Federal Register / Vol. 51. No. 88 / Wednesday, May 7, 1986 / Notices 
16983 
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 reccmb'nar.t DNA 
molecules. 
Appendix K-l/I-C. 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 
in a manner which prevents the release 
of aerosols or contamination of exposed 
surfaces. 
Appendix K-llI-D. Exhaust g3ses 
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 . incineration) 
to prevent the release of viable 
organisms containing recombinant DNA 
molecules to the environment. 
Appendix K-UI-E. 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-tll-F. Rotating seals and 
other mechanical devices directly 
associated with a closed system used 
for the propagation and growth of viabie 
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 HEPA filters or through 
other equivalent treatment devices. 
Appendix K-11I-C. A closed system 
used for the propagation and grow th 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-II’-H. A closed system 
used for the propagation and grow th of 
viable organisms containing the 
recombinant DNA molecules shall be 
tested for integrity of the containment 
features using the organism that will 
Recombinant DNA Research, 
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 
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-lll-I. 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-Iil-J. The universal 
biohazard sign shall be posted on each 
clcsed system and primary containment 
equipment when used to contain viable 
organisms containing recombinant DNA 
molecules. 
Appendix K-II1-K. Emergency plans 
required by Section IV-B~3-f shall 
include methods and procedures for 
handling large losses of culture on an 
emergency basis. 
Appendix K-IV—BL3-LS Level 
Appendix K-1V-A. 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 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 
provided all physical containment 
requirements specified in Appendix G- 
II— C of the Guidelines are met. 
Appendix K-IV-B. Culture fluids 
(except as allowed in Appendix K-1V- 
C) 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 organisms that will serve as the host 
for propagating the recombinant DNA 
molecules. 
Appendix K-IV-C. Sample collection 
from a closed system, the addition of 
materials to a dosed system, and the 
Volume 1 1 
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. 
Appendix K-IV-D. 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., incineration) 
to prevent the release of viable 
organisms containing recombinant DNA 
molecules to the environment. 
Appendix K-IV-E. 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-IV-F. 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 
w ith equipment design, in order to 
maintain the integrity of containment 
features. 
Appendix K-IV-G. 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 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 recombinant DNA 
molecules shall include monitoring or 
sensing devices that monitor the 
integrity of containment during 
operations. 
Appendix K-lV-1. 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 hest for propagating the 
recombinant DNA molecules. Testing 
shall be accomplished prior to the 
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