[Title 32 CFR H]
[Code of Federal Regulations (annual edition) - July 1, 2002 Edition]
[Title 32 - NATIONAL DEFENSE]
[Chapter V - DEPARTMENT OF THE ARMY]
[Subchapter H - SUPPLIES AND EQUIPMENT]
[Part 627 - THE BIOLOGICAL DEFENSE SAFETY PROGRAM, TECHNICAL SAFETY REQUIREMENTS (DA PAMPHLET 385-69)]
[Subpart H - Engineering Controls]
[From the U.S. Government Printing Office]
32NATIONAL DEFENSE32002-07-012002-07-01falseEngineering ControlsHSubpart HNATIONAL DEFENSEDEPARTMENT OF THE ARMYSUPPLIES AND EQUIPMENTTHE BIOLOGICAL DEFENSE SAFETY PROGRAM, TECHNICAL SAFETY REQUIREMENTS (DA PAMPHLET 385-69)
Subpart H--Engineering Controls
Sec. 627.49 Introduction.
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.
Sec. 627.50 Class I biological safety cabinet.
(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.
Sec. 627.51 Class II biological safety cabinet.
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.
[[Page 566]]
(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 Sec. 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.
[[Page 567]]
(4) Additional certifications or requirements. None.
Sec. 627.52 Class III biological safety cabinet.
(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.
Sec. 627.53 Fume hood.
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.
Sec. 627.54 Glove box.
(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
[[Page 568]]
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.
Sec. 627.55 Ventilated balance enclosures.
(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.
Sec. 627.56 Ventilated cage enclosures.
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
m\3\ /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 m\3\ /min per
cage is required for a 0.24 m\3\ 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 m\3\ /min per cage is required for a 0.24 m\3\ unit.
Ventilation rates may vary with the size of the cage, and the number and
type of animals being housed.
[[Page 569]]
Sec. 627.57 Ventilated cage areas.
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.
Appendix A to Part 627--References
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 Secs. 627.31(h)(2)
and 627.31(h)(4).)
AR 40-5
Preventive Medicine. (Cited in Sec. 627.8.)
AR 40-10
Health Hazard Assessment Program in Support of the Army Materiel
Acquisition Decision Process. (Cited in Sec. 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 Sec. 627.40(a).)
AR 40-66
Medical Records and Quality Assurance Administration. (Cited in
Sec. 627.9.)
AR 40-400
Patient Administration. (Cited in Sec. 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 Secs. 627.36(a)(6) and 627.40(b).)
AR 385-10
Army Safety Program. (Cited in Secs. 627.6 and 627.31(h)(4).)
AR 385-69
Biological Defense Safety Program. (Cited in Secs. 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 Sec. 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.
[[Page 570]]
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.
[[Page 571]]
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.
[[Page 572]]
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.
[[Page 573]]
Appendix C to Part 627--Laboratory Safety Inspection Checklist
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--
[[Page 574]]
(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?
[[Page 575]]
(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?
Appendix D to Part 627--Packaging and Labeling Requirements for Shipment
of Etiologic Agents
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.
Appendix E to Part 627--Permits for Importation and Shipment of
Etiologic Agents
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.
Appendix F to Part 627--Drawings, Biological Safety Cabinets
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Appendix G to Part 627--Glossary
Abbreviations
AIHA American Industrial Hygiene Association
AMC United States Army Materiel Command
ANSI American National Standards Institute
AR Army Regulation
ATCC American Type Culture Collection
ASHRAE American Society of Heating, Refrigerating, and
Air Condition Engineers, Inc.
BDP Biological Defense Program
BL biosafety level
CDC Centers for Disease Control
CFR Code of Federal Regulations
DA PAM Department of Army Pamphlet
DHEW Department of Health, Education, and Welfare
DOD Department of Defense
DOT Department of Transportation
DNA deoxyribonucleic acid
EPA Environmental Protection Agency
EtO ethylene oxide
FDA Food and Drug Administration
fpm feet per minute
HEPA high efficiency particulate air
[[Page 579]]
HHS Health and Human Services
IATA International Air Transport Association
IBC Institutional Biosafety Committee
ICAO International Civil Aviation Organization
lfpm linear feet per minute
LS large-scale
m meter
min minute
MSDS Material Safety Data Sheets
MSHA Mine Safety and Health Administration
NCCLS National Committee for Clinical Laboratory
Standards
NCI National Cancer Institute
NEPA National Environmental Policy Act
NFPA National Fire Protection Association
NIH National Institutes of Health
NIOSH National Institute for Occupational Safety and
Health
NRC Nuclear Regulatory Commission
NSF National Sanitation Foundation
OSHA Occupational Safety and Health Administration
pH the negative logarithm of hydrogen ion
concentration
PHS Public Health Service
PPE personal protective equipment
ppm parts per million
psi pounds per square inch
RCRA-Listed Resource Conservation Recovery Act of 1976
Listed Hazardous Waste
RDTE research, development, test, and evaluation
RPO Radiation Protection Officer
SALS Subcommittee on Arbovirus Laboratory Safety
SAR supplied-air respirator
SCBA self-contained breathing apparatus
SOP Standing Operating Procedure
TD to deliver
TLV threshold limit value
USDA United States Department of Agriculture
UV ultraviolet
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
[[Page 580]]
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.
[[Page 581]]
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\
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\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.