33179 
Federal Register / VoL 56, No. 138 / Thursday. July 18, 199i / Notices 
accordance with applicable 
governmental environmental 
regulations. 
Appendix K-II-F. Addition of 
materiala to a system, sample collection, 
transfer of culture fluids tvithin/between 
systems, and processing of culture fluids 
shall be conducted In a manner that 
maintains employee exposure to viable 
orga n i sm s containing recombinant DNA 
molecules at a level that does not 
adversely affect the health and safety of 
employees. 
Appendix K-H-G. The facility's 
e m erg en cy response plan shall include 
provisions for handling spills. 
Appendix K-m. BLl-LS Level 
Appendix K-DI-A Spills and 
accidents which result in overt 
exposures to organisms containing 
recombinant DNA molecules are 
i mm ediately reported to the laboratory 
director. Medical evaluation, 
surveillance, and treatment are provided 
as appropriate and written records are 
maintained. 
Appendix K-III-B. Cultures of viable 
organisms containing recombinant DNA 
molecules shall be handled in a closed 
83 rstem (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 contairunent equipment 
proWded all physical containment 
requirements specified in Appendix G- 
II-A of the Guidelines are met 
Appendix K-HI-C. Culture fluids 
(except as allowed in Appendix K-III-D) 
shall not be removed from a closed 
system or other primary contairunent 
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. 
Appendix K-lII-D. Sample collection 
from a closed system, the addition of 
materials to a dosed system, and the 
transfer of culture fluids from one dosed 
system to another shall be done in a ' 
manner which minimi7.es the release of 
aerosols or contamination of exposed ■- 
surfaces. • .• 
Appendix K-III-E. Exhaust gases 
removed from a closed system or other 
primary containment equipment shall be 
treated by filters which have effidendes 
equivalent to HEPA filters or by other 
equivalent procedures (e.g., indneration) 
to minimize the release of viable 
organisms containing recombinant DNA 
molecules to the environment 
Appendix K-III-F, A dosed system or 
other primary contairunent 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 wiU serve as the host for 
propagating ffie recombinant DNA 
molecules. 
Appendix K-4II-G. Emdgency plans 
required by Section IV-B-8^ shall 
indude methods and prodedmres for 
handling large losses of culture on an 
emergency basis. 
Appendix K-IV. BL2-LS Level 
^pendix K-TV-A. Cultures of viable 
organisms containing recombinant DNA 
molecules shall be h^dled in a dosed 
system (e.g„ dosed vessel used for the 
propagation and growffi of cultures) or 
other primary contairunent equipment 
(eg„ Class m biological safety cabinet 
containing a centrifi^e used to process 
culture fluids) which is designed to 
prevent the escape of viable organisms. 
Volimxes less than 10 liters may be 
handled outside of a dosed system or 
other primary containment equipment 
provided all physical containment 
requirements spedffed in Appendix G- 
n-B of the Guidelines are met 
Appendix K-IV-B. Culture fluids 
(except as allowed in Appendix K-IV- 
C) shall not be removed a dosed 
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. 
Appendix K-TV-C. Sample collection 
from a dosed system, the addition of . 
materials to a dosed system, and the 
transfer of cultures fluids from one 
closed system to another shall be done 
in a manner which prevents the release 
of aerosols or contamination of exposed 
smfaces. 
Appendix K-IV-D. Elxhaust gases 
removed from a dosed system or other 
primary containment equipment shall be 
treated by filters which have effidendes 
equivalent to HEPA filters dr by other ... 
equivalent procedures (e.g., indneration) 
to prevent the release of viable 
organisms containing recombinant DNA 
molecules to the environment. 
Appenc^ K-lV-E. A dosed system or 
other prunary 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 n^g the 
organisms that will serve as the host for 
propagating the recombinant DNA 
molecules. ‘ • 
Appendix K-4V-^. Rotating seals airf 
other mechanical devices dire^y ' 
assod^ied with a dosed systeni us^ 
for the propagation and growth of viable 
organisms containing recoiiibinant DNA 
molectiles shall be designed to prevent 
leakage or shall be fully endosed in 
ventilated housings that are exhausted 
through filters which have effidendes 
equivalent to HEPA filters or through 
o^er equivalent treatment devices. 
Appendix K-IV-G. A dosed 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-TV-H. A dosed 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 molecules 
and following modification or 
replacement of essential containment 
featmes. Procedures and methods used 
in the testing shall be appropriate for the 
equipment design and for recovery and 
demonstration of the test oiganisnL 
Records of tests and results shall be 
maintained on file. ' 
Appendix K-IV-L A dosed 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 contauung • 
recombinant DNA molecules. „ r 
Appendix K-IV-j. The universal v' 
biohazard sign shall be posted on eac 
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Recombinant DNA Research, Volume 14 
