Federat Register / Vol. 46, No. 233 / Friday, December 4, 1981 / Notices 
59403 
(i.e., from the controlled access area to 
the laboratory area). If the ventilation 
system provides positive pressure 
supply air, the system shall operate in a 
manner that prevents the reversal of the 
direction of air movement or shall be 
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. 
n-B-4. P4 Level. 
U-B-4-a. Laboratory Practices. 
II-B-4-a-(l). Laboratory doors shall 
be kept closed while experiments are in 
progress. 
lI-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. 
U-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. 
ll-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. 
lI-B-4-a-(6). Persons shall wash their 
hands after handling organisms 
containing recombinant DNA molecules 
and when they leave the laboratory. 
Il-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. 
lI-B-4-a-(8). Biological materials to 
be removed from the P4 facility in a 
viable or intact state shall be 
trahsferred to a nonbreakable sealed 
container, which is then removed from 
the P4 facility through a pass-through 
disinfectant dunk tank or fumigation 
chamber. 
Il-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. 
n-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 
instructions and all other required 
procedures. 
n-B-4-a-(12). Persons under 18 years 
of age shall not enter the P4 facility. 
II-B-4-a-(13). Personnel shall enter 
into and exit from the P4 facility only 
through the clothing change and shower 
rooms. Personnel shall shower at each 
egress from the P4 facility. Air locks 
shall not be used for personnel entry or 
exit except for emergencies. 
II-B-4-a-(14). Street clothing shall be 
removed in the outer side of the 
clothing-change area and kept there. 
Complete laboratory clothing, including 
undergarments, head cover, shoes, and 
either pants and shirts or jumpsuits, 
shall be used by all persons who enter 
the P4 facility. Upon exit, personnel 
shall store this clothing in lockers 
provided for this purpose or discard it 
into collection hampers before entering 
the shower area. 
II-B-4-a-(15). The universal 
biohazard sign is required on the P4 
facility access doors and on all interior 
doors to individual laboratory rooms 
where experiments are conducted. The 
sign shall also be posted on freezers, 
refrigerators, or other units used to store 
organisms containing recombinant DNA 
molecules. 
II-B-4-a-(16). An insect and rodent 
control program shall be instituted. 
n-B-4-a-(17). Animals and plants not 
related to the experiment shall not be 
permitted in the laboratory in which the 
experiment is being conducted. 
II-B-4-a-(18). Vacuum outlets shall be 
protected by filter and liquid 
disinfectant traps. 
U-B-4-a-(19). Use of the hypodermic 
needle eind syringe shall be avoided 
when alternate methods are available. 
II-B- 4 -a-(20). The laboratory shall be 
kept neat and clean. 
n-B-4-a-(21). If experiments 
involving other organisms which require 
lower levels of containment are to be 
conducted in the P4 facility concurrently 
with experiments requiring P4-level 
containment, they shall be conducted in 
accordance with all P4-level laboratory 
practices specified in this section. 
II-B-4-b. Containment Equipment. 
II-B-4-b-(l). Experimental procedures 
involving organisms that require P4- 
level physical containment shall be 
conducted either in (i) a Class HI cabinet 
system or in (ii) Class 1 or Class II 
cabinets that are located in a specially 
designed area in which all personnel are 
required to wear one-piece positive- 
pressure isolation suits. 
II-B-4-b-(2). Laboratory animals in 
experiments requiring P4-level physical 
containment shall be housed either in 
cages contained in Class III cabinets or 
in partial containment caging systems 
(such as Horsfall units [19A], open cages 
placed in ventilated enclosures, or solid- 
wall and -bottom ca,ges covered by filter 
bonnets, or solid-wall and -bottom cages 
placed on holding racks equipped with 
ultraviolet irradiation lamps and 
reflectors) that are loated in a specially f 
designed area in which all personnel arej 
required to wear one-piece positive- I 
pressure suits. j 
II-B-4-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 HI can be conducted in 
the P4 facility using containment 
equipment requirements specified for 
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