17182 
Federal Register / Vol. 47. No. 77 / Wednesday. April 21. 1982 / Notices 
which is closed before removal from the 
laboratory. 
ll-B-2-a-{9). Only persons who have 
been advised of the nature of the 
research being conducted shall enter the 
laboratory. 
Il-B-2-a-{10). The universal 
biohazard sign shall be posted on all 
laboratory access doors when 
experiments requiring P2 containment 
are in process. Freezers and 
refrigerators or other units used to store 
organisms containing recombinant DNA 
molecules shall also be posted with the 
universal biohazard sign. 
n-B-2-a-{ll). An insect and rodent 
control programs shall be instituted. 
n-B-2-a-(12). The use of laboratory 
gowns, coats, or uniforms is required. 
Laboratory clothing shall not be worn to 
the lunch room or outside of the building 
in which the laboratory is located. 
n-B-2-a-(13). Animals not related to 
the experiment shall not be permitted in 
the laboratory. 
H-B-2-a-(14). Use of the hypodermic 
needle and syringe shall be avoided 
when alternative methods are available. 
D-B-2-a-{15). The laboratory shall be 
kept neat and clean. 
D-B-2-a-{16). Experiments of lesser 
biohazard potential can be carried out 
concurrently in carefully demarcated 
areas of the same laboratory. 
U-B-2-b. Containment Equipment 
Biological safety cabinets (20) shall.be 
used to contain aerosol-producing 
equipment, such as blender, lyophilizers. 
sonicators. and centrifuges, when used 
to process organisms containing 
recombinant DNA molecules, except 
where equipment design provides for 
containment of the potential aerosol. For 
example, a centrifuge may be operated 
In the open if a sealed head or safety 
centrifuge cups are used. 
D-B-2-C. Special Laboratory Design. 
An autoclave for sterilization of wastes 
and contaminated materials shall be 
available in the same building in which 
organisms containing recombinant DNA 
molecules are used. 
n-B-3. P3 Uvel. 
D-B-3-a. Laboratory Practices. 
U-B-3-a-{l). Laboratory doors shall 
be kept closed while experiments are in 
progress. 
U-B-3-a-{2). Work surfaces shall be 
decontaminated following the 
completion of the experimental activity, 
and immediately following spills of 
organisms containing recombinant DNA 
molecules. 
n~B-3-a-{3). All laboratory wastes be 
steam-sterilized (autoclaved) before 
disposal. Other contaminated materials, 
such as glassware, animal cages, 
laboratory equipment, and radioactive 
wastes, shall be decontaminated by a 
method demonstrated to be effective 
before washing, reuse, or disposal. 
lI-B-3-a-{4). Mechanical pipetting 
devices shall be used; pipetting by 
mouth is prohibited. 
Il-B-3-a-{5). Eating, drinking, 
smoking, and storage of food are not 
permitted in the laboratory area in 
which recombinant DNA materials are 
handled. 
Il-B-3-a-(6). Persons shall wash their 
hands after handling organisms 
containing recombinant DNA molecules 
and when they leave the laboratory. 
Il-B-3-a-(7). Care shall be exercised 
to minimize the creation of aerosols. For 
example, manipulations such as 
inserting a hot inoculating loop or 
needle into a culture, flaming an 
inoculation loop or needle so that it 
splatters, and forceful ejection of fluids 
from pipettes or syringes shall be 
avoided. 
II-B-3-a-(8). Contaminated materials 
that are to be steam-sterilized 
(autoclaved) or decontaminated at a site 
away from the laboratory shall be 
placed in a durable leak-proof container, 
which is closed before removal from the 
laboratory. 
II-B-3-a-(9). Entry Into laboratory 
shall be through a controlled access 
area. Only persons who have been 
advised of the nature of the research 
being conducted shall enter the 
controlled access area. Only persons 
required on the basis of program or 
support needs shall be authorized to 
enter the laboratory. Such persons shall 
be advised of the nature of the research 
being conducted before entry, and shall 
comply with all required entry and exit 
procedures. 
Il-B-3-a-(10). Persons under 16 years 
of age shall not enter the laboratory. 
II-B-3-a-(ll). The universal 
biohazard sign shall be posted on the 
controlled access area door and on all 
labortory doors when experiments 
requiring P3-level containment are in 
progress. Freezers and refrigerators or 
other units used to store organisms 
containing recombinant DNA molecules 
shall also be posted with the universal 
biohazard sign. 
Il-B-3-a-(12). An insect and rodent 
control program shall be instituted. 
U-B-3-a-{13). Laboratory clothing that 
protects street clothing (e g., long-sleeve 
solid-front or wrap-around gowns, no- 
button or slipover jackets) shall be worn 
in the laboratory. Front-button 
laboratory coats are unsuitable. 
Laboratory clothing shall not be worn 
outside the laboratory and shall be 
decontaminated before it is sent to the 
laundry. 
II-&^a-(14). Raincoats, overcoats, 
topcoats, coats, hats. caps, and such 
street outer-wear shall not be kept in the 
laboratory. 
lI-B-3-a-(15). Gloves shall be worn 
when handling materials requiring P3 
containment. They shall be removed 
aseptically immediately after the 
handling procedure and 
decontaminated. 
Il-B-3-a-(16). Animals and plants are 
not related to the experiment shall not 
be permitted in the laboratory. 
II-B-3-a-(17). Vacuum outlets shall be 
protected by filter and liquid 
disinfectant traps. 
II-B-3-a-(18). Use of hypodermic 
needle and syringe shall be avoided 
when alternative methods are available. 
II-B-3-a-(19). The laboratory shall be 
kept neat and clean. 
Il-B-3-a-(20). If experiments 
involving other organisms which require 
lower levels of containment are to be 
conducted in the same laboratory 
concurrently with experiments requiring 
P3-leveI physical containment, they 
shall be conducted in accordance with 
all P3-level laboratory practices. 
U-B-3-b. Containment Equipment 
II-B-3-b-(l). Biological safety 
cabinets [20] shall be used for all 
equipment and manipulations that 
produce aerosols— e.g., pipetting, 
dilutions, transfer operations, plating, 
flaming, grinding, blending, drying 
sonicating, shaking, centrifuging — where 
these procedures involve organisms 
containing recombinant DNA molecules, 
except where equipment design 
provides for containment of the 
potential aerosol. 
II-B-3-b-{2). Laboratory animals held 
in a P3 area shall be housed in partial- 
containment caging systems, such as 
Horsfall units (19A], 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 radiation lamps and 
reflectors. 
(Note. — Conventional caging systems may 
be used, provided that ail 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.) 
Il-B-3-b-(3). Alternative Selection of 
Containment Equipment Experimental 
procedures involving a host-vector 
system that provides a one-step higher 
level of biological containment than that 
specified can be conducted in the P3 
laboratory using containment equipment 
specified for the P2 level of physical 
containment. Experimental procedures 
involving a host-vector system that 
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