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Federal Register / Vol. 59, No. 127 / Tuesday, )uly 5, 1994 / Notices 
uniforms, laboratory coats) shall be 
provided that are appropriate for the 
risk of exposure to viable organisms 
containing recombinant DNA 
molecules. Eating, drinking, smoking, 
applying cosmetics, and mouth 
pipetting shall be prohibited in the work 
area. 
Appendix K-II-D. Cultures of viable 
organisms containing recombinant DNA 
molecules shall be handled in facilities 
intended to safeguard health during 
work with microorganisms that do not 
require containment. 
Appendix K-II-E. Discharges 
containing viable reconibinant 
organisms shall be handled in 
accordance with applic.able 
governmental environmental 
regulations. 
Appendix K-U-F. Addition of 
materials to a system, sample collection, 
transfer of culture fluids within/ 
between systems, and processing of 
cuhure fluids shall be conducted in a 
manner that maintains employee's 
exposure to viable organisms containing 
recombinant DNA molecules at a level 
that does not adversely affect the health 
and safety of employees. 
Appendix K-II-G. The facility’s 
emergency response plan shall include 
provisions for handling spills. Spills 
and accidents which result in overt 
exposures to organisms containing 
recombinant DNA molecules Eire 
immediately reported to the Biological 
Safety Officer, Institutional Biosafety 
Committee, NIH/ORDA, and other 
appropriate authorities (if applicable). 
Reports to NTH/ORDA shall sent to 
the Office of Recombinant DNA 
Activities, National Institutes of Health, 
Building 31, room 4B11, Bethesda, 
Maryland 20892, (301) 496-9838. 
Appendix K-IU. Biosafety Level 1 
(BLl) — Large Scale 
Appendix K-IIT-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 dther 
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 20692, (301) 496-9838. 
Medical evaluation, surveillance, and 
treatment are provided as appropriate 
and written records are maintained. 
Appendix K-UI-B. 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) w'hich 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 
provided all physical containment 
requirements specified in Appendix G- 
Il-A are met. 
Appendix K-llI-C. Culture fluids 
(except as allowed in Appendix K-III- 
D) 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. 
Appendix K— IIl-D. Sample collection 
from a closed system, the addition of 
materiab to a closed system, and the 
transfer of cuhure fluids from one 
closed system to another shall be 
conducted in a manner which 
minimizes the release of aerosols or 
contamination of exposed surfaces. 
Appendix K-UI-E. Exhaust gases 
removed from a closed system or other 
primary containment equiprment shall 
be treated by filters which have 
efficiencies equivalent to high efficiency 
particulate air/HEPA filters or by other 
equivalent procedures (e.g., 
incineration) to minimize the release of 
viable organisms containing 
recombinant DNA molecules to the 
environment. 
App>endix K-llI-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 
organism that will serve as the host for 
propagating the recombinant DNA 
molecules. 
Appendix K-IIl-G. Emergency plans 
required by Sections IV-B-2-b-(6) and 
IV-B-3-c-(3) shall include methods 
and procedures for handling large losses 
of culture on an emergency basis. 
Appendix K-IV. Biosafety Level 2 
(BL2) — Large Scale 
Appendix K-IV-A. Spills and 
accidents w'hich 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-FV-B. Cuhures 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 ni biological safety cabinet 
containing a centri^ge used to process 
culture fluids) which is designed to 
prevent the escape of viable organisms. 
Volumes less than 10 liters may be 
hemdled outside of a closed system or 
other primary containment equipment 
provided all physical containment 
requirements specified in Appendix G— 
II-B are met. 
Appendix K-IV-C. Culture fluids 
(except as allowed in Appendix K-IV- 
D) 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. 
Appendix K-IV-D. 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 
conducted in a manner which prevents 
the release of aerosols or contamination 
of exposed surfaces. 
Appendix K-IV-E. Exhaust gases 
removed from a closed system or other 
primary containment equipment shall 
be treated by filters which have 
efficiencies equivalent to high efficiency 
particulate air/HEPA filters or by other 
equivalent procedures (e.g., 
incineration) to prevenf the release of 
viable organisms containing 
recombinant DNA molecules to the 
environment. 
Appendix K— IV-F. A closed system or 
other primary contaiiunent 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 
procedme is one which has been 
demonstrated to be effective using the 
Recombinant DNA Research, Volume 19 
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