Federal Register / Vol. 45, No. 72 / Friday, April 11, 1980 / Notices 
24969 
are made for large-scale research or 
production of viable organisms 
containing recombinant DNA molecules 
which require P4 containment at the 
laboratory scale. If necessary, these 
requirements will be established on an 
individual case basis. 
VII-B. Pl-LS Level 
VII-V-1. Cultures of viable organisms 
containing recombinant DNA molecules 
shall be handled in a closed system (e.g., 
closed vessel used for the propagation 
and growth of cultures) or other primary 
containment equipment (e.g., biological 
safety cabinet containing a centrifuge 
used to process culture fluids) which is 
designed to reduce the potential for 
escape of viable organisms. Volumes 
less than 10 liters may be handled 
outside of a closed system or other 
primary containment equipment 
providing all physical containment 
requirements specified in Section 1I-B-1, 
of the Guidelines are met. 
VH-B-2. Culture fluids (except as 
allowed in VII-B3.) shall not be removed 
from a closed system or other primary 
containment equipment unless the 
viable organisms containing 
recombinant DNA molecules have been 
inactivated by a validated inactivation 
procedure. A validated inactivation 
procedure is one which has been 
demonstrated to be effective using the 
organism that will serve as the host for 
propagating the recombinant DNA 
molecules. 
VH-B-3. Sample collection from a 
closed system, the addition of materials 
to a closed system and the transfer of 
culture fluids from one closed system to 
another shall be done in a manner 
which prevents the release of aerosols 
or contamination of exposed surfaces. 
VII-B-4. Exhaust gases removed from 
a closed system or other primary 
containment equipment shall be treated 
by Biters which have efficiencies 
equivalent to HEPA filters or by other 
equivalent procedures (e.g., incineration) 
to prevent die release of viable 
organisms containing recombinant DNA 
molecules to the environment. 
VII-B-5. A closed system or other 
primary containment equipment that has 
contained viable organisms containing 
recombinant DNA molecules shall not 
be opened for maintenance or other 
purposes unless it has been sterilized by 
a validated sterilization procedure. A 
validated sterilization procedure is one 
which has been demonstrated to be 
effective using the organism that will 
serve as the host for propagating the 
recombinant DNA molecules. 
VII-B-6. Emergency plans required by 
Section IV-D-3-d. shall include methods 
and procedures for handling large losses 
of culture on an emergency basis. 
VII-C. P2-LS Level. 
VII-tC- 1. Cultures of viable organisms 
containing recombinant DNA molecules 
shall be handled in a closed system (e.g., 
closed vessel used for the propagation 
and growth of cultures) or other primary 
containment equipment (e.g., Class III 
biological safety cabinet containing a 
centrifuge used to process culture fluids) 
which is designed to prevent the escape 
of viable organisms. Volumes less than 
10 liters may be handled outside of a 
closed system or other primary • 
containment equipment providing all 
physical containment requirements 
specified in Section II-B-2. of the 
Guidelines are met. 
VII-C-2. Culture fluids (except as 
allowed in VII-C-3.) shall not be 
removed from a closed system or other 
primary containment equipment unless 
the viable organisms containing 
recombinant DNA molecules have been 
inactivated by a validated inactivation 
procedure. A validated inactivation 
procedure is one which has been 
demonstrated to be effective using the 
organism that will serve as the host for 
propagating jthe recombinant DNA 
molecules. 
VII-C-3. Sample collection from a 
closed system, the addition of materials 
to a closed system, and the transfer of 
cultures fluids from one closed system 
to another shall be done ip a manner 
which prevents the release of aerosols 
or contamination of exposed surfaces. 
Vn-C-4. Exhaust gases removed from 
a closed system or other primary 
containment equipment shall be treated 
by filters which have efficiencies 
equivalent to HEPA filters or by other 
equivalent procedures (e.g. 
incinereation) to prevent the release of 
viable organisms containing • 
recombinant DNA molecules to the 
environment. 
VII-C-5. A closed system or other 
primary containment equipment that has 
contained viable organisms containing 
recombinant DNA molecules shall not 
be opended for maintenance or other 
purposes unless it has been sterilized by 
a validated sterilization procedure. A 
validated sterilization procedure is one 
which has been demonstrated to be 
effective using the organism that will 
serve as the host for propagating the 
recombinant DNA molecules. 
VII-C-6. Rotating seals and other 
mechanical devices directly associated 
with a closed system used for the 
propagation and growth of viable 
organisms containing recombinant DNA 
molecules shall be designed to prevent 
leakage or shall be fully enclosed in 
ventilated housings that are exhasted 
through filters which have efficiencies 
equivalent to HEPA filters or through 
other equivalent treatment devices. 
VII-C-7. A closed system used for the 
propagation and growth of viable 
organisms containing recombinant DNA 
molecules and other primary 
containment equipment used to contain 
operations involving viable organisms 
containing recombinant DNA molecules 
shall include monitoring or sensing 
devices that monitor the integrity of 
containment during operations. 
VII-C-8. A closed system used for the 
propagation and growth of viable 
organisms containing the recombinant 
DNA molecules shall be tested for 
integrity of the containment features 
using the organism that will serve as the 
host for propagating recombinant DNA 
molecules. Testing shall be 
accomplished prior to the introduction 
of viable organisms containing 
recombinant DNA molecues, and 
following modification or replacement of 
essential containment features. 
Procedures and methods used in the 
testing shall be appropriate for the 
equipment design and for recovery and 
demonstration of the test organism. 
Records of tests and results shall be 
maintained on file. 
VD-C-9. A closed system used for the 
propagation and growth of viable 
organisms containing recombinant DNA 
molecules shall be permanently 
identified. This identification shall be 
used in all records reflecting testing, 
operation, and maintenance and in all 
documentation relating to the use of this 
equipment for research or production 
activities involving viable organisms 
containing recombinant DNA molecules. 
VII-C-10. The universal biohazard 
sign shall be posted on each closed 
system and primary containment 
equipment when used to contain viable 
organisms containing recombinant DNA 
molecules. 
VII-C-11. Emergency plans required 
by Section IV-D-3-d shall include 
methods and procedures for handling 
large losses of culture on an emergency 
hfl Ql a 
VII-D. P3-LS Level. 
VU-D-1. Cultures of viable organisms 
containing recombinant DNA molecules 
shall be handled in a closed system (en- 
closed vessels used for the propagation 
ajid growth of cultures) or other primary 
containment equipment (e.g., Class III 
biological safety cabinet containing a 
centrifuge used to process culture fluids) 
which is designed to prevent the escape 
of viable organisms. Volumes less than 
10 liters may be handled outside of a 
closed system providing all physical 
containment requirements specified in 
Section II-B-3 of the Guidelines are met. 
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