NOTICES 
49599 
from the controlled access area Into the 
laboratory environment. The exhaust air 
•hall not be recirculated to any other areas 
of the building. Recirculation of air within 
the laboratory room, however, may be pro- 
vided. The exhaust air from the laboratory 
shall be discharged to the outdoors so that 
It la 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. 
•(8) The treated exhaust air from Class I 
and Class n biological safety cabinets ' may 
be discharged either directly to the labora- 
tory or to the outdoors The treated exhaust 
air from a Class ni cabinet shall be dis- 
charged 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 con- 
nected to this system as to avoid any 
interference with the air balance of the cabi- 
net or building exhaust air system. 
4. P4 Level — a. Laboratory Practices (1) 
Laboratory doors shall be kept closed while 
experiments are in progress. 
(3) Work surfaces shall be decontaminated 
following the completion of the experimental 
activity and Immediately following spills of 
organisms containing recombinant DNA 
molecules. 
(8) All biological wastes shall be decon- 
taminated before disposal. Other contami- 
nated materials such as glassware, animal 
cagea and laboratory equipment shall be 
decontaminated before washing, reuse, or 
dlipocii. 
(4) Mechnlcal pipetting devices shall be 
used: pipetting by mouth Is prohibited. 
• (8) Eating, drinking, smoking, and stor- 
age of food are not permitted In the P4 
facility. 
(8) Persons shall wash their hands after 
handling organisms containing recombinant 
DNA molecules and when they leave the 
laboratory. 
(7) Care shall be exercised to minimise the 
creation of aerosols, hr example, manipula- 
tions such ss 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. 
*<8) Biological materials to be removed 
from the P4 facility In a viable or Intact 
state, shall bo transferred to a nonbreak able 
sealed contained which la then removed from 
the P4 facility through a pass-through dis- 
infectant dunk tank or fumigation chamber. 
*(8) No materials, except for biological 
materials that are to remain In a viable or 
Intact state, shall be removed from the P4 
faellitv unless they have been sterilised or 
decontaminated as they pass out of the P4 
facility. All wastes and other materials as 
well as equipment not damaged by high 
temper a ture or steam shall be sterlltred 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 though a pass-through fumi- 
gation chamber. 
•(10) Materials within the Oats III 
cabinets shall be removed from the cabinet 
system only after being sterilized In an at- 
tached double-door autoclave or after being 
contained In a non-breakable seaaled con- 
tainer which la then passed through a disin- 
fectant dunk tank or a fumigation chamber. 
• (11) Only persona whose entry Into the 
P4 facility la required on the basis of program 
or support needs shall be authorized to en- 
ter Such persona shall be advised of the na- 
ture of the research being conducted and In- 
structed aa to the appropriate safeguarda to 
ensure their safety before entry. Such per- 
sons shall comply with Instructions and all 
other required procedures. 
•(12) Persons under 18 years of age shall 
not enter the P4 facility. 
•(13) Personnel shall enter Into and exit 
from the P4 facility only through the cloth- 
ing change and shower rooms. Personnel 
shall shower at each exit from the P4 fa- 
cility. Air locks shall not be used for per- 
sonnel entry or exit except for emergenclea 
•(14) 8treet clothing shall be removed In 
the outer side of the P4 facility clothing 
change area and kept there. Complete labora- 
tory clothing Including undergarments, 
pants and shirts or Jumpsuits, shoes, head 
cover, and gloves shall be provided and used 
by all persons who enter Into the P4 facility. 
Upon exit, this clothing shall be stored In 
lockers provided for this purpose or discarded 
Into collection hampers before personnel 
enter Into the shower area. 
•(18) The universal biohazard sign Is re- 
quired on the P4 facility access doors and all 
m tenor doors to Individual laboratory rooms 
where experiments are conducted. 
(18) An insect and rodent control program 
shall be instituted. 
(17) Animals and plants not related to the 
experiment shall not be permitted In the 
laboratory In which the experiment Is being 
conducted. 
(18) Use of the hypodermic needle and 
syringe shall be avoided when alternate 
methods are available. 
(10) The laboratory shall be kept neat, 
dean, and free of materials not pertinent to 
the research. 
*(20) If experiments Involving other or- 
ganisms which require lower levels of con- 
tainment are to be conducted In the P4 fa- 
cility concurrently with experiments requlr- 
•flse aw. II, D. tor description of biological containment. 
c. Special laboratory design. *(1) The lab- 
oratory shall be located In a restricted sc- 
enes facility which Is either a separate build- 
ing or a clearly demarcated and Isolated sons 
erlthln a building. Clothing change areas and 
shower rooms shall be provided for personnel 
entry and egress These rooms shall be ar- 
ranged so that personnel egress Is through 
the shower area to the change room. A dou- 
ble door ventilated vestibule or ultraviolet 
air lock shall be provided for passage of ma- 
terials. supplies, and equipment which are 
not brought Into the P4 facility through the 
change room area. 
• (3) Walls, floors, and ceilings of the P4 
facility are constructed to form an Internal 
•hell, which readily allows vapor phase de- 
contamination and la animal- and Insect- 
proof. All penetrations through these struc- 
tures 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 air- 
tight ) 
• (3) A foot, elbow, or automatically op- 
erated hand washing facility ah all be pro- 
vided near the door within each laboratory 
In which experiments Involving recombinant 
DNA are conducted In open -face biological 
safety cabinets. 
•(4) Where a central vacuum system 
Is provided. It shall not serve areas out- 
ing P4 level containment, they shall be con- 
ducted In accordance with all P4 level labo- 
ratory practices specified In this section. 
b. Containment equipment. »(1) Experi- 
mental procedures Involving organisms which 
require P4 level physical containment shall 
be conducted either In (a) a Class III cab- 
inet system or In (b) Class I or Class n 
cabinets that are located in a specially de- 
signed area In which all personnel are re- 
quired to wear one-piece positive pressure 
Isolation suits. 
• (2) Laboratory animals Involved In ex- 
periments requiring P4 level physical con- 
tainment shall be housed either In cages con- 
tained In Class in cabinets or In partial 
containment caging systems (such as Hors- 
fall units, open cages placed In ventilated 
enclosures, or solid wall and bottom cages 
covered by Alter 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 re- 
quired to wear one-piece positive pressure 
suits. 
•(3) Alternative re lection of containment 
equipment. Experimental procedures Involv- 
ing a host-vector system which provides a 
one-step higher level of biological contain- 
ment than that specified In Section III can 
be conducted In the P4 facility using con- 
tainment equipment requirements specified 
for the P3 level of physical containment. Al- 
ternative combinations of containment safe- 
guards are shown In Table n on following 
page. 
aide of the P4 facility. The vacuum sys- 
tem shall Include ln-llne HEP A filters as near 
as practicable to each use point or service 
cock. The filters shall be Installed so as to 
permit In -place decontamination and re- 
placement. Water supply and liquid and gas- 
eous services provided to the P4 facility shall 
be protected by devices that prevent back 
flow 
•(8) Food-operated water fountains are 
permitted In the corridors of the P4 facility. ■ 
The water service provided to the fountain 
•hall be independent of the water services to 
the laboratory areas of the P4 facility. 
(6) Laboratory doors shall be self-closing. 
• (7) A double door autoclave shall be pro- 
vided for sterilization of material passing out 
of the P4 facility. The autoclave doors shall 
be Interlocked so that both doors will not 
be open at the same time. 
•(8) A pass-through dunk tank or fumi- 
gation chamber shall be provided for removal 
of material and equipment that cannot be 
heat sterilized from the P4 facility. 
• (0) All liquid affluents from the P4 fa- 
cility shall be collected and decontaminated 
before disposal. Liquid effluents from bio- 
logical safety cabinets and laboratory sinks 
shall be sterilized by heat. Liquid effluents 
from the shower and hand washing facilities 
may be Inactivated by chemical treatment. 
HEPA Alters shall be Installed In all effluent 
drain vent lines. 
Table II. — Combination! of containment tafe/juardt 
Chat Station of expert 
to guidelli 
m«nl arcortlinf 
Combinations of physical and biological containment 
Physical 
Biological* 
Laboratory 
Laboratory 
Containment 
Biological 
containment 
containment 
dettlfn 
*p*rlfWd for — 
practices 
rpectllrd lor— 
equipment 
specified lor— 
containment 
p* 
nvi 
Pi 
Pi 
Pi 
HVl 
Pi 
HVl 
Pi 
Pi 
PI 
HV2 
FEDERAL REGISTER, VOL 43, NO. 107— TUESDAY, SEPTEMBER 27, 1*77 
[ 167 ] 
