32 C.F.R. Subpart H—Engineering Controls
Title 32 - National Defense
As required by the OSHA and recommended by the American Industrial Hygiene Association (AIHA) and the CDC, engineering controls and proper microbiological techniques are the primary means of protecting personnel who work with potentially hazardous biological materials. In situations of potentially higher hazard, these engineering controls are supplemented by personal protective clothing and equipment. Thus, the engineering controls discussed in this chapter will be the primary means of personnel and environmental protection when working with etiologic agents. Because of the importance of these engineering controls, this chapter contains not only requirements for the engineering and construction of these controls, but also requirements for their certification and continuous satisfactory performance. These will be described for each engineering control. (a) Description. The Class I biological safety cabinet (figure H-I in appendix F to this part) is a ventilated cabinet for personnel protection only. The cabinet provides an uncirculated inward flow of air away from the operator. The exhaust is passed through a HEPA filter. It may be discharged into the laboratory or vented out of the laboratory and dispersed away from occupied spaces or air intakes. When the exhaust is recirculated in a BL–2 or BL–3 facility, the cabinet must be tested and certified annually. In a BL–4 facility, if the exhaust is recirculated, the cabinet must be tested and certified semiannually. (b) Uses. These cabinets are used if personnel protection against the microorganisms is required; for modest quantities of volatile, toxic, or radioactive chemicals (in concentrations and quantities associated with biological systems) if vented to the outside; and when sterility is not required. They are commonly used for housing tabletop centrifuges, in the necropsy of small animals, and for changing animal bedding. (c) Prohibitions. This class of cabinet is not to be used when sterility must be maintained. In addition, volatile, toxic, or radioactive materials can not be used in this class of cabinet when the exhaust air is not exhausted to the exterior. (d) Certifications and requirements. (1) The inward air velocity on these cabinets will be an average of 100 plus or minus 20 linear feet per minute (lfpm). Each cabinet must be certified before use and semiannually thereafter by a face velocity test. Additionally, smoke tests will be performed annually to verify containment. (2) The exhaust system will have a HEPA filter, which will be tested initially upon installation, after repair or replacement, and every 2 years thereafter (except when required more often). Filters will be certified to be 99.97 percent effective in capturing particulate matter by a leakage test using mineral oil or other appropriate aerosol dispersed as 0.3 micron droplets. All Class II biological safety cabinets (figure H-II in appendix F to this part) are ventilated cabinets for personnel and product protection, having an open front with inward air flow for personnel protection. (a) Operating standards. (1) All of these cabinets must conform and be certified to meet National Sanitation Foundation (NSF) Standard No. 49 revised, June 1987, for the applicable type of cabinet. (2) After installation and before use, and annually thereafter, the cabinets will be tested in accordance with NSF Standard No. 49 (latest revision June 1987) as follows: (i) Primary (required) tests— (A) Velocity profile test. (B) Work access opening airflow (face velocity) test. (C) HEPA filter leak test. (D) Cabinet integrity test (soap bubble test) for cabinets with positive pressure internal plenums. (ii) Secondary (optional) tests— (A) Vibration test. (B) Electrical leakage and ground circuit resistance tests. (C) Noise level test. (D) Lighting intensity test. (E) UV light intensity test. (3) After repairs or alterations to the cabinetry or ventilation system that affect the cabinet, the tests listed in §627.51(a)(2) will be performed for the relevant parameters. (4) The work access opening airflow (face velocity) test, as specified in NSF Standard No. 49 (latest revision, June 1987), will be performed to check that the cabinet is within specifications on an annual basis for BL–1 and BL–2 and toxin use. This test will be performed semiannually on cabinets used for BL–3 and BL–4 as well as for work with dry forms of toxins. (5) When the exhaust is recirculated in a BL–4 facility, the cabinet must be tested and certified semiannually. (b) Class IIA biological safety cabinets—(1) Description. A Class IIA biological safety cabinet is one in which typically 70 percent of the air is recirculated within the cabinet and the exhaust passes through a HEPA filter before discharge. The exhaust may be exhausted into the room and positive-pressure contaminated ducts and plenums within the cabinet are allowed. Type A cabinets shall have a minimum calculated face velocity of 75 feet per minute (fmp). (2) Uses. These cabinets are for working with low-to-moderate risk biological samples and for protecting personnel against biological material while providing a sterile atmosphere in which to handle the material. (3) Prohibitions. Materials that are toxic or volatile must not be used in these cabinets. (c) Class IIB1 biological safety cabinets—(1) Description A Class IIB1 biological safety cabinet is one that maintains a minimum average inflow of air of 100 plus or minus 20 lfpm and in which typically 30 percent of the air is recirculated. All recirculated and exhausted air passes through two HEPA filters in series. All contaminated internal ducts and plenums are under negative pressure. Type B cabinets shall have a minimum calculated face velocity of 100 fpm. (2) Uses. When ultra-sterility is needed, these are the cabinets of choice. The double filtration achieves a cleaner atmosphere. Minute quantities of volatile, toxic, or volatile radioactive materials coincidental to use in biological systems may also be used in these cabinets. (3) Prohibitions. More than minute quantities of toxic, volatile, or radioactive materials must not be used in these cabinets. (4) Additional certifications or requirements. None. (d) Class IIB2 biological safety cabinets—(1) Description. A Class IIB2 biological safety cabinet is one that maintains a minimum average of 100 plus or minus 20 lfpm inward flow and in which all air is exhausted directly from the cabinet through a HEPA filter without recirculation within the cabinet. All contaminated ducts and plenums are under negative pressure. Type B cabinets shall have a minimum calculated face velocity of 100 fpm. (2) Uses. These cabinets are recommended when small quantities of volatile, flammable, or toxic chemicals must be used coincidentally with items requiring sterility. (3) Prohibitions. While these cabinets do offer the greatest degree of safety for volatile, toxic, and flammable chemical handling in a sterile environment, they are not to be used in place of a fume hood to prepare stock solutions of hazardous chemicals. (e) Class IIB3 biological safety cabinets—(1) Description. A Class IIB3 biological safety cabinet is one that meets all of the requirements of a Class IIB2 biological safety cabinet except that it recirculates most (typically 70 percent) of the air inside the cabinet. Type B cabinets shall have a minimum calculated face velocity of 100 fpm. (2) Uses. Minute amounts of nonflammable chemicals can be used coincidentally with low-to-moderate risk biological agents. (3) Prohibitions. Flammable materials and more than minute amounts of toxic, radioactive, or volatile chemicals must not be used in these cabinets. (4) Additional certifications or requirements. None. (a) Description. These cabinets (figure H-III in appendix F to this part) are totally enclosed, ventilated cabinets of gas-tight construction. Operations are conducted through attached rubber gloves. The supply of air is drawn into the cabinet through HEPA filters. The exhaust air is treated by double HEPA filtration, or by HEPA filtration followed by incineration, and is not allowed to recirculate within the room. (b) Uses. These cabinets provide the ultimate protection for personnel. They are suitable for low, moderate, and high-risk etiologic agents. (c) Prohibitions. More than minute amounts of flammables must not be used in these cabinets. (d) Certifications and requirements. (1) These cabinets will have a manometer or magnehelic gauge that indicates the negative pressure that is maintained inside the cabinet. The pressure inside the cabinet should be a minimum of 0.5 inches water gauge negative to the surrounding room. (2) These cabinets will be pressure tested by the soap bubble or halogen leak test as prescribed in NSF Standard No. 49, appendix B1 (latest revision, June 1987), and certified, when the HEPA filter units are serviced. Fume hoods in which etiologic agents are handled must use proven technologies to provide optimal containment. Fume hood placement, design, and capture testing requirements for use in designing new laboratories can be found in the latest edition of Industrial Ventilation, A Manual of Recommended Practices, published by the American Conference of Governmental Industrial Hygienists. (a) Description. Fume hoods are common chemical laboratory furnishings designed to capture fumes from chemicals that are used within them. Air is drawn through the opening and vented to the exterior without recirculation. (b) Uses. Fume hoods provide excellent containment for handling hazardous chemicals. (c) Prohibitions. Moderate risk biologicals and open containers of dry forms of toxins must not be used in a fume hood without HEPA filtration. Fume hoods should never be used when sterility is required. (d) Certification and requirements. (1) Inward air flow will be an average of 100 plus or minus 20 lfpm as measured at the face of the fume hood. Proper function of laboratory hoods is not only a function of face velocity. An evaluation of the total operating environment is necessary. (2) When filters are required, they will be certified by the mineral oil droplet (HEPA) or Freon (Charcoal) leak test as appropriate. Leakage through the filters will be less than 0.05 percent for Freon and 0.03 percent for oil droplets when initially installed. (3) Fume hoods will be provided with indicator devices to give a warning should the ventilation system fail or if the hood face velocity falls below an average of 80 lfpm (4) Hood air flow will be certified when installed, when maintenance is performed on the ventilation system, and semiannually thereafter. (a) Description. A glove box is an enclosure that provides a positive barrier from liquids, solids, and chemical vapors. A glove box has viewing ports and glove ports for access. The box maintains personnel protection through solid barriers and maintenance of a negative pressure relative to its surroundings. (b) Uses. Glove boxes are used when extreme containment is needed for highly toxic chemicals, especially for dry chemicals that can be swept out of containers by the airflow in hoods. (c) Prohibitions. Unventilated boxes must not be used with volatile flammable materials and should be used with volatile toxic materials unless dilution ventilation is provided. (d) Additional certifications and requirements. (1) The glove box will be maintained at a pressure of at least 0.25 inches water guage less than its surroundings. (2) The pressure differential will be indicated by a manometer or magnehelic guage. Indicator devices will display a loss of pressure below 0.25 inches water guage. (3) Gloves will be changed at appropriate intervals (dependent on the box contents) to ensure they provide the protection needed. (4) Inlets that provide dilution air will be protected by HEPA filters. (a) Description. A ventilated balance enclosure is a box that surrounds a balance and has a small open area for access and handling material in the front. Air is exhausted out the rear of the enclosure. (b) Uses. A ventilated balance enclosure is used when containment of a balance is required to weigh hazardous materials that have a low vapor pressure (such as toxins). These enclosures are also used when it is best to use the balance in other than a fume hood (due to the turbulence and vibration) and when biological safety cabinets or glove boxes are inappropriate or unavailable. Dry forms of toxins may be weighed in these enclosures. (c) Prohibitions. Very volatile or highly toxic volatile materials must not be handled in ventilated balance enclosures unless they are placed in closed containers in a properly functioning fume hood before being transferred to the balance enclosure. (d) Additional certifications or requirements. (1) The flow through the openings in the enclosure will be at least 60 lfpm and must average between 60 and 80 lfpm. (2) Containment will be certified prior to first use and annually thereafter by smoke tubes. (3) The air flow will be certified initially and semiannually by averaging readings taken from the face of the opening. There are a number of cage-ventilated enclosures in which infected animals may be housed at levels corresponding to the various classes of biological safety cabinets. A brief description of four different types of animal ventilated cages is given below. This is not a complete description of all the different animal ventilated cages available. The proper functioning of these will be tested initially, upon each connection to exhaust sources, and at least annually. The inward flow rates on the partial containment systems and pressure checks on the total containment cages will be performed. Prior to selecting such equipment, an evaluation of the function and the equipment should be made, and the methods for testing and decontamination should be analyzed and documented. (a) Filter-top cages. Small laboratory animal polystyrene or polycarbonate cage bottoms are fitted with a dome shaped glass fiber or polyester filter cage cover. The dome shaped filters help reduce the dissemination of aerosols, and the spread of infectious agents. Adequate ventilation around cages fitted with a dome shaped filter is essential since they may contain elevated ammonia and carbon dioxide levels, and high temperature and humidity. Ventilation recommendations in the NIH publication 86–23, 1985 “Guide for the Care and Use of Laboratory Animals” will be followed. (b) Forced ventilation cages. This is a small HEPA-filtered cage connected to a centralized exhaust system. A minimum airflow of 0.03 m3 /min per cage is required. Ventilation rates may vary with the size of the cage, and the number and type of animals being housed. (c) Cubicle-type isolation cage. This is a partial containment unit which holds several animal cages. This unit is a negative pressure HEPA-filtered stainless steel cage. A minimum airflow of 0.3 m3 /min per cage is required for a 0.24 m3 unit. Ventilation rates may vary with the size of the cage and the number and type of animals being housed. (d) Total containment cage. This unit is a negative pressure or positive pressure HEPA-filtered stainless steel cage which has the filters incorporated into the design. It is halogen gas-leak tight and can be considered a Class III biological safety cabinet. A minimum airflow of 0.3 m3 /min per cage is required for a 0.24 m3 unit. Ventilation rates may vary with the size of the cage, and the number and type of animals being housed. Ventilated cage areas within a room that are solid-walled and bottomed areas for containing multiple cages housing infected animals. The containment for these areas is equivalent to the Class I biological safety cabinet. For testing purposes, they will be treated the same as a Class I biological safety cabinet. Publications referenced in this part can be obtained from the National Technical Information Services, U.S. Department of Commerce, 5285 Port Royal Road, Springfield, VA 22161. Required Publications AR 11–34 Army Respiratory Protection Program. (Cited in §§627.31(h)(2) and 627.31(h)(4).) AR 40–5 Preventive Medicine. (Cited in §627.8.) AR 40–10 Health Hazard Assessment Program in Support of the Army Materiel Acquisition Decision Process. (Cited in §627.7(a)(8).) AR 40–12 Medical and Agricultural Foreign and Domestic Quarantine Regulations for Vessels, Aircraft, and Other Transports of the Armed Forces. (Cited in §627.40(a).) AR 40–66 Medical Records and Quality Assurance Administration. (Cited in §627.9.) AR 40–400 Patient Administration. (Cited in §627.8(e).) AR 70–65 Management of Controlled Substances, Ethyl Alcohol, and Hazardous Biological Substances in Army Research, Development, Test, and Evaluation Facilities. (Cited in §§627.36(a)(6) and 627.40(b).) AR 385–10 Army Safety Program. (Cited in §§627.6 and 627.31(h)(4).) AR 385–69 Biological Defense Safety Program. (Cited in §§627.6, 627.7(a), 627.7(a)(8), 627.7(d), 627.11(c), 627.18(a) and 627.18(f)(1).) AR 740–32 Responsibilities for Technical Escort of Dangerous Materials. (Cited in §627.39.) Related Publications A related publication is merely a source of additional information. The user does not have to read it to understand this pamphlet. AR 40–14 Control and Recording Procedures for Exposure to Ionizing Radiation and Radioactive Materials. ANSI Z86.1–1973 Breathing Air ASHRAE Standard 62 Bacterial Toxins: A Table of Lethal Amounts, Gill, D.M., Microbiological Reviews, Volume 46, Number 1; March 1982, pages 86–94. Biohazards Reference Manual American Industrial Hygiene Association, 1985, Clinical Medicine Branch, Division of Host Factors, Center for Infectious Disease, Centers for Disease Control, Atlanta, GA 30333, telephone: (404) 639–3356, Compressed Gas Association Pamphlet G–7.1 Grade D Breathing Air Dangerous Goods Regulations, International Air Transport Association (IATA), Publications Section, 2000 Peel Street, Montreal, Quebec, Canada H3A 2R4, Tel (514) 844–6311. DHEW Pub. No. (NIH) 76–1165 Biological Safety Manual for Research Involving Oncogenic Viruses, Executive Order 12196 Safety and Health Programs for Federal Employees, 26 February 1980 Guide for Adult Immunizations, Published by the American College of Physicians, Guide for Transportation of Hazardous Materials, Vol. 4(1) February 10, 1975. (Copies may be obtained from the Office of Research Grants Inquiries, NIH, Department of Health and Human Services, 5333 Westbard Avenue, Bethesda, MD 20205.) Guidelines for Laboratory Design, Health and Safety Considerations, L. DiBerardinis, et al., John Wiley and Sons, 1987 Guidelines for Prevention of Herpesvirus Simiae (B Virus) Infection in Monkey Handlers, Kaplan, J.E., et al., Mortality and Morbidity Weekly Report, Volume 36, Number 41; October 23, 1987, pages 680–689. HHS Publication No. (NIH) 88–8395, Biosafety in Microbiological and Biomedical Laboratories Industrial Ventilation, A Manual of Recommended Practice Published by the American Conference of Governmental Industrial Hygienists. Laboratory Safety for Arboviruses and Certain Other Viruses of Vertebrates, The American Journal of Tropical Medicine and Hygiene, 29:1359–1381, 1980. NIH Guidelines for Research involving Recombinant DNA Molecules (51 FR 16958, May 7, 1986). NIH publication 86–23, Guide for the Care and Use of Laboratory Animals NSF Standard #49, National Sanitation Foundation Standard Number 49, Class II (Laminar Flow) Biohazard Cabinetry Packaging and Shipping of Biological Materials at ATCC, The American Type Culture Collection (ATCC). (Copies may be obtained from the ATCC, 12301 Parklawn Drive, Rockville, MD 20852. Telephone (301) 881–2600.) Postal Bulletin No. 21246, International Mail-Hazardous Materials Procedures for the Domestic Handling and Transport of Diagnostic Specimens and Etiologic Agents, National Committee for Clinical Laboratory Standards (NCCLS), (H5–A2), Second edition. Vol. 5, No. 1. (Copies may be obtained from the NCCLS, 771 East Lancaster Avenue, Villanova, PA 19085.) Restricted Articles Tariff 6–D, Air Transport Association Technical Instructions for the Safe Transport of Dangerous Goods by Air, International Civil Aviation Organization (ICAO) Intereg Group, 5724 Pulaski Road, Chicago, IL 60646, Tel. (312) 478–0900. The Centers for Disease Control, Office of Biosafety, 1600 Clifton Road NE., Atlanta, Georgia 30333. Telephone (404) 639–3883, or FTS: 236–3883. 9 CFR Parts 102 Through 104, 122 Animals and Animal products. 10 CFR Chapter 1 Nuclear Regulatory Commission. 21 CFR Parts 312, 600 Through 680 Food and drugs. 29 CFR Part 1910 Occupational Health and Safety Administration Safety and Health Standards. 39 CFR Part 111 Postal Service. 40 CFR Parts 1500 Through 1508 Protection of environment. 42 CFR Parts 71 and 72 Public Health Service Foreign Quarantine Regulations. 49 CFR Parts 172 and 173 The Department of Transportation. C–1. The checklist that follows is not an exhaustive list of the items to consider when inspecting facilities where etiologic agents are used. It does provide some basic guidelines to remind safety and nonsafety professionals of the things that need to be considered in the laboratories they manage. The checklist should be used as follows: All area should be inspected using the general list in C–2. Certain items are optional, such as radiation safety. If no radioactive material is present in the room, then this would not be applicable. For BL–1 facilities the list in C–2 is adequate, while BL–2, BL–3, and BL–4 facilities must use the list in C–2 together with the appropriate list in C–3 to C–5. C–2. Basic checklist (a) Housekeeping (1) Is the room free of clutter? (2) Are all aisles from the work areas to the available exits maintained clear of obstructions? (3) Are all safety equipment items unobstructed and ready for use? (4) Is the room clean? (b) Fire safety (1) Is the fire extinguisher hung in its proper place, ready for use, and unobstructed? (2) Are there excess flammables located outside National Fire Protection Association (NFPA) approved cabinetry? (3) Are all Class IA flammables that are in breakable containers in pint or smaller containers? (4) Are all Class IB flammables that are in breakable containers in liter or smaller containers? (c) Chemical safety (1) Are the chemicals stored with compatible materials? (2) Have the chemical fume hoods been certified in the last 6 months? (3) Are the eyewash and deluge shower unobstructed and ready for use? (4) Is the eyewash and deluge shower tested regularly to document proper operation? (5) Is the organic waste container maintained in a closed position? (6) Are all reagents and solutions properly labeled? (7) Is a spill kit within a reasonable distance from the work areas? (8) Is appropriate protective clothing available for the chemical hazards present? (9) Is there a written hazard communication program? (10) Have the personnel in the laboratory been trained in the provisions and principles of the hazard communication program? (11) Are MSDSs located where they are available to the laboratory workers? (12) Is there a written chemical hygiene plan? (d) Radiation safety (1) Are the radioactive materials stored double-contained? (2) Is the containment for the radiation waste container adequate to preclude the spread of radiation? (3) Are all containers appropriately labeled with radiation labels? (4) Are all entrances to the room appropriately labeled? (e) Electrical safety (1) Are excess extension cords being utilized? (2) Are there any frayed cords in the room? (3) Are there any cords on the floor across normal traffic patterns in the room? (f) General laboratory safety (1) Are sharps discarded and destroyed in a safe manner? (2) Are work surfaces decontaminated daily and after a spill? (3) Is the appropriate attire worn by everyone in the room? (4) Is there evidence that personnel eat, drink, smoke, or store food, drinks, or tobacco in the room? (5) Was mouth pipetting observed? (6) Are all gas cylinders secured and are all cylinders not in use capped? (7) Are cylinders of oxidizers stored at least 20 feet from cylinders of flammable gases in the same room? (8) Are the contents of the cylinders clearly labeled? (9) Are the cylinders transported on appropriate dollies or hand trucks? (10) Is there a written respiratory protection program where respirators are used? (g) Etiologic agents (1) Are all containers of etiologic agents appropriately labeled? (i) Are freezers, refrigerators, and similar storage units labeled with the biohazard warning sign? (ii) Are the storage and shipping containers adequate and properly labeled? (2) Have all personnel been adequately trained in general microbiological techniques? (3) Are laboratory doors kept closed when experiments are in progress? (4) Are all operations conducted over plastic-backed absorbent paper or spill trays? C–3. Biosafety level 2 supplemental checklist (a) Are all floor drains filled with water or suitable disinfectant? (b) Is the SOP for an etiologic agent spill signed by all personnel who work with etiologic agents in the room? (c) If biological safety cabinets are used, have they been certified within the last year? (d) Are the appropriate decontaminants available? (e) Are all entrances to the laboratory posted with— (1) The appropriate special provisions for entry? (2) The universal biohazard symbol? (3) The name and telephone number of the laboratory director or other responsible person? (f) Is entry limited and restricted? (g) Are gloves being worn when handling infected animals or infectious or toxic materials? (h) Is eye and respiratory protection being worn in rooms where nonhuman primates are present? (i) If materials are being transported off-site for decontamination, is the containment adequate? C–4. Biosafety level 3 supplemental checklist (a) Is laboratory clothing decontaminated before being sent to the laundry? (b) Are all windows and penetrations through the walls and ceilings sealed? (c) If biological safety cabinets are used, have they been certified within the last year? (d) Are the appropriate decontaminants available? (e) Are all entrances to the facility posted with— (1) The appropriate special provisions for entry? (2) The universal biohazard symbol? (3) The name and telephone number of the laboratory director or other responsible person? (f) Is entry limited and restricted? (g) Are gloves being worn when handling infected animals or infectious or toxic materials? (h) Is eye and respiratory protection being worn in rooms where nonhuman primates are present? (i) Do the monitors indicate that the room is under negative pressure relative to all entrances? (j) Are all vacuum lines protected with HEPA filters and liquid disinfectant traps? (k) Is the autoclave being properly maintained and certified? (l) Is the foot, elbow, or automatic handwash sink operating properly? (m) Are all operations with etiologic agents being conducted inside biological safety cabinets or other approved engineering controls? (n) Are all infected animals housed using appropriate primary containment systems? (o) Do all personnel who enter rooms housing infected animals wear appropriate respiratory protection? (p) Do personnel who exit rooms having infected animals leave their protective clothing in the animal and laboratory rooms? (q) If available, has the UV pass box ouput been certified within the last 3 months? C–5. Biosafety level 4 supplemental inspection checklist (a) Precautions for all areas. (1) Are all penetrations through the walls and ceilings sealed? (2) Are the appropriate decontaminants available and used properly? (3) Are all entrances to the facility posted with— (i) The appropriate special provisions for entry? (ii) The universal biohazard symbol? (iii) The name and telephone number of the laboratory director or other responsible person? (4) Is access to the laboratory controlled strictly and documented? (5) Do the monitors indicate that the room is under negative pressure relative to all entrances? (6) Are all vacuum lines protected with HEPA filters and liquid disinfectant traps? (7) Is the autoclave being properly maintained and certified? (8) Is the foot, elbow, or automatic handwash sink operating properly? (9) Do the self-closing doors to the facility operate properly? (10) Do personnel completely exchange street clothing for laboratory clothing before entry and shower upon exiting? (11) Is the dunk tank disinfectant fresh and appropriate for the agents in use? (b) Suit areas. (1) Are all operations with etiologic agents conducted in Class I or II biological safety cabinets? (2) Do the procedures in place ensure that, as much as possible, the contamination remains inside the cabinets (such as ensuring that everything removed from within the cabinets, such as gloves being worn, instruments, glassware, or similar items, are decontaminated or properly packaged first)? (3) Are the Class I or II cabinets in the facility certified every 6 months? (4) Does the suit decontamination shower have adequate appropriate decontaminant available? (5) Has the suit decontamination shower been used or tested in the last month? (6) Is the ventilated suit air supply and emergency air supply adequate and working properly? (7) Is the emergency alarm system working properly? (8) Are all of the one-piece positive pressure suits available for use in serviceable condition? (9) Are infected animals housed in appropriate primary containment systems? (10) Is the static pressure in the suit area negative to all surrounding areas? (c) Nonsuit areas. (1) Are all operations with etiologic agents conducted inside Class III biological safety cabinets? (2) Were the Class III biological safety cabinets certified before initiating the current operation? (3) Are all infected animals housed in Class III cabinet containment caging systems? D–1. Packaging and Labeling of Etiologic Agents, from HHS publication No. (NIH) 88–8395. D–2. Guidelines for the Air Shipment of Diagnostic Specimens, from the Air Transport Association of America, Cargo Services Division, 1709 New York Ave., NW., Washington, DC 20006. E–1. Permit Application to Import or Transport Agents or Vectors of Human Disease. Department of Health, Education and Welfare, PHS, CDC, Office of Biosafety, Atlanta, Georgia 30333. E–2. Permit Application to Import Controlled Material; Import or Transport Organisms or Vectors. U.S. Department of Agriculture, Animal and Plant Health Inspection Service, Veterinary Services, Federal Building, Hyattsville, Maryland 20782. Terms Approved respiratory protection Equipment which is tested and listed as satisfactory according to standards established by a competent authority (such as NIOSH, Mine Safety and Health Administration (MSHA), or host country agency) to provide respiratory protection against the particular hazard for which it is designed. For military agent protection, DA and Department of Defense (DOD) are the approval authorities. (Approval authority may be specified by law.) Biocontainment area An area which meets the requirements for a BL–3 or BL–4 facility. The area may be an entire building or a single room within a building. See subpart G for details. Biological Safety Cabinets Engineering controls designed to enable laboratory workers to handle infectious etiologic agents and to provide primary containment of any resultant aerosol. There are three major classes of cabinets (I, II, and III) and several subclasses of class II cabinets. Each type of cabinet provides a different degree of protection to personnel and to the products handled inside them. The various classes of cabinets are described in detail in subpart H. Biosafety Level 1 The facilities, equipment, and procedures suitable for work involving agents of no known or of minimal potential hazard to laboratory personnel and the environment. Biosafety Level 2 The facilities, equipment, and procedures applicable to clinical, diagnostic, or teaching laboratories, and suitable for work involving indigenous agents of moderate potential hazard to personnel and the environment. It differs from BL–1 in that (1) laboratory personnel have specific training in handling pathogenic agents, (2) the laboratory is directed by scientists with experience in the handling of specific agents, (3) access to the laboratory is limited when work is being conducted, and (4) certain procedures in which infectious aerosols could be created are conducted in biological safety cabinets or other physical containment equipment. Biosafety Level 3 The facilities, equipment, and procedures applicable to clinical, diagnostic, research, or production facilities in which work is performed with indigenous or exotic agents where potential exists for infection by aerosol, and the disease may have serious or lethal consequences. It differs from BL–2 in that (1) more extensive training in handling pathogenic and potentially lethal agents is necessary for laboratory personnel; (2) all procedures involving the manipulation of infectious material are conducted within biological safety cabinets, other physical containment devices, or by personnel wearing appropriate personal protective clothing and devices; (3) the laboratory has special engineering and design features, including access zones, sealed penetrations, and directional airflow; and (4) any modification of BL–3 recommendations must be made only by the commander. Biosafety Level 4 The facilities, equipment, and procedures required for work with dangerous and exotic agents which pose a high individual risk of life-threatening disease. It differs from BL–3 in that (1) members of the laboratory staff have specific and thorough training in handling extremely hazardous infectious agents; (2) laboratory personnel understand the primary and secondary containment functions of the standard and special practices, containment equipment, and laboratory design characteristics; (3) access to the laboratory is strictly controlled by the institute director; (4) the facility is either in a separate building or in a controlled area within a building, completely isolated from all other areas of the building; (5) a specific facility operations manual is prepared or adopted; (6) within work areas of the facility, all activities are confined to Class III biological safety cabinets or Class I or Class II biological safety cabinets used in conjunction with one-piece positive pressure personnel suits ventilated by a life support system; and (7) the maximum containment laboratory has special engineering and design features to prevent microorganisms from being disseminated to the environment. Building A structure that contains the requisite components necessary to support a facility that is designed according to the required biosafety level. The building can contain one or more facilities conforming to one or more biosafety level. Confirmed Exposure Any mishap with a BDP agent in which there was direct evidence of an actual exposure such as a measurable rise in antibody titer to the agent or a confirmed diagnosis of intoxication or disease. Etiologic Agents Any viable microorganism, or its toxin which causes or may cause human disease, including those agents listed in 42 CFR 72.3 of the Department of Health and Human Services regulations, and any agent of biological origin that poses a degree of hazard similar to those agents. Facility An area within a building that provides appropriate protective barriers for persons working in the facility and the environment external to the facility, and outside of the building. HEPA Filter A filter which removes particulate matter down to submicron sized particles from the air passed through it with a minimum efficiency of 99.97 percent. While the filters remove particulate matter with great efficiency, vapors and gases (for example, from volatile chemicals) are passed through without restriction. HEPA filters are used as the primary means of removing infectious agents from air exhausted from engineering controls and facilities. Human Lethal Dose The estimated quantity of a toxin that is a minimum lethal dose for a 70 kilogram individual based upon published data or upon estimates extrapolated from animal toxicity data. Commander or Institute Director The commander or institute director of an Army activity conducting RDTE with BDP etiologic agents, or the equivalent, at a research organization under contract to the BDP. Institution An organization such as an Army RDTE activity (institute, agency, center, and so forth) or a contract organization such as a school of medicine, or research institute that conducts RDTE with BDP etiologic agents. Laboratory An individual room or rooms within a facility that provide space in which work with etiologic agents can be performed. It contains all of the appropriate engineering features and equipment required at a given biosafety level to protect personnel working in it and the environment external to the facility. Large-Scale Operations Research or production involving viable etiologic agents in quantities greater than 10 liters of culture. Maximum Containment Area An area which meets the requirements for a BL–4 facility. The area may be an entire building or a single room within the building. See chapter 7 for details. Molded Masks Formed masks that fit snugly around the mouth and nose and are designed to protect against a nontoxic nuisance level of dusts and powders. These do not require approval by NIOSH or MSHA. Masks made of gauze do not qualify. Potential Accidental Exposure Any accident in which there was reason to believe that anyone working with a BDP agent may have been exposed to that agent, yet no measurable rise in antibody titer or diagnosis of intoxication or disease was made. However, the high probability existed for introduction of an agent through mucous membranes, respiratory tract, broken skin, or the circulatory system as a direct result of the accident, injury, or incident. Resource Conservation Recovery Act of 1976 Listed Hazardous Waste The waste materials listed by the Environmental Protection Agency under authority of the RCRA for which the agency regulates disposal. A description and listing of these wastes is located in 40 CFR part 261. Suite An area consisting of more than one room, designed to be a functional unit in which entire operations can be facilitated. Suites may contain a combination of laboratories or animal holding rooms and associated support areas within a facility that are designed to conform to a particular biosafety level. There may be one or more suites within a facility. Toxin Toxic material of etiologic origin that has been isolated from the parent organism.1 1 The publication “Bacterial Toxins: a Table of Lethal Amounts,” (Gill, D.M. (1982) Microbiological Reviews, 46:86–94) contains a useful table of mammalian toxicities of numerous toxins.
Title 32: National Defense
PART 627—THE BIOLOGICAL DEFENSE SAFETY PROGRAM, TECHNICAL SAFETY REQUIREMENTS (DA PAMPHLET 385–69)
Subpart H—Engineering Controls
§ 627.49 Introduction.
§ 627.50 Class I biological safety cabinet.
§ 627.51 Class II biological safety cabinet.
§ 627.52 Class III biological safety cabinet.
§ 627.53 Fume hood.
§ 627.54 Glove box.
§ 627.55 Ventilated balance enclosures.
§ 627.56 Ventilated cage enclosures.
§ 627.57 Ventilated cage areas.
Appendix A to Part 627—References
Appendix B to Part 627—Resource List for Immunoprophylaxis of Personnel at Risk
B-1. Recommendations for Immunoprophylaxis of Personnel at Risk---------------------------------------------------------------------------------------------------------------- Description of disease Product Recommended for use in Source of product----------------------------------------------------------------------------------------------------------------Anthrax.......................... Inactivated vaccine. Personnel working regularly with USAMRIID.\1\ cultures, diagnostic materials, or infected animals.Botulism......................... Pentavalent toxoid Personnel working regularly with CDC.\3\ (A,B,C,D,E) cultures or toxin. (IND).\2\.Cholera.......................... Inactivated vaccine. Personnel working regularly with Commercially large volumes or high available. concentrations of infectious materials.Diphtheria Tetanus (Adult)....... Combined toxoid..... All laboratory and animal care Commercially personnel irrespective of agents available. handled.Eastern equine encephalitis (EEE) Inactivated vaccine Personnel who work directly and USAMRIID.\1\ (IND) \2\. regularly with EEE in the laboratory.Hepatitis A...................... Immune Serum Animal care personnel working Commercially Globulin [ISG directly with chimpanzees available. (Human)]. naturally or experimentally infected with Hepatitis A virus.Hepatitis B...................... Serum-derived or Personnel working regularly with Commercially recombinant vaccine. human blood and blood components. available.Influenza........................ Inactivated vaccine. (Vaccines prepared from earlier Commercially isolated strains may be of little available. value in personnel working with recent isolates from humans or animals).Japanese Encephalitis............ Inactivated vaccine Personnel who work directly and CDC.\3\ (IND) \2\. regularly with JE virus in the laboratory.Measles.......................... Live attenuated Measles-susceptible personnel Commercially virus vaccine. working with the agent or available. potentially infectious clinical materials.Meningococcal Meningitis......... Purified Personnel working regularly with Commercially polysaccharide large volumes or high available. vaccine. concentrations of infectious materials (does not protect against infection with group B meningococcus).Plague........................... Inactivated vaccine. Personnel working regularly with Commercially cultures of Yersinia pestis or available. infected rodents or fleas.Poliomyelitis.................... Inactivated (IPV) Polio-susceptible personnel Commercially and live attenuated working with the virus or available. (OPV) vaccines. entering laboratories or animal rooms where the virus is in use.Pox viruses (Vaccinia, Cowpox, or Live (lyophilized) Personnel working with orthopox CDC.\3\ Monkey Pox viruses). vaccinia virus. viruses transmissible to humans, with animals infected with these agents, and persons entering areas where these viruses are in use.Q Fever (Phase II) vaccine....... Inactivated (IND) Personnel who have no demonstrable USAMRIID.\1\ \2\. sensitivity to Q fever antigen and who are at high risk of exposure to infectious materials or animals.Rabies........................... Human diploid line Personnel working with all strains Commercially cell inactivated of rabies virus, with infected available. vaccine. animals, or persons entering areas where these activities are conducted.Rift Valley Fever................ Inactivated virus All laboratory and animal care USAMRIID.\1\ vaccine (IND) \2\. personnel working with the agent or infected animals and all personnel entering laboratories or animal rooms when the agent is in use.Rubella.......................... Live attenuated Rubella-susceptible personnel, Commercially virus vaccine. especially women, working with available. ``wild'' strains or in areas where these viruses are in use.Tuberculosis..................... Live, attenuated BCG vaccine ordinarily is not used Commercially (BCG) bacterial in laboratory personnel in the available. vaccine. U.S.Tularemia........................ Live attenuated Personnel working regularly with USAMRIID.\1\ bacterial vaccine cultures or infected animals or (IND).\2\. persons entering areas where the agent of infected animals are in use.Typhoid.......................... Inactivated vaccine. Personnel who have no demonstrated Commercially sensitivity to the vaccine and available. who work regularly with cultures.Venezuelan equine (VEE) Live attenuated Personnel working with VEE and the USAMRIID.\1\ encephalitis. (TC83) viral Equine Cabassou, Everglades, vaccine (IND).\2\. Mucambo, and Tonate viruses, or who enter areas where these viruses are in use.Western equine encephalitis (WEE) Inactivated vaccine Personnel who work directly and USAMRIID.\1\ (IND) \2\ with WEE regularly in the laboratory. virus.Yellow Fever..................... Live attenuated Personnel working with virulent Commercially (17D) virus vaccine. and avirulent strains of Yellow available. Fever virus.----------------------------------------------------------------------------------------------------------------\1\ For information, contact: United States Army Medical Materiel Development Activity, Fort Detrick, Frederick, MD 21701, telephone: (301) 663-7661.\2\ Investigational New Drug (IND).\3\ Clinical Medicine Branch, Division of Host Factors, Center for Infectious Disease, Centers for Disease Control, Atlanta, GA 30333, telephone: (404) 639-3356.Source: Adapted from recommendations of the PHS Immunization Practices Advisory Committee and Biosafety in Microbiological and Biomedical Laboratories.
Appendix C to Part 627—Laboratory Safety Inspection Checklist
Appendix D to Part 627—Packaging and Labeling Requirements for Shipment of Etiologic Agents
Appendix E to Part 627—Permits for Importation and Shipment of Etiologic Agents
Appendix F to Part 627—Drawings, Biological Safety Cabinets
Appendix G to Part 627—Glossary
Abbreviations AIHA American Industrial Hygiene AssociationAMC United States Army Materiel CommandANSI American National Standards InstituteAR Army RegulationATCC American Type Culture CollectionASHRAE American Society of Heating, Refrigerating, and Air Condition Engineers, Inc.BDP Biological Defense ProgramBL biosafety levelCDC Centers for Disease ControlCFR Code of Federal RegulationsDA PAM Department of Army PamphletDHEW Department of Health, Education, and WelfareDOD Department of DefenseDOT Department of TransportationDNA deoxyribonucleic acidEPA Environmental Protection AgencyEtO ethylene oxideFDA Food and Drug Administrationfpm feet per minuteHEPA high efficiency particulate airHHS Health and Human ServicesIATA International Air Transport AssociationIBC Institutional Biosafety CommitteeICAO International Civil Aviation Organizationlfpm linear feet per minuteLS large-scalem metermin minuteMSDS Material Safety Data SheetsMSHA Mine Safety and Health AdministrationNCCLS National Committee for Clinical Laboratory StandardsNCI National Cancer InstituteNEPA National Environmental Policy ActNFPA National Fire Protection AssociationNIH National Institutes of HealthNIOSH National Institute for Occupational Safety and HealthNRC Nuclear Regulatory CommissionNSF National Sanitation FoundationOSHA Occupational Safety and Health AdministrationpH the negative logarithm of hydrogen ion concentrationPHS Public Health ServicePPE personal protective equipmentppm parts per millionpsi pounds per square inchRCRA-Listed Resource Conservation Recovery Act of 1976 Listed Hazardous WasteRDTE research, development, test, and evaluationRPO Radiation Protection OfficerSALS Subcommittee on Arbovirus Laboratory SafetySAR supplied-air respiratorSCBA self-contained breathing apparatusSOP Standing Operating ProcedureTD to deliverTLV threshold limit valueUSDA United States Department of AgricultureUV ultraviolet
