Federal Register / Vol. 45, No. 227 / Friday, November 21, 1980 / Notices 
77387 
II-B-2-a-(16). Experiments of lesser 
biohazard potential can be carried out 
concurrently in carefully demarcated 
areas of the same laboratory. 
H-B-2-b. Containment Equipment. 
Biological safety cabinets (20) shall be 
used to contain aerosol-producing 
equipment, such as blenders, 
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. 
II-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. 
Il-B-3. P3 Level. 
Il-B-3-a. Laboratory Practices. 
II— B— 3— a— (1). Laboratory doors shall 
be kept closed while experiments are in 
progress. 
II-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. 
Il-B-3-a-(3). All laboratory wastes 
shall 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. 
II— B— 3— a— (4). Mechanical pipetting 
devices shall be used; pipetting by 
mouth is prohibited. 
II— 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. 
II-B-3-a-(6). Persons shall wash their 
hands after handling organisms 
containing recombinant DNA molecules 
and when they leave the laboratory. 
II-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 the 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. 
II-B-3-a-(10). Persons under 16 years 
of age shall not enter the laboratory. 
II— B— 3— a— (11). The universal 
biohazard sign shall be posted on the 
controlled access area door and on all 
laboratory 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. 
II-B-3-a-(12). An insect and rodent 
control program shall be instituted. 
II— 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— B— 3— 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. 
II— B— 3— a— (16). Animals and plants 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 springe shall be avoided 
when alternative methods are available. 
II— B— 3— a— (19). The laboratory shall be 
kept neat and clean. 
II— 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-level physical containment, they 
shall be conducted in accordance with 
all P3-level laboratory practices. 
II-B-3-b. Containment Equipment. 
II— B— 3— b— (1 ) 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 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.) 
II— 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 in Part 
III 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 provides a one- 
step lower level of biological 
containment than that specified in Part 
III can be conducted in the P3 laboratory 
using containment equipment specified 
for the P4 level of physical containment. 
Alternative combinations of 
containment safeguards are shown in 
Table I. 
Table I . — Combinations ot Containment Safeguards 
Alternate combinations of physical and biological containment 
Classification of experiment 
According to guidelines Physical containment 
Physical 
containment 
Biological 1 
containment 
Laboratory 
design 
specified for— 
Laboratory 
practices 
specified for— 
Containment 
equipment 
specified for— 
Biological 
containment 
P3 
HV3 
P3 
P3 
P3 
HV3 
P3 
HV3 
P3 
P3 
P4 
HV2 
P3 
HV2 
P3 
P3 
P3 
HV2 
P3 
HV2 
P3 
P3 
P2 
HV3 
[ 6 ] 
