Federal Register / Vol. 47, No. 167 / Friday, August 27, 1982 / Notices 
38061 
followed if an accident contaminates 
personnel or the environment. The 
principal investigator must ensure that 
everyone in the laboratory is familiar 
with both the potential hazards of the 
work and the emergency plan. (See 
Sections IV-B-3— d and fV-B-S^.) If a 
research group is working with a known 
pathogen where there is an elective 
vaccine it should be made available to 
all workers. Where serological 
monitoring is clearly appropriate it shall 
be provided. (See Section IV-B-l-f.) 
The "Laboratory Safety Monograph", 
available from ORDA. describes 
practices, equipment, and facilities in 
detail. 
Appendix C-II. Phytica! Containment 
Leveii. The objective of physical 
containment is to confine organisms 
containing recombinant DNA molecules, 
and thus to reduce the potential for 
exposure of the laboratory worker, 
persons outside of the laboratory, and 
the environment to organisms containing 
recombinant DNA molecules. Physical 
containment is achieved through the use 
of laboratory practices, containment 
equipment, and special laboratory 
design. Emphasis is placed on primary 
means ef physical containment which 
are provided by laboratory practices 
and containment equipment. Special 
laboratory design provides s secondary 
means of protection against the 
accidental release of organisms outside 
the laboratory or to the environment. 
Special laboratory design is used 
primarily in facilities in which 
experiments of moderate to high 
potential hazards are perform^. 
Combinations of laboratory practices, 
containment equipment, and special 
laboratory design can be made to 
achieve different levels of physical 
containment Four levels of physical 
containment which are designated as 
Pi. P2, P3, snd P4. are described. It 
should be emphasized that the 
descriptions and assignments of 
physical contaiiunent detailed below are 
based on existing approaches to 
containment of pathogenic organisms. 
For example, the "Classification of 
Etiologic Agents on the Basis of 
Hazard." (2| prepared by the Centers for 
Disease Control describes four general 
levels which roughly correspond to our 
descriptions for Pi, P2. P3. and P4: and 
the National Cancer Institute describes 
three levels for research on oncogenic 
viruses which roughly correspond to our 
P2. P3. and P4 levels. (3) 
It is recognized that several different 
combinations of laboratory practices, 
containment equipment, and special 
laboratory design may be appropriate 
for containment of specific research 
activities. The Guidelines, therefore. 
allow alternative selections of primary 
containment equipment within facilities 
that have been designed to provide P3 
and P4 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 the experiment. 
Consideration will also be given by the 
Director, NIH, «vith the advice of the 
Recombinant DNA Advisory Committee 
to other combinations which achieve an 
equivalent level of containment. (See 
Section IV-C-l-b-(2Hb).) 
Appendix G-II-A. Pi Level. 
Appendix C-U-A-1. Laboratory 
PracUcee. 
Appendix C-II-A-l-a. Laboratory 
doors shall be kept closed while 
experiments are in progress. 
Appendix C-II-A-l-b. Work surfaces 
shall be decontaminated daily, and 
immediately following spills of 
organisms containing recombinant DNA 
molecules. 
Appendix C-Il-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 C-ll-A-l-d. Mechanical 
ipetting devices shall be used: pipetting 
y mouth is prohibited. 
Appendix C-D-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-fl-A-l-f. Persons shall 
wash their hands after handling 
organisms containing recombinant DNA 
molecules and when they leave the 
laboratory. 
Appendix C-H-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-Il-A-l-i. An insect and 
rodent control program shall be 
instituted. 
Appendix G-O-A-l-j. The use of 
laboratory gowns, coats, or uniforms is 
discretionary with the laboratory 
supervisor. 
Appendix G-II-A-l-k. Use of the 
hyp^ermic needle and syringe shall be 
avoided when alternative methods are 
available. 
Appendix C-U-A-l-l. The laboratory 
shall be kept neat and clean. 
Appendix C-II-A-2. Containment 
Equipment. Special containment 
equipment is not required at the Pi level. 
Appendix G-II-A-3. Special 
Laboratory Design. Special laboratory 
design is not required at the Pi level. 
Appendix G-Il-B. P2 Level. 
Appendix G-II-B-1. Laboratory 
Practices. 
Appendix G-II-B-l-a. Laboratory 
doors shall be kept closed while 
experiments are in progress. 
Appendix G-II-B-1 -b. Work surfaces 
shall be decontaminated daily, and 
Immediately following spills of 
organisms containing recombinant DNA 
molecules. 
Appendix G-II-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-II-B-l-d. Mechanical 
pipetting devices shall be used: pipetting 
by mouth is prohibited. 
Appendix G-II-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 G-II-B-l-f. Persons shall 
wash their hands after handling 
organisms contaiiUng recombinant DNA 
molecules and when they leave the 
laboratory. 
Appendix G-Il-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 Q-U-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 Mfore removal from the 
laboratory. 
Appendix G-Il-B-l-i. Only persons 
who have been advised of the nature of 
the research being conducted shall enter 
the laboratory. 
Appendix G-U-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. 
( 15 ] 
