33072 
NO f ICES 
lory equipment shall be decontaminat- 
ed before washing, reuse, or disposal. 
H-B-2-a-<4). Mechanical pipetting 
devices shall be used: pipetting by 
mouth is prohibited. 
II-B-2-a-<5). Eating, drinking, smok- 
ing and stcr&ge of food are not per- 
mitted in the laboratory area. 
II-B-2-a-<8). Persons shall wash 
their hands after handling organisms 
containing recombinant DNA mole- 
cules and when they leave the labora- 
tory. 
Il-B-2-a-<7). ‘Care shall be exer- 
cised to minimize the creation of aero- 
sols. For example, manipulations such 
as Inserting a hot Inoculating loop or 
needle Into a culture. Taming an in- 
oculation loop or needle so that it 
splatters, and forceful ejection of 
fluids from pipettes or syringes shall 
be avoided. 
II-B-2-a-<8). Contaminated materi- 
als that are to be decontaminated at s 
site away from the laboratory shall be 
placed in a durable leak proof contain- 
er which is closed before removal from 
the laboratory. 
II-B-2-a-<9i. *On’y persons who 
have been advised of the nature of the 
research be.ng conducted shall enter 
the laboratory. 
II-B-2-a-<10). ‘Children under 12 
years of age shall not enter the labora- 
tory . 
I1-B 2-a-tll). The universal bioha- 
zard sign shall be posted on all labora- 
tory access doors when experiments 
requiring P2 containment are in pro- 
gress. Freezers and refrigerators used 
to store organisms containing recom- 
binant DNA molecules shall also be 
pasted with the universal biohazard 
sign. 
lJ-B-2-a-<12). An insect and rodent 
control program shall be instituted. 
II 3-2-a-< 13'. The use of laboratory 
gtrens, coats, o* uniforms is required. 
Laboratory clothing shall not be wem 
to the lunch room or outside of the 
building in which the laboratory is lo- 
cated. 
II-B-2-*-<M>. ‘Animals not related 
to the experiment shall not be permit- 
ted in the laboratory. 
n-B-2-a-tl5'. Use of the hypoder- 
mic needle and syringe shall be avoid- 
ed when alternative methods are avail- 
able. 
II-B-2-a-t 18). The laboratory shall 
be kept neat end clean. 
II-B-2-a-« 17). ‘Experiments of lesser 
biohazard potential can be carried cut 
concurrently in carefully demarcated 
areas of the same laboratory. 
II-B-2-b. Contairvrient Equipment 
‘Biological safety cabinets (20) shall 
be used to contain aerosol-producing 
equipment such as blenders, lyophi- 
‘Deraoles laboratory practices, contain- 
ment equipment, or special laboratory 
design which were not required at the next 
lower level of containment. 
lizers, sonicators. and centrifuges 
when used to process organisms con- 
taining recombinant DNA molecules, 
except where equipment design pro- 
vides 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. 
II-B-2-c. SpecicU Laboratory Derim. 
•An autoclave for sterilization of 
wastes and contaminated materials 
shall be available In the same building 
in which organisms containing recom- 
binant DNA molecules are used. 
II-B-3. P3 Level. 
II-B-3-a. Laboratory Practice*. 
II-B-3-a-< 1 ). Laboratory doors shall 
be kept closed while experiments are 
in progress. 
ri-B-3-a-(2). ‘Work surfaces shall be 
decontaminated following the comple- 
tion of the experimental activity, and 
Lmmedia'ely folowlng spills of organ- 
isms contain ng recombinant DNA 
molecules. 
II-B-3-a-<3). All biological wastes 
shall be decontaminated before dispos- 
al. Other contaminated materials such 
as glassware, ar.lmtl cages, and labora- 
tory equipment shall be decontaminat- 
ed before washing, reuse, or disposal. 
II-B-3-a-<4). Mechanical pipetting 
devices shall be used: pipetting by 
month is prohibited. 
II-B-3-a-< 5 >. Ea'ing. drinking, smok- 
ing. and storage of food are not per- 
mitted in the laboratory area. 
II-B-3 a-< 6). Persons shall wash 
their hands after handling organisms 
containing recombinant DNA mole- 
cules ar.d when they leave the labora- 
tory. 
lI-B-3-a-<7). 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 in- 
oculation loop or needle so that it 
splatters, and forceful ejection of 
fluids from pipettes or synnges shall 
be avoided. 
!I-B-3-a-t8>. Contaminated materi- 
als that are to be decontaminated at a 
site away from the laboratory shall be 
placed in a durable leak-proof contain- 
er which is closed before removal from 
the iabora'ory. 
Il-B-3-a-(9). ‘Entry into the labora- 
tory shall be through a controlled 
access area. Only persons who have 
been advised of the nature of the re- 
search 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. 
II-B-3-a-<10/. Children under 12 
years of age shall not enter the labora- 
tory. 
II-B-3-a 11). The universal bioha- 
zard s.jn shall be pasted on the cen- 
tre/ led access area door and on all lab- 
oratory doers when experiments re- 
qu.r.rg P3-level containment are in 
progress. Freezers and refrigerators 
used to store organisms containing re- 
combinant DNA molecules shall also 
be ported with the universal biohazard 
s’gn. 
II-3-3-a-<12). An insect and rodent 
control program shall be instituted. 
II-B-3-a-<13). ‘Laboratory clothing 
tnat pro'ects street ciothing tLe.. long- 
sleeve solid- front or wrap-around 
gowns, no-button or slipover jackets, 
etc.) shall be worn in the laboratory. 
Front-button laboratory coats are un- 
su'taoie. Laboratory clothing shall not 
be worn ouU.de the laboratory and 
shah be decontaminated before it is 
sent to the laundry. 
II-B-3-a-tl4). ‘Raincoats. overcoaU. 
topcoats, coats, hats. caps, and such 
street outer- wear shall not be kept in 
the laboratory. 
U-B-3-a-t 15 ). “Gloves shall be worn 
when hardlmg materials requiring P3 
containment. They shall be removed 
asepticaliy immediately after the han- 
dling procedure and decontaminated. 
II-3-3-a-<16). ‘Animals end plants 
not related to the experiment shall 
net ber permitted in the laboratory. 
II-B-3-a-( 17). ‘Vacuum outlets shall 
be protected by filter and liquid disin- 
fectant traps. 
II-B-3-a-< 18). Cse of hypodermic 
needle and syringe shall be avoided 
when alternative methods are availa- 
ble. 
I!-B-3-a-<19). The laboratory shall 
be kept neat and clean. 
Il-B-3-a-<20). ‘If experiments in- 
volving other organ .sms which require 
lower levels of containment are to be 
conducted in the same laboratory con- 
currently with experiments requiring 
P3-ieve! physical containment, they 
shall be conducted in accordance with 
all P3-Ievel laboratory prac tices. 
II-B-3-o. CorMunment Equipment 
II-B-3-b-< 1 ). ‘Biological safety cabi- 
nets 20 shall be used tor all equipment 
and manipulations that produce aero- 
sols — e.g.. pipetting, dilutions, transfer 
operations, plating, flaming, grinding, 
blending, drying, sonicating, shaking, 
centrifuging— where these procedures 
Involve organisms containing recom- 
binant DNA molecules, except where 
equipment design provides for contain- 
ment of the potential aerosoL 
II-E-3-b-2). ‘Laboratory animals 
held in a P3 area shall be housed in 
partial containment caging systems 
such as Eorsfali units, open cages 
placed in ventilated enclosures, soiid- 
wall-and-bottom cages covered by 
filter bonnets, cr solid-wall and 
FEDERAL REGISTER. VOL 4j NO. 14Z — FRIC AT, JULY 28 1978 
[ 33 ] 
