Federal Register / Vol. 47. No. 167 / Friday, August 27. 1982 / Notices 
38063 
conducted in the P3 laboratory using Alternative combinations of 
containment equipment specified for the . containment safeguards are shown in 
P4 level of physical containment. Table 1. 
Table I.— Possible Combinations Of Containment Safequaaos 
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Laboratory daav< 
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Labcratary 
pracaoaa apactaad 
tar- 
Cootaavnara 
aqutamara 
■QioSaO tar 
PI 
PI 
P* 
Pi 
KV3. 
M 
P% 
Pi 
P4 ... . 
HV1 
PI 
PI 
Pi 
Pi 
KV1 
P9 
P* 
Pi 
Pi 
HV2 
' 8m App<nfti I lor Oooor^ptton Of 0WiD9l co l oorOMniioro 
Appendix G-D-C-3. Special 
Laboratory Design. 
Appendix G-U-C-3-«. The laboratory 
shall be separated by a controlled 
access area from areas that are open to 
unrestricted traffic flow. A controlled 
access area is an anteroom, a chanM 
room, an air lock or any other double- 
door arrangement that separates the 
laboratory from areas open to 
unrestricted tragic flow. 
Appendix C-U-C-9^. Tha surfaces of 
walls, floors, and ceilings shall be 
readily cleanable. Penetrations through 
these surfaces shall be sealed or 
capable of being sealed to facilitate 
space decontamination. 
Appendix G-n-C-3-c A foot-, elbow-, 
or automatically-operated hand-washing 
facility shall be provided near each 
primary laboratory exit area. 
Appendix C-Il-C-3-d. Windows in 
the laboratory shall be sealed. 
Appendix C-O-C-3-e. An autoclave 
for sterilization of wastes and 
contaminated materials shall be 
available in the same building (and 
preferably within the controlled 
laboratory area) in which organisms 
containing recombinant DNA molecules 
are used. 
Appendix C-II-C-3-f. The laboratory 
shall have a ventilation system that is 
capable of controlling air movement. 
The movement of air shall be from areas 
of lower contamination potential to 
areas of higher contamination potential 
(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 moveoMot 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 
Altered by HEPA Alters or equivalent. 
The exhaust air from the labwatory area 
can be discharged to the outdoors 
without flltration or other mecms for 
effectively reducing an accidental 
aerosol burden provided that it can be 
dispersed clear of occupied buildings 
and air intakes. 
Appendix C-Il-C-3-g. The treated 
exhaust-air from Class I and Class U 
biological safety cabinets [12] may be 
discharged kither 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. 
Appendix C-II-D. P4 Level. 
Appendix C-II-D-1. Laboratory 
Practices. 
Appendix G-O-D-l-a. Laboratory 
doors shall be kept closed while 
experiments are in progress. 
Appendix C-D-O-l-b. Work surfaces 
shall be decontaminated following the 
completion of the experimental activity 
and inrmediately follo%ving spills of 
organisms containing recombinant DNA 
molecules. 
Appendix C-Il-D-l-c All laboratory 
wastes shall be steam-sterilized 
(autoclaved) before disposal. Other 
contaminated materials such as 
glassware, animal cages, laboratory 
equipmenL and radioactive wastes shall 
be decontaminated by a method 
demonstrated to be effective before 
washing, reuse, or dispose. 
Appendix G-Il-D-l-d. Mechanical 
pipetting devices shall be used: pipetting 
by mouth Is prohibited. 
Appendix G-Il-D-l-e. Eating, 
drinking, smoking, and storage of food 
are not permitted in the P4 facility. 
Appendix G-II-D-l-f. Persons shall 
wash their hands after handling 
organisms containing rerombiant DNA 
molecules and when they leave the 
laboratory. 
Appendix G-II-D-l-g. 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. 
Appendix G-II-D-l-h. 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 them removed from 
the P4 facility through a passthrough 
disinfectant dunk tank or fumigation 
chamber. 
Appendix G-II-D-l-i. 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 passthrough 
fumigation chamber. 
Appendix G-II-D-l-j. 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. 
Appendix G-D-D-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 requir^ 
procedures. 
Appendix G-U-D-1-1. Persons under 
18 years of age shall not enter the P4 
facility. 
Appendix G-II-D-l-m. Personnel 
shall enter into and exit from the P4 
facility only through the clothing chang< 
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. 
[ 17 ] 
