Biohazards are biological agents or substances present in the work environment that may present a hazard to the health and well-being of the worker and the campus community. Biological agents include infectious and parasitic agents, noninfectious microorganisms (fungi, yeast, algae), plants and animals that cause occupational disease. The University's Biosafety program is established to protect individuals from exposure to biohazards through the application of administrative and engineering controls.
Information for Principal Investigators
Principal investigators (PIs) must apply for research authorizations to work with regulated materials or human or animal subjects. The following is a list of those authorizations before beginning research at Cal State Fullerton.
|What to do||How to do it||Contact for questions|
Apply for an animal use protocol.
Fill out a Request for Approval of Animal Care and Use
See the Animal Care and Use Occupational Health Program. For more information, please call the Office of Grants and Contracts at 657-278-2106.
Apply for a Biohazard Use Authorization
|Fill out a Biohazard Use Authorization (under development)||
See the University's Biosafety Program. For more information, contact the EHS Office at 657-278-7233.
Apply for a Controlled Substance Use Authorization
|Fill out a Controlled Substance Use Authorization||See the University Controlled Substance Program. Call EHS at 657-278-7233 for more information.|
Apply for an Institutional Review Board application.
|See the IRB website.||For more information, contact the Office of Loans and Grants at 657-278-2106.|
Apply for a Laser Use Authorization.
|Fill out a Laser Use Authorization.||See the University Laser Safety Program or call EHS at 657-278-7233.|
Apply for use of nanoparticles in research.
Apply for a Radioactive Use Authorization
|See How to Fill out a Radioactive Use Authorization.||See the Radiation Safety Program. Contact the Radiation Safety Officer in EHS at 657-278-7233 or email email@example.com|
Apply for a Biohazard Use Authorization
|Fill out a Biohazard Use Authorization (under development)||See the University Select Agents Program or call EHS at 657-278-7233 for more information.|
Apply for use of stem cells in research.
|Fill out a Biohazard Use Authorization (under development)||For more information, contact EHS at 657-278-7233.|
Animal Care and Use
The purpose of this program is to establish a uniform set of guidelines for the handling and use of laboratory and wild animals at California State University, Fullerton. It will assist in safeguarding the overall health and safety of the employees that may come in contact with laboratory and wild animals.
Animal Care and Use Occupational Health Program
The purpose of the Animal Care and Use Occupational Health Program is to establish a uniform set of guidelines for the handling and use of laboratory and wild animals at California State University, Fullerton.
Animals on Campus
There are significant health and safety hazards and nuisances created by animals (primarily dogs) on campus. Accordingly, the following campus policy relating to animals is in effect:
Animals, even though leashed, shall not be brought onto California State University, Fullerton property, including all buildings, except for:Seeing-eye dogs, which must have a valid dog license as evidence of current rabies vaccinations.Animals involved in authorized university research.Animals securely confined in a properly ventilated vehicle for a period not to exceed 30 minutes duration.
Animals found on campus in violation of this policy are subject to being picked up and turned over to the Orange County Animal shelter. Call the University Police (ext. 2515) to report infractions.Exceptions to these regulations must be approved in advance by the University Police (ext. 2515) and Environmental Health and Safety (ext. 7233).
Handling Animal Reservoirs of Hantavirus Guidelines
These guidelines are based on practices used by Center for Disease Control personnel in areas known to have fatal human cases of Hantavirus infection.
Prevention of Salmonella Exposure from Reptiles
Salmonellosis is a zoonotic form of gastroenteritis that is most commonly contracted through oral ingestion (e.g., contaminated chicken products). However, a high proportion of reptiles are asymptomatic carriers of Salmonella. Attempts to eliminate Salmonella with antibiotics are usually unsuccessful and produce strains that are resistant to the antibiotic. Transmission to humans can occur through open cuts, splashing of contaminated material into the eyes or through inhalation of sprayed contaminated solutions. Symptoms include muscle aches, headache, nausea, vomiting and diarrhea, and usually last for 2-4 days. Antibiotics are not normally used to limit the duration or severity of the disease in humans. In some people, Salmonellosis can be quite severe, leading to septicemia, death or spontaneous abortion.
The following categories of people should avoid all contact, direct or indirect, with any reptile as the risk of serious, symptomatic infection is greatly increased:
- Women who are pregnant (due to the threat to the fetus)
- Anyone with HIV/AIDS or other immunodeficiency disorders
- Anyone who has had transplant surgery or is on anti-rejection therapy
- Anyone who is on any drug which suppresses or alters immune function including: steroids, cancer chemotherapy, biological response modifiers and others
- Anyone receiving radiation treatment
- Infants and children up to five years of age
- Those who are elderly, frail, or with poor nutritional status
What to do to avoid becoming infected:
- Visitors should not touch the reptile cages and should avoid touching any surfaces that might be contaminated.
- Wash your hands with soap and hot water for at least 30 seconds after handling reptiles or participating in a class in the MH-207-217 area. Antibacterial soap is preferable.
- Do not handle any reptiles or their caging if you have open cuts or sores on your hands (rubber gloves are recommended).
- Avoid splashing when washing reptile enclosures (consider goggles or face mask when washing).
- Do not share caging or temporary housing containers, wash tubs, etc., among different species or individuals unless they have been disinfected.
If in doubt about any condition or treatment you or a household member is undergoing, or about any disease or disorder you or a household member may have with respect to its effect on immune status, please consult your physician. Also, if you have any of the above-mentioned symptoms, you should contact your physician.
If you have any questions, please contact the Animal Care Supervisor (657-278-5388) or the Biology Department at (657-278-3614).
CSUF thanks California State Polytechnic University, Pomona for this information.
Related Links+Expand All
Anthrax Information - Centers for Disease Control
Anthrax is an acute disease caused by the bacterium Bacillus anthracis. Most forms of the disease are lethal, and it affects both humans and other animals. There are effective vaccines against anthrax, and some forms of the disease respond well to antibiotic treatment. For more information, please review the Centers for Disease Control Emergency Preparedness and Response to Anthrax.
Autoclaves use high temperatures and pressure to inactivate biologically active material to ensure it is non-viable prior to waste disposal. See below for directions on how to safely operate the autoclave. If you still have questions or need help with autoclave operations contact the College of Natural Sciences and Mathematics x2638 for the DBH autoclaves or Biology x3614 for the MH autoclaves. You can call EHS x7233 or email firstname.lastname@example.org for safety related issues. Contact Physical Plant x3494 for maintenance.
- Ensure no items have been left by previous users, especially sharps or other items that could pose a hazard. Remove these items or contact NSM or the Biology technical staff to have items removed.
- Clean the strainer before loading the autoclave.
- Ensure the material to be autoclaved does not contain any hazardous or radioactive material. Contact EHS at x7233 for instructions on how to dispose of hazardous or radioactive material.
- Loosen bottle caps before loading to prevent them from shattering; DO NOT over-fill liquids; ensure containers are free of cracks.
- Ensure any plastic materials are compatible with the autoclave, so that they maintain their integrity with autoclave temperatures and pressure.
- Place solids into a labeled "autoclave bag"; ensure that solids have at least 100 ml of liquid present with the load to effectively sterilize them.
- Place a metal drip pan underneath items to catch spills.
- Place autoclave indicator tape on every item autoclaved and check the tape to see if a color change has occurred after the run is complete. The indicator tape confirms that the run achieved the proper temperature and pressure to inactivate biological material.
- Close the autoclave door before using.
- Autoclave when there are fewer people around to minimize odor complaints.
- Set the proper time for the material that is being autoclaved. Most liquids and dry goods can be autoclaved in 15 minutes. Check the protocol.
- Autoclave waste liquids or solids for 45 minutes to properly inactivate biological material.
- Use thermal protective gloves, apron, and face shield to protect against heat.
- Handle carefully: liquids can "bump" or suddenly erupt and spill when the container is moved.
- Use the drip pan when removing the bags to avoid spills. Clean up all spilled liquid inside or outside the autoclave. If over a liter, obtain a spill kit from one of the labs. Spill kits contain absorbent material that will assist in the cleanup.
- Clean the filters/traps using tap water. Traps are located on the floor of the autoclave.
Dispose of autoclaved waste properly
- Liquids can be disposed down the drain as long as no hazardous materials are present.
- Do not drain dispose agar or any material that can solidify and clog the drain. Solidify agar in a beaker and put in the regular trash.
- Place the autoclave bags into regular opaque plastic bags and take directly to the outside dumpsters.
Specific instructions to turn on the autoclaves in DBH
- Press the Controls 'ON' switch. The green indicator in the upper left corner of the switch will light.
- Open the door. The Unsealed Door Indicator will remain lit until the door is closed and sealed. If the Unsealed Door Indicator is not lit, the door will not open, press Unseal Door Switch.
- Load the autoclave, and then close the door.Select Processing Cycle. Wrapped (looks like a burrito), Unwrapped (scissors) or Liquids (flask).
- Select Exposure Temperature (in C). Usually this is 121 ⁰ C. Note: Check the temperature. Never autoclave things over 130⁰ C.
- Select Exposure Time Hours are displayed at the left of the color; minutes at the right. For example, 02:30 means two hours and thirty minutes. The time selection will remain displayed until it is changed.
- Select Exhaust (Drying) If no drying time is desired, set the Exhaust (Drying) Time Display to 00:00. If you are autoclaving liquids, you cannot change the Exhaust time. It is preset.
- Use care in opening the door. Open slightly to vent steam then open fully to remove items while wearing PPE.
The University's Biosafety Program has been established to protect individuals from exposure to biohazards through the application of administrative and engineering controls.
Biosafety Cabinet Guidelines
What is a Biosafety Cabinet?
A biosafety cabinet (BSC) is not chemical fume hood. Fume hoods are designed to remove chemical fumes and aerosols away from the work area. BSCs are designed to provide both a clean work environment and protection for employees who work with biological hazards. BSCs use vertical laminar airflow to create a barrier to airborne particles, such as microorganisms. They use High Efficiency Particulate Air (HEPA) filters to clean air going into the work area and out to the environment. The air in most BSCs is re-circulated over the work area through the HEPA filter. The HEPA filter removes airborne particles from the air, but does not remove chemical fumes. A single exception is a specific special model of Class II Type B2 BSC that is UL classified as a fume hood.
What is a Laminar Flow Clean Bench?
A laminar flow clean bench provides a space to work with a product or specimen where it will be protected from contamination by particulates such as microorganisms. This is accomplished by the laminar flow of clean air from a HEPA filter, which is blown across the workspace and out toward the user and the lab. Thus the basic laminar flow clean bench provides no protection for the user from chemically hazardous or infectious materials, including particulate or volatile hazards, and aerosols.
When Should I Use a Biosafety Cabinet?
Use a BSC for manipulations of human pathogens that are likely to create aerosols (such as vortexing open tubes, pipetting, opening caps after centrifuging, sonicating, aspirating with a syringe, etc.). Use for all manipulation of airborne transmitted pathogens (such as Brucella abortus, Mycobacterium tuberculosis, etc.).
Open Flames in a Biosafety Cabinet
Using open flames, such as Bunsen burners, in a BSC disrupts the air flow, compromising protection of both the worker and the work. In addition, if the flame of the burner is too large, or ignites a wash bottle of ethanol (often found in a BSC), the excessive heat may melt the adhesive holding the HEPA filter together or burn holes in the filter media. Alternative methods such as electric incinerators or disposable inoculating loops are recommended.
Ultraviolet Lights in a Biosafety Cabinet
Remember that the ultraviolet (UV) light in your BSC is only useful as an extra precaution in keeping the work area decontaminated between uses, because UV light has very little power to penetrate, even through a dust particle. Always clean and decontaminate the work area thoroughly using a chemical disinfectant after each use. If installed, UV lamps should be cleaned weekly to remove any dust or dirt which may block the germicidal effectiveness of the UV light. UV lamps must be turned off when the room is occupied to protect laboratory occupants’ skin and eyes from UV exposure. Exposure to UV light can cause burns to the corneas and skin cancer. Never work in a laboratory with a UV light on and always turn off UV lights when working in the BSC.
Annual Certification Testing
Improper airflow or filter leaks in a BSC (and laminar hoods) could expose laboratory personnel to bio-hazardous materials. To ensure that BSCs on campus are providing necessary protection to workers and the environment, it is essential that the BSCs be routinely inspected. EHS contracts with a qualified service company to annually service all BSCs and laminar hoods on campus that are used to contain biological hazards. You will be notified when annual testing will be performed. Testing is done according to the nationally accepted standards of NSF International. Your BSC should have a label on it stating the date it was last tested.
Moving or Repairs
In addition to annual testing, BSCs must also be re-tested whenever they are moved or have filters changed. This must be done by a qualified servicing company. Call EHS at 7233 to schedule testing other than annual.
Purchasing a New Biosafety Cabinet
If you plan to purchase a new BSC, notify EHS at x7233 for assistance in choosing the appropriate BSC for your needs and to get the BSC on the schedule for annual certification testing. Use the following guidelines when purchasing a BSC:
- The BSC should be certified by according to NSF Standard 49. Work with any infectious agents or recombinant DNA classified as requiring Biosafety Level 2 or higher containment will not be permitted in a BSC that does not pass certification testing for containment.
- Verify with the Biosafety Officer that the BSC type (Class II Type A, Class II Type B2, etc.) is appropriate for the type of work that it will be used for (type of biological agents to be contained and any chemicals to be used.)
- If the BSC is a Class II Type B3, the connection to the exhaust preferably should be a thimble connection and not a gas tight connection.
- Installation of BSCs must allow access to both supply and exhaust filters for annual certification testing and filter changes:
- Top of cabinet must be far enough below the ceiling (at least 18") to allow field testing of exhaust flow according to NSF Standard 49.
- Any connections to exhaust ductwork must allow access for field testing of exhaust flow according to NSF Standard 49.
- Any outlets inside the work area of the BSC should be ground fault circuit interrupter (GFCI) outlets.
Biological Waste Procedures
Biohazardous (Infectious) Waste
|Type of Waste||Containment||Autoclave Treatment||Disposal|
|Animal blood, body fluids||Labeled autoclave container||Yes||Place in opaque bag and place in refuse bin at the loading dock.|
|Animal carcasses and parts preserved in formaldehyde or other preservative.||Drain all liquids and place into labeled containers.
Place carcasses and parts in opaque plastic bag.
|No||Call the Safety Office to arrange for disposal of liquids and freezer storage of carcasses in MH121B.|
|Contaminated disposable plastics and other trash||Labeled autoclave bag||Yes||Trash. If any broken glass or glass pipettes present, place bag in cardboard box before disposal in trash.|
|Human/medical Waste - human liquid blood or tissue (infected or not)||Place in red biohazard bags||No||Call EHS for disposal. Must not be stored more than 5 business days prior to disposal.|
|Pathogenic bacteria cultures, tissue/cell cultures, viruses, parasites, biologically produced toxins||Labeled autoclave container. Do not put sharps into autoclave bags.||Yes||
Liquid parts: drain, flushed with copious amounts of water.
Solid parts: Remove solids, place in trash bag, and dispose in the refuse bin at the loading dock.
|Sharps (contaminated syringes, needles, razor blades, scalpels, glass slides, cover slips, Pasteur pipettes)||Approved Biohazard sharps container (Obtain from the Safety Office)||No||Call EHS for pick up. X7233|
Non-Biohazardous (Non-Infectious) Waste
|Type of Waste||Containment||Disposal|
|All animals (invertebrate and vertebrate) carcasses and parts||Put in opaque plastic bags. NO plastic, paper, needles, foil, etc. should be put in with the animals.||Contact EHS to arrange for freezer storage in MH121B.|
|Animal blood, body fluids||Labeled container||Flush down drain using garbage disposal, if possible; flush sink with copious amounts of water.|
|Disposable pipettes and other disposable glassware (NO LIQUIDS)||Heavy cardboard boxes, seams sealed with tape to prevent shards from coming out.||Seal tightly with tape, label and place in trash.|
|Human and animal urine||Flush down the lab drain; use copious amounts of water; rinse the sink afterwards.|
|Sharps||Request a non-infectious sharps container from EHS||Seal tightly, label and place in trash.|
Dispose of animals and animal parts into a clear plastic bag with no markings. Zip-lock bags work well. DO NOT use bags with the biohazard symbol on them. DO NOT place any gloves, paper towels, bench paper, syringes, aluminum foil or any other materials into the bag.
As soon as a carcass is ready for disposal:
Call EHS at x7233 to gain access to the carcass disposal freezer in MH 121 B2.Store the carcasses in a freezer prior to bringing them to the disposal area.If a carcass cannot be stored in a cold place and/or needs immediate disposal, contact EHS.Let EHS know if the carcasses are contaminated with radioactive materials prior to disposal.Remember, frozen animals are not bendable. Do not pack animals into spaces where removing them is difficult when they freeze. Freezer drawers and ice-filled compartments are the biggest problems.
Exception: Very small animal parts can be disposed of in the normal trash if they are very small, sealed in a plastic bag and do not create an odor problem.
If you have any questions, please call EHS at x7233.
A laboratory centrifuge can be an important tool in the university lab. It can also be a dangerous instrument if used or maintained improperly. Most hazards associated with centrifugation stem from one of two sources: mechanical conditions, and processing hazardous materials. This Safety instruction addresses both of these categories and presents methods for controlling the risks associated with them.
- Balance all samples as closely as possible.
- Rotors are rated for certain maximum speeds: KNOW THEM!
- Always put on the lid and secure it.
- Never open a centrifuge until the rotor has stopped.
- Clean all spills immediately.
- With infectious/biologically hazardous material: WAIT 10 MINUTES AFTER THE ROTOR HAS STOPPED before it is opened. Allow aerosols to settle, then wipe the rotor and centrifuge interior thoroughly.
- Look at rotors for corrosion and fatigue.
- Close lid when not in use especially if the centrifuge is refrigerated!!
- Sign the log books every time you use the centrifuge. Make notes on the centrifuge condition.
- Check the adapters and use them when needed; be sure to use the correct ones.
- Check the seals on the lids and report ones that are cracked or missing.
- Always ensure that loads are evenly balanced before a run.
- Always observe the manufacturer's maximum speed and sample density ratings.
- Always observe speed reductions when running high density solutions, plastic adapters, or stainless steel tubes.
Even when manufacturers' recommendations are closely followed, metal rotors will suffer fatigue. Repeated cyclical stretching and relaxation will cause changes in the metal's microstructure, resulting in eventual cracks and failure. Centrifuge manufacturers typically give both an expiration date (beyond which the rotor should not be used under any circumstance) and a maximum number of runs. To prevent mechanical rotor failure due to fatigue, observe the following:
- Never use a rotor past the manufacturer's expiration or safe-service date.
- Keep a rotor-use log to prevent overuse. (Note: some newer equipment may have data-logging capability. Consult the manufacturer's instructions for specific record keeping requirements.)
Many rotors are made from either titanium or aluminum alloy, chosen for their advantageous mechanical properties. While titanium alloys are quite corrosion-resistant, aluminum alloys are not. Note that although a rotor may be made of titanium alloy, other centrifuge components may be made from aluminum due to design considerations. When corrosion occurs, the metal is weakened and less able to bear the stress from the centrifugal force exerted during operation. The combination of stress and corrosion causes the rotor to fail more quickly and at lower stress levels than an uncorroded rotor. To prevent corrosion, observe the following:
- Select titanium-alloy or comparable rotors for areas where corrosive solutions, like KBr, will be used regularly.
Centrifugal force puts a load on the rotor, causing stretching or a change in the dimensions of the metal. Each rotor is designed to withstand a certain amount of stress and return to its original dimensions. However, if that amount of stress is exceeded, the rotor will not return to its original shape and size. This causes minute cracks and other wear that will deteriorate the rotor over time, leading to possibly dangerous consequences. To prevent stress, the following practices are strongly recommended:
- Never clean rotors or associated parts with abrasive wire brushes.
- Avoid using alkaline detergents or cleaning solutions on aluminum parts. (Note: most solutions designed for radioactive decontamination are highly alkaline. See the Hazardous Samples section below for more detail.)
- If corrosive or alkaline materials have been run or spilled, be sure to wash affected parts of the centrifuge immediately and allow them to air dry.
- Store the rotor away from the centrifuge in a dry area, with all cavities facing downward to prevent the accumulation of moisture.
- Use only rotors compatible with your centrifuge. Consult the operating manual for a list of compatible rotors for each centrifuge.
- Never attempt to open the door while the rotor is spinning or attempt to stop the rotor by hand.
- Do not attempt to move the centrifuge while it is in operation.
- Inspect the rotor before use and any time the rotor may have been subject to damage (i.e. dropped). Do not use the rotor if any cracks, rough spots, pitting, discolorations, or other abnormalities are present.
- Contact the manufacturer for details and service.Consider maintaining a service contract with a manufacturer's representative.
Centrifuging Hazardous Materials
Centrifugation of hazardous samples may result in exposure to chemical, biological, or radioactive agents. Careful consideration must be given to work practices to avoid hazards. The following is a list of practices that should be followed whenever hazardous materials are centrifuged:
- When possible, samples should be aspirated rather than poured from centrifuge tubes.
- Load and unload hazardous samples in ventilated enclosures (biosafety cabinet for biological specimens, fume hoods, etc.)
- Centrifuges used with hazardous aerosols and under a vacuum should be fitted with an appropriate in-line filter to protect the vacuum pump. (Contact the manufacturer for retrofits.)
- When hazardous samples are centrifuged, contain samples in safety cups, sealed tubes, or safety rotors.
- When safety containers are not available, centrifuge in a ventilated enclosure or evacuate the chamber before opening the lid via a vacuum port.
- Wait at least 10 minutes after the centrifuge has stopped to allow any aerosols generated in the chamber to settle.
- Clean and decontaminate all parts after each use, according to the manufacturer's instructions. (Note: some rotors may be autoclaved. Check with the manufacturer.)
- Develop standard operating procedures (SOPs) to minimize contamination and exposure. This is especially important when using radioactive materials, as many commercially prepared radioactive decontamination agents are highly alkaline and may subject metals to corrosion.
Spills and Leaks
Occasionally samples may spill or leak inside the centrifuge due to failure of the rotor or associated centrifuge parts. Anytime a sample containing hazardous materials has leaked inside the centrifuge this should be treated as hazmat spill and the following steps should be followed:
- Close the centrifuge lid immediately with the samples remaining inside and turn the centrifuge off.
- It may be necessary to vacate the lab depending on the nature of the spilled material and its ability to generate an aerosol. When the lab must be vacated, secure the room such that others cannot gain access. Post signage indicating restricted entry.
- All personnel should decontaminate themselves and seek medical attention as necessary.
- Notify principal investigators, lab supervisors, and coworkers as quickly as possible about the issue. Notify Environmental Heath and Safety for assistance - x7233.
A controlled substance is generally a drug or chemical whose manufacture, possession, or use are regulated by a government and subject to legislative control. Please see the University's Controlled Substances Program.
There is evidence that hantavirus is the cause of a potentially fatal Hantavirus Pulmonary Syndrome. Hantavirus is carried in the urine, saliva and feces if rodents, particularly rats and field mice. The greatest risk exposure is from breathing an aerosol containing the virus. For more information on Hantavirus, please go to the Centers for Disease Control website.
Controlling Rodent Exposure in Occupied Work Areas
- Reduce the amount of food and water that is available to rodents.
- Keep food covered or in a refrigerator.
- Do not leave dirty dishes out for long periods or allow them to soak in water.
- Keep all bulk grains and animal foods outside in secure containers.
- Do not overfeed birds.
- Improve housekeeping in work spaces and storage areas to limit the availability of nesting areas.
- Place garbage in rodent-proof containers, and empty the containers regularly (preferably daily).
- Seal, cover, or screen all openings that are large enough for mice to enter (anything over inch). This includes areas where pipes and wires enter the building.
- If you discover animal nests, droppings, or carcasses in your work area, contact Facilities Service Center at x3494.
Trapping and Clean-up Methods
Employees are strongly encouraged to avoid contact with rodents and rodent materials. Employees should contact the Service Control Center at x3494 for renewal and rodent-proofing.
However, if any employee chooses to clean-up rodents or droppings before the appropriate personnel can arrive, the following precautions should be taken.
- Wear rubber gloves when handling any rodent material.
- Use one of the following disinfectants to kill the virus. Apply the solution liberally (poured or sprayed on the material) before sweeping or mopping.Dilute bleach (1 part bleach plus 9 parts water, make fresh daily), Lysol, or 70% Alcohol
- If reusable rubber gloves are used, wash them with disinfectant, and them with soap and water. Disinfect any utensils that were used.
- DO NOT USE RODENT POISONS.
- Use spring-loaded traps that kill the rodent. The traps should be placed on newspapers and baited appropriately.
- After catching a mouse, sprinkle the newspaper with the recommended amount of flea insecticide. The insecticide will kill fleas to prevent the transmission of plague. Commercially available insecticides include Sevin (carbaryl powder) or pyrethrin sprays.
- Wear rubber gloves, saturate the mouse with disinfectant, and wait five minutes for the virus to be inactivated.
- Wrap the newspaper around the mouse and the trap, deposit these materials in a double plastic bag, and place the bag in the dumpster.
- If traps are reused, spray them thoroughly with disinfectant as described in the "clean-up methods."
- Vacuuming is not an acceptable for cleaning up rodent droppings. Fecal material should be saturated with disinfectant and collected in a plastic bag.
- Carpets and upholstered furniture can be cleaned with disinfectant or with commercial steam cleaners.
- Contaminated clothing should be laundered with detergent and hot water.
Disposal of Contaminated Materials
Contaminated materials should be soaked with the disinfectant mentioned above and then double-bagged in plastic for refuse collection.
At CSUF: Please contact EHS at x7233 for more information.
Safe Handling of Infectious Agents
Primary and Continuous Cell Cultures
Cell cultures, in general, present few biohazards in the laboratory, as evidenced by their extremely wide usage and the rare cases of transmitted infections to laboratory personnel. Primary cell cultures initiated with tissues from infected humans or animals are recognized hazards.
Continuous cell cultures present no real documented risk in the laboratory unless they are carelessly contaminated with an infectious agent. All continuous cell lines should be regularly monitored for contamination with infectious agents, and it should be emphasized that all nutrient media or other reagents that may contain ingredients of biologic origin must be treated as though they contain potentially infectious agents.
The Eight Basic Rules of Biosafety
The most common means of exposure can be essentially eliminated as occupational hazards by following the eight basic rules of biosafety:
- Do not mouth pipette.
- Manipulate infectious fluids carefully to avoid spills and the production of aerosols and droplets.
- Restrict the use of needles and syringes to those procedures for which there are no alternatives; use needles, syringes, and other "sharps" carefully to avoid self-inoculation; and dispose of "sharps" in leak- and puncture-resistant containers.
- Use protective laboratory coats, eye protection, closed toed shoes, and gloves.
- Wash hands following all laboratory activities, following the removal of gloves, and immediately following contact with infectious materials at the end of an operational at the end of the day.
- Do not wear gloves outside the lab area.
- Decontaminate work surfaces before and after use, and immediately after spills.
- Do not eat, drink, store food, or apply cosmetics in the laboratory.
Labeling of Specimens With the Laboratory
Some form of labeling is necessary to maintain the identity of specimens in the laboratory and to ensure that the analytical results obtained are properly recorded and reported. In addition, it is the practice in many cases that special hazard warning labels be affixed to specimens that are known to be hazardous (e.g., specimens obtained from patients known to be infected with hepatitis B virus (HBV) or human immunodeficiency virus (HIV).
The need for such special labeling is concerned more with ethical or regulatory issues (e.g., workers' right-to-know) than with laboratory safety. All clinical material must be considered to be infectious, and must be handled with exactly the same precautions as are used for processing specimens with hazard warning labels.
The risk of exposure of laboratory personnel can be minimized by the use of carefully selected safety equipment. A primary objective of containment is to control aerosols, but in a broader sense safety equipment should serve effectively to isolate the worker from the toxic or infectious material being processed. In many situations, however, the need is just the reverse: i.e., to protect the product or the work from contamination originating with the worker or the environment. Finally, there is often the need to protect both the worker and the product, as in handling cell cultures.
Biological Safety Cabinets
Most laboratory procedures generate aerosols that may spread infectious material in the work area and pose a risk of infection to the worker. Biological safety cabinets are used extensively to prevent the escape of aerosols or droplets and to protect materials from airborne contamination. There are three major types of this very useful safety device, referred to as Class I, Class II, and Class III. These instruments are distinct from horizontal or vertical laminar flow "clean benches," which should never be used for handling infectious, toxic, or sensitizing material.
The type used at California State University, Fullerton is the Class II biological safety cabinet, which provides protection to the worker, the environment, and the products. The airflow velocity at the face of the work opening is at least 75 linear feet per minute (lfpm), and both the supply and the exhaust air are HEPA-filtered. These cabinets are partial containment devices, which, if used in conjunction with good laboratory practices, can dramatically reduce the risk of exposure of operators to infectious aerosols and droplets.
It is emphasized that biological safety cabinets are not chemical fume hoods. Some of the air (30 to 70 percent) drawn in through the work opening of these cabinets is re-circulated within the cabinet. Accordingly, users should be aware of the possible buildup of hazardous concentrations within the cabinet if toxic, flammable, or explosive materials are used. In addition, users of Class IIA type cabinets should know that non-particulate toxic, flammable, or explosive materials are not removed by HEPA filters, and are thus discharged back into the laboratory room.
The operational efficiency of each biological safety cabinet should be specifically tested and the system certified before the instrument is placed in operation after installation, and subsequently on an annual basis. Recertification is also required if the unit is relocated or if maintenance that may affect performance is done. Maintenance work on biological safety cabinets should be performed by trained service personnel only. In addition, cabinet users should understand the operation of the equipment, its limitations, and the proper procedures to be followed. Laboratory directors are responsible for providing such training.
Clothing, Masks, and Face Shields
Laboratory coats, gowns, safety glasses, face shields, masks, closed toed shoes, and gloves offer some personal protection and are often used in combination with other safety devices such as biological safety cabinets. Special laboratory clothing protects street wear from contamination. It should not be worn outside of the laboratory. Each of these items has a particular use in protecting the worker and should be used when circumstances require. Gloves are especially important when handling any potentially infectious material such as blood or other biological specimens. Safety glasses, face shields, and masks may protect mucous membranes of the eye, nose, and mouth from splash or droplet hazards during operations performed outside of a biological safety cabinet.
The primary responsibility for the safe handling and disposal of infectious waste resides with the generator of the waste. This responsibility extends to the ultimate point of disposal even when there are other parties involved in handling the waste. No waste management program is functional unless all appropriate personnel are cognizant of the aims of the program and trained in the procedures for handling the waste. Training should be a continuing process.
Persons who generate laboratory waste are responsible for preparing the waste so that potential occupational exposures and environmental contamination are minimized. Cell culture and infectious wastes need to be segregated by the generator from all other waste streams.
Containment and treatment
After use, disposable Petri plates, flasks and other containment growth containers must be placed in autoclavable bags which are clearly marked with the biohazard symbol. Autoclave bags should be placed inside leak proof containers while they are being filled. When full, bags should be lightly closed with tape or a rubber band, then autoclaved. Open bags slightly or puncture the bag with several holes to allow venting of steam during autoclaving. Place waste bags in metal pans to catch any leakage that occurs during autoclaving. Autoclave on the "liquids" cycle for 45 minutes at 121 degrees C (250 degrees F) and at least 15 pounds of pressure. The autoclaved bags must then be taken directly outside to the trash dumpsters for immediate disposal.
Liquids should be treated with 0.01% to 0.05% (final concentration) bleach. Household bleach solutions are 5-7% sodium hypochlorite, so diluting them 100 fold in the liquid to be treated is sufficient to destroy most cultures. For example, add 1 ml of bleach for every 100 ml of culture. Leave the solution for at least ten minutes for maximum effectiveness. This solution can then be safely dumped in the laboratory drain, but it must be flushed with copious amounts of water to flush the solution out of the drain trap.
Large volumes of cultures, or cultures which are pathogenic or toxic, must be inactivated by autoclaving. Place culture containers in secondary containers whenever a chance of spillage may occur. Autoclave on the "liquids" cycle at the parameters described above.
DO NOT LET FULL BAGS (AUTOCLAVED OR NOT) OR OLD CULTURES SIT AROUND. TAKE CARE OF THEM IMMEDIATELY AND DISPOSE OF THEM IN THE OUTSIDE DUMPSTERS!
If you have any questions about cell cultures, or any other infectious agents, please contact the Environmental Health & Safety Office in T-1475, x7233.
Most of this information is taken directly from Biosafety in the Laboratory, National Academy Press, Washington, DC. 1989.
Select agents are biological agents or biological toxins which have been declared by the U.S. Department of Health and Human Services or by the U.S. Department of Agriculture to have the "potential to pose a severe threat to public health and safety". The University's use of select agents in biomedical research is controlled by the University's Select Agents Program.
For further information, the following information is available: