NOTICES 
33073 
bottom-cages placed on holding racks 
equipped with ultraviolet radiation 
lamps and reflectors. (Note: Conven- 
tional caging systems may be used, 
provided that all personnel wear ap- 
propriate personal protective devices. 
These shall include at a minimum wra- 
paround gowns, head covers, gloves, 
shoe covers, and respirators. All per- 
sonnel shall shower on exit from areas 
where these devices are required.) 
II-B-3-b-(3). * Alternative Selection 
of Containment Equipment Experi- 
mental procedures involving a host- 
vector system which provides a one- 
step higher level of biological contain- 
ment than that specified in Part III 
can be conducted in the P3 laboratory 
using containment equipment speci- 
fied for the P2 level of physical con- 
tainment. Experimental procedures in- 
volving a host-vector system which 
provides a one-step low T er level of bio- 
logical containment than that speci- 
fied 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 safe- 
guards are shown in table I. 
Table I 
CCMBINATICNS OF SAFEGUARDS 
Classification of 
experiment 
accord ire to Guidelines 
Alter rate 
canbinations cf phvsical ar.d biological containment 
Fhvsical Containment 
Laooracorv Lsooratcry 
Containment 
Physical 
Biological* 
design practices 
equipment 
Biological 
cents inrnerrt 
ccntairre.ot 
soecii lec 
fer: scecified for: 
scecified for: 
containment 
Pi 
H73 
P3 
P3 
P3 
HV3 
P3 
HV3 
P3 
P3 
P4 
HV2 
Pi 
HV2 
P3 
P3 
P3 
HV2 
P3 
HV2 
P3 
?3 
P2 
HV3 
P3 
KV2 
F3 
P3 
P4 
HV1 
P3 
KVl 
P3 
P3 
?3 
HVl 
P3 
KVl 
P3 
P3 
P2 
HV2 
’See Section II-D for description of biological containment. 
II-B-3-c. Special Laboratory Design. 
II-B-3-c-(l). ’The laboratory shall 
be separated from areas which are 
open to unrestricted traffic flow by a 
controlled access area. A controiled 
access area is an anteroom, a change 
room, an air lock or any other double- 
door arrangement which separates the 
laboratory from areas w'hich are open 
to unrestricted traffic flow. 
II-B-3-c-(2). ‘The surfaces of walls, 
floors, and ceilings shall be readily 
cleanable. Penetrations through these 
surfaces shall be sealed or capable of 
being sealed to facilitate space decon- 
tamination. 
II-B-3-c-(3). *A foot, elbow, or auto- 
matically operated handwashing facili- 
ty shall be provided near each primary 
laboratory exit area. 
II-B-3-c-(4). ’Windows in the labo- 
ratory shall be sealed. 
II-B-3-c-(5). ’Laboratory doors shall 
be self-closing. 
II-B-3-c-(6). ’An autoclave for steril- 
ization of wastes and contaminated 
materials shall be available in the 
same building (and preferably within 
the controlled laboratory area) in 
w-hich organisms containing recombin- 
ant DNA molecules are used. 
II-B-3-c-(7). ’An exhaust air ventila- 
tion system shall be provided. This 
system shall be balanced so that the 
direction of airflow is from the con- 
trolled access area into the laboratory 
environment. The exhaust air shall 
not be recirculated to any other areas 
of the building. Recirculation of air 
within the laboratory room, however, 
may be provided. The exhaust air 
from the laboratory shall be dis- 
charged to the outdoors so that it is 
dispersed clear of occupied buildings 
and air intakes, the exhaust air from 
the laboratory can be discharged to 
the outdoors without filtration or 
other treatment. 
II-B-3-c-(8). ’The treated exhaust 
air from class I and class II biological 
safety cabifiets (20) may be discharged 
either directly to the laboratory or to 
the outdoors. The treated exhaust air 
from a class III cabinet shall be dis- 
charged directly to the outdoors. If 
the treated exhaust air from these 
cabinets is to be discharged to the out- 
doors through a building exhaust air 
system, it shall be connected to this 
system so as to avoid any interference 
with the air balance of the cabinet or 
building exhaust air system. 
II-B-4. P4 Level 
FEDERAL REGISTER, VOL. 43, NO. 146— FRIDAY, JULY 28, 
II-B-4-a. Laboratory Practices. 
II-B-4-a-(l). Laboratory doors shall 
be kept closed while experiments are 
in progress. 
II-B-4-a-(2). Work surfaces shall be 
decontaminated following the comple- 
tion of the experimental activity and 
immediately following spills of organ- 
isms containing recombinant DNA 
molecules. 
II-B-4-a-(3). All biological wastes 
shall be decontaminated before dispos- 
al. Other contaminated materials such 
as glassware, animal cages, and labora- 
tory equipment shall be decontaminat- 
ed before washing, reuse, or disposal. 
II-B-4-a-(4). Mechanical pipetting 
devices shall be used; pipetting by 
mouth is prohibited. 
II-B-4-a-(5). ’Eating, drinking, 
smoking, and storage of food are not 
permitted in the P4 facility. 
II-B-4-a-(6). Persons shall wash 
their hands after handling organisms 
containing recombinant DNA mole- 
cules and when they leave the labora- 
tory. 
II-B-4-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 in- 
oculation loop or needle so that is 
splatters, and forceful ejection of 
fluids from pipettes or syringes shall 
be avoided. 
II-B-4-a-(8). ’Biological materials to 
be removed from the P4 facility in a 
viable or intact state, shall be trans- 
ferred to a nonbreakable sealed con- 
tainer w'hich is then removed from the 
P4 facility through a passthrough dis- 
infectant dunk tank or fumigation 
chamber. 
II-B-4-a-(9). ’No materials, except 
for biological materials that are to 
remain in a viable or intact state, shall 
be removed from the P4 facility unless 
they have been sterilized or decon- 
taminated as they pass out of the P4 
facility. All wastes and other materials 
as well as equipment not damaged by 
high temperature or steam shall be 
sterilized in the double-door autoclave 
of the P4 facility. Other materials 
which may be damaged by tempera- 
ture or steam shall be removed from 
the P4 facility through a passthrough 
fumigation chamber. 
II-B-4-a-(10). ’Materials within the 
class III cabinets shall be removed 
from the cabinet system only after 
being sterilized in an attached double- 
door autoclave or after being con- 
tained in a nonbreakable sealed con- 
tainer which is then passed through a 
disinfectant dunk tank or a fumigation 
chamber. 
1978 
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