Federal Register / Vol. 59, No. 127 / Tuesday, July 5, 1994 / Notices 
34525 
organisms that will serve as the host for 
propagating the recombinant DNA 
molecules. 
Appendix K-FV-G. 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 exhausted 
through filters which have efficiencies 
equivalent to high efficiency particulate 
air/HEPA filters or through other 
equivalent treatment devices. 
Appendix K-IV-H. 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 sensing devices 
that monitor the integrity of 
containment during operations. 
Appendix K-FV-I. 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 accompUshed prior to the 
introduction of viable organisms 
containing recombinant DNA molecules 
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. 
Appendix K-IV-J. 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 use of this equipment for 
research or production activities 
involving viable organisms containing 
recombinant DNA molecules. 
Appendix K-IV-K. The universal 
biosafety sign shall be posted on each 
closed system and primary containment 
equipment when used to contain viable 
organisms containing recombinant DNA 
molecules. 
Appendix K-IV-L. Emergency plans 
required by Sections IV-B-2-b-(6) and 
rV-B-3-c-{3) shall include methods 
and procedures for handling large losses 
of culture on an emergency basis. 
Appendix K-V. Biosafety Level 3 
(BL3) — Large Scale 
Appendix K-V-A. Spills and 
accidents which result in overt 
exposures to organisms containing 
recombinant DNA molecules are 
immediately reported to the Biological 
Safety Officer, Institutional Biosafety 
Committee, NIH/ORDA, and other 
appropriate authorities (if applicable). 
Reports to NIH/ORDA shall be sent to 
the Office of Recombinant DNA 
Activities, National Institutes of Health, 
Building 31, Room 4B11, Bethesda, 
Maryland 20892, (301) 496-9838. 
Medical evaluation, surveillance, and 
treatment are provided as appropriate 
and written records are maintained. 
Appendix K-V-B. Cultures of viable 
organisms containing recombinant DNA 
molecules shall be handled in a closed 
system (e.g., closed vessels used for the 
propagation and growth of cultures) or 
other primary containment equipinent 
(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 
provided all physical containment 
requirements specified in Appendix G- 
II-C are met. 
Appendix K-V-C. Culture fluids 
(except as allowed in Appendix K-V-D) 
shall not be removed from a closed 
system or other primary containment 
equipment unless the viable oiganisms 
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 organisms that will serve as the host 
for propagating the recombinant DNA 
molecules. 
Appendix K-V-D. 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 
conducted in a manner which prevents 
the release of aerosols or contamination 
of exposed surfaces. 
Appendix K-V-E. Exhaust gases 
removed from a closed system or other 
rimary containment equipment shall 
e treated by filters which have 
efficiencies equivalent to high efficiency 
particulate air/HEPA filters or by other 
equivalent procedures (e.g., 
incineration) to prevent the release of 
viable organisms containing 
recombinant DNA molecules to the 
environment. 
Appendix K-V-F. 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 
organisms that will serve as the host for 
propagating the recombinant DNA 
molecules. 
Appendix K-V-G. A closed system 
used for the propagation and growth of 
viable organisms containing 
recombinant DNA molecules shall be 
operated so that the space above the 
culture level will be maintained at a 
pressure as low as possible, consistent 
with equipment design, in order to 
maintain the integrity of containment 
features. 
Appendix K-V-H. Rotating seals and 
other mechanical devices directly 
associated with a closed system used to 
contain viable organisms containing 
recombinant DNA molecules shall be 
designed to prevent leakage or shall be 
fully enclosed in ventilated housings 
that are exhausted through filters which 
have efficiencies equivalent to high 
efficiency particulate air/HEPA filters or 
through other equivalent treatment 
devices. 
Appendix K-V-I. 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. 
Appendix K-V-J. A closed system 
used for the propagation and growth of 
viable organisms containing 
recombinant DNA molecules shall be 
tested for integrity of the containment 
features using the organisms that will 
serve as the host for propagating the 
recombinant DNA molecules. Testing 
shall be accomplished prior to the 
introduction of viable organisms 
containing recombinant DNA molecules 
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. 
Appendix K-V-K. A closed system 
used for the propagation and grovrih of 
viable organisms containing 
recombinant DNA molecules shall be 
permanently identified. This 
identification shall be used in all 
records reflecting testing, operation, 
maintenance, and use of this equipment 
for research production activities 
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