when working with a small amount of chemicals and there is a low probability of splash.

BSL-1

Suitable for work involving well characterized agents not known to cause disease in healthy human adults. These agents present a minimal potential hazard to lab personnel and the environment. Low/No risk agents (Baker's and Brewer's yeast) Examples: Non-pathogenic bacterial species Yeast Bacillus subtilus Access to the laboratory is limited or restricted at the discretion of the Principal Investigator when experiments are in progress. The laboratory is designed so that it can be easily cleaned. Bench tops are impervious to water and resistant to acids, alkalis, organic solvents, and moderate heat. Laboratory furniture is sturdy. Spaces between benches, cabinets, and equipment are accessible for cleaning. Chairs used in laboratory work must be covered with a non-porous material that can be easily cleaned and decontaminated with disinfectant. Each laboratory contains a sink for hand washing. An insect and rodent control program is in effect. A program shall be implemented to control undesired species (e.g., weed, rodent, or arthropod pests and pathogens), by methods appropriate to the organisms and in accordance with applicable state and Federal laws. If you have pests in your lab, submit a service request with IPF. Examples of some pests include: wild rodents, cockroaches, flies, and ants. If the laboratory has windows that open, they are fitted with fly screens.

Examples of autoclave use

Surgical Instruments: hemostats, forceps, scalpels, and scissors, et cetera. Equipment: animal cages, beakers, flasks, media or broth prep, petri dishes, test tubes, certain lab equipment. Greenhouse and Plant Applications: transgenic soil, plants, plant pathogens. Biohazardous Waste: includes several categories of materials to be autoclaved.

__________ __________ are how the pathogens are transmitted through the environment from the reservoir to the susceptible host. Pathogens enter a host through entry portals such as mucous membranes (eyes, nose, and mouth), the GI tract, open wounds, non-intact skin, respiratory tract, surgical incisions or any type of vascular access from invasive procedures.

TRANSMISSION MODES

The Chemical Hygiene Plan

The Chemical Hygiene Plan is a chemical safety manual for laboratory workers that contains: Hazards of chemicals Appropriate work practices and procedures Controls to protect all workers Basic standard operating procedures (SOPs) (lab-specific SOPs must be created)

EPA Fines

The EPA and Michigan Department of Environment, Great Lakes, and Energy (EGLE) have the power to fine the University for non-compliance with waste regulations. On average, the fines amount to about $1000 per container violation; however, the EPA can fine up to $70,117 per container, per day that the violation is not corrected. This cost would be paid by the Department or PI. Willful neglect on the part of hazardous waste generators can result in jail time or fines.

Non-RCRA Regulated Waste

The State of Michigan passed PA 451, Part 121 which oversees the proper disposal of liquid industrial by-products. If a generator determines that their waste is not listed and does not exhibit a characteristic as identified by RCRA, it may still be regulated by Michigan Department of Environment, Great Lakes, and Energy (EGLE) Part 121 regulations. The most common non-RCRA wastes on MSU campus are ethidium bromide gels, acrylamide gels, formalin, and laboratory debris contaminated with carcinogens. The generator will still need to collect those wastes and submit a pickup request to EHS for waste disposal.

EPA and Disinfectants

The U.S. Environmental Protection Agency (EPA) defines disinfect/cold sterilants as pesticides. All requirements on the pesticide product label must be followed when using the product. Not following the requirements given by the manufacturer are against the law. When selecting a disinfectant, keep in mind the following: -It should be EPA-registered disinfectant and appropriate against the material being disinfected. (As defined on the EPA's website.) -Follow the manufacturer's instructions for dilution and contact time. -It may have hazardous properties that require special handling - refer to the MSDS.

Dose-Response

The dose-response relationship is important in understanding the cause and effect relationship between chemical exposure and health effects. All chemicals are toxic at a high enough dose...our goal is to reduce workplace exposures to the level where no adverse effects are observed, into the safe region of the dose-response relationship.

Sharps

The following items are considered sharps and must be placed in sharps containers and disposed of through the EHS: -Needles -Syringes with or without needles -Scalpels -Anything that is sharp enough to puncture the skin AND is contaminated with biological materials Broken or discarded glass that is not biologically or chemically contaminated should be discarded in a broken glass container.