Federal Register / Vol. 48, No. 44 / Friday, March 4, 1983 / Notices 
9439 
• Appendix K shall replace Appendix 
C when quantities in excess of 10 liters 
of culture are involved in research or 
production. 
• The institution shall appoint a 
biological safety officer if it engages In 
large-scale research or production 
activities involving viable organisms 
containing recombinant DNA molecules. 
The duties of the bological safety officer 
shall include those specified in Section 
IV-B-4 of the Guidelines. 
• The institution shall establish and 
maintain a health surveillance program 
for personnel engaged in large-scale 
research or production activities 
involving viable organisms containing 
recombinant DNA molecules which 
require P3 containment at the laboratory 
scale. The program shall include: 
preassignment and periodic physical 
and medical examinations; collection, 
maintenance and analysis of scrum 
specimens for monitoring serologic 
changes that may result from the 
employee’s work experience; and 
provisions for the investigation of any 
serious, unusual or extended illness of 
employees to determine possible 
occupational origin. 
Appendix K-A. Selection of Physical 
Containment Levels. The selection of 
the physical containment levels required 
for recombinant DNA research or 
production involving more than 10 liters 
of culture is based on the containment 
guidelines established in Part III of the 
Guidelines. For purposes of large-scale 
research or production, three physical 
containment levels are established. 
These are referred to as Pl-LS, P2-LS, 
and P3-LS. The Pl-LS level of physical 
containment is required for large-scale 
research or production of viable 
organisms containing recombinant DNA 
molecules which require Pi containment 
at the laboratory scale. The P2-LS level 
of physical containment is required for 
large-scale research or production of 
viable organisms containing 
recombinant DNA molecules wnich 
require P2 containment at the laboratory 
scale. The P3-LS level of physical 
containment is required for large-scale 
research or production of viable 
organisms containing recombinant DNA 
molecules which require P3 containment 
at the laboratory scale. No provisions 
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 basis. 
Appendix K-B. Pl-LS Level. 
Appendix K-B-l. Cultures of viable 
organisms containing recombinant DNA 
molecules shall be handled in a closed 
system (e.g., closed vessel used for the 
propogation and growth of cultures) or 
other primary containment equipment 
(eg.. 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 30 liters may be 
handled outside of a closed system or 
other primary containment equipment 
provided all physical containment 
requirements specified in Appendix G- 
1 1— A of the Guidelines are met. 
Appendix K-B-2. Culture fluids 
(except as allowed in Appendix K-B-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 the recombinant DNA 
molecules. 
Appendix K-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. 
Appendix K-B-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., incineration) 
to prevent the release of viable 
organisms containing recombinant DNA 
molecules to the environment. 
Appendix K-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. 
Appendix K-B-6. 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-C. P2-LS Level. 
- Appendix K-C-l. 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 
provided all physical containment 
requirements specified in Appendix G- 
1 1— B of the Guidelines are met. 
Appendix K-C-2. Culture fluids 
(except as allowed in Appendix K-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 the recombinant DNA 
molecules. 
Appendix K-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 
in a manner which prevents the release 
or aerosols or contamination of exposed 
surfaces. 
Appendix K-C-4. Exhaust gases 
removed from a closed system or other 
primary containment equipment shall be 
treated by filters which have efficiences 
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-C-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. 
Appendix K-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 exhausted 
through filters which have efficiencies 
equivalent to HEPA filters or through 
other equivalent treatment devices. 
Appendix K-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 
