Federal Register / Vol. 48, No. 100 / Wednesday, June 1, 1983 / Notices 
24571 
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. 
Appendix 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-II-C-l-q. Vacuum 
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-II-C-l-s. The laboratory 
shall be kept neat and clean. 
Appendix G-II-C-l-t. If experiments 
involving other organisms which require 
lower levels of containment are to be 
conducted in the same laboratory 
concurrently with expermiments 
requiring P3-leve! 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 aerosol. 
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 filter bonnets, or solid-wall 
and -bottom cages placed on holding 
racks equipped with ultraviolet 
radiation lamps and relectors. 
Note. — Conventional caging systems may 
he used, provided that all personnel wear 
appropriate personal protective devices. 
These 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 specified can be 
conducted in the P3 laboratory using 
containment equipment specified for the 
1 See Appendix I for description of biological containment. 
Appendix G-II-C-3. 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 traffic 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-Il-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 handwashing 
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-lI-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 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 
P2 level of 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 specified for the 
P4 level of physical containment. 
Alternative combinations of 
containment safeguards are shown in 
Table I. 
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 other 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. 
Appendix G-II-D-1. Laboratory 
Practices. 
Appendix G-II-D-l-a. Laboratory 
doors shall be kept closed while 
experiments are in progress. 
Appendix G-II-D-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-II-D-l-c. All laboratory 
wastes shall be steam-sterilized 
(autoclaved) before disposal. Other 
Table I.— Possible Combinations of Containment Safeguards 
Classification of experiment 
Alternate combinations of physical and biological containment 
Physical Containment 
Physical containment 
Bioogical 
containment 1 
Laboratory design 
specified for 
Laboratory 
practices 
specified for 
Containment 
equipment 
specified for 
Biotogical 
containment 
P3 
HV2 
P 3 
P3 
P3 
HV2 
PI 
HV2 
P3 
P3 
P4 
HV1 
P3 
HV1 
P3 
P3 
P3 . ... 
HV1 
P3 
HV1 
P3 
P3 
P 2 
HV2 
[1511 
