60112 
NOTICES 
balance of the cabinet and the build- 
ing 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 comple- 
tion of the experimental activity and 
immediately following spills of organ- 
isms 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 ra- 
dioactive wastes shall be decontami- 
nated by a method demonstrated to be 
effective before washing, reuse, or dis- 
posal. 
II-B-4-a-(4). Mechanical pipetting 
devices shall be used; pipetting by 
mouth is prohibited. 
II-B-4-a-(5). Eating, drinking, smok- 
ing, and storage of food are not per- 
mitted 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 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 trans- 
ferred to a nonbreakable sealed con- 
tainer, 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 (auto- 
claved) 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 equip- 
ment not damaged by high tempera- 
ture 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. 
n-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 enter- 
ing, such persons shall be advised of 
the nature of the research being con- 
ducted and shall be instructed as to 
the appropriate safeguards to ensure 
their safety. They shall comply with 
instructions and all other required 
procedures. 
II-B-4-a-(12). Persons under 18 
years of age shall not enter the P4 fa- 
cility. 
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, includ- 
ing undergarments, head cover, shoes, 
and either pants and shirts or jump- 
suits, shall be used by all persons who 
enter the P4 facility. Upon exit, per- 
sonnel shall store this clothing in lock- 
ers provided for this purpose or dis- 
card it into collection hampers before 
entering the shower area. 
II-B-4-a-(15). The universal bioha- 
zard 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, refrig- 
erators, or other units used to store or- 
ganisms containing recombinant DNA 
molecules. 
II-B-4-a-(16). An insect and rodent 
control program shall be instituted. 
II-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 con- 
ducted. 
II-B-4-a-(18). Vacuum outlets shall 
be protected by filter and liquid disin- 
fectant traps. 
II-B-4-a-(19). Use of the hypoder- 
mic needle and syringe shall be avoid- 
ed when alternate methods are availa- 
ble. • 
II-B-4-a-(20). The laboratory shall 
be kept neat and clean. 
II-B-4-a-(21). If experiments involv- 
ing other organisms which require 
lower levels of containment are to be 
conducted in the P4 facility concur- 
rently with experiments requiring P4- 
level containment, they shall be con- 
ducted in accordance with all P4-level 
laboratory practices specified in this 
section. 
II-B-4-b. Containment Equipment 
II-B-4-b-(l). Experimental proce- 
dures involving organisms that require 
P4-level physical containment shall be 
conducted either in (i) a Class III cabi- 
net system or in (ii) Class I or Class II 
cabinets that are located in a specially 
designed area in which all personnel | 
are required to wear one-piece posi- 
tive-pressure isolation suits. 
II 
U-B-4-b-<2). Laboratory animals in- 
volved in experiments requiring P4- 
level physical containment shall be 
housed either in cages contained in 
Class III cabinets or in partial-contain- 
ment caging systems (such as Horsfall 
units [19A], open cages placed in ven- 
tilated enclosures, or solid-wall and - 
bottom cages covered by filter bon- 
nets, or solid-wall and -bottom cages 
placed on holding racks equipped with 
ultraviolet irradiation lamps and re- j 
Electors) that are located in a specially 
designed area in which all personnel i 
are required to wear one-piece posl- ad 
tive-pressure suits. • i 
II-B-4-b-(3). Alternative Selection of 
Containment Equipment Experimen- >' 
tal procedures involving a host-vector 1 
system that provides a one-step higher "■ 
level of biological containment than 5 
that specified in Part III can be con- il 
ducted in the P4 facility using contain- ji 
ment equipment requirements sped- ■ 
fied for the P3 level of physical con- 
tainment. Alternative combinations of 
containment safeguards are shown in 
Table H. 
Table XI.— Combinations of Containment Safeguards 
Classification of experiment Alternate combinations of physical and biological containment 
according to Guidelines 
Physical containment 
Physical Biological * Laboratory Laboratory Containment Biological 
containment containment design specified practices equipment containment 
for specified for. specif ted'for: : 
. . 
EM HV1 P4 P4 P4 HV1 
P4 HV1 P4 P4 ** P3 HVZ ji 
* See Section U-D for description of biological containment** In this case gloves shall be worn, in addi- 
tion to the clothing requirements specified in ri-B-4-a-(I41. 
FEDERAL REGISTER, VOL 43, NO. 247— FRIDAY, DECEMBER 22, 1978 
