Federal Register / Vol. 46, No. 233 / Friday, December 4, 1981 / Notices 
59371 
available in the same building in which 
organisms containing recombinant DNA 
molecules are used. 
II-B-3. P3 Level. 
II-B-3-a. Laboratory Practices. 
II-B-3-a-(l). Laboratory doors shall 
be kept closed while experiments are in 
progress. 
II-B-3-a-(2). Work surfaces shall be 
decontaminated following the 
completion of the experimental activity, 
and immediately following spills of 
organisms containing recombinant DNA 
molecules. 
II-B-3-a-(3). 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. 
II-B-3-a-(4). Mechanical pipetting 
devices shall be used; pipetting by 
mouth is prohibited. 
n-B-3-a-(5). Eating, drinking, 
smoking, and storage of food are not 
permitted in the laboratory area in 
which recombinant DNA materials are 
handled. 
II-B-3-a-(6). Persons shall wash their 
hands after handling organisms 
containing recombinant DNA molecules 
and when they leave the laboratory, 
II-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 
inoculation loop or needle so that it 
splatters, and forceful ejection of fluids 
from pipettes or syringes shall be 
avoided. 
II-B-3-a-(8). 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. 
II-B-3-a-(9). 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. 
II-B-3-a-(10). Persons under 16 years 
of age shall not enter the laboratory. 
II-B-3-a-(ll). 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. 
II-B-3-a-(12). An insect and rodent 
control program shall be instituted. 
II-B-3-a-(13). 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. 
II-B-3-a-(14). Raincoats, overcoats, 
topcoats, coats, hats, caps, and such 
street outer-wear shall not be kept in the 
laboratory. 
II-B-3-a-(15). Gloves shall be worn 
when handling materials requiring P3 
containment They shall be removed 
aseptically immediately after the 
handling procedure and 
decontaminated. 
II-B-3-a-(16). Animals and plants not 
related to the experiment shall not be 
permitted in the laboratory. 
II-B-3-a-(17). Vacmun outlets shall be 
protected by filter and liquid 
disinfectant traps. 
II-B-3-a-(18). Use of hypodermic 
needle and syringe shall be avoided 
when alternative methods are available. 
II-B-3-a-(19). The laboratory shall be 
kept neat and clean. 
II-B-3-a-(20). If experiments 
involving other organisms which require 
lower levels of containment are to be 
conducted in the same laboratory 
concurrently with experiments requiring 
P3-level physical containment, they 
shall be conducted in accordance with 
all P3-level laboratory practices. 
U-B-3-b. Containment Equipment. \ 
lI-B-3-b- (1). Biological safety 
cabinets[20] 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. 
II-B-3-b-(2). Laboratory animals held 
in a P3 area shall be housed in partial- 
containment caging systems, such as 
Horsfall units[19A], 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 
reflectors. 
Note. — Conventional caging systems may ' 
be 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. H | 
II-B-3-b-(3). Alternative Selection of '■ j 
Containment Equipment. Experimental S •, 
procedures involving a host-vector M 
system that provides a one-step higher ¥ | 
level of biological containment can be j ! 
conducted in the P3 laboratory using v 
containment equipment specified for the A i 
P2 level of physical containment. - \ 
Experimental procedures involving a { 
host-vector system that provides a one- I 
step lower level of biological ^ I 
containment can be conducted in the P3 
laboratory using containment equipment | | 
specified for the P4 level of physical u j 
containment. Alternative combinations |. i 
of containment safeguards are shown in 
Table I. |! 
Table I.— Combinations of Containment Safeguards 
Classification of experiment 
Alternate combinations of physical and biological containmsrrt 
Physical 
containment 
Biological* containment 
Physical Containment 
Biological ] 
containment f 
Laboratory 
design 
spedfiea lor— 
Laboratory 
practices 
specified for— 
Containment 
equipment 
specified for— 
P3 
HV3 
P3 
P3 
HV3. 
HVZ 
HV2. j 
HV3. ' 
HVI. 
HV1. 
HV2. J 
P3 
HV3 
P3 
P3 
P4 
P3 
HV2 
P3 
P3.^ 
P3 
P3 
HV2.^ 
P3 
P3 
P2 
HV9 
P3« 
P3 
P4 : 
P3 
HV1 
P3 
P3 
P3 
P3 
HV1 
P3., 
P3 
P2 
' See Section ll-O tor description of biological containment 
II-B-3-C. Special Laboratory Design. 
U-B-3-c(l). 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 
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