23128 
Federal Register / Vol. 47, No. 102 / Wednesday. May 26, 1962 / Notices 
to all worker*. Where terologicel monitoring 
ia clearly appropriate it ahall be provided. 
(See Section IV-B-l-f.) 
The "Laboratory Safety Monograph", 
available from ORDA. describe* practices, 
equipment, and facilities in detail 
Appendix G-fl. Phytica! Containment 
LeveJs. Tha objective of physical containment 
ia to coofina organisms containing 
recombinant DNA molecules, and thus to 
reduce the potential for expoaure of the 
laboratory worker, persons outside of the 
laboratory, and the environment to organisms 
containing recombinant DNA molecules. 
Physical containment ia achieved through the 
use of laboratory practices, containment 
equipmenl and *{wcial laboratory design. 
Emphasis ia plac^ on primary means of 
physical containment which are provided by 
la^ratory practices and containment 
equipment Special laboratory design 
provides a secondary meatu of protection 
against the accidental release of organisms 
outside the laboratory or to the envlroiunent 
Special laboratory design la used primarily in 
facilities in which experiments of moderate to 
high potential hazards are performed. 
Combinations of laboratory practices, 
oontalnment equipment and special 
laboratory design can be made to achieve 
different levels of physical containment Four 
levels of physical containment, which are 
designate as Pi, P2. P3. and P4. are 
described. It should be emphasized that the 
descriptions and assignments of physical 
containment detailed below are based on 
existing approachee to containment of 
pathogenic organisms. For example, the 
"Qasaificatioo of Etlologlc Agents on the 
Basis of Hazard." (2| prepared by the Center* 
for Disease Control describes four general 
levels which roughly correepond to our 
descriptions for Pi. P2. P3. and Pi: and the 
National Cancer Institute describes three 
levels for research on oncogenic viruses 
which roughly correspond to our P2. P3. and 
Pi levels. (3) 
It Is recognized that several different 
combinations of laboratory practices, 
contaiiunent equipment a^ special 
laboratory design may be appropriate for 
contairunent of speciflc research activities. 
The Guidelines, therefore, allow alternative 
selections of primary contalmeni equipment 
arithln fadlit^ that have been designed to 
provide P3 and Pi levels of physical 
containment. The selection of alternative 
methods of primary containment is 
dependent however, on the level of 
biological containment provided by the host- 
vector system used In t^ expenment. 
Consideration will also be given by the 
Director, NIK with the advice of tha 
Recombinant DNA Advisory Committee to 
other combinations which achieve an 
eqoivalenl level of containment (See Section 
IV-C-l-b-<2Hb).) 
Appendix G-Il-A. P! Level 
Appendix G-U-A-1. Laboratory Practice*. 
Appendix G-O-A-l-a. Laboratory doors 
shall be kept closed while experiments are in 
progre s s. 
Appendix G-U-A-l-b. Work surfaces shall 
be decontaminated daily, and immediately 
following spills of organisms containing 
recombinant DNA molecules. 
Appendix G-D-A-l-c. All biological 
wastes shall be decontaminated before 
disposal Other contaminated materials, such 
as glassware, animal cages, and laboratory 
equipment shall be decontaminated before 
washing, reuse, or disposal. 
Appendix G-O-A-l-d. Mechanical 
pipetting devices shall be used: pi|>etting by 
mouth is prohibited. 
Appen^x G-U-A-l-e. Eating, drinking, 
smoking, and storage of foods are not 
permitted in the laboratory area in which 
recombinant DNA materials are handled. 
Appendix G-Il-A-l-f. Persons shall wash 
their hands after handling organisms 
containing recombinant DNA molecules and 
when they leave the laboratory. 
Appendix G-O-A-l-g. Care shall be taken 
in the conduct of all procedures to minimize 
the creation of aerosols. 
Appendix G-D-A-l-h. Contaminated 
materials that are to be decontaminated at a 
site away from the laboratory shall be placed 
in a durable leakproof container, which is 
closed before removal from the laboratory. 
Appendix C-O-A-t-4. An insect and lodent 
control program shall be Instituted. 
Appendix C-D-A-l-(. The use of 
laboratory gowns, coats, or uniforms is 
discretionary with the laboratory supervisor. 
Appendix G-O-A-I-k. Use of hypodermic 
needle and syringe ahall be avoided when 
alternative methods are available. 
Appendix G-Q-A-l-l The laboratory ahall 
be kept neat and clean. 
Appendix G-U-A-2. Containment 
Equipment Special containment equipment is 
not required at the Pi level 
Appendix G-ll-A-3. Special Laboratory 
Deeign. Special laboratory design is not 
required at the Pi level 
Appendix G-Il-B. P2 Level. 
Appendix G-II-B-1. Laboratory Practice*. 
Appendix G-ll-B-l-a. Laboratory doors 
shall be kept closed while experiments are in 
progress. 
Appendix G-Il-B-l-b. Work surfaces shall 
be decontamiitated dally, and Immediately 
following spills of organisms containing 
recombinant DNA molecules. 
Appendix G-D-B-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 means 
demonstrated to be effective before washing, 
reuse, or disposal 
Appendix G-U-B-l-d. Mechanical 
pipetting devices shall be used: pipetting by 
mouth is prohibited. 
Appendix C-U-B-l-e. Eating, drinking, 
smoking, and storage of food are not 
permitted in the laboratory area in which 
recombinant DNA materials are handled. 
Appendix C-H-B-l-f. Persons shall wash 
their hands after handling organisms 
containing recombinant DNA molecules and 
when they leave the laboratory. 
Appendix G-B-B-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-D-B-l-h. Contaminated 
materials that are to be steam sterilized 
(autoclaved) or decontaminated at a site 
away from the laboratory shall be placed in a 
durable leak-proof container, which is closed 
before removal from the laboratory. 
Appendix G-II-B-l-i. Only persons who 
have been advised of the nature of the 
research being conducted shall enter the 
laboratory. 
Appendix C-ll-B-l-j. The universal 
biohazard sign shall be posted on all 
laboratory access doors when experiments 
requiring P2 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-lI-B-l-k. An insect and 
rodent control program shall be instituted. 
Appen4ix G-II-B-1-1. The use of 
laboratory gowns, coats, or uniforms is 
required. Laboratory clothing shall not be 
worn to the lunch room or outside of the 
building in which the laboratory is located. 
Appendix C-Q-B-l-m. Animals not related 
to the experiment shall not be permitted in 
the laboratory. 
Appendix G— il-B-l-n. Use of tho 
hypodermic needle and syrirtge shall be 
avoided when alternative methods are 
available. 
Appendix G-Il-B-l-o. The laboratory shall 
be kept neat and clean. 
Appendix G-lI-B-l-p. Experiments of 
lesser biohazard potential can be carried out 
conctirrently in carefully demarcated areas of 
the same laboratory. 
Appendix G-U-^2. Containment 
Equipment Biological safety cabinets (12) 
shall be used to contain aerosol-producing 
equipment such as blenders, lyophilizers, 
sonicators. and centrifuges, when used to 
process organisms containing recombinant 
DNA molecules, except where equipment 
design provides for containment of the 
potential aerosol. For example, a centrifuge 
may be operated in the open if a sealed head 
or safety centrifuge cups are used. 
Appendix G-Il-B-3. Special Laboratory 
Deeign. An autoclave for sterilization of 
wastes and contaminated materials shall be 
available in the same building in which 
organisms containing recombinant DNA 
molecules are used. 
Appendix G-Il-C P3 Level. 
Appendix G-U-C-1. Laboratory Practice*. 
Appendix G-Il-C-l-a. Laboratory doors 
shall be kept closed while experiments are in 
progress. 
Appendix G-ll-C-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-C-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 
[ 451 ] 
