Table of Contents Biological Safety Manual
INTRODUCTION ............................................................................................................................... 2 BIOLOGICAL HAZARDS AND CONTROLS ....................................................................... 4 A. B. C. D. E. F.
CLASSIFICATION OF BIOLOGICAL HAZARDS .................................................................................... 4 BIOLOGICAL SAFETY GUIDELINES ................................................................................................... 5 LABORATORY BIOSAFETY LEVELS................................................................................................... 6 LABORATORY EQUIPMENT ............................................................................................................... 7 PERSONAL PROTECTIVE EQUIPMENT ............................................................................................. 10 EMERGENCY PROCEDURES FOR THE SPILL OR RELEASE OF AN INFECTIOUS AGENT....................... 11
INFECTIOUS WASTE MANAGEMENT ............................................................................... 12 A. B. C. D. E. F.
INFECTIOUS WASTES...................................................................................................................... 12 INFECTIOUS WASTE TREATMENT ................................................................................................... 13 RECORD KEEPING .......................................................................................................................... 14 DISPOSAL METHODS OF TREATED WASTE:.................................................................................... 14 LABELING WASTE .......................................................................................................................... 16 OFF-SITE TREATMENT AND MANIFESTING..................................................................................... 16
OCCUPATIONAL HEALTH ....................................................................................................... 17 TRANSPORT AND ACQUISITION OF INFECTIOUS AGENTS ............................... 19 A.
SHIPPING AND RECEIVING .............................................................................................................. 19
SECURITY OF SELECT AGENTS AND APHIS CONTROLLED AGENTS ......... 20 A. B.
PRELIMINARY APPROVAL .............................................................................................................. 20 REGISTRATION ............................................................................................................................... 21
OTHER INSTITUTIONAL POLICIES, PUBLICATIONS AND REFERENCES .. 22 APPENDICES ..................................................................................................................................... 23 APPENDIX I:CLASSIFICATION OF INFECTIOUS AGENTS ON THE BASIS OF HAZARD ................................ 23 APPENDIX II:.......................................................................................................................................... 27 APPENDIX III: ........................................................................................................................................ 29 APPENDIX IV: ........................................................................................................................................ 33 APPENDIX V........................................................................................................................................... 34
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Introduction Personnel who work in biological laboratories at the University of Texas – El Paso may handle or come into contact with hazardous biological agents. Over the years, there have been many documented cases of lab personnel acquiring diseases, some of which are fatal, due to their work with biological agents. Only approximately 20% of these reported cases have been attributed to a specific known incident, the rest are assumed to be related to careless work practices in the lab, specifically exposure to infectious aerosols. Effective risk assessment, proper handling based upon the conclusions of the risk assessment, and final disposal of biohazardous materials after adequate disinfection treatment, greatly reduce the potential for exposure to infectious or harmful agents. Therefore, whenever work with biological agents is performed, all appropriate steps must be taken to protect personnel and the environment. This Biological Safety Manual presents general information and biosafety practices as recommended by the Center for Disease Control and Prevention (CDC) and National Institutes of Health (NIH). The guidance presented in this manual should be used and may be customized for specific applications and protocols applicable in each individual bioscience laboratory, and can be used in conjunction with other available scientific information on risk assessment, to further minimize the potential for laboratoryassociated infections. The HHS Publication, Biosafety in Microbiological and Biomedical Laboratories (BMBL), Fourth Edition, is a very good source for agent specific information to aid in the risk assessment process. This Biological Safety Manual is supplemental to the University’s Laboratory Safety Manual and the University’s Chemical Hygiene Program, both of which address safe working practices in the lab and Industrial Hygiene aspects of laboratory work up to and including medical prophylaxis and monitoring for laboratory worker protection. Responsibilities Principal Investigators are responsible for ensuring that all work with infectious agents is conducted according to CDC and NIH guidelines, as outlined in this manual, including the universal precautions when handling blood and blood products. The Principal Investigators are responsible to perform a risk assessment to ensure all the appropriate equipment, including safety equipment, is available and maintained in the laboratory. They are to have all laboratory staff who work with infectious agents attend a biological safety class and a laboratory safety class provided by EH&S. The Principal Investigators are also responsible for training all laboratory staff both on the specific hazards of the infectious material they will be working with and on the proper usage and maintenance of laboratory safety equipment. The Principal Investigator is also responsible to initiate consultative discussions with the biological safety manager on security issues, opportunities for risk reduction, the potential need for medical monitoring or prophylaxis, and other issues surrounding the well-being of the laboratory staff. Laboratory staff is responsible to only use infectious materials for which they have been trained, and only utilizing the practices and procedures adopted in their specific laboratory. Laboratory staff are expected to follow the Principal Investigator’s instructions regarding the use of infectious materials in the lab and to observe the written Standard Operating Procedures for the Laboratory and the guidelines in this manual. Laboratory staffs are encouraged to openly inquire about questions or concerns they may have regarding activities within their lab.
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EH&S staff are responsible to provide biosafety services to the laboratories and the University. Those services include collection of hazardous wastes, spill response services, periodic monitoring of practices and procedures utilized in the laboratories, radioactive materials services, periodic inspection of chemical and biological materials storage, periodic inspection and testing of eyewashes, safety showers, chemical fumehoods and biological safety cabinets, and reporting noted deficiencies to laboratory management as necessary. The biological safety manager, as a member of EH&S staff, shall be available at all times to offer suggestions and opinions on security and safe practices to the Principal Investigators and the laboratory staff. The biological safety manager is responsible to the University as a monitor to reassure continued adequacy of practices and conditions within the biosciences, to report noted inconsistencies with accepted standards, to identify developmental and facility needs to further enhance the University’s capacity to meet its academic mission. To satisfactorily achieve the responsibilities of the EH&S department, it will during its routine visits to the bioscience laboratories, initiate dialog, offer assistance where appropriate, and follow-up where necessary. Twice annually the laboratories will be inspected to document that all aspects of laboratory operations are meeting recognized standards. If necessary, follow-up visits to laboratories will be scheduled with the Principal Investigators, with copies of the re-inspection reports being forwarded up management channels as warranted.
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Biological Hazards and Controls This chapter will discuss biological hazards and their classification on the basis of hazard type; biological safety guidelines as recommended by CDC/NIH; laboratory biosafety levels; laboratory equipment, including the biological safety cabinet; personal protective equipment; and emergency procedures for responding to biological spills and releases. A.
Classification of Biological Hazards The NIH defines biological hazards as "agents presenting a risk or potential risk to the wellbeing of man, or other animals, either directly through infection or indirectly through disruption of the environment." Biological hazards include materials or organisms known or suspected to contain infectious agents, recombinant DNA molecules, and oncogenic viruses. Infectious Agents (Etiologic Agents) An infectious agent is a viable microorganism, or its toxin, which causes or may cause disease in humans or animals. This classification includes bacteria, viruses, parasites, and fungal agents that have been assigned to Classes 1 through 4 on the basis of the hazards they present. Examples of infectious agents include Salmonella (Class II Bacterial Agent) and Influenza viruses (Class II Viral Agent). See Appendix I for the Classification of Etiologic Agents on the Basis of Hazard. A class of infectious agents of particular concern is Bloodborne Pathogens. A human bloodborne pathogen is a pathogenic microorganism present in human blood and body fluids, such as semen, vaginal fluid, amniotic fluid, saliva, and urine that can cause disease in humans. The bloodborne pathogens of greatest concern are hepatitis B (HBV) and human immunodeficiency virus (HIV). Whenever work with any bloodborne pathogen is performed, the CDCs "Universal Precautions" should be practiced. (Universal Precautions are discussed in the Biological Safety Guidelines section.) Recombinant DNA Molecules Recombinant DNA molecules are molecules constructed outside living cells by joining natural or synthetic DNA segments to DNA molecules that can replicate in a living cell, or DNA molecules that result from the replication described above. Oncogenic Virus An oncogenic virus is a virus that is believed to cause tumors in humans or animals. See Appendix I for a listing of oncogenic viruses.
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B.
Biological Safety Guidelines The following are general guidelines for the safe handling of biological agents in the laboratory. More detailed guidelines are available from CDC/NIHs "Biosafety in Microbiological and Biomedical Laboratories" and NIHs "Guidelines for Research Involving Recombinant DNA Molecules." All work involving biological agents at UTEP should follow CDC/NIH guidelines. 1.
All work involving infectious agents must be performed in properly functioning Biological Safety Cabinets (BSC) appropriate for the agent. Containment is very important to minimize exposure to bioaerosols.
2.
Wear disposable gloves when working with infectious agents to protect against exposure by contact. Other ways to control contact exposure include using absorbent paper on work surfaces and frequently disinfecting work surfaces.
3.
Wash your hands thoroughly after working with any biological agents and before leaving the lab to minimize chances of exposure due to ingestion.
4.
Exercise extreme caution when using "sharps," such as syringes, razor blades, and glass pipettes, to minimize inoculation hazards. Handle lab animals carefully as inoculation can also occur through infected animal bites.
5.
After using a needle, do not re-cap, bend, break, remove it from the syringe, or manipulate it in any other way, as many people have accidentally inoculated themselves while doing so. All sharps should promptly be placed as is into an appropriate sharps container to prevent any injuries.
6.
Be sure to disinfect work surfaces when finished with an experiment.
7.
All contaminated waste must be handled and stored properly, including disinfection, to prevent contact exposure of other lab personnel as well as housekeeping staff and waste handlers.
Universal Precautions Under Universal Precautions, all human blood and certain body fluids are considered potentially infectious for HIV, HBV, and other bloodborne pathogens. Universal Precautions apply to blood, blood contaminated body fluids, semen, and vaginal secretions. Universal Precautions also apply to tissues and to the following fluids: cerebrospinal, synovial, pleural, peritoneal, pericardial, and amniotic fluids. Universal Precautions do not apply to feces, saliva, nasal secretions, sputum, sweat, tears, urine, and vomitus unless they are visibly contaminated with blood.
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The Universal Precautions are summarized below and should be practiced whenever work is performed with human blood or body fluids listed above.
C.
1.
Use protective barriers such as gloves, gowns, aprons, masks, or protective eyewear to reduce the risk of skin or mucous membrane exposure to potentially infectious material. Remove all protective clothing before leaving the laboratory.
2.
Wash hands and other skin surfaces immediately if contaminated with blood and after the removal of gloves.
3.
Take precautions to prevent injuries caused by needles, scalpels, and other sharp instruments or devices. After using a needle, do not re-cap, bend, break, remove it from the syringe, or manipulate it in any other way. All sharps should promptly be placed as is into an appropriate sharps container to prevent any injuries.
4.
Keep all specimens of blood or body fluids in well-constructed containers with a secure lid to prevent leakage.
5.
Use biological safety cabinets whenever procedures are conducted that have a high potential for generating aerosols.
6.
Never pipette by mouth.
7.
Decontaminate work surfaces promptly after a spill and when work activities are completed.
Laboratory Biosafety Levels
The CDC and NIH describe four biosafety levels (BSL) for activities involving infectious agents. The levels are designated in ascending order by degree of protection provided to lab personnel, the environment, and the community. BSL1 is for work with infectious agents that pose minimal or no hazards while BSL4 is for work with infectious agents which pose the greatest hazard. Each level recommends facility design, lab practices, and safety equipment appropriate for working with the infectious agent involved. BSL1 through BSL4 are discussed briefly below. A more exhaustive discussion of biosafety level criteria can be found in CDC/NIHs "Biosafety in Microbiological and Biomedical Laboratories." Biosafety Level 1 BSL1 practices, safety equipment, and facilities are appropriate for undergraduate teaching laboratories using microorganisms not known to cause disease in healthy adult humans. BSL1 represents a basic level of containment that relies on standard microbiological practices with no special primary or secondary barriers recommended. Biosafety Level 2 BSL2 practices, safety equipment, and facilities are recommended for clinical, diagnostic, research, or teaching laboratories involving moderate risk agent associated with human disease of varying severity. The primary hazards to lab personnel working with these agents include accidental skin or mucous membrane exposures, or ingestion of infectious materials.
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BSL2 is appropriate when work is done with any human-derived blood, body fluids, or tissues where the presence of an infectious agent may be unknown. Primary barriers recommended include biological safety cabinets (BSC) and personal protective equipment (PPE). Secondary barriers recommended include waste decontamination facilities. Biosafety Level 3 BSL3 practices, safety equipment, and facilities are recommended for clinical, diagnostic, research, or teaching laboratories involving indigenous or exotic agents with a potential for respiratory transmission, and which may cause serious and potentially lethal infection. Primary hazards to lab personnel working with these agents include autoinoculation, ingestion, and exposure to infectious aerosols. Primary barriers recommended include BSCs or other enclosed equipment. Secondary barriers for this level include controlled access to the laboratory, a specialized ventilation system, and waste decontamination facilities. Biosafety Level 4 BSL4 represents maximum containment and is required for dangerous and exotic agents which pose a high risk of life-threatening disease, which may be transmitted via the aerosol route, and for which there is no available vaccine or therapy. See Appendix II for a summary of recommended biosafety levels for infectious agents. D.
Laboratory Equipment
A general understanding of biological laboratory equipment and how it works is essential to working safely with infectious agents. The following is a discussion of typical biological laboratory equipment, including biological safety cabinets centrifuges, sonicators, homogenizers, and blenders, and guidelines for their proper use. Biological Safety Cabinets Biological safety cabinets (BSC) are among the most effective, as well as the most commonly used, primary containment devices in laboratories working with infectious agents. The BSC is designed to capture and contain any infectious particulates or aerosols generated within the BSCs interior and exhaust them through a high-efficiency particulate air (HEPA) filter either into the laboratory, or to the outside. The three general types of BSCs available (Class I, II, and III) are discussed below. More detailed information on BSCs can be found in CDC/NIHs "Biosafety in Microbiological and Biomedical Laboratories." Class I The Class I BSC is a negative-pressure, ventilated cabinet usually operated with an open front. All of the air from the cabinet is exhausted through a HEPA filter either into the laboratory, or to the outside. The Class I BSC is designed for general microbiological research with low- and moderate-risk agents (Biosafety Level 1 and 2) and is useful for the containment of mixers, blenders, and other equipment.
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These cabinets are not appropriate for handling research materials that are vulnerable to airborne contamination, since the inward flow of unfiltered air from the laboratory can carry microbial contaminants into the cabinet. (Note: Class I BSCs are no longer being manufactured on a regular basis; many have been replaced by Class II BSCs.) Class II The Class II BSC is similar to the Class I BSC except Class II BSCs have an increased face velocity and the additional advantage of providing protection to the research material by HEPA filtration of the air flow into the cabinet across the work surface. This type of cabinet will protect the user, environment, and the research material and is suitable for work with moderate- to high-risk agents (Biosafety Level 2 and 3). Class II BSCs are classified into two types: A and B. Basically, type A cabinets exhaust the cabinet air into the laboratory. Since the air is recirculated with in the laboratory, volatile or toxic chemicals and radionuclides should not be used inside this type of cabinet. Type B cabinets are further divided into sub-types B1, B2, and B3. Type B cabinets are ducted to the exhaust system that is maintained under negative pressure, thus allowing work to be done with volatile or toxic chemicals and radionuclides. Class III The Class III BSC is a negative pressure, totally enclosed and gas-tight ventilated cabinet that offers the highest degree of protection to personnel, environment, and research materials. The Class III cabinet is suitable for work with extremely high-risk agents (Biosafety Level 3 or 4 agents.) All operations in the work area of the cabinet are performed through rubber gloves attached to entry portals. Supply air is HEPA-filtered, and the cabinet exhaust air is filtered by two HEPA filters in series, or HEPA filtration followed by incineration, before discharge to the outside of the facility. See Appendix III diagrams of the different types of Biological Safety Cabinets. Proper Use As with any other piece of laboratory equipment, personnel must be trained in the proper use of the biological safety cabinets. Of particular note are those activities that may disrupt the inward directional airflow through the work opening of Class I and II cabinets. Repeated insertion and withdrawal of the workers' arms in and from the work chamber, opening and closing doors to the laboratory or the isolation cubicle, improper placement or operation of materials or equipment within the work chamber, or brisk walking past the BSC while it is in use are demonstrated causes of the escape of aerosolized particles from within the cabinet. Class I and II cabinets should be located away from traffic patterns and doors. Fans, heating and air-conditioning registers, and other air handling devices can also disrupt airflow patterns if located adjacent to the BSC. Strict adherence to recommended practices for the use of BSCs and proper placement in the laboratory are important in attaining the maximum containment capability of the equipment and maintaining mechanical performance of the equipment itself. The following are additional safety guidelines for proper use of the BSC: 1.
Disinfect the work surface of the BSC before and after use. A 70% ethanol or 1:10 bleach solution is a suitable disinfectant.
2.
Be careful not to place any objects on the air intake or exhaust grills as this would disrupt the airflow.
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3.
A sign can be posted on any doors around the cabinet stating that the cabinet is in use and thereby prevent unnecessary opening and closing of doors which disrupts the BSC air flow.
4.
Always wear a lab coat while using the cabinet and conduct your work at least four inches past the BSC opening.
5.
If a burner is necessary, place it towards the back of the cabinet because it will cause some air turbulence.
6.
It is a good idea to keep a disinfectant handy in the event that a spill might occur.
7.
Operate the cabinet for five minutes after completing any work inside the cabinet to purge any air-borne contaminants.
8.
Thoroughly wash your hands and arms before leaving the lab.
It is imperative that Class I and II BSCs are tested and certified at the time of installation within the laboratory, at any time the BSC is moved, and at least annually thereafter.
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Centrifuges, Sonicators, Homogenizers, and Blenders These instruments deserve special consideration because of their potential to create aerosols. If carcinogens, toxic chemicals, or infectious agents are going to be used in any of these instruments, then precautions must be taken to prevent the exposure of lab personnel to generated aerosols. Centrifuges Safety features for the centrifuge include sealed buckets, safety trunnion cups, or sealed heads to prevent the escape of aerosols and liquids. Additional safety guidelines for proper centrifuge use are: 1.
Make sure the lid is on and secured before operating the centrifuge.
2.
Always balance the load in the centrifuge; if you are not filling the entire centrifuge rack, position the tubes opposite one another. If you have an odd number of tubes, use a tube filled with an appropriate amount of water to equal the weights of the other tubes.
3.
If vibration occurs, stop the centrifuge and check the load balance. Never operate an unbalanced centrifuge as the excessive vibration could result in breaking the tubes inside and generating hazardous aerosols.
4.
Keep the rotors and buckets clean and promptly clean any breakage or spills.
5.
Routinely inspect your centrifuge to ensure leakage is not occurring. An indicator, such as Fluorescein, can be used to detect leaks. Fluorescein can be added to water and then centrifuged. A UV fluorescent light can then be used to detect fluorescein on the work surface, floors, and walls.
Sonicators, Homogenizers, and Blenders The use of any of these instruments with infectious agents should be conducted inside a biological safety cabinet to contain any hazardous aerosols that are generated. Blenders should have leak-proof bearings and a tight fitting gasket lid. Inspect the lid and gaskets routinely to ensure they are in good condition. Household blenders are not appropriate as they do not prevent the spread of aerosols. E.
Personal Protective Equipment
The type of personal protective clothing or equipment recommended depends on the Biosafety Level of the laboratory (see Section C.) For BSL1 and BSL2 laboratories, standard protective clothing includes a lab coat and latex gloves. Additional protection, such as a face shield, safety glasses or goggles, may be necessary where there is a splash potential and when handling human blood and body products. BSL3 laboratories have more specific requirements for protective clothing including solid-front or wrap around lab gowns (typical button-down-the-front lab coats are not acceptable), puncture resistant gloves, and respirators. BSL4 laboratories have the most stringent requirements for protective clothing including positive pressure suits that totally enclose and isolate the user from the surrounding lab environment. Whenever protective clothing or equipment becomes contaminated it should be removed and either decontaminated or replaced. Do not reuse disposable gloves, discard after use or when torn. Wearing double layers of gloves provides extra protection in the event the outer gloves becomes contaminated and needs to be removed and discarded without exposing the skin. Leave all protective clothing in the lab; do not wear it outside the lab. rev 08/2012
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F.
Emergency Procedures for the Spill or Release of an Infectious Agent
Biological laboratories must be prepared for spills or releases involving infectious agents. Response to a spill or release of an infectious agent must be thorough and prompt to prevent further injury and contamination. The following are only general guidelines; each lab should design their own response plan based on their unique hazards and the location of the laboratory. In the Event of a Biological Spill . . . 1.
Notify the people in the immediate area and if necessary evacuate the lab. The decision to evacuate is a judgment call based on the properties and hazards of the spilled material. Notify EH&S and UTEP Police if an evacuation is necessary.
2.
Always attend to injured persons before attending to the spill. Seek medical help if necessary.
3.
Try to contain the spill to keep it from spreading.
The following spill clean-up procedures are based on the biosafety level of the agent involved. Spills or releases involving BSL1 Agents: 1.
Wear a lab coat and disposable gloves.
2.
Soak a paper towel(s) in an appropriate disinfectant, such as a 1:10 bleach solution or 70% ethanol, and place over the spill area.
3.
Place the paper towel(s) and gloves into a biohazard bag for disposal.
Spills or releases involving BSL2 Agents: 1.
If infectious aerosols or droplets are generated from the spill or release, evacuate and close the lab. Allow 30 minutes for the droplets to settle and the aerosol concentration to decrease.
2.
Wear appropriate protective clothing and equipment including gloves, lab coat, and an approved respirator equipped with HEPA filters.
3.
Cover the spill area with paper towels, pour a 1:10 bleach solution around the edges of the spill and then into the spill area. Allow 20 minutes contact time for proper disinfection.
4.
Use paper towels to clean the area, working from the outer edges to the center. Clean the area with fresh towels soaked in a disinfectant.
5.
Place all clean-up materials and gloves into a biohazard bag for disposal or decontamination. Wash hands and arms thoroughly.
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6.
A small spill of material that did not result in the generation of aerosols or significant contamination can be cleaned using steps 2-5 above.
Spills or releases involving BSL3 Agents: 1.
If the spill occurs inside a biological safety cabinet, keep the cabinet running and clean the spill following the steps outlined in Spills or releases involving BSL2 agents, with the exception that protective clothing appropriate for BSL3 be worn. If the spill inside the cabinet is substantial, it may be necessary to decontaminate the cabinet's fan, filters, and airflow plenums. Contact EH&S for assistance. 2.
If a minor spill occurs outside of a biological safety cabinet, clean the spill following the steps outlined in Spills or releases involving BSL2 agents, with the exception that protective clothing appropriate for BSL3 be worn.
3.
If a substantial spill occurs outside the biological safety cabinet, evacuate the lab and notify the appropriate personnel, including UTEP Police and EH&S.
Infectious Waste Management Infectious waste from biological laboratories and medical facilities is regulated by The Texas Commission on Environmental Quality (TCEQ) and the Texas Department of Health (TDH). These infectious materials are considered as “special” waste. A special waste is any solid waste that is not regulated as hazardous waste but because of its quantity, concentration, physical and/or chemical characteristics, or biological properties requires special handling and disposal to protect human health and the environment. Types of infectious waste will be discussed in the following sections along with proper treatment and disposal methods, and record keeping requirements. A.
Infectious Wastes
Infectious waste includes waste from microbiology and pathology labs, laboratory animal facilities, blood and blood products, and sharps; that is known or suspected to contain viable infectious microorganisms. Potential sources of infectious wastes from microbiology laboratories are discarded cultures and stocks of infectious agents and associated biologicals; discarded cultures of specimens from medical, pathological, pharmaceutical, research, clinical, commercial, and industrial laboratories; discarded live and attenuated vaccines, but not the empty containers; used disposable culture dishes; and used disposable devices used to transfer, inoculate, or mix cultures. Potential sources of infectious wastes from pathology laboratories are any human materials such as tissues and body parts, laboratory specimens of blood and tissue after completion of laboratory analysis, and anatomical remains. Potential source of infectious wastes from laboratory animal facilities are the bedding of animals exposed to pathogens, animal carcasses or body parts, and animal blood and blood products. Blood and blood products include all human blood, serum, plasma, and other blood components. As required by the Universal Precautions, all blood and blood products should be handled as if it is known to contain pathogens. rev 08/2012
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"Sharps" is a broad term used to describe any sharp object that has the potential to puncture the skin if handled improperly, thus presenting an inoculation hazard if the "sharp" is contaminated. Sharps include syringes, razor and scalpel blades, glass pipettes, and microscope slides. B.
Infectious Waste Treatment
The most common infectious waste treatment and disposal methods are chemical disinfection, steam sterilization, and incineration. These methods will be discussed below with some guidelines. Chemical Disinfection Liquid and gaseous chemicals are used routinely for decontaminating infectious waste. Common liquid disinfectants include alcohol (ethyl or isopropyl), formaldehyde, and chlorine compounds (bleach) while ethylene oxide is a common gas disinfectant. See Appendix IV for a table of liquid and gas decontaminants and their use in infectious waste management. All waste that has been immersed in a liquid disinfectant must be thoroughly drained before disposal. Steam Sterilization (Autoclaving) Steam sterilization, or autoclaving, usually is considered to be the treatment method of choice for decontaminating non-disposable cultures, laboratory glassware, pipettes, syringes, or other small items known to have infectious agents. It provides a technically sound treatment method for rendering infectious material safe. Autoclaving is not recommended for treating large volumes of waste or insulated materials such as animal carcasses or human body parts. Autoclave temperature, pressure, and time settings are very important to ensure adequate decontamination. The temperature and pressure settings differ depending on the type of autoclave used and the time setting varies according to load conditions. Gravity displacement autoclaves operate at 121°C and 15 psi for a recommended minimum of 60 minutes under normal conditions while vacuumtype autoclaves operate at 132°C and 27 psi for a recommended minimum of 10 minutes. The shorter time period for this type of autoclave is due to the higher temperatures and pressures attained therefore resulting in greater steam penetration. Once a waste has been autoclaved and is no longer infectious, it can be disposed of through the general trash. However, do not place "Biohazard" bags into the general trash, instead, attach a "waste treated" tag (available from EH&S) to the biohazard bag then place it into a general trash bag before discarding. Autoclaves should be tested periodically to ensure adequate disinfection. Incineration Incineration is the method of choice for treating large volumes of infectious waste, animal carcasses, contaminated bedding, and human anatomical parts. The main advantage of incineration is the significant reduction in waste volume and the unobjectionable end product, ash. Complete combustion of waste material is crucial to proper incineration. The factors involved in complete combustion are time, temperature, and turbulence. The waste should be retained in the combustion chamber(s) for a long enough time and at a high enough temperature to allow for mixing (turbulence) with excess oxygen, so that the combustion reactions can go to completion.
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A deficiency in any one or more of these critical combustion parameters can result in smoke or odor production, excessive emissions of harmful gaseous by-products, and the discharge of incompletely burned waste residue. Another important consideration is how the waste is introduced into the incinerator, or feed rate. Overfeeding an incinerator can result in smoke and odors and incomplete combustion. For those who do not have access to the incinerator located on campus, commercial treatment and disposal is available. Contact EH&S for more information. To request a biological waste pick-up from EH&S, complete a "Hazardous Material Pick-Up and Disposal Request" form (see Appendix IV) and contact EH&S. Waste must be disinfected/decontaminated prior to any pick-ups. C.
Record Keeping
All lab personnel who treat and dispose of infectious waste on site must keep the following records. A lab that generates 50 pounds or less per calendar month of infectious waste must record the following: 1.
Date of treatment;
2.
Amount of waste treated;
3.
Treatment method used and conditions of treatment; and
4.
Name Printed and initials of person(s) performing treatment.
A lab that generates more than 50 pounds per calendar month of infectious waste must record the following: All of the above, plus: A written procedure for the operation and testing of any equipment used and a written procedure for the preparation of any chemicals used in treatment. If wastes are treated off-site then other record keeping will apply to document that the waste was treated in accordance with applicable rules. See sub-section F below for more details. D.
Disposal Methods of Treated Waste:
Chemical Disinfection Treated waste maybe disposed of as general waste in the municipal landfill. All waste material must be thoroughly drained before disposed. Chemical liquid waste material, used, as the disinfectant must be characterized as hazardous or nonhazardous waste, prior to disposal. Contact EH&S for assistance on properly characterizing the waste. If chlorine compounds (bleach) are used, this substance may be disposed of through the sanitary sewer.
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Steam Sterilization (Autoclaving) Waste treated through this process, may be disposed of in the municipal landfill. Incineration The ash generated through this treatment process may also be disposed of in the municipal sanitary landfill. Sharps All sharps materials treated from any of the above methods may be disposed of in the municipal landfill. All broken glassware and pipettes shall be placed in puncture-resistant packaging and discarded with routine municipal solid waste. All hypodermic needles, syringes with attached needles, scalpel blades, and/or razors shall be placed in containers designed for sharps. An encapsulating agent may be added to the container to solidify and encase the sharps. The agent must completely fill the container. The container and solidified contents must withstand an applied pressure of 40 pounds per square inch without disintegration. The container shall be identified as containing sharps which have been encapsulated in accordance with 30 TAC 330.1004(d)(4)(c), and may be discarded with routine municipal solid waste. If the sharps container has not been encapsulated, then the container must be segregated from the regular municipal trash and shall be collected and transported without compaction to the solid waste Landfill.
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E.
Labeling Waste
All treated waste material must be identified by the use of a label which states that the contents of the disposable container have been treated in accordance with the provisions of 25 TAC 1.136(a). All other markings on the package must be covered or removed, prior to disposed. Contact EH&S for appropriate labels. F.
Off-Site Treatment and Manifesting
If waste is not to be treated on site, it shall be released only to a registered medical waste transporter for disposal. The registered transporter shall provide a signed manifest for each shipment using a form approved by TCEQ. The generator must maintain all shipping/treatment manifests for a period of three years following the date of shipment. Within 30 days of each off-site shipment a treatment record must be returned by the transporter. This record should be attached to the shipping manifest as proof that all shipments have been properly disposed of. Any failure of the transporter to return a treatment record within 30 days should be reported immediately to EH&S for further action and resolution.
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Occupational Health As a matter of practice it is always a primary consideration to remove through engineering controls any risk associated with an action. Many research protocols provide for the direct use of hazardous chemicals, infectious agents and toxins, or perhaps even the use of vertebrate animals in the production of the research findings. As such the performance of the research protocols require engineering controls (such as BSCs and chemical fume-hoods), the use of personal protective equipment (such as gloves, eyewear and respiratory protection), and aggressive diligent hygiene practice of Universal Precautions. The engineering controls, PPE, and hygiene practices used in the laboratory are in combination critical to the protection of laboratory workers. Likewise, the risk assessment performed by the Principal Investigator in consultation with the Environmental Health and Safety office is an essential element to determine to what extent these measures should be employed in the laboratory performance of the protocol. In addition to the preventative measures taken by the laboratory personnel, adequate knowledge of the chemical hazards present in the laboratory is necessary to ensure personnel are safe. As such the Principal Investigator must keep of listing of hazardous materials available for the lab personnel to review and should instruct the personnel on those hazards and how to avoid them. Knowledge of the emergency procedures and available emergency equipment is crucial to the safety of the laboratory personnel. The location of exits and meeting places, the sounds of relevant alarms, and the whereabouts of emergency eyewashes, alarm pull stations, fire extinguishers, prepared disinfectants and spill kits all must be addressed in the laboratory specific training. Despite all of the engineering controls and the personal working knowledge of a laboratory and its inherent hazards, medical prophylaxis, vaccination or other intervention is sometimes warranted. It is the responsibility of the Principal Investigator during the risk assessment to identify if a vaccine, other medical procedures, or monitoring are called for regarding the risks inherent to the research. To be considered during the risk assessment are zoonotic diseases, infectious agent exposures, sharps exposures, chemical sensitivities, physical hazards, and any treatments or procedural methodologies that could effectively minimize the risk associated with those exposures. Where available, licensed vaccines for which the benefits clearly outweigh the risk should be required to be offered for all personnel identified as at risk. Other treatments may be available or recommended if the benefits do not clearly outweigh the risk. Specific information on a variety of agents and available treatment is available in Section VII of the BMBL, fourth edition. One should not undertake any inoculation or treatment without the proper advisement by an attending Medical Physician. All cost associated with the medical treatments should be at the expense of the University and shall not be charged back to the treated individual. In the event of an accident or injury that occurs in the performance of laboratory functions or field work done as part of the regular activities of the University or its research, notification of the accident time, date, persons involved, severity, etc., must be reported back through appropriate channels to both the Human Resources office and the Environmental Health and Safety office. In all cases the immediacy of the injury should be dealt with first, with any required reporting occurring only after the injury has been stabilized. The reports must be done preferably on the same business day but at a minimum within 24 hours of the event.
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Environmental Health &Safety and Human Resources keep records involving injuries. Records shall be maintained by the department in order to document the risk assessment, the decision to require or offer medical intervention, and proof as to the administration of such or the declination, whichever may be the case. Decisions regarding medical treatments shall be a topic of discussion at both the Institutional BioSafety Committee and the Institutional Animal Care and Use Committee meetings, as appropriate and applicable.
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Transport and Acquisition of Infectious Agents In recent years it has become increasingly necessary to be vigilant in our activities relating to shipping, receiving and storing of infectious agents. Many infectious materials must now be tracked very closely and in a quantifiable way from the date of shipping from the supplier, through the final use, and even through the destruction. The intent of this section of the Biological Safety Manual is to describe for the researchers how they must comply, at what point different diligent levels of administrative controls apply. A.
Shipping and Receiving
The Public Health Service, in 1996, published 42 CFR, part 72, to regulate the interstate shipment of Etiological Agents. Contained within these rules at Section 72.3 is a list of infectious agents, which must be packaged, labeled and shipped in accordance with 72.3(a) through (f).
A sublist found at Section 72.3(f) must additionally be shipped by registered mail or an equivalent system. The Purchasing department at UTEP has been provided with the lists found within 42 CFR, part 72, and is instructed to notify the Environmental Health and Safety office for guidance and approval prior to initiating a purchase of the listed items.
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Security of Select Agents and APHIS Controlled Agents A Select Agent is an infectious material that is listed by the Health and Human Service, Centers for Disease Control (CDC) as a possible terrorist weapon for use against humans. The U.S. Department of Agriculture, Animal and Plant Health Inspection Service (APHIS) has a similar list of materials which are recognized as potential terrorist weapons because of the impact they could have upon food supplies in the nation. For the purposes of this manual the term “Select Agent” is intended to be synonymous with APHIS Controlled Agent and the controls mentioned apply to both. A current listing of the Select Agents and APHIS listed agents may be found at 42 CFR part 73 and 7 CFR part 331, respectively. Currently the University of Texas at El Paso doesn’t have any listed Select Agents, nor does it have a registration to acquire Select Agents. This being said we must still recognize that as the University grows, and its research becomes even more diverse with the addition of specialized containment systems, it is inevitable that the use of Select Agents in research will become so in the not too distant future. Therefore, it is important in this manual to describe the processes and requirements that will become part of standard operations. A.
Preliminary Approval
Select Agents and APHIS listed agents both require significant containment and security measures. As such, these materials are strictly controlled by the Federal Government. Each laboratory planning to propose use of Select Agents in research must first draft a research protocol and request a security risk assessment from a joint team of both UTEP Police and the EH&S office. This request shall be routed through the EH&S office. The Biological Safety Manager will determine what level of compliance requirements will be applicable, depending upon the Select Agent and laboratory proposed. The purpose of this screening inspection and security risk assessment is to determine the ability of the lab in question and its assigned personnel to meet the requirements for “registration” with the CDC (or APHIS) for use of the agent(s) in question. UT System policies regarding background investigations have been revised and are now available. All UT System component institutions are required to implement these policies as they are applicable to our operations. These policies are intended to complement the USA PATRIOT Act and the recently added 42 CFR Part 73, which define ‘restricted persons’. If it appears that the security and containment capabilities of the laboratory, the proposed level of safety training and occupational health, and the laboratory personnel are all suitable to the proposed project, the Principal Investigator must then submit the proposed protocol, with EH&S and PD comments, to a work group assigned by the Office of Research and Sponsored Projects and the Dean of Science to review such matters. If the proposal is consistent with ORSP policies and the Dean of Science agrees with the space and resource allocation necessary for the proposed research, it may then be presented for approval to the Institutional Biosafety Committee (IBC). The IBC may only indorse the project as meeting safety and health requirements. If animals are proposed as part of the protocol then it must additionally be presented to and accepted by the Institutional Animal Care and Use Committee (IACUC).
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B.
Registration
Only after the Preliminary Approval is performed and all mentioned campus departments have endorsed their aspects of the project, a registration application may then be drafted and sent to the affect agency, CDC or APHIS. The University President has assigned the Director of Environmental Health and Safety as the “Responsible Facility Official” who would be responsible to file the registration application on behalf of the University and the laboratory involved. Although UT System has specific requirements and methods stated regarding background investigations, all persons tentatively approved by UTEP or any other component institution for future access to select agents must be ultimately approved by the U.S. Attorney General (USAG) before a registration will be granted. The USAG’s final approval for access to select agents will be contingent upon a thorough background investigation performed either by the Federal Government, or an assigned contracted agency. CDC and APHIS as well will have their own investigation teams who would be assigned to review the adequacy of the proposed protocol and the institutional policies that would support the successful safe and secure handling of the select agent(s) while in use on the campus. Only after this review process is done would a registration be granted to the specific laboratory and protocol proposed. See Appendix V for more information regarding procedures and controls to be implemented when Select Agents and APHIS agents are to be employed.
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Other Institutional Policies, Publications and References 1.
Laboratory Safety Manual, University of Texas at El Paso.
2.
Chemical Hygiene Program, University of Texas at El Paso.
3.
Radiation Safety Manual, University of Texas at El Paso.
4.
Emergency Response Guide, University of Texas at El Paso.
5.
Hazardous Materials Handling and Disposal Policy and Procedures, University of Texas at El Paso.
6.
Biosafety in Microbiological and Biomedical Laboratories. The Centers for Disease Control and Prevention and the National Institutes of Health, Fourth Edition, 1999.
2.
Biosafety in the Laboratory: Prudent Practices for the Handling and Disposal of Infectious Materials. National Research Council, Committee on Hazardous Biological Substances in the Laboratory, 1989.
3.
Guide for the Care and Use of Laboratory Animals. National Research Council, 1996.
4.
Occupational Health and Safety in the Care and Use of Research Animals. Nation Research Council, 1997.
5.
Occupational Health and Safety Administration Standards, 29 CFR 1910.1450. Occupational exposure to hazardous chemicals in laboratories.
6.
AAALAC International Position Statements
7.
Public Health Service Policy on Humane Care and Use of Laboratory Animals. Reprinted 2000.
8.
Occupational Health and Safety Administration Standards, 29 CFR 1910.1030. Bloodborne pathogens.
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Appendices Appendix I:Classification of Infectious Agents on the Basis of Hazard Class 1 Agents All bacterial, parasitic, fungal, viral, rickettsial, and chiamydial agents not included in higher classes. Class 2 Agents Bacterial Agents Acinetobacier calcoacelicits Aciiizobacilltis-spp. Aeromoiias hydrofoil Arizona hinshawii-all serotypes Bacillus anthracis Bordetella-spp. Borrelia recurrentis, B. vincentii Campylobacter fetus Campylobacter jejuni Chlamydia psittaci Chlamydia trachomatis Clostridium botulinum Cl. chuvoei, Cl. haemolyticum, Cl. histolyticum, Cl. novyi, Cl. septicum, Cl. tetani Corynebacterium diptheriae, C. equi, C. haemolyticum, C. pseudotuberculosis, C. pyogenes, C. renale Edwardsiella tarda Erysipelothrix insidiosa Escherichia coli - all enteropathogenic, enterotoxigenic, enteroinvasive, and stains bearing K1 antigen Haemophilus ducreyi, H. influenzae Klebsiella-spp. and serotypes Legionella pneumophila Leptospira interrogans-spp. Listeria-spp. Moraxella-spp. Mycobacterium-spp. except those listed in Class 3 Mycoplasma-spp. except Mycoplasma mycoides and M. agalactiae, which are forbidden Neissaria gonorrhoea, N. meningitides Nocardia-spp. Pasterella-spp. except those listed in Class 3 Salmonella-spp. and all serotypes Shigella-spp. and all serotypes Sphaerophorus necrophorus Staphylococcus aureus Streptobacillus moniliformis Streptococcus pneumoniae, S. pyogenes Treponema crateum, T. pallidum, T. pertenue Vibrio cholerae, V. parahaemolyticus Yersinia enterocolitica
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Appendix I:Classification of Infectious Agents on the Basis of Hazard Continued Fungal Agents Actinomycetales (including Actinomyces spp. and Arachnia propionica) Blastomyces dermatitdis Cryptococcus neoformans Paracoccidioides brasiliensis Parasitic Agents Entamoeba histolytica Leishmania-spp. Naeglaria gruberi, N. fowleri Schistosoma mansoni Toxoplasma gondii Toxocara canis Trichinella spiralis Trypanosoma cruzi Viral, Rickettsial, and Chlamydial Agents Adenoviruses-human-all types Cache Valley virus Corona viruses Coxsackie A and B viruses Cytomegaloviruses Echoviruses-all types Encephalomyocarditis virus (EMC) Flanders virus Hart Park virus Hepatitis-associated antigen material Herpesvirus-associated antigen material Herpesviruses-except Herpesvirus simiae (Monkey B virus) which is Class 4 HTLV I/II Human Immunodeficiency virus (except large volumes or high concentrations which require BL 3) Influenza viruses-all types except A/PR8/34, which is Class 1 Langat virus Measles virus Mumps virus Parainfluenza viruses-all types except Parainfluenza virus 4, SF 4 strain, which is Class 1 Polioviruses-all types, wild and attenuated Poxviruses-all types, except Alastrim, Smallpox, and Whitepox, which are forbidden, and Monkey pox, which depending on the experiment is Class 3 or 4 Rabies virus-all strains except Rabies street virus, which is Class 3 or 4 Reoviruses-all types Respiratory syncytial virus Rhinoviruses-all types Rochalitmaea vinsonii Rubella virus Simian viruses-all types except Herpesvirus simiae (Monkey B virus) and Marburg virus, which are Class 4.
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Appendix I:Classification of Infectious Agents on the Basis of Hazard Continued Class 4 Sindbis virus Tensaw virus Turlock virus Vaccinia virus Varicella virus Vesicular stomtitis virus Yellow fever virus, 17d vaccine strain Class 3 Agents Bacterial Agents Bartonella-spp. Brucella-spp. Francisella tularensis Mycobacterium avium complex, M. bovis, M. tuberculosis Pasteurella multocida type B ("buffalo" and other foregn virulent strains) Yersinia pestis Fungal Agents Coccidioides immitis Histoplasma capsulatum Histoplasma capsulatum var. duboisii Parasitic Agents None Viral, Rickettsial, and Chlamydial Agents Arboviruses-all strains, except those in Class 2 and 4 Coxiella burnettii Ehrlichia-spp. Lymphocytic choriomeningitis virus (LMC) Monkey pox virus, when used in vitro Rabies street virus Rickettsia-spp. (except R. ruminantium) West Nile and Semliki Forrest viruses, depending on conditions of use and geographical location of the laboratory Yellow fever virus-wild, when used in vitro Class 4 Agents Bacterial, Fungal, and Parasitic Agents None Viral, Rickettsial, and Chlamydial Agents Ebola fever virus Hemorrhagic fever agents, including Crimean hemorrhagic fever, Congo, Junin, and Machupo viruses Herpesvirus simiae (Monkey B virus)
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Appendix I:Classification of Infectious Agents on the Basis of Hazard Continued Lassa fever virus (Mastomys natalensis) Marburg virus (Cercopithecus spp.) Monkey pox, when used for transmission or animal inoculation experiments Tick-borne encephalitis virus complex, including Russian spring-summer encephalitis, Kyasanur forest disease, Omsk hemorrhagic fever and Central European encephalitis viruses Venezuelan equine encephalitis virus, epidemic strains, when used for transmission or animal inoculation experiements Yellow fever virus-wild, when used for transmission or nimal inoculation experiements Low-Risk Oncogenic Viruses AD7-SV40 Adenovirus Avian Leukosis Bovine Leukemia Bovine Papilloma CELO Dog Sarcoma Guinea Pig Herpes Hamster Leukemia HTLV I/Ii Lucke (Frog) Marek's Mason-Pfizer Monkey Virus Mouse Mammary Tumor Murine Leukemia Murine Sarcoma Polyoma Rat Leukemia Rat Mammary Tumor Rous Sarcoma Shope Fibroma Shope Papilloma SV-40 Moderate-Risk Oncogenic Viruses Ad2-SV40 EBV FeLV FeSV GaLV HV Ateles HV Saimiri SSV-1 Yaba
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Appendix II: Summary of Recommended Biosafety Levels for Infectious Agents BSL
Agents
Practices
Safety Equipment (Primary Barriers)
Facilities (Secondary Barriers)
1
Not known to consistently cause disease in healthy human adults.
Standard animal care and management practices, including appropriate medical surveillance programs
As required for normal care of each species.
Standard animal facility • •
•
2
3
Associated with human disease. Hazard: percutaneous exposure, ingestion, mucous membrane exposure.
Indigenous or exotic agents with potential for aerosol transmission; disease may have serious health effects.
• • • • •
• •
•
Dangerous/exotic agents that pose high risk of life threatening disease; aerosol transmission, or related agents with unknown risk of transmission.
•
ABSL-1 facility plus: • •
•
ABSL-2 equipment plus:
Controlled access Decontamination of clothing before laundering Cages decontaminated before bedding removed Disinfectant foot bath as needed
ABSL-3 practices plus:
•
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Limited access Biohazard warning signs Sharps precautions Biosafety manual Decontamination of all infectious wastes and of animal cages prior to washing
ABSL-2 practices plus:
•
4
ABSL-1 equipment plus primary barriers: containment equipment appropriate for animal species; PPES: laboratory coats, gloves, face and respiratory protection as needed.
ABSL-1 practices plus:
•
•
Containment equipment for housing animals and cage dumping activities Class I or II BSCs available for manipulative procedures (inoculation, necropsy) that may create infectious aerosols . PPEs: appropriate respiratory protection
ABSL-3 equipment plus:
Entrance through change room where personal clothing is removed and laboratory clothing is put on; shower on exiting All wastes are decontaminated before removal from the facility
•
Maximum containment equipment (i.e., Class III BSC or partial containment equipment in combination with full body, air-supplied positive-pressure personnel suit) used for all procedures and activities
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No recirculation of exhaust air Directional air flow recommended Handwashing sink recommended
Autoclave available Handwashing sink available in the animal room. Mechanical cage washer used
ABSL-2 facility plus: • • • •
•
Physical separation from access corridors Self-closing, double-door access Sealed penetrations Sealed windows Autoclave available in facility
ABSL-3 facility plus: • •
•
Separate building or isolated zone Dedicated supply and exhaust, vacuum and decontamination systems Other requirements outlined in the text
BSL
1
2
3
Agents
Practices
Safety Equipment (Primary Barriers)
Facilities (Secondary Barriers)
Not known to consistently cause disease in healthy adults
Standard Microbiological Practices
None required
Open bench top sink required
Associated with human disease, hazard = percutaneous injury, ingestion, mucous membrane exposure
BSL-1 practice plus:
Primary barriers = Class I or II BSCs or other physical containment devices used for all manipulations of agents that cause splashes or aerosols of infectious materials; PPEs: laboratory coats; gloves; face protection as needed
BSL-1 plus: Autoclave available
Indigenous or exotic agents with potential for aerosol transmission; disease may have serious or lethal consequences
BSL-2 practice plus:
Primary barriers = Class I or II BCSs or other physical containment devices used for all open manipulations of agents; PPEs: protective lab clothing; gloves; respiratory protection as needed
BSL-2 plus:
• • • •
• • • •
Limited access Biohazard warning signs "Sharps" precautions Biosafety manual defining any needed waste decontamination or medical surveillance policies
Controlled access Decontamination of all waste Decontamination of lab clothing before laundering Baseline serum
• • • •
4
Dangerous/exotic agents which pose high risk of life-threatening disease, aerosol-transmitted lab infections; or related agents with unknown risk of transmission
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BSL-3 practices plus: • • •
Clothing change before entering Shower on exit All material decontaminated on exit from facility
Primary barriers = All procedures conducted in Class III BSCs or Class I or II BSCs in combination with full-body, air-supplied, positive pressure personnel suit
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Physical separation from access corridors Self-closing, double-door access Exhausted air not recirculated Negative airflow into laboratory
BSL-3 plus: • • •
Separate building or isolated zone Dedicated supply and exhaust, vacuum, and decon systems Other requirements outlined in the text
Appendix III: Diagrams of Biological Safety Cabinets
Figure 1. Class I Biological Safety Cabinet. A. Front opening B. Work surface C. Window D. Exhaust plenum E. HEPA filter
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Figure 2(b). Class II, Type B1 BSC. A. Blowers B. Supply HEPA filters C. Sliding sash D. Positive pressure plenums E. Additional supply HEPA filter or back-pressure plate F. Exhaust HEPA filter G. Negative pressure exhaust plenum, H. work surface
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Figure 2(d). Table-top model of a Class II, Type B3 BSC. A. Front opening B. Sliding sash C. Light D. Supply HEPA filter E. Positive pressure plenum F. Exhaust HEPA filter G. Control panel H. Negative pressure plenum I. Work surface
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Figure 3. Class III BSC. A. Stand B. Glove ports C. O-ring for attaching arm-length gloves to cabinet D. Sloped glass viewing window E. Supply HEPA filter F. Exhaust HEPA filter (Note that the second exhaust HEPA filter required for Class III cabinets is not depicted in this diagram) G. Double-ended autoclave
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Appendix IV: University of Texas at El Paso Hazardous Material Pick-up and Disposal Request A copy of this form is required for each container. Make sure all materials are stored in chemically compatible containers and properly labeled. Materials should be segregated according to the type of hazard Do not use the wording “hazardous waste” as the waste name. Secure a label to the container identifying the material, location and log sheet number. The work area is responsible for the identification of the material. If the material cannot be positively identified, the material will be sent it to a contract laboratory for analysis and the work area billed for the analytical costs. When the material requires disposal, complete this form. Attach the pink copy of the form and the log sheet to the container. For pickup, technical assistance, containers, labels, waste “pick-up” forms, and log sheets call the EH&S office at 747-7124. Log No. ________________ Material Name: _______________________________ Dept. ___________________Requested by: ______________________ Ext: _________ Located in: Bldg _____________________Room _____________ MATERIAL IDENTIFICATION Quantity: ___________ Container: Size________ Type __________________ Physical Form: Solid [ ] Liquid [ ] Gas [ ] Solution [ ] Mixed Phase [ ] Chemical and Physical Hazards: Carcinogenic [ ]
Combustible
[]
Flammable [ ]
Oxidant
[]
Combustible [ ] Etiological [] Irritant [] Poison [] Corrosive [] Explosive [ ] Mutagenic [ ] Reactive [ ] Special Hazards __________________________________________ Radioactive? Yes [ ] No [ ] If yes, Isotope:_________ Activity: _______milliCuries pH _________ Contains halogenated solvent? Yes [ ] No [ ] SAFETY AND HANDLING PRECAUTIONS Gloves [ ] Goggles [ ] Apron [ ] Respirator [ ] Other _________________________ I hereby certify that the above information is complete and accurate to the best of my knowledge and ability to determine and that there is no deliberate or willful omission. Name: (Print)____________________________________ Title: ___________________ Signature: _________________________________________ Date:_________________
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Appendix V Laboratory Security and Emergency Response Guidance for Laboratories Working with Select Agents December 6, 2002 Prepared by Jonathan Y. Richmond, Ph.D.1 Shanna L. Nesby-O’Dell, D.V.M.2 1Office of the Director Office of Health and Safety (Retired) 2Office of the Director Office of Health and Safety Summary In recent years, concern has increased regarding use of biologic materials as agents of terrorism, but these same agents are often necessary tools in clinical and research microbiology laboratories. Traditional biosafety guidelines for laboratories have emphasized use of optimal work practices, appropriate containment equipment, well-designed facilities, and administrative controls to minimize risk of worker injury and to ensure safeguards against laboratory contamination. The guidelines discussed in this report were first published in 1999 (U.S. Department of Health and Human Services/CDC and National Institutes of Health. Biosafety in microbiological and biomedical laboratories [BMBL]. Richmond JY, McKinney RW, eds. 4th ed. Washington, DC: US Department of Health and Human Services, 1999 [Appendix F]). In that report, physical security concerns were addressed, and efforts were focused on preventing unauthorized entry to laboratory areas and preventing unauthorized removal of dangerous biologic agents from the laboratory. Appendix F of BMBL is now being revised to include additional information regarding personnel, risk assessments, and inventory controls. The guidelines contained in this report are intended for laboratories working with select agents under biosafety-level 2, 3, or 4 conditions as described in Sections II and III of BMBL. These recommendations include conducting facility risk assessments and developing comprehensive security plans to minimize the probability of misuse of select agents. Risk assessments should include systematic, site-specific reviews of 1) physical security; 2) security of data and electronic technology systems; 3) employee security; 4) access controls to laboratory and animal areas; 5) procedures for agent inventory and accountability; 6) shipping/transfer and receiving of select agents; 7) unintentional incident and injury policies; 8) emergency response plans; and 9) policies that address breaches in security. The security plan should be an integral part of daily operations. All employees should be well trained and equipped, and the plan should be reviewed annually, at least. Introduction Traditional laboratory biosafety guidelines have emphasized use of optimal work practices, appropriate containment equipment, well-designed facilities, and administrative controls to minimize risks of unintentional infection or injury for laboratory workers and to prevent contamination of the outside environment (1). Although clinical and research microbiology laboratories might contain dangerous biologic, chemical, and radioactive materials, to date, only a limited number of reports have been published of materials being used intentionally to injure laboratory workers or others (2–7).
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However, recently, concern has increased regarding possible use of biologic, chemical, and radioactive materials as terrorism agents (8,9). In the United States, recent terrorism incidents (10) have resulted in the substantial enhancement of existing regulations and creation of new regulations governing laboratory security to prevent such incidents. The Public Health Security and Bioterrorism Preparedness and Response Act of 2002* (the Act) required institutions to notify the U.S. Department of Health and Human Services (DHHS) or the U.S. Department of Agriculture (USDA) of the possession of specific pathogens or toxins (i.e., select agents†), as defined by DHHS, or certain animal and plant pathogens or toxins (i.e., high-consequence pathogens), as defined by USDA. The Act provides for expanded regulatory oversight of these agents and a process for limiting access to them to persons who have a legitimate need to handle or use such agents. The Act also requires specified federal agencies to withhold from public disclosure, among other requirements, site-specific information regarding the identification of persons, the nature and location of agents present in a facility, and the local security mechanisms in use. In addition, the Uniting and Strengthening America by Providing Appropriate Tools Required To Intercept and Obstruct Terrorism (USA PATRIOT) Act of 2001§ prohibits restricted persons from shipping, possessing, or receiving select agents. Violation of either of these statutes carries criminal penalties. Appendix F of the 4th edition of the CDC/National Institutes of Health, Biosafety in Microbiological and Biomedical Laboratories (BMBL) was the first edition to address laboratory security concerns (1). However, that publication primarily addressed physical security concerns (e.g., preventing unauthorized entry to laboratory areas and preventing unauthorized removal of dangerous biologic agents from the laboratory). The guidelines presented here are provided to assist facility managers with meeting the regulatory mandate of 42 Code of Federal Regulation (CFR) 73 and, therefore, include information regarding personnel, risk assessments, and inventory controls. These guidelines are intended for laboratories where select agents are used under biosafety levels (BSL) 2, 3, or 4 as described in Sections II and III of BMBL. Appendix F of BMBL is being revised to include consideration of the following biosecurity policies and procedures: • risk and threat assessment; • facility security plans; • physical security; • data and electronic technology systems; • security policies for personnel; • policies regarding accessing the laboratory and animal areas; • specimen accountability; • receipt of agents into the laboratory; • transfer or shipping of select agents from the laboratory; • emergency response plans; and • reporting of incidents, unintentional injuries, and security breaches. Definitions Biosafety: Development and implementation of administrative policies, work practices, facility design, and safety equipment to prevent transmission of biologic agents to workers, other persons, and the environment.
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Biosecurity: Protection of high-consequence microbial agents and toxins, or critical relevant information, against theft or diversion by those who intend to pursue intentional misuse. Biologic Terrorism: Use of biologic agents or toxins (e.g., pathogenic organisms that affect humans, animals, or plants) for terrorist purposes. Responsible official: A facility official who has been designated the responsibility and authority to ensure that the requirements of Title 42, CFR, Part 73, are met. Risk: A measure of the potential loss of a specific biologic agent of concern, on the basis of the probability of occurrence of an adversary event, effectiveness of protection, and consequence of loss. Select agent: Specifically regulated pathogens and toxins as defined in Title 42, CFR, Part 73, including pathogens and toxins regulated by both DHHS and USDA (i.e., overlapping agents or toxins). Threat: The capability of an adversary, coupled with intentions, to undertake malevolent actions. Threat assessment: A judgment, based on available information, of the actual or potential threat of malevolent action. Vulnerability: An exploitable capability, security weakness, or deficiency at a facility. Exploitable capabilities or weaknesses are those inherent in the design or layout of the biologic laboratory and its protection, or those existing because of the failure to meet or maintain prescribed security standards when evaluated against defined threats. Vulnerability assessment: A systematic evaluation process in which qualitative and quantitative techniques are applied to arrive at an effectiveness level for a security system to protect biologic laboratories and operations from specifically defined acts that can oppose or harm a person’s interest. Risk Assessment Recommendation: Conduct a risk assessment and threat analysis of the facility as a precursor to the security plan. Background: In April 1998, the General Accounting Office issued a report regarding terrorism (11). A key finding of that report was that threat and risk assessments are widely recognized as valid decisionsupport tools for establishing and prioritizing security program requirements. A threat analysis, the first step in determining risk, identifies and evaluates each threat on the basis of different factors (e.g., the capability and intent to attack an asset, the likelihood of a successful attack, and the attack’s probable lethality). Risk management is the deliberate process of understanding risk (i.e., the likelihood that a threat will harm an asset with certain severity of consequences) and deciding on and implementing actions to reduce that risk. Risk management principles are based on acknowledgment that 1) although risk usually cannot be eliminated, it can be reduced by enhancing protection from validated and credible threats; 2) although threats are possible, certain threats are more probable than others; and 3) all assets are not equally critical. Therefore, each facility should implement certain measures to enhance security regarding select agents. The following actions should assist decision-makers in implementing this recommendation:
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•
Each facility should conduct a risk assessment and threat analysis of its assets and select agents. The threat should be defined against the vulnerabilities of the laboratory to determine the necessary components of a facility security plan and system (12,13).
•
The risk assessment should include a systematic approach in which threats are defined and vulnerabilities are examined; risks associated with those vulnerabilities are mitigated with a security systems approach (12,13).
•
Ensure the security plan includes collaboration between senior management, scientific staff, human resource officials, information technology (IT) staff, engineering officials, and security officials. This coordinated approach is critical to ensuring that security recommendations provide a reasonable and adequate assurance of laboratory security without unduly impacting the scientific work.
Facility Security Plans Recommendation: Establish a facility security plan. •
Each facility should develop a comprehensive security plan that complies with 42 CFR Part 73 and reviews the need for policies in —physical security; —data and IT system security; —security policies for personnel; —policies for accessing select agent areas; —specimen accountability; —receipt of select agents into the laboratory; —transfer or shipping of select agents from the laboratory; —emergency response plans; and —reporting of incidents, injuries, and breaches.
•
Develop security policies based on site-specific assessments. Security plans should include measures that address physical security of building and laboratory areas. Policies should also address concerns associated with access, use, storage, and transfer of sensitive data. If sensitive electronic data are present, IT specialists should assess the security of hardware and software products in addition to the security of local area networks.
•
Review safety, security, and IT policies and procedures at least annually for consistency and applicability. These procedures should also be reviewed after any incident or change in regulations. Necessary changes should be incorporated into the revised plans and communicated to all.
•
Laboratory supervisors should ensure that all laboratory workers and visitors understand security requirements and that all employees are trained and equipped to follow established procedures. The security plan should be an integral part of daily operations. New employees should receive training when they first begin work, and all employees should receive training at least annually thereafter. Training should be updated as policies and procedures change. All training should be documented by maintaining records of training schedules and employee attendance.
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•
Security plans should receive periodic performance testing to determine their effectiveness. Test procedures can vary from a simple check of keys, locks, and alarms to a full-scale laboratory or facility exercise.
Security Policies for Personnel Recommendation: Establish security-related policies for all personnel. •
Honest, reliable, and conscientious workers represent the foundation of an effective security program. Facility administrators and laboratory directors should be familiar with all laboratory workers.
•
Establish a policy for screening employees who require access to select agent areas to include full- and part-time employees, contractors, emergency personnel, and visitors. Additional screening might be necessary for employees who require access to other types of sensitive or secure data and work areas. These screening procedures should be commensurate with the sensitivity of the data and work areas (e.g., federal security clearances for government employees and contractors).
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Ensure that all workers approved for access to select agents (e.g., students, research scientists, and other short-term employees) wear visible identification badges that include, at a minimum, a photograph, the wearer’s name, and an expiration date. Facility administrators should consider using easily recognizable marks on the identification badges to indicate access to sensitive or secure areas.
Access Control Recommendation: Control access to areas where select agents are used or stored. •
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Consolidate laboratory work areas to the greatest extent possible to implement security measures more effectively. Separate select agent areas from the public areas of the buildings. Lock all select agent areas when unoccupied. Use keys or other security devices to permit entry into these areas. Methods of secure access and monitoring controls can include key or electronic locking pass keys, combination key pad, use of lock-boxes to store materials in freezers or refrigerators, video surveillance cameras, or other control requirements. Protocols for periodically changing combination keypad access numbers should be developed. Assess the need for graded levels of security protection on the basis of site-specific risk and threat analysis. This security can be accomplished through card access systems, biometrics, or other systems that provide restricted access. Lock all freezers, refrigerators, cabinets, and other containers where select agents are stored when they are not in direct view of a laboratory worker. Limit access to select agent areas to authorized personnel who have been cleared by the U.S. Department of Justice as indicated in 42 CFR Part 73. All others entering select agent areas must be escorted and monitored by authorized personnel. Record all entries into these areas, including entries by visitors, maintenance workers, service workers, and others needing one-time or occasional entry. Limit routine cleaning, maintenance, and repairs to hours when authorized employees are present and able to serve as escorts and monitors.
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Establish procedures and training for admitting repair personnel or other contractors who require repetitive or emergency access to select agent areas. Ensure visitors are issued identification badges, including name and expiration date, and escorted and monitored into and out of select agent areas. Such visits should be kept to a minimum. Ensure procedures are in place for reporting and removing unauthorized persons. These procedures should be developed through collaboration among senior scientific, administrative, and security management personnel. These procedures should be included in security training and reviewed for compliance at least annually.
Select Agent Accountability Recommendation: Establish a system of accountability for select agents. •
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Establish an accounting procedure to ensure adequate control of select agents and maintain upto-date inventory of seed stocks, toxins, and agents in long-term storage. Records should include data regarding the agent’s location, use, storage method, inventory, external transfers (sender/receiver, transfer date, and amount), internal transfer (sender/receiver, transfer date, amount), further distribution, and destruction (method, amount, date, and a point of contact). Establish procedures that maintain accurate and up-to-date records of authorizations for entry into limited access areas (i.e., a current list of persons who possess door keys and those who have knowledge of keypad access numbers or the security system).
Receiving Select Agents Recommendation: Develop procedures for bringing select agent specimens into the laboratory. • • •
A centralized receiving area for select agents is recommended to maximize safety and minimize security hazards associated with damaged or unknown packages. Facilities should establish procedures for inspecting all packages (i.e., by visual or noninvasive techniques) before they are brought into the laboratory area. Suspicious packages should be handled as prescribed by federal and state law enforcement agencies. Biologic safety cabinet or other appropriate containment device should be used when opening packages containing specimens, bacterial or virus isolates, or toxins. Packages should be opened by trained, authorized personnel.
Transfer or Shipping of Select Agents Recommendation: Develop procedures for transferring or shipping select agents from the laboratory. •
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Package, label, and transport select agents in conformance with all applicable local, federal, and international transportation and shipping regulations, including U.S. Department of Transportation (DOT) regulations. Materials that are transported by airline carrier should also comply with packaging and shipping regulations set by the International Air Transport Association (IATA). Personnel who package, handle, and ship these agents (including import and export) should be subject to all applicable training. The responsible facility official should be notified of all select agent transfers, internal or external. Ensure required permits (e.g., granted by the U.S. Public Health Service, USDA, DOT, U.S. Department of Commerce, and IATA) are obtained before select agents are prepared for transport. Standard operating procedures should be in place for import and export activities.
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Decontaminate contaminated or possibly contaminated materials before they leave the laboratory area. Avoid hand-carrying select agents when transferring them to other external facilities. If select agents are to be hand carried on common carriers, all applicable packaging, transport, and training regulations should be followed. Develop and follow a protocol for intra-facility transfer of all select agents.
Emergency Response Plans Recommendation: Implement an emergency response plan. • • • • • •
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Limiting access to select agent laboratory and animal areas can make implementing an emergency response more difficult. This should be considered as emergency plans are developed. Evaluate select agent laboratory and animal areas for safety and security concerns before an emergency plan is developed. Develop and integrate laboratory emergency plans with facility-wide plans. These plans should also include such adverse event assessments as bomb threats, severe weather (e.g., hurricanes or floods), earthquakes, power outages, and other natural or man-made disasters. Include facility administrators, scientific directors, principal investigators, laboratory workers, maintenance and engineering support staff, facility safety officers, and facility security officials in emergency planning. Include provisions for immediate notification of and response by laboratory and animal directors, laboratory workers, safety office personnel, or other knowledgeable persons when an emergency occurs. Establish advance coordination with local police, fire, and other emergency responders to assist community emergency responders in planning for emergencies in select agent laboratory and animal areas. Discussion should address security concerns associated with sharing of sensitive information regarding secure work areas. Consider circumstances that might require the emergency relocation of select agents to another secure location. Reevaluate and train employees and conduct exercises of the emergency response plan at least annually.
Incident Reporting Recommendation: Establish a protocol for reporting adverse incidents. •
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Ensure that laboratory directors, in cooperation with facility safety, security, and public relations officials, have policies and procedures in place for reporting and investigating unintentional injuries, incidents (e.g., unauthorized personnel in restricted areas, missing biologic agents or toxins, and unusual or threatening phone calls), or breaches in security measures. DHHS or USDA should be notified immediately if select agents are discovered to be missing, released outside the laboratory, involved in worker exposures or infections, or misused. Additionally, all incidents involving select agents (e.g., occupational exposure or breaches of primary containment) should be reported to local and state public health authorities.
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Acknowledgments CDC is grateful to the members of the Select Agent Interagency Workgroup, Biosecurity Subcommittee, and recognizes the contributions of Rachel E. Levinson, M.A., Chairman Biosecurity Subcommittee and Jonathan Y. Richmond, Ph.D., Assistant Chairman, Biosecurity Subcommittee. References 1. US Department of Health and Human Services/CDC and National Institutes of Health. Biosafety in microbiological and biomedical laboratories. Richmond JY, McKinney RW, eds. 4th ed. Washington, DC: US Department of Health and Human Services, 1999. 2. Török TJ, Tauxe RV, Wise RP, et al. Large community outbreak of salmonellosis caused by intentional contamination of restaurant salad bars. JAMA 1997;278:389–95. 3. Kolavic SA, Kimura A, Simons SL, Slutsker L, Barth S, Haley CE. Outbreak of Shigella dysenteriae type 2 among laboratory workers due to intentional food contamination. JAMA 1997;278:396–8. 4. US Nuclear Regulatory Commission. Report to Congress on abnormal occurrences July– September 1995; dissemination of information. Federal Register 1996;61:7123–4. 5. US Nuclear Regulatory Commission. Incident investigation report: ingestion of phosphorus-32 at Massachusetts Institute of Technology, Cambridge, Massachusetts, identified on August 19, 1995 [NUREG- 1535]. Washington, DC: US Nuclear Regulatory Commission, 1995. 6. US Nuclear Regulatory Commission. Preliminary notification of event or unusual occurrence PNO-1-98-052. Subject: intentional ingestion of iodine-125 tainted food (Brown University), November 16, 1998. Washington, DC: US Nuclear Regulatory Commission, 1998. 7. US Nuclear Regulatory Commission. National Institutes of Health issuance of director’s decision under 10 CFR Sec. 2.206. Federal Register 1997;62:50018–33. 8. Atlas RM. Biological weapons pose challenge for microbiology community. ASM News 1998;64:3839. 9. Ruys T. Laboratory design principles. In: Handbook of facilities planning. Ruys T, ed. New York, NY: John Wiley & Sons, 1990;257–64. 10. CDC. Update: investigation of anthrax associated with intentional exposure and interim public health guidelines, October 2001. MMWR 2001;50:889–93. 11. US General Accounting Office. Combating terrorism: threat and risk assessments can help prioritize and target program investments. Washington, DC: US General Accounting Office, 1998. Publication no. GAO/NSIAD-98-74. 12. Johnson B. Understanding, assessing, and communicating topics related to risk in biomedical research facilities [Chapter 10]. In: Richmond JY, ed. Anthology of biosafety: IV. Issues in public health, Mundelein, IL: American Biological Safety Association, 2001;149–166. 13. Royes C, Johnson B. Security considerations for microbiological and biomedical facilities [Chapter 6]. In: Richmond JY, ed. Anthology of biosafety: V. BSL–4 laboratories. Mundelein, IL: American Biological Safety Association, 2002;131–148.
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