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Federal Register / Vol. 51, No. 88 / Wednesday, May 7, 1986 / Notices 
instructions on practices and procedures 
and to follow them. 
Appendix G-II-B-3 — Containment 
Equipment 
Appendix C-II-B-3-a. Biological 
safety cabinets (Class I or II) (see 
Appendix G — III— 12) or other appropriate 
personal protective or physical 
containment devices are used whenever: 
Appendix G-II-B-3-a-(lJ. Procedures 
with a high potential for creating 
aerosols are conducted (15). These may 
include centrifuging, grinding, blending, 
vigorous shaking or mixing, sonic 
disruption, opening containers of 
materials whose internal pressures may 
be different from ambient pressures, 
inoculating animals intranasally, and 
harvesting infected tissues from animals 
or eggs. 
Appendix G-II-B-3-a-(2J. High 
concentrations or large volumes of 
organisms containing recombinant DNA 
molecules are used. Such materials may 
be centrifuged in the open laboratory if 
sealed heads or centrifuge safety cups 
are used and if they are opened only in 
a biological safety cabinet. 
Appendix G-II-B-4 Laboratory 
Facilities 
Appendix G-II-B—L-a. The laboratory 
is designed so that it can be easily 
cleaned. 
Appendix G-II-B—4-b. Bench tops are 
impervious to water and resistant to 
acids, alkalis, organic solvents, and 
moderate heat. 
Appendix G-II-B-4-c. Laboratory 
furniture is sturdy and spaces between 
benches, cabinets, and equipment are 
accessible for cleaning. 
Appendix G-II-B-4-d. Each 
laboratory contains a sink for hand- 
washing. 
Appendix G-II-B-4-e. If the 
laboratory has windows that open, they 
are fitted with fly screens. 
Appendix G-II-B-4-f. An autoclave 
for decontaminating laboratory wastes 
is available. 
Appendix G-Il-C — Biosafety Level 3 
(BL3) [16] 
Appendix G-II-C-1. Standard 
Microbiological Practices. 
Appendix G-II-C-l-a. Work surfaces 
are decontaminated at least once a day 
and after any spill of viable material. 
Appendix G-II-C-l-b. All 
contaminated liquid or solid wastes are 
decontaminated before disposal. 
Appendix G-ll-C-l-c. Mechanical 
pipetting devices are used; mouth 
pipetting is prohibited. 
Appendix G-II-C-l-d. Eating, 
drinking, smoking, storing food, and 
applying cosmetics are not permitted in 
the work area. 
Appendix G-U-C-l-e. Persons wash 
their hands after handling materials 
involving organisms containing 
recombinant DNA molecules, and 
animals, and when they leave the 
laboratory. 
Appendix G-II-C-l-f. All procedures 
are performed carefully to minimize the 
creation of aerosols. 
Appendix G-ll-C-l-g. Persons under 
16 years of age shall not enter the 
laboratory. 
Appendix G-II-C-l-h. If experiments 
involving other organisms which require 
lower levels of containment are to be 
conducted in the same laboratory 
concurrently with experiments requiring 
BL3 level physical containment, they 
shall be conducted in accordance with 
all BL3 level laboratory practices. 
Appendix G-II-C-2 — Special Practices 
Appendix G-II-C-2-a. Laboratory 
doors are kept closed when experiments 
are in progress. 
Appendix G-II-C-2-b. Contaminated 
materials that are to be decontaminated 
at a site away from the laboratory are 
placed in a durable leakproof container 
which is closed before being removed 
from the laboratory. 
Appendix G-Il-C-2-c. The laboratory 
director controls access to the 
laboratory and restricts access to 
persons whose presence is required for 
program or support purposes. The 
director has the final responsibility for 
assessing each circumstance and 
determining who may enter or work in 
the laboratory. 
Appendix G-II-C-2-d. The laboratory 
director establishes policies and 
procedures whereby only persons who 
have been advised of the potential 
biohazard, who meet any specific entry 
requirements (e.g., immunization), and 
who comply with all entry and exit 
procedures enter the laboratory or 
animal rooms. 
Appendix G-Il-C-2-e. When 
organisms containing recombinant DNA 
molecules or experimental animals are 
present in the laboratory or containment 
module, a hazard warning sign 
incorporating the universal biohazard 
symbol is posted on all laboratory and 
animal room access doors. The hazard 
warning sign identifies the agent, lists 
the name and telephone number of the 
laboratory director or other responsible 
person(s), and indicates any special 
requirements for entering the laboratory, 
such as the need for immunizations, 
respirators, or other personal protective 
measures. 
Appendix G-II-C-2-f. All activities 
involving organisms containing 
recombinant DNA molecules are 
conducted in biological safety cabinets 
or other physical containment devices 
within the containment module. No 
work in open vessels is conducted on 
the open bench. 
Appendix G-II-C-2-g. The work 
surfaces of biological safety cabinets 
and other containment equipment are 
decontaminated when work with 
organisms containing recombinant DNA 
molecules is finished. Plastic-backed 
paper toweling used on nonperforated 
work surfaces within biological safety 
cabinets facilitates clean-up. 
Appendix G-U-C-2-h. An insect and 
rodent program is in effect. 
Appendix G-II-C-2-i. Laboratory 
clothing that protects street clothing 
(e.g., solid front or wTap-around gowns, 
scrub suits, coveralls) is worn in the 
laboratory. Laboratory clothing is not 
worn outside the laboratory, and it is 
decontaminated before being laundered. 
Appendix G-II-C-2-j. Special care is 
taken to avoid skin contamination with 
contaminated materials: gloves should 
be worn when handling infected animals 
and when skin contact with infectious 
materials is unavoidable. 
Appendix G-II-C-2-k. Molded 
surgical masks or respirators are worn 
in rooms containing experimental 
animals. 
Appendix G-II-C-2-1. Animals and 
plants not related to the work being 
conducted are not permitted in the 
laboratory. 
Appendix G-Il-C-2-m. Laboratory 
animals held in a BL3 area shall be 
housed in partial-containment caging 
systems, such as Horsfall units [11], 
open cages placed in ventilated 
enclosures, solid-wall and -bottom cages 
covered by filter bonnets, or solid-wall 
and -bottom cages placed on holding 
racks equipped with ultraviolet in 
radiation lamps and reflectors. 
Note. — Conventional caging systems may 
be used provided that all personnel wear 
appropriate personal protective devices. 
These shall include at a minimum wrap- 
around gowns, head covers, gloves, shoe 
covers, and respirators. All personnel shall 
shower on exit from areas where these 
devices are required. 
Appendix G-II-C-2-n. All wastes 
from laboratories and animal rooms are 
appropriately decontaminated before 
disposal. 
Appendix G-II-C-2-o. Vacuum lines 
are protected with high efficiency 
particulate air (HEPA) filters and liquid 
disinfectant traps. 
Appendix G-lI-C-2-p. Hypodermic 
needles and syringes are used only for 
parenteral injection and aspiration of 
fluids from laboratory animals and 
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Recombinant DNA Research, Volume 1 1 
