32 C.F.R. Subpart C—Operational Requirements
Title 32 - National Defense
(a) Medical. Before to assignment to work with etiologic agents, personnel will be evaluated by the appropriate medical personnel with respect to their assignment and will be evaluated in the medical surveillance program described in §627.8. (b) Training. All personnel directly or indirectly involved with containment or handling of known and potentially biohazardous material shall receive instruction that adequately prepares them for their assigned duties. Training will be given by occupationally qualified personnel as determined by the commander. This training will be documented and will include— (1) General training— (i) Personal hygiene related to laboratory work. (ii) Laboratory practices. (iii) Personal protective equipment. (iv) Effective use of engineering controls. (v) Packaging, transportation, and shipment of etiologic agents (when applicable). (vi) Hazardous and infectious waste disposal, handling, and minimization procedures. (2) Training conducted specifically for the facilities that the individual will be working in, including— (i) Procedures for the facility. (ii) Reporting incidents and accidents. (iii) Labeling and posting of signs. (iv) Biohazardous waste handling, approaches to minimizing the volume of waste, decontamination, packaging, and disposal. (v) Emergency procedures. (3) Additional general training required for work in facilities where viable etiologic agents are present. (i) Aseptic technique and procedures to include hands-on instruction and demonstration of proficiency. (ii) Concept and definition of biosafety levels. (iii) Disinfection and sterilization. (iv) Safe use of workplace equipment, for example autoclave and centrifuge. (v) Monitoring and auditing requirements. (vi) Precautions for handling blood, tissues, and body fluids (when applicable). (vii) The infectivity, pathogenicity, mode(s) of transmission, and medical surveillance requirements of specific agents. (viii) Training for all new employees will include a period of supervised orientation in the facilities by a scientist or technician with specific training in the procedures and properties of the etiologic agents in use. During the training period, new laboratory personnel will be under the constant supervision of appropriately trained personnel. (ix) Personnel who are assigned tasks in BL–2, BL–3, or BL–4 facilities will also have specific training in handling pathogens. (x) Personnel assigned duties in a BL–4 facility will also have specific and thorough training in handling extremely hazardous infectious agents, the primary and secondary containment functions of standard and special practices, use of personal protective equipment, containment equipment, and laboratory design characteristics. (4) Additional general training for handling toxins will include relevant items from §627.10 plus— (i) The availability of reference material on the hazards and safe handling of toxic substances. (ii) The biological effects of the toxin(s) in use. (a) Evaluation of the risks. The risk assessment of laboratory activities involving the use of etiologic agents is ultimately a subjective process. Those risks associated with the agent, as well as with any adjunct elements of the activity to be conducted, (chemicals, radioisotopes, end-products, and so forth) must be considered in the assessment. The appropriate biosafety level for work with a particular agent or animal study depends on the virulence, pathogenicity, biological stability, route of transmission, and communicability of the agent; the nature of the laboratory; the procedures and manipulations to be used; the quantity and concentration of the agent; and the availability of effective vaccines or therapeutic measures. (b) The characteristics of etiologic agents, primary laboratory hazards of working with the agent, and recommended biosafety levels are described by CDC-NIH (HHS publication No. (NIH) 88–8395), the considerations for recombinant DNA molecules are described by NIH, and those for oncogenic viruses are described by NCI-NIH (sources listed below). The commander or institute director will assign work with given etiologic agents to the appropriate biosafety level. A risk assessment should take into account not only the NIH Guidelines for Research Involving Recombinant DNA Molecules, but also potential hazards associated with the organism and the product of the experimentation. (1) When established guidelines exist, these will be followed. The primary source guidelines are— (i) HHS Publication No. (NIH) 88–8395, Biosafety in Microbiological and Biomedical Laboratories, as amended, and updates published in Morbidity and Mortality Weekly Report. (ii) NIH Guidelines for Research Involving Recombinant DNA Molecules (FR 51: 16958–16985 and updates). (iii) The publication by the American Committee on Arthropod-Borne Viruses Subcommittee on Arbovirus Laboratory Safety (SALS) entitled Laboratory Safety for Arboviruses and Certain Other Viruses of Vertebrates in the American Journal of Tropical Medicine and Hygiene, 29(6), 1980, pp. 1359–1381. (iv) The Department of Health and Human Services Publication No. (NIH) 76–1165 by the National Cancer Institute (NCI) entitled Biological Safety Manual for Research Involving Oncogenic Viruses. (2) When samples with unidentified viable agents are obtained, a knowledgeable and qualified scientist will evaluate the risks and make recommendations to the safety officer, who will add recommendations for review and approval by the commander or institute director. When guidelines for a specific organism are not established, in addition to these steps, the CDC or SALS or both will be consulted. Their recommendations will be documented and provided to the commander or institute director before approval. (c) Selection of facilities. The facility requirements identified by the risk assessment will be adhered to. Any variations and compensatory measures will be approved by the IBC (when recombinant DNA molecules are involved), the safety officer, and the commander or institute director before a request for an exception or waiver is submitted as stated in AR 385–69. (d) Policies and procedures. Policies in the form of a laboratory safety manual, regulations, memorandums, or SOPs are required for work with etiologic agents in the BDP. Before beginning a new procedure, the policies and procedures will be reviewed to ascertain that the intended operations are described and to determine the requirements that apply to the operation. If procedures exist for the intended operation, personnel will be trained to follow them; if procedures do not exist, then a detailed SOP will be written, reviewed, and approved before beginning the operation. SOPs will conform to the requirements stated in §627.7(d), and be signed by all personnel who are required to follow the procedures, thus acknowledging that they have read and understood the contents. All SOPs that pertain to a specific area (room, laboratory, or suite) will be available at the worksite. The general requirements for use of etiologic agents are composed of two sets of requirements, with the requirements for toxins being a subset of the requirements for handling viable etiologic agents. These requirements are as follows— (a) General techniques applicable to etiologic agents. (1) A fully fastened long-sleeved laboratory coat, gown, uniform, or coveralls will be worn in laboratories or animal rooms. (2) Eating, drinking, smoking, and applying cosmetics are not permitted in the work areas. (3) Personnel must wash their hands after they handle etiologic agents or animals, and before leaving the laboratory area. (4) Mouth pipetting is strictly prohibited. Mechanical pipetting aids must be used. (5) Gloves— (i) Will be worn when manipulating etiologic agents and handling containers of etiologic agents. Gloves are not required when materials are packaged appropriately for shipment. (ii) Will be selected based on the hazards. (iii) Will be changed frequently (or decontaminated frequently), and will be decontaminated or discarded into a labeled biohazard container after each use and immediately upon observable direct contact with an etiologic agent. (iv) Will be removed at the work-space (workbench or hood) after handling etiologic agents to ensure that doorknobs and other surfaces are not contaminated. (6) Good housekeeping will be maintained. This includes— (i) Work areas free of clutter. (ii) Work environment free of tripping hazards, with adequate access to exits, emergency equipment, controls, and such. (iii) Benches and general work areas will be cleaned regularly using a wet sponge or similar method with disinfectant as appropriate. Methods that stir up dust such as sweeping or using vacuum cleaners, (except for HEPA-filtered vacuum cleaners) are unacceptable. (iv) Specific work areas will be cleaned and decontaminated immediately following each use of an etiologic agent (at least once a day) and after any spill of viable material. (v) Hallways and stairways will not be used for storage. (7) All solutions, reagents, and chemicals will be labeled. (8) All contaminated liquid or solid wastes will be inactivated before disposal. (9) Work will be conducted over spill trays or plastic-backed absorbent paper. The paper will be removed, decontaminated, or disinfected, and the general area wiped with decontaminant at the end of each day or at the end of the experiment, whichever occurs first. (10) Etiologic agents will be kept in closed containers when not in use. Cultures, solutions, or dried etiologic agents in glass vessels transported or incubated within a room or suite will be handled in nonbreakable, leak-proof pans, trays, pails, carboys, or other secondary containers large enough to contain all the material, if the glass vessel leaks or breaks. Etiologic agents removed from a room or suite for transport to another approved area within the same building will be placed in a closed unbreakable secondary container before removal from the laboratory. The secondary container will be labeled on the exterior with a biohazard symbol and identification of the contents, including the required biosafety level, the scientific name, the concentration (if applicable), and the responsible individual. The secondary containers will be wiped with suitable disinfectant before removal from the laboratory or area. (11) Working stocks of etiologic agents will be stored in double containers. The primary and secondary containers will provide a positive seal and the secondary container will be unbreakable. The secondary container will be labeled as stated in §627.12 (a)(10) and with the date stored. (12) Storage units (for example, freezers, refrigerators, cabinets, and hoods) will be labeled with the universal biohazard sign and indicate the classes of etiologic agents contained in them. Storage units will be secured when not in use. (13) All contaminated materials, containers, spills, and solutions will be decontaminated or disinfected by approved methods before disposal. (14) After injection of an etiologic agent into animals, the site of injection will be swabbed with a decontaminant. (15) Syringes. (i) Reusable or disposable syringes will be of the fixed needle or LUER-LOK type (or equivalent) to assure that the needle cannot separate during use. (ii) After use, nondisposable glass syringes with attached needles contaminated with etiologic agents will be submerged in a container of decontaminant. Disposable syringes will be discarded with needles attached in puncture-proof rigid containers. Needles will not be recapped after use. (iii) Sterilized or decontaminated containers marked “Syringes and/or Needles” may be deposited in appropriate refuse containers after proper packaging and destruction of the contents. [Note: Many States, especially those on the Eastern seaboard, have implemented strict requirements for the disposal of medical wastes. For example, Maryland has designated all waste from a microbiological laboratory as hazardous waste with licensing requirements for generators of 50 kilograms per month or more of waste, while all medical waste released for transport off-site must be manifested to a State licensed medical waste hauler with the destination specified. Additionally, in some cases, the local government (for example, a city) regulates the disposal of these wastes. These requirements will be identified and followed.] Needles or syringes may not be destroyed by clipping. A mechanical shear may be used to smash or sheer needles after or concurrently with sterilization or decontamination. (16) Refrigerators, deep freezers, and dry ice chests should be checked, cleaned out, and defrosted periodically to remove any ampules, tubes, and so forth, containing etiologic agents that may have broken during storage. Rubber gloves and respiratory protection appropriate to the materials in storage should be worn during cleaning. Do not store flammable solutions in nonexplosion proof refrigerators. (b) Additional techniques applicable to work with viable etiologic agents. The major objective of these techniques is to assist in protection against laboratory acquired infections. Air sampling studies have shown that aerosols are generated from most of the manipulations of bacterial and viral cultures common to research laboratories. The generation of aerosols during routine laboratory manipulations must be considered when evaluating the individual degree of risk, keeping in mind the four main factors governing infection: dosage, virulence of the organism, route of infection (for example, skin, eyes, mouth, lungs), and host susceptibility (for example, state of health, natural resistance, previous infection, response to vaccines and toxoids). The requirements stated below are minimum handling requirements to prevent accidental infection created by incidental aerosols. (1) All procedures are performed carefully to minimize the creation of aerosols. (2) No infectious mixtures will be prepared by bubbling air through a liquid. (3) Pipettes. (i) No infectious material will be forcibly ejected from pipettes. Only to deliver (TD) pipettes will be used. (ii) Pipettes used with infectious or toxic materials will be plugged with cotton unless they are used exclusively in a gas-tight cabinet system. (iii) Contaminated pipettes will be placed horizontally in a rigid container containing enough disinfectant for complete immersion. Cylinders used for vertical discard are not recommended. The container and pipettes must be autoclaved as a unit and replaced by a clean container containing fresh disinfectant. (iv) Pipetting devices must be used. Under no circumstances is mouth pipetting permitted. (4) Syringes. (i) Using syringes and needles for making dilutions of etiologic agents is not recommended. (ii) When removing a syringe and needle from a rubber stopper bottle containing viable etiologic agents, an alcohol soaked pledget around the stopper and needle will be used. (iii) Excess fluid and bubbles should be expelled from syringes vertically into a cotton pledget soaked with disinfectant or into a small bottle containing disinfectant-soaked cotton. (iv) The site of injection of an animal will be swabbed with a disinfectant before and after injection. (v) After use, syringes contaminated with residual infectious fluid will be submerged in a container of disinfectant in a safety cabinet prior to removal for autoclaving. To minimize accidental injection of infectious material, the removable needles should remain on such syringes until after autoclaving. When possible, syringes with attached needles should be placed in a pan separate from that holding other discarded materials. (vi) Caps will not be placed over needles until after disinfection. During recapping, procedures to prevent personal injuries will be used. (5) Centrifuges and shakers. (i) Before centrifuging, tubes, rotors, seals, and gaskets will be checked for cleanliness and integrity. In low speed clinical-type centrifuges, a germicidal solution may be added between the tube and trunnion cup to disinfect the outer surfaces of both and to cushion against shocks that might break the tube. Metal or plastic tubes (other than nitro-cellulose) will be used. (ii) Decanting from centrifuge tubes will be avoided. If decanting is necessary, the outer rim will be wiped with a disinfectant after decanting so that material on the lip cannot spin off as an aerosol. Centrifuge tubes will not be filled byond the level the manufacturer recommends. (iii) Broth cultures will be shaken in a manner that avoids wetting the plug or cap. (6) Water baths in which viable etiologic agents are incubated must contain a disinfectant. For cold water baths, 70 percent propylene glycol is recommended. The disinfectant should be changed frequently. (7) When a laboratory vacuum is used to manipulate viable etiologic agents, a secondary reservoir containing disinfectant and a HEPA filter must be employed to ensure that the laboratory vacuum lines do not become contaminated. (8) Test tubes. (i) Tubes containing viable etiologic agents should be manipulated with extreme care. Studies have shown that simple procedures, such as removing a tube cap or transferring an inoculum, can create a potentially hazardous aerosol. (ii) Manipulation of biohazardous test tubes will be conducted in biological safety cabinets. Tubes and racks of tubes containing biohazardous material should be clearly marked. The individual employee must ensure that tubes containing biohazardous material are properly sterilized prior to disposal or glassware washing. Safety test tube trays should be used in place of conventional test tube racks to minimize spillage from broken tubes. When safety test tube trays are not used, the conventional test tube racks will be placed in a tray large enough to contain any potential spill. A safety test tube tray is one having a solid bottom and sides deep enough to hold all liquids, should a test tube break. (9) Care should be exercised when using membrane filters to obtain sterile filtrates of viable etiologic agents. Due to the fragility of the membranes and other factors, such filtrates cannot be considered noninfectious until laboratory culture or other tests have proven their sterility. (10) The preparation, handling, and use of dry powders of viable etiologic agents in open containers presents unusual hazards. The slightest manipulation of such powders can cause the generation of aerosols containing a high concentration of etiologic agents. Therefore, work with dry powders of etiologic agents in open containers should be carried out in gas-tight biological safety cabinets. (a) Requirements beyond those for all etiologic agents. BL–1 operations follow the general techniques described in §§627.12(a) and 617.12(b). (b) Additional laboratory requirement. Contaminated materials that are to be decontaminated at a site away from the laboratory are placed in a durable leak-proof container which is closed before being removed from the laboratory. Examples of suitable containers are metal tubs with lids or plastic bags that are sealed and then placed inside a rigid container for transport. (c) Additional animal requirements. (1) Bedding materials from animal cages will be removed in such a manner as to minimize the creation of aerosols and disposed of in compliance with applicable institutional or local requirements. (2) Cages are washed manually or in a cagewasher. Temperature of final rinse water will be a minimum of 180 °F. (3) Laboratory coats, gowns, or uniforms worn in animal rooms shall not be worn in other areas. (a) Additional requirements. In addition to the general microbiological techniques stated in §627.13, BL–2 operations include the following requirements: (1) When etiologic agents are in use, a hazard warning sign incorporating the universal biohazard symbol is posted on the access door of the work area. The hazard warning sign identifies the etiologic agent, lists the name and telephone number of the institute director or other responsible person(s), and indicates the special requirement(s) for entering the laboratory. (2) Animals not involved in the work being performed are not permitted in the laboratory. (3) Special care is taken to avoid skin contamination with the etiologic agents; gloves will be worn when handling etiologic agents or infected animals. (4) All wastes from laboratories and animal rooms are decontaminated before disposal. (5) Hypodermic needles and syringes are used only for parenteral injection and aspiration of fluids from laboratory animals and diaphragm bottles. (6) Spills and accidents which result in a potential exposure to etiologic agents will be reported immediately to the safety officer, the project leader, and the institute director. (7) Biological safety cabinets (Class I or II) will be used when: (i) Procedures with a high potential for creating infectious aerosols are conducted. (ii) High concentrations or large volumes of etiologic agents are used. (8) Laboratory coats, gowns, smocks, or uniforms will be removed before leaving the animal facility or laboratory area. (b) Additional animal requirements. (1) Cages must be decontaminated, preferably by autoclaving, before they are cleaned and washed. (2) Approved molded masks are worn by all personnel entering animal rooms housing nonhuman primates. (3) If floor drains are provided, the drain traps will be kept filled with water or a suitable disinfectant. (a) Additional requirements. In addition to the requirements stated in §§627.13 and 627.14, the following requirements apply— (1) Approved molded masks or respirators with HEPA filters are worn by all personnel in rooms housing infected animals. (2) Protective clothing worn in a laboratory or animal room will be removed before exiting the laboratory or animal room. (3) Clothing worn in laboratories and animal areas to protect street clothing will be decontaminated before being laundered. (b) Additional laboratory requirements. (1) Laboratory doors will be kept closed. (2) All activities involving etiologic agents will be conducted in biological safety cabinets (Class I, II, or III) or other physical containment devices within the containment module. No work in open vessels is conducted outside a biological safety cabinet. (3) The work surfaces of biological safety cabinets and other containment equipment will be decontaminated after work with etiologic agents. Plastic-backed paper toweling should be used on nonperforated work surfaces within biological safety cabinets to facilitate clean-up. (c) Additional animal requirements. (1) Cages are autoclaved before bedding is removed and before they are cleaned and washed. (2) Gloves are removed aseptically and autoclaved with other wastes before being disposed of or reused. (3) Boots, shoe covers, or other protective footwear and disinfectant foot baths must be available and used when indicated. (4) Personal protective clothing and equipment and other physical containment devices are used for all procedures and manipulations of etiologic agents or infected animals. The risk of infectious aerosols from infected animals or their bedding shall be reduced by housing animals in partial containment caging systems as described in §627.56. (d) Work with BL–3 etiologic agents that require additional secondary containment. Facilities in which work with certain viruses, for example, Rift Valley fever, yellow fever, and Venezuelan equine encephalitis, is conducted require HEPA filtration of Xallexhaust air prior to discharge from the laboratory. All persons working with those agents for which a vaccine is available should be immunized. Laboratory work at BL–4 must follow the requirements stated in §§627.13, 627.14 and 627.15 as well as the following: (a) All activities are conducted in Class III biological safety cabinets or in Class I or II biological safety cabinets in conjunction with a one-piece positive pressure personnel suit ventilated by a life-support system. (b) Biological materials to be removed from the Class III cabinet or from the maximum containment laboratory in a viable or intact state must be transferred to a sealed nonbreakable primary container, enclosed in a nonbreakable sealed secondary container, and removed from the facility through a disinfectant dunk tank, fumigation chamber, or an airlock designed for this purpose. (c) No materials, except for biological materials that are to remain in a viable or intact state, are removed from the maximum containment laboratory unless they have been autoclaved or decontaminated before they leave the facility. Equipment or material which might be damaged by high temperature or steam is decontaminated by gaseous or vapor methods in an airlock or chamber designed for this purpose. (d) Personnel may enter and leave the facility only through the clothing change and shower rooms. Personnel must shower each time they leave the facility. Personnel may use the airlocks to enter or leave the laboratory only in an emergency. (e) Street clothing must be removed in the outer clothing change room and kept there. Complete laboratory clothing, including undergarments, pants and shirts or jumpsuits, shoes, and gloves, will be provided and must be used by all personnel entering the facility. Head covers are provided for personnel who do not wash their hair during the shower. When leaving the laboratory and before proceeding into the shower area, personnel must remove their laboratory clothing and store it in a locker or hamper in the inner change room. (f) When etiologic agents or infected animals are present in the laboratory or animal rooms, a hazard warning sign incorporating the universal biohazard symbol must be posted on all access doors. The sign must identify the agent, list the name of the commander or institute director or other responsible person(s), and indicate any special requirements for entering the area (for example, the need for immunizations or respirators). (g) Supplies and materials needed in the facility are brought in by way of the double-doored autoclave, fumigation chamber, or airlock which is appropriately decontaminated after each use. After securing the outer doors, personnel within the facility retrieve materials by opening the interior doors of the autoclave, fumigation chamber, or airlock. These doors are secured after materials are brought into the facility. (h) Materials (for example, animals and clothing) not related to the experiment being conducted are not permitted in the facility. (i) Whenever possible, avoid using any glass items. The laboratory facilities, equipment, and procedures appropriate for work with toxins of biological origin must reflect the intrinsic level of hazard posed by a particular toxin as well as the potential risks inherent in the operations performed. All toxins must be considered to pose a hazard in an aerosol form. However, most toxins exert their effects only after parenteral exposure or ingestion, and a few toxins present a dermal hazard. In general, toxins of biological origin are not intrinsically volatile. Thus, the laboratory safety precautions appropriate for handling these materials closely parallel those for handling infectious organisms. The requirements in this section for the laboratory use of toxins of biological origin include the requirements in §627.12(a) and the following: (a) Vacuum lines. When vacuum lines are used with systems containing toxins, they will be protected with a HEPA filter to prevent entry of toxins into the lines (or sink drains when water aspirators are used). (b) Preparation of concentrated stock solutions and handling closed primary containers of dry toxins. Preparation of primary containers of toxin stock solutions and manipulations of closed primary containers of dry forms of toxins will be conducted— (1) In a chemical fume hood, a glove box, or a biological safety cabinet or equivalent containment system approved by the safety officer. (2) While wearing eye protection if using an open-fronted containment system. (3) Ensuring that gloves worn when handling toxins will be disposed of as toxin waste, with decontamination if required. (4) With the room door closed and posted with a universal biohazard sign, or other sign, indicating that toxin work is in progress. Extraneous personnel shall not be permitted in the room during operations. (5) Ensuring that toxins removed from hoods or biological safety cabinets are double-contained during transport. (6) After verification of hood or biological safety cabinet inward airflow is made by the user before initiating work. (7) Within the operationally effective zone of the hood or biological safety cabinet. (8) Ensuring that nondisposable laboratory clothing is decontaminated before release for laundering. (9) Ensuring that all individuals who handle toxins wash their hands upon each exit from the laboratory. (10) With two knowledgeable individuals present whenever more than an estimated human lethal dose is handled in a syringe with a needle. Each must be familiar with the applicable procedures, maintain visual contact with the other, and be ready to assist in the event of an accident. (c) Manipulations with open containers of dry forms of toxins. Handling dry forms of toxins in uncovered containers (for example, during weighing) will be performed following the requirements stated in §§627.12(a), 627.17 (a) and (b), and the following: (1) Manipulations will be conducted in a HEPA filtered chemical fume hood, glove box, or biological safety cabinet. In addition the exhaust may be charcoal filtered if the material is volatile. (2) When using an open-fronted fume hood or biological safety cabinet, protective clothing, including gloves and a disposable long-sleeved body covering (gown, laboratory coat, smock, coverall, or similar garment) will be worn so that hands and arms are completely covered. Eye and approved respiratory protection is also required. The protective clothing will not be worn outside of the laboratory and will be disposed of as solid toxin waste. (3) Before containers are removed from the hood, cabinet, or glove box, the exterior of the closed primary container will be decontaminated and placed in a clean secondary container. (4) When toxins are in use, the room will be posted to indicate “Toxins in Use—Authorized Personnel Only.” Any special entry requirements will be posted on the entrance(s) to the room. (5) All operations will be conducted with two knowledgeable individuals present. Each must be familiar with the applicable procedures, maintain visual contact with the other, and be ready to assist in the event of an accident. (6) Individuals handling toxins will wash their hands upon leaving the laboratory. (d) Additional considerations of specific toxin properties. The following requirements are in addition to the requirements stated in the paragraphs above. Determine whether the material fits §627.17 (b) or (c), and complies with the appropriate section and the following when applicable: (1) When handling dry forms of toxins that are electrostatic— (i) Do not wear gloves (such as latex) that help to generate static electricity. (ii) Use glove bag within a hood or biological safety cabinet, a glove box, or a class III biological safety cabinet. (2) When handling toxins that are percutaneous hazards (irritants, necrotic to tissue, or extremely toxic from dermal exposure)— (i) Gloves will be selected that are known to be impervious to the toxin and the diluent (when applicable) for the duration of the manipulations. (ii) Disposable laboratory clothing will be worn, left in the laboratory upon exit, and disposed of as solid toxin waste. (e) Aerosol exposures. The requirements found in §627.17 (a) and (b) will be complied with plus the following: (1) Chambers, nose-only exposure apparatus, and generation system must be placed inside a fume hood, glove box, or a Class III biological safety cabinet. Glove boxes and Class III biological safety cabinets will have HEPA filters on both inlet and outlet air ports. (2) The atmosphere from within the exposure chamber will be HEPA filtered before release inside the hood, glove box, or cabinet. (3) All items inside the hood, glove box, or Class III biological safety cabinet will be decontaminated upon removal. Materials such as experimental samples that cannot be decontaminated directly will be placed in a closed secondary container, the exterior of which will be decontaminated and labeled appropriately. Animals will have any areas exposed to toxin wiped clean after removal from the exposure apparatus. (4) The interior of the hood, glove box, or cabinet containing the chamber and all items will be decontaminated periodically, for example, at the end of a series of related experiments. Until decontamintated, the hood, box, or cabinet will be posted to indicate that toxins are in use, and access to the equipment and apparatus restricted to necessary, authorized personnel. (a) Introduction. All laboratories will establish specific emergency plans for their facilities. Plans will include liaison through proper channels with local emergency groups and with community officials. These plans will include both the building and the individual laboratories. For the building, the plan must describe evacuation routes, facilities for medical treatment, and procedures for reporting accidents and emergencies. The plans will be reinforced by drills. Emergency groups and community officials must be informed of emergency plans in advance of any call for assistance. See AR 385–69. (b) General emergency procedures. The following emergency procedures will be followed for laboratory accidents or incidents— (1) Using appropriate personal protection, assist persons involved, remove contaminated clothing if necessary, decontaminate affected areas, and remove personnel from exposure to further injury if necessary; do not move an injured person not in danger of further harm. Render immediate first aid if necessary. (2) Warn personnel in adjacent areas of any potential hazards to their safety. (3) In case of fire or explosion, call the fire department or community fire brigade immediately. Follow local rules for dealing with incipient fire. Portable fire extinguishers will be made available with instructions for their use. Fire fighters responding to the fire scene will be advised to wear a self-contained positive pressure breathing appartus to protect themselves from toxic combustion by-products. (4) Laboratories must be prepared for problems resulting from severe weather or loss of a utility service. In the event of the latter, most ventilation systems not supplied with emergency power will become inoperative. All potentially hazardous laboratory work must stop until service has been restored and appropriate action has been taken to prevent personnel exposure to etiologic agents. (5) In a medical emergency, summon medical help immediately. Laboratories without a medical staff must have personnel trained in first aid available during working hours. (6) For small-scale laboratory accidents, secure the laboratory, leave the area, and call for assistance. (7) When handling mixed hazards (for example, a substance or mixture that may be infectious and radioactive, or infectious and chemically toxic), respond with procedures addressing the greater hazard first, and then follow through with those for the lesser hazards to ensure that all appropriate steps have been taken. (c) Evacuation procedures. Building and laboratory evacuation procedures will be established and communicated to all personnel. (1) Emergency alarm system. (i) There will be a system to alert personnel of an emergency that requires evacuation of the laboratory or building. Laboratory personnel must be familiar with the location and operation of alarm equipment. (ii) Isolated areas (for example, cold, warm, or sterile rooms) will be equipped with an alarm or communication system that can be used to alert others outside to the presence of a worker inside, or to warn workers inside of an emergency that requires evacuation. (2) Evacuation routes will be established and an outside assembly area for evacuated personnel must be designated. All individuals should be accounted for. (3) Shut-down and start-up procedures. (i) Guidelines for shutting down operations during an emergency evacuation will be available in writing. Those guidelines will include procedures for handling any power failure emergency. (ii) Written procedures will also be provided to ensure that personnel do not return to the laboratory until the emergency is ended. Those procedures must also contain start-up operations for the laboratory. (iii) All shut-down and start-up procedures will be available to personnel and reviewed semiannually. (4) All aspects of the building evacuation procedur will be tested semiannually with practice drills. (d) Spills. (1) All areas where work with etiologic agents is performed will have designated personnel to respond to a spill and provide protective apparel, safety equipment, and materials necessary to contain and clean up the spill. Protective clothing requirements are described in §627.21. Also, there will be supplies on hand to deal with the spill consistent with the hazard and quantities of the spilled substance. (2) The safety officer will be notified immediately of all spills. The first line supervisor will ensure that proper clean-up techniques are employed. (3) Etiologic agents. (i) A program for responding to spills of etiologic agents will be developed and implemented. This program will contain emergency response procedures for a biological spill, which will be tailored to the potential hazard of the material being used, the associated laboratory reagents involved, the volume of material, and the location of the materials within the laboratory. Generally, the spill should be confined to a small area while minimizing the substance's conversion to an aerosol. The spill will be chemically decontaminated or neutralized, followed by a cleanup with careful disposal of the residue. If the spilled material is volatile and noninfectious, it may be allowed to evaporate but must be exhausted by a chemical hood or ventilation system. (ii) When a mishap occurs that may generate an aerosol of etiologic agents requiring BL–2 (or higher) containment, the room must be evacuated immediately, the doors closed, and all clothing decontaminated, unless the spill occurs in a class II or class III biological safety cabinet. Sufficient time must be allowed for the droplets to settle and the aerosols to be reduced by the air changes of the ventilation system before decontaminating the area. The area will then be decontaminated to prevent exposure to the infectious agents or toxic substances. Reentry procedures to perform the decontamination will conform to §627.18(e). (iii) A spill of biohazardous material within a biological safety cabinet requires a special response and cleanup procedure. Cleanup will be initiated while the cabinet continues to operate, using an effective chemical decontaminating agent. Aerosol generation during decontamination and the escape of contaminants from the cabinet must be prevented. Caution must be exercised in choosing the decontaminant, keeping in mind that fumes from flammable organic solvents, such as alcohol, can reach dangerous concentrations within a biological safety cabinet. (4) Combined radioactive and biological spills. (i) Both the radiation protection officer (RPO) and the safety officer must be notified immediately whenever there is a spill of radioactive biological material, regardless of its size. Laboratory personnel may be expected to clean up the spill. The RPO will direct the cleanup, in accordance with the NRC license for the facility. (ii) The spill will be cleaned up in a way that minimizes the generation of aerosols and spread of contamination. All items used in cleaning up the spill must be disposed of as radioactive waste. (iii) Following cleanup, the area, affected protective clothing, and all affected equipment and supplies must be surveyed for residual radioactive contamination. All potentially affected areas and items that are not disposable will be wipe-tested to verify that unfixed radioactive contamination has been removed. If fixed contamination is found, the RPO will determine the requirements for additional cleanup. (e) Reentry procedures. This section applies when reentry is necessary to clean up a spill outside of a hood or biological safety cabinet, or to decontaminate or service engineering controls that have failed or malfunctioned so that they do not provide the required containment. (1) When agents requiring BL–1 or BL–1 LS containment are involved, the clothing requirements stated in §627.30 (a) or (b) as appropriate will be followed. Individuals will remove the required protective clothing when finished and wash their hands before proceeding to other tasks. (2) When agents requiring BL–2, BL–2 LS, or toxin procedures and containment are involved, personnel will be required to wear the clothing described in §627.30 (c) or (d) as appropriate. Outer protective clothing will be removed and left in the room before exiting and personnel will wash their hands before proceeding on to other activities. (3) When agents requiring BL–3, or BL–3 LS containment are involved, containers for sealing up inner protective clothing and decontaminant will be placed at the room exit. Personnel will be required to wear the clothing described in paragraph 4–10e. When exiting the area after decontamination procedures, individuals will remove their outer layer of protective clothing just before exiting the room. Once outside the room, the inner layer of protective clothing (for example, coverall) will be removed and placed in the container and the inner gloves will be decontaminated before being removed and placed in the container. Personnel will proceed directly to the shower facility to take a complete shower before exiting the facility. (4) When agents requiring BL–4 containment are involved, the following applies as appropriate to the type of BL–4 facility: (i) When a spill requiring clean-up is in an area designed for use with personal positive pressure suits, the entry and exit procedures will be those normally required to enter or exit the area. (ii) When entering a nonsuit area where a spill of etiologic agent has occurred outside the containment of a Class III biological safety cabinet, personnel will wear the clothing as described in §627.30(f). Before entry, decontamination areas will be established. To accomplish this, two step-in decontamination pans with the appropriate disinfectant will be set up [one just inside the room (where the contamination exists) and the second immediately outside the room]. Immediately outside the room, there will also be a sealable container suitable for sealing up the suit and any air lines (if used). (iii) When exiting the room, suited individuals will place all equipment and other items in autoclaves or disinfectant, step into the disinfectant pan, and wash down the exterior of their suits with appropriate disinfectant. When completed, the door to the room will be opened and the individual will step through the doorway into the second disinfectant pan. The suit will be thoroughly rinsed with disinfectant again before moving toward the exit from the facility. The suit (but not the respirator) will be placed in the provided container. The individual will proceed through another doorway before removing the respirator and placing it in a closed container for decontamination. The individual will then proceed directly to the shower area and take a full shower before exiting the area. In case they are needed, personnel will be standing by ready to render assistance. Suited individuals will be visually observed, if possible. When visual observation is not possible, a communications system is required. (f) Mishap reports and investigations. (1) Each institution must have a defined system for reporting laboratory injuries, illnesses, and mishaps, as well as for investigating them. These events will be documented and reported to the appropriate safety, supervisory, and occupational health personnel. Those organizations subject to the regulations promulgated by the OSHA will follow the specific requirements for reporting injuries in the work place contained in those regulations. The requirements stated in AR 385–69, State, and local government requirements for similar reporting will be followed. (2) Form(s) for recording mishaps will be available and completed for all laboratory mishaps. Those reports must include a description of the mishap and any factors contributing to it. In addition, a description of any first aid or other health care given to the employee will be included. Responsibility for completing these forms must be clearly defined in the facility safety manual. Mishaps will be reviewed periodically by the safety officer, the safety committee, the employee health unit, or other appropriate personnel. Individual reports or a summary must be sent, along with recommended changes in laboratory procedure or policy, to the commander or institute director. Policy or procedural changes must be implemented if deemed necessary by the commander or institute director. (3) Any mishaps with etiologic agents used under sponsorship of the BDP that result in sero-conversion or a laboratory-acquired illness will be reported. (a) Large-scale. In addition to the requirements stated in §627.13, the following applies to research or production activities involving viable etiologic agents in quantities greater than 10 liters: (1) All large-scale operations will be conducted in facilities described in §627.47. (2) Cultures will be handled in a closed system. (3) Sample collection, the addition of materials, and the transfer of culture fluids shall be done in a manner which minimizes the release of aerosols or contamination of exposed surfaces. (4) A closed system or other primary containment equipment that has contained viable organisms shall not be opened for maintenance or other purposes unless it has been sterilized. (5) SOPs will include a section describing and requiring a validation of the process equipment's proper function. (6) Scientists, technicians, equipment workers, and support personnel with access to the large-scale production area during its operation will be included in the medical surveillance program. (b) BL–2—LS. In addition to the requirements stated in §§627.19(a) and 627.14, the following procedures will be employed for BL–2—LS: (1) Rotating seals and other mechanical devices directly associated with the closed system used for the propagation and growth of viable organisms shall be designed to prevent leakage or shall be fully enclosed in ventilated housings that are exhausted through filters which have efficiencies equivalent to HEPA filters or through other equivalent treatment devices. (2) A closed system used for the propagation and growth of viable organisms and other primary containment equipment used to contain operations involving viable organisms shall include monitoring or sensing devices that monitor the integrity of containment during operations. (3) Systems used to propagate and grow viable organisms shall be permanently identified. This identification shall be used in all records reflecting testing, operation, and maintenance and in all documentation relating to the use of this equipment. (c) BL–3—LS. In addition to the requirements stated in §§627.19(a) and 617.14, the following procedures apply: (1) Personnel entry into the controlled area shall be through the entry area specified in §627.47(c)(1). (2) Persons entering the controlled area shall exchange or cover their personal clothing with work garments such as jumpsuits, long sleeved laboratory coats, pants and shirts, head cover, and shoes or shoe covers. On exit from the controlled area, the work clothing may be stored in a locker separate from that used for personal clothing, or discarded for laundering. Clothing shall be decontaminated before laundering. (3) Entry into the controlled area during periods when work is in progress shall be restricted to those persons required to meet program support needs. (4) Prior to entry, all persons shall be informed of the operating practices, emergency procedures, and the nature of the work conducted. (5) The universal biohazard sign shall be posted on entry doors to the controlled area and all internal doors. The sign posted on the entry doors to the controlled area shall include a statement of agents in use and personnel authorized to enter. (6) Equipment and materials required for the management of accidents involving viable organisms shall be available in the controlled area. (d) BL–4—LS. Guidelines for these operations are not established. If these are needed, they must be established by the United States Army Surgeon General or the NIH on an individual basis. Operations that combine etiologic agents with radioactive material present unique problems. When this is the case, the following apply: (a) Radiation program. A radiation program meeting the requirements of AR 385–11 and NRC licensing that allows the particular isotope and its use are required. The requirements for acquisition, handling procedures, labeling, storage, training, monitoring, and disposal will be described in an organization policy document. (b) Procedure approval. In addition to the required approvals for work with etiologic agents, the RPO will approve all SOPs involving the use of radioactive materials. Laboratory operators must be fully trained, with annual training updates as required by the existing license. (c) Special situations. (1) The laboratory waste must be segregated as radioactive waste and disposed of as such after it has been decontaminated. Do not mix nonradioactive waste with radioactive waste as the disposal of radioactive waste is much more complex and expensive. When RCRA-listed chemicals are mixed with radioactive waste, it becomes “mixed waste” for which there is currently no means of disposal. (2) Activities conducted with radioisotopes should be confined to the smallest number of areas or rooms consistent with requirements. (3) Decontamination methods specific to etiologic agents will not always remove radioactivity. Other methods, such as specialized detergents and solvents designed for this use, should be employed to remove residual radioactivity.
Title 32: National Defense
PART 627—THE BIOLOGICAL DEFENSE SAFETY PROGRAM, TECHNICAL SAFETY REQUIREMENTS (DA PAMPHLET 385–69)
Subpart C—Operational Requirements
§ 627.10 Personnel prerequisites.
§ 627.11 Operational prerequisites.
§ 627.12 General laboratory techniques.
§ 627.13 Biosafety level 1.
§ 627.14 Biosafety level 2.
§ 627.15 Biosafety level 3.
§ 627.16 Biosafety level 4.
§ 627.17 Toxins.
§ 627.18 Emergencies.
§ 627.19 Large-scale operations.
§ 627.20 Operations with radioactive material.
