69214 
Federal Register / Vol. 44, No. 232 / Friday, November 30, 1979 / Notices 
Table I 
COMBINATIONS OF CONTAINMENT SAFEGUARDS 
Classification of 
experiment Alternate combinations 
according to Guidelines of physical and biological containment 
Physical 
containment 
Biological* 
containment 
Physical Containment 
Biological 
containment 
Laboratory 
design 
specified for: 
Laboratory 
practices 
specified for: 
Containment 
ecjuipment 
specified for: 
P3 
HV3 
P3 
P3 
P3 
HV3 
P3 
HV3 
P3 
P3 
P4 
'HV2 
P3 
HV2 
P3 
P3 
P3 
HV2 
P3 
HV2 
P3 
P3 
P2 
HV3 
P3 
HV2 
P3 
P3 
P4 
HV1 
P3 
HV1 
P3 
P3 
P3 
HV1 
P3 
HV1 
P3 
P3 
P2 
HV2 
*See Section TI-D for description of biological containment. 
equipped with an alarm that would be 
actuated in the event that reversal in the 
direction of air movement were to occur. 
The exhaust air from the laboratory area 
shall not be recirculated to other areas 
of the building unless the exhaust air is 
filtered by HEPA filters or equivalent. 
The exhaust air from the laboratory area 
can be discharged to the outdoors 
without filtration or other means for 
effectively reducing an accidental 
aerosol burden provided that it can be 
dispersed clear of occupied buildings 
and air intakes. 
II-B-3-c-(7). The treated exhaust-air 
from Class I and Class II biological 
safety cabinets [20] may be discharged 
either to the laboratory or to the 
outdoors. The treated exhaust-air from a 
Class III cabinet shall be discharged 
directly to the outdoors. If the treated 
exhaust-air from these cabinets is to be 
discharged to the outdoors 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 and the building ventilation 
system. 
II-B-4. P4 Level. 
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 
completion of the experimental activity 
and immediately following spills of 
organisms containing recombinant DNA 
molecules. 
II-B-4-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-^l-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 molecules 
and when they leave the laboratory. 
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 
inoculation loop or needle so that it 
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 
transferred to a nonbreakable sealed 
container, which is then removed from 
the P4 facility through a pass-through 
disinfectant 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 steam-sterilized (autoclaved) or 
decontaminated by a means 
demonstrated to be effective 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 steam 
sterilized in the double-door autoclave 
of the P4 facility. Other materials which 
may be damaged by temperature or 
steam shall be removed from the P4 
facility through a pass-through 
fumigation chamber. 
II-B-4-a-(10). Materials within the 
Class III cabinets shall be removed from 
the cabinet system only after being 
steam-sterilized in an attached double- 
door autoclave or after being contained 
in a nonbreakable sealed 
container, which is then passed through 
a disinfectant dunk tank or a fumigation 
chamber. 
II-B-4-a-(ll). Only persons whose 
entry into the P4 facility is required to 
meet program or support needs shall be 
authorized to enter. Before entering, 
such persons shall be advised of the 
nature of the research being conducted 
and shall be instructed as to the 
appropriate safeguards to ensure their 
safety. They shall comply with 
[200] 
