Federal Register / Vol. 47, No. 102 / Wednesday, May 26, 1982 / Notices 
23129 
Appendix G-D-C-l-d. Mechanical 
pipetting devices shall be used; pipetting by 
mouth is prohibited. 
Appendix G-II-C-l-e. Eating, drinking, 
smoking, and storage of food are not 
permitted in the laboratory area in which 
recombinant DNA materials are handled. 
Appendix G-II-C-l-f. Persons shall wash 
their hands after handling organisms 
containing recombinant DNA molecules and 
when they leave the laboratory. 
Appendix G-II-C-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-Il-C-l-h. Contaminated 
materials that are to be steamsterilized 
(autoclaved] or decontaminated at a site 
away from the laboratory shall be placed in 
durable leak-proof container, which is closed 
before removal from the laboratory. 
Appendix G-II-C-l-i. Entry into the 
laboratory shall be through a controlled 
access area. Only persons who have been 
advised of the nature of the research being 
conducted shall enter the controlled access 
area. Only persons required on the basis of 
program or support needs shall be authorized 
to enter the laboratory. Such persons shall be 
advised of the nature of the research being 
conducted before entry, and shall comply 
with all required entry and exit procedures. 
Appendix G-U-C-l-j. Persons under 16 of 
age shall not enter the laboratory. 
Appendix G-II-C-l-k. The universal 
biohazard sign shall be posted on the 
controlled access area door and on all 
laboratory doors when experiments requiring 
P3-level containment are in progress. 
Freezers and refrigerators or other units used 
to store organisms containing recombinant 
DNA molecules, shall also be posted with the 
universal biohazard sign. 
Appendix G-II-C-1-1. An insect and rodent 
control program shall be instituted. 
Appendix G-II-C-l-m. Laboratory clothing 
that protects street clothing (e.g., long-sleeve 
solid-front or wrap-around gowns, no-button 
or slipover jackets) shall be worn in the 
laboratory. Front-button laboratory coats are 
unsuitable. Laboratory clothing shall not be 
worn outside the laboratory and shall be 
decontaminated before it is sent to the 
laundry. 
Appendix G-II-C-l-n. Raincoats, 
overcoats, topcoats, coats, hats, caps, and 
such street outer-wear shall not be kept in the 
laboratory. 
Appenix G-II-C-l-o. Gloves shall be 
worn when handling materials requiring P3 
containment. They shall be removed 
aseptically immediately after the handling 
procedure and decontaminated. 
Appendix G-II-C-l-p. Animals and plants 
not related to the experiment shall not be 
permitted in the laboratory. 
Appendix G-Il-C-l-q. Vaccum outlets 
shall be protected by filter and liquid 
disinfectant traps. 
Appendix G-II-C-l-r. Use of hypodermic 
needle and syringe shall be avoided when 
alternative methods are available. 
Appendix G-Il-C-l-s. The laboratory shall 
be kept neat and clean. 
Appendix G-U-C-l-t. If experiments 
involving other organisms which require 
lower levels of containment are to be 
conducted in the same laboratory 
concurrently with experiments requiring PS- 
level physical containment, they shall be 
conducted in accordance with all P3-level 
laboratory practices. 
Appendix G-II-C-2. Containment 
Equipment. 
Appendix G-II-C-2-a. Biological safety 
cabinets [12] shall be used for all equipment 
and manipulations that produce aerosols — 
e.g., pipetting, dilutions, transfer operations, 
plating, flaming, grinding, blending, drying, 
sonicating, shaking, centrifuging — where 
these procedures involve organisms 
containing recombinant DNA molecules, 
except where equipment design provides for 
containment of the potential aerosal. 
Appendix G-II-C-2-b. Laboratory animals 
held in a P3 area shall be housed in partial- 
containment caging systems, such as Horsfall 
units [11], open cages placed in ventilated 
enclosures, solid-wall and -bottom cages 
covered by Hlter bonnets, or solid-wall and 
Appendix G-II-C-33. Special Laboratory 
Design. 
Appendix G-II-C-3-a. The laboratory shall 
be separated by a controlled access area 
from areas that are open to unrestricted 
traffrc flow. A controlled access area is an 
anteroom, a change room, an air lock or any 
other double-door arrangement that 
separates the laboratory from areas open to 
unrestricted traffic flow. 
Appendix G-II-C-3-b. The 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-II-C-3-c. A foot-, elbow-, or 
automatically-operated hand-washing facility 
shall be provided near each primary 
laboratory exit area. 
Appendix G-II-C-3-d. Windows in the 
laboratory shall be sealed. 
Appendix G-Il-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 G-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 
bottom cages placed on holding racks 
equipped with ultraviolet radiation lamps 
and reflectors. 
Note. — Conventional caging systems may 
be used, provided that all personnel wear 
appropriate personal protective devices, 
liiese shall include, at a.minimum, wrap- 
around gowns, head covers, gloves, shoe 
covers, and respirators. All personnel shall 
shower on exit from areas where these 
devices are required. 
Appendix G-II-C-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 
specihed can be conducted in the P3 
laboratory using containment equipment 
specified for the P2 level or physical 
containment. Experimental procedures 
involving a host-vector system that provides 
a one-step lower level of biological 
containment than that specified can be 
conducted in the P3 laboratory using 
containment equipment specihed for the P4 
level of physical containment. Alternative 
combinations of containment safeguards are 
shown in Table I. 
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 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 ther means for 
effectively reducing an accidental aerosol 
burden provided that it can be dispersed 
clear of occupied buildings and air intakes. 
Appendix G-II-C-3-g. The treated exhaust- 
air from Class I and Class II biological safety 
cabinets [12] 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. 
Appendix G-II-D. P4 Level. 
Table I— Possible Combinations of Containment Safeguards 
Classification of experiment 
Alternate combinations of physical and biological containment 
Physical containment 
Physical containment 
Biological ^ 
containment 
Laboratory 
design specified 
for 
Laboratory 
practices, 
specified for 
Containment 
equipment 
specified for 
Biological 
containment 
P3 
HV2 
P3 
P3 
P3 
HV2. 
P3 
HV2 
P3 
P3 
P4 
HV1. 
P3 
HV1 
P3 
P3 
P3 - 
HV1. 
P3 
HV1 
P3 
P3 
P2....' 
HVi 
* See Appendix I for description of biological containment 
[ 452 ] 
