BIOSAFETY LEVEL LABORATORIES

Transcription

1 BIOSAFETY LEVEL LABORATORIES A. BIOSAFETY LEVEL 1 Biosafety Level 1 (BSL-1) is suitable for work involving well-characterized agents not known to consistently cause disease in immunocompetent adult humans, and present minimal potential hazard to laboratory personnel and the environment. BSL-1 laboratories are not necessarily separated from the general traffic pattern in the building. Work is typically conducted on open bench tops using standard microbiological practices. Special containment equipment is not required, but may be used as determined by appropriate risk assessment. Laboratory personnel must have specific training in the procedures conducted in the laboratory and must be supervised by a scientist with training in microbiology or related science. 1. BSL-1 Standard Microbiological Practices Enforce institutional policies that control access to the laboratory; Wash hands after working with potentially hazardous materials and before leaving the laboratory; Eating, drinking, smoking, handling contact lenses, applying cosmetics and storing food for human consumption is not permitted in the laboratory. Food must be stored outside of the laboratory; Mouth pipetting is prohibited; Develop and implement policies for handling sharps, such as needles, scalpels, pipettes, and broken glassware. Whenever practical, laboratory supervisors should adopt improved engineering practice controls that reduce the risk of sharp injuries. Precautions must always be taken with sharp items and include: o Needles must not be bent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated by hand before disposal; o Used disposable needles and syringes must be carefully placed in puncture resistant containers used for sharps disposal; o Non-disposable sharps must be placed in a hard walled container for transport to a processing area for decontamination, preferably autoclaving; and o Broken glassware must not be handled directly. Instead it must be removed using a brush and dustpan, tongs, or forceps. Plastic ware should be substituted for glassware whenever possible. Perform all procedures to minimize the creation of splashes and/or aerosols; Decontaminate work surfaces with appropriate disinfectant after completion of work and after any spill or splash of potentially infectious material; Decontaminate all cultures, stocks, and other potentially infectious materials before disposal using an effective method; Post a sign incorporating the universal biohazard symbol at the entrance to the laboratory when infectious agents are present. Posted information must include: o Laboratory biosafety level; o Supervisor s name; o Telephone number; and o Procedures for entering and exiting the laboratory. BSL-1

2 Ensure an effective integrated pest management program is in place; and Ensure laboratory personnel receive appropriate training regarding their duties, the necessary precautions to prevent exposures, and exposure evaluating procedures. Personnel must receive annual updates or additional training when procedural or policy changes occur. Laboratory personnel should be provided with information regarding immune competence and conditions that may predispose them to infection. Individual having these conditions should be encouraged to self-identify to UMSL s healthcare provider for appropriate counseling and guidance. 2. BSL-1 Safety Equipment (Primary Barriers and PPE) Special containment devices or equipment, such as Biosafety Cabinets (BSC) are not generally required; Protective laboratory coats, gowns, smocks, or uniforms designated for laboratory use are recommended to prevent contamination of personal clothing; Protective eyewear must be worn when conducting procedures that have the potential to create splashes of microorganism or other hazardous materials. Persons who wear contact lenses in laboratories should also wear eye protection; and Gloves must be worn to protect hands from exposure to hazardous materials. Glove selection should be based on appropriate risk assessment. Alternatives to latex gloves should be available. In addition, BSL-1 laboratory workers should: o Change gloves when contaminated, glove integrity is compromised or when otherwise necessary; o Remove gloves and wash hands when work with hazardous materials has been completed and before leaving the laboratory; and o Do not wash or reuse disposable gloves. Dispose of used gloves with other contaminated laboratory waste. Hand washing protocols must be rigorously followed. 3. BSL-1 Laboratory Facilities (Secondary Barriers) Laboratories should have doors for access control; Laboratories should have sinks for hand washing; The laboratory should be designed so that it can be easily cleaned. Carpets and rugs in laboratories are not appropriate; Laboratory furniture must be capable of supporting anticipated loads and uses. Spaced between benches, cabinets, and equipment should be accessible for cleaning: o Bench tops must be impervious to water and resistant to heat, organic solvents, acids, alkalis, and other chemicals; and o Chairs used in laboratory work must be covered with a non-porous material that can be easily cleaned and decontaminated with appropriate disinfectant. Laboratory windows that open to the exterior should be fitted with screens. BSL-2

3 B. BIOSAFETY LEVEL 2 Biosafety Level 2 (BSL-2) builds on BSL-1. BSL-2 is suitable for work involving agents that pose moderate hazards to personnel and the environment. It differs from BSL-1 in that laboratory personnel have specific training in handling pathogenic agents and are supervised by scientists competent in handling infectious agents and procedures, access to the laboratory is restricted when work is being conducted, and all procedures in which infectious aerosols or splashes may be created are conducted in BSC or other physical containment equipment. According to NIH Guidelines, all forms of research utilizing recombinant or synthetic nucleic acid molecules must be reviewed and overseen by the Institutional Biosafety Committee. 1. BSL-2 Standard Microbiological Practices Enforce institutional policies that control access to the laboratory; Wash hands after working with potentially hazardous materials and before leaving the laboratory; Eating, drinking, smoking, handling contact lenses, applying cosmetics and storing food for human consumption is not permitted in the laboratory. Food must be stored outside of the laboratory; Mouth pipetting is prohibited; Develop and implement policies for handling sharps, such as needles, scalpels, pipettes, and broken glassware. Whenever practical, laboratory supervisors should adopt improved engineering practice controls that reduce the risk of sharp injuries. Precautions must always be taken with sharp items and include: o Needles must not be bent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated by hand before disposal; o Used disposable needles and syringes must be carefully placed in puncture resistant containers used for sharps disposal; o Non-disposable sharps must be placed in a hard walled container for transport to a processing area for decontamination, preferably autoclaving; and o Broken glassware must not be handled directly. Instead it must be removed using a brush and dustpan, tongs, or forceps. Plastic ware should be substituted for glassware whenever possible. Perform all procedures to minimize the creation of splashes and/or aerosols; Decontaminate work surfaces with appropriate disinfectant after completion of work and after any spill or splash of potentially infectious material; Decontaminate all cultures, stocks, and other potentially infectious materials before disposal using an effective method; Post a sign incorporating the universal biohazard symbol at the entrance to the laboratory when infectious agents are present. Posted information must include: o Laboratory biosafety level; o Supervisor s name; o Telephone number; and o Procedures for entering and exiting the laboratory Ensure an effective integrated pest management program is in place; and BSL-3

4 Ensure laboratory personnel receive appropriate training regarding their duties, the necessary precautions to prevent exposures, and exposure evaluating procedures. Personnel must receive annual updates or additional training when procedural or policy changes occur. Laboratory personnel should be provided with information regarding immune competence and conditions that may predispose them to infection. Individual having these conditions should be encouraged to self-identify to UMSL s healthcare provider for appropriate counseling and guidance. 2. BSL-2 Special Practices All persons entering the laboratory must be advised of the potential hazards and meet specific entry/exit requirements; Laboratory personnel must be provided medical surveillance, as appropriate, and offered available immunizations for agents handled or potentially present in the laboratory; Policies and procedures must be established for collecting and storing serum samples from at-risk personnel; A laboratory-specific biosafety manual must be prepared and adopted as policy. The biosafety manual must be made available and accessible; The laboratory supervisor must ensure that laboratory personnel demonstrate proficiency in standard and special microbiological practices before working with BSL-2 agents; Potentially infectious materials must be placed in a durable, leak-proof container during collection, handling, processing, storage, or transport within the facility; Laboratory equipment should be routinely decontaminated, as well as after spills, splashes, or other potential contamination: o Spills involving infectious materials must be contained, decontaminated, and cleaned up by staff properly trained and equipped to work with infectious material; o Equipment must be decontaminated before repair, maintenance, or removal from the laboratory; and o The method of decontamination must be appropriate for the agents in use. Incidents that may result in exposure to infectious materials must be immediately evaluated and treated according to procedures described in the laboratory biosafety manual. All such incidents must be reported to the laboratory supervisor. Medical evaluation, surveillance, and treatment should be provided and appropriate records maintained; Animals and plants not associated with the work being performed must not be permitted in the laboratory; and All procedures involving the manipulation of infectious materials that may generate an aerosol should be conducted in a Biosafety Cabinets or other physical containment devices. BSL-4

5 3. BSL-2 Safety Equipment (Primary Barriers and PPE) Properly maintained Biosafety Cabinets, other appropriate PPE, or other physical containment devices must be used whenever: o Procedures with a potential for creating infectious aerosols or splashes are conducted. These may include pipetting, centrifuging, grinding, blending, shaking, mixing, sonicating, opening containers or infectious materials, inoculating animals intranasally, and harvesting infection tissue from animals or eggs; or o High concentrations or large volumes of infectious agents are used. Such materials may be centrifuged in the open laboratory using sealed rotor heads or centrifuge safety cups. Protective laboratory coats, gowns, smocks, or uniforms designated for laboratory use must be worn while working with hazardous materials. Remove protective clothing before leaving for non-laboratory areas. Dispose of protective clothing appropriately, or deposit it for laundering. It is recommended that laboratory clothing not be taken home; Eye and face protection (goggles, mask, face shield or other splatter guard) is used for anticipated splashes or sprays of infectious or other hazardous materials when the microorganism must be handled outside the BSC or other containment device. Eye and face protection must be disposed of with other contaminated laboratory waste or decontaminated before reuse. Persons who wear contact lenses in laboratories should also wear eye protection; Gloves must be worn to protect hands from exposure to hazardous materials. Glove selection should be based on appropriate risk assessment. Alternatives to latex gloves should be available. In addition, BSL-2 laboratory workers should: o Change gloves when contaminated, glove integrity is compromised or when otherwise necessary; o Remove gloves and wash hands when work with hazardous materials has been completed and before leaving the laboratory; and o Do not wash or reuse disposable gloves. Dispose of used gloves with other contaminated laboratory waste. Hand washing protocols must be rigorously followed. Eye, face and respiratory protection should be used in rooms containing infected animals as determined by the risk assessment. 4. BSL-2 Laboratory Facilities (Secondary Barriers) Laboratory doors should be self-closing and have locks in accordance with UMSL standards; Laboratories must have a sink for hand washing. The sink may be manually, handsfree, or automatically operated. It should be located near the exit door; The laboratory should be designed so that it can be easily cleaned and decontaminated. Carpets and rugs in laboratories are not permitted; Laboratory furniture must be capable of supporting anticipated loads and uses. Spaces between benches, cabinets, and equipment should be accessible for cleaning: BSL-5

6 o Bench tops must be impervious to water and resistant to heat, organic solvents, acids, alkalis, and other chemicals; and o Chairs used in the laboratory must be covered with non-porous material that can be easily cleaned and decontaminated with appropriate disinfectant. Laboratory windows that open to the exterior are not recommended. However, if a laboratory does have windows that open to the exterior, they must be fitted with screens; BSCs must be installed so that fluctuations of the room air supply and exhaust do not interfere with proper operation. BSCs should be located away from doors, windows that can be opened, heavily traveled laboratory areas, and other possible airflow disruptions; Vacuum lines should be protected with liquid disinfectant traps; An eyewash station must be readily available; HEPA filtered exhaust air from a Class II BSC can be safely recirculated back into the laboratory environment if the cabinet is tested and certified at least annually and operated according to the manufacturer s recommendations. Each department/pi is responsible for ensuring that this certification is performed. BSCs can also be connected to the laboratory exhaust system by either a thimble (canopy) connection or directly exhausted to the outside through a hard connection. Provisions to assure proper safety cabinet performance and air system operation must be verified; and A method for decontaminating all laboratory wastes should be available (i.e., autoclave, chemical disinfection, incineration, or other validated decontamination method). BSL-6

7 C. PLANT BIOSAFETY LEVELS (BL-P) A plant biosafety level is a combination of physical and biological containment practices to reduce the public health threat to humans or animals, and avoid an unintentional transmission or release of regulated plant material into the environment. The BL-P levels of containment were specifically devised to describe containment for transgenic plants. There are four BL-Ps; BL1-P to BL4-P. As the BL-P increases, so does the level of protection and stringency of physical and/or biological containment practices used to minimize the impact on natural ecosystems outside the facility. UMSL only has a BL1-P facility. 1. BL1-P BL1-P is a low level containment that is designated for experiments that are deemed a low risk to the environment. This designation also applies to plant associated microorganisms that are considered to have a minimal impact on the environment and are not easily disseminated. Some examples include: plants that are not noxious weeds, plants with no potential for out-crossing with related species, and Agrobacterium-mediated transfer of innocuous genetic material. Work involving other organisms that require a containment level of BL1-P or lower may be conducted concurrently in the greenhouse bay as long as all work is conducted using BL1-P practices. 2. General BL1-P Practices Access to the greenhouse shall be limited or restricted, when experiments are in progress; Personnel shall be required to read and follow instructions on BL1-P greenhouse practices and procedures prior to entering the greenhouse; All procedures shall be performed in accordance with accepted greenhouse practices that are appropriate to the experimental organism; Experimental organisms shall be rendered biologically inactive by appropriate methods before disposal outside of the greenhouse facility; A program shall be implemented to control undesired species (e.g., weed, rodent, or arthropod pests and pathogens), by methods appropriate to the organisms and in accordance with applicable state and Federal laws; If macroorganisms (e.g., flying arthropods or nematodes) are released within the greenhouse, precautions shall be taken to minimize escape from the greenhouse facility; The greenhouse floor may be composed of gravel or other porous material. At a minimum, impervious (e.g., concrete) walkways are recommended; and Windows and other openings in the walls and roof of the greenhouse facility may be open for ventilation as needed for proper operation and do not require any special barrier to contain or exclude pollen, microorganisms, or small flying animals (e.g., arthropods and birds); however, screens are recommended BSL-7