23130 
Federal Register / Vol. 47, No. 102 / Wednesday, May 26, 1982 / Notices 
Appendix G-D-D-1. Laboratory Practices. 
Appendix G-U-D-l-e. Laboratory doors 
shall be kept closed while experiments are io 
progress. 
Appendix C-Il-O-l-b. Work surfaces shall 
be decontaminated following the completion 
of the experimental activity and immediately 
following spills of organisms containing 
recombinant DNA molecules. 
Appendix G-U-D-l-c 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. 
Appendix G-D-O-l-d. Mechanical 
pipetting devices shall be' used: pipetting by 
mouth is prohibited. 
Appendix C-D-O-l-e. Eating, drinking, 
smoking, and storage of food are not 
permitted in the P4 facility. 
Appendix C-O-O-l-f. Arsons shall wash 
their hands after handling organisms 
containing recombinant DNA molecules and 
when they leave the laboratory. 
Appendix C-O-D-l-g. Care shall be 
exendaed to minimize the creation of 
aerosols. For example, manipulations such as 
inserting a hot inoculating loop or needle Into 
a culture, flaming an inoculation lop or 
needle so that it splatters, and forceful 
election of fluids from pipettes or syringes 
shall be avoided. 
Appendix C-0-4>-l-h. Biological materials 
to be removed from the P4 faciUty in a viable 
or Intact state shall be transferred to a 
oonbreakable sealed container, which Is then 
removed from the P4 facility through a pass- 
through disinfectant dunk tank or fumigetion 
chamber. 
Appendix C-O-D-l-i. No materials, except 
for biological materials that are to remain in 
a viable or intact state, shall be removed 
from the Pe faciUty 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 
aquipment 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 damag^ by 
temperature or steam shall be removed from 
the P4 facUlty through a pasethrough 
fumigation chamber. 
Appendix C-D-D-1-). Materials within the 
Oass 01 cabinets shall be removed from the 
cabinet system only after being steam- 
sterilized In an attached double-door 
autoclave or after beiitg contained in a 
nonbraakabla sealed container, which ia then 
passed through a disinfectant dunk tank or a 
fumigation chamber. 
Appendix C-ll-O-l-k. 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. 
Appendix C-H-D-l-L Persons under 18 
yeers of age shall not enter the P4 facility. 
Appendix G-U-D-l-m. Personnel shall 
enter into and exit fron the P4 facility only 
through the clothing ohange 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. 
Appendix G-II-D-l-n. 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. 
Appendix G-D-D-l-o. 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 unit* used to store organisms 
containing recombinant DNA molecules. 
Appendix C-O-D-l-p. An insect and 
rodent control program shall be instituted. 
Appendix G-U-D-l-q. Animals and plants 
not related to the experiment shall not be 
permitted in the laboratory in which the 
experiment is being conducted. 
Appendix C-II-D-l-r. Vacuum outlets 
shall be protected by filter and liquid 
disinfectant traps. 
Appendix C-U-O-l-s. Use of the 
hypodermic needle and syringe shall be 
avoided when alternate methods are 
available. 
Appendix C-lI-O-l-L The Laboratory shall 
be kept neat and clean. 
Appendix C-U-D-3. Special Laboratory 
Design. 
Appendix G-D-O-8-a. The laboratory shall 
be located in a restricted-access facility 
which is either a separate building or a 
clearly demart»ted and isolated zone within 
a building. Clothing-change areas and shower 
rooms shall be provided for personnel entry 
and egress. These rooms shall be arranged so 
that personnel leave through the shower area 
to the change room. A double-door ventilated 
vestibule or ultraviolet air lock shall be 
provided for passage of materials, supplies, 
and equipment which are not brought into the 
P4 facility through the change room area. 
Appendix G-0-D-3-b. Walls, floors, and 
ceilings of the P4 facility are constructed to 
form an internal shell which readily allows 
vapor-phase decontamination and is animal- 
Appendix G-II-D-l-u. 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. . 
Appendix G-Il-D-2. Containment 
Equipment 
Appendix G-U-D-2-a. Experimental 
procedures involving organisms that require 
P4-level physical containment shall be 
conducted either in (i) a Class UI cabinet 
system or in (ii) Gass I 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. 
Appendix C-U-B-4-b. Laboratory animals 
involved tn experiments requiring P4-level 
physical containment shall be housed either 
in cages contained in Gass 111 cabinets or in 
partial containment caging systems (such as 
Horsfall units [11], open cages placed in 
ventilated enclosures, or solid-wall and- 
bottom cages covered by filter bonnets, or 
solid-wall and -bottom cages placed on 
holding racks equipped with ultraviolet 
irradiation lamps and reflectors) that are 
located in a specially designed area in which 
all personnel are required to wear one-piece 
positive-pressure suits. 
Appendix G-n-D-2-c. 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 can be conducted in the P4 facility 
using containment equipment requirements 
specified for the P3 level of physical 
containment. Alternative combinations of 
containment safeguards are shown in Table 
U. 
and insect-proof. All penetrations through 
these structures and surfaces are sealed. (The 
Integrity of the walls, floors, ceilings, and 
penetration seals should ensure adequate 
containment of a vapor-phase decontaminant 
under static pressure conditions. This 
requirement does not imply that these 
surfaces must be airtight] 
Appendix G-n-D-3-c. A foot-, elbow-, or 
automatically-operated handwashing facility 
shall be provided near the door within each 
laboratory in which experiments involving 
recombinant DNA are conducted in openface 
biological safety cabinets. 
Appendix G-II-D-S-d. Central vacuum 
systems are permitted. The system, if 
provided, shall not serve areas outside the P4 
facility. The vacuum system shall include in- 
line HEPA filters near each use point or 
Table H.— Possible Combinations or Containment Safeguards 
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