18984 
Federal Register J Vol. 51, No. 88 / Wednesday, May 7, 1986 / Notices 
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 the use of this equipment for 
research production activities involving 
viable organisms containing 
recombinant DNA molecules. 
Appendix K-IV-K. The universal 
biohazard 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 Section IV-B-3-f shall 
include methods and procedures for 
handling large losses of culture on an 
emergency basis. 
Appendix K-IV-M. Closed systems 
and other primary containment 
equipment used in handling cultures of 
viable organisms containing 
recombinant DNA molecules shall be 
located within a controlled area which 
meets the following requirments: 
Appendix K-IV-M-1. The controlled 
area shall have a separate entry area. 
The entry area shall be a double-doored 
space such as an air lock, anteroom, or 
change room that separates the 
controlled area from the balance of the 
facility. 
Appendix K-F/-M-2. The surfaces of 
walls, ceilings, and floors in the 
controlled area shall be such as to 
permit ready cleaning and 
decontamination. 
Appendix K-IV-M-3. Penetrations 
into the controlled area shall be sealed 
to permitTiquid or vapor phase space 
decontamination. 
Appendix K-IV-M— i. All utilities and 
service or process piping and wiring 
entering the controlled area shall be 
protected against contamination. 
Appendix K-IV-M-5. Hand-w'ashing 
facilities equipped with foot, elbow, or 
automatically operated valves shall be 
located at each major work area and 
near each primary exit. 
Appendix K-IV-M-6. A shower 
facility shall be provided. This facility 
shall be located in close proximity to the 
controlled area. 
Appendix K-IV-M-7. The controlled 
area shall be designed to preclude 
release of culture fluids outside the 
controlled area in the event of an 
accidental spill or release from the 
closed systems or other primary 
containment equipment. 
Appendix K-IV-M-8. The controlled 
area shall have a ventilation system that 
is capable of controlling air movement. 
The movement of air shall be from areas 
of lower contamination potential to 
area3 of higher contamination potential. 
If the ventilation system provides 
positive pressure supply air, the system 
shall operate in a manner that prevents 
the reversal of the direction of air 
movement or shall be equipped with an 
alarm that would be actuated in the 
event that reversal in the direction of air 
movement were to occur. The exhaust 
air from the controlled area shall not be 
recirculated to other areas of the 
facility. The exhaust air from the 
controlled area may be discharged to 
the outdoors without filtration or other 
means for effectively reducing an 
accidental aerosol burden provided that 
it can be dispersed clear or occupied 
buildings and air intakes. 
Appendix K-IV-N. The following 
personnel and operational practices 
shall be required: 
Appendix K-IV-N-1. Personnel entry 
into the controlled area shall be through 
the entry area specifed in Appendix K- 
IV-M-I. 
Appendix K-IV-N-2. Persons entering 
the controlled area shall exchange or 
cover their personal clothing with work 
garments such as jumpsuits, laboratory 
coats, pants and shirts, head cover, and 
shoes or shoe covers. On exit from the 
controlled area the work clothing may 
be stored in a locker separate from that 
used for personal clothing or discarded 
for laundering. Clothing shall be 
decontaminated before laundering. 
Appendix K-IV-N-3. Entry into the 
controlled area during periods when 
work is in progress shall be restricted to 
those persons required to meet program 
or support needs. Prior to entry all 
persons shall be informed of the 
operating practices, emergency 
procedures, and the nature of the work 
conducted. 
Appendix K-IV-N-4. Persons under 18 
years of age shall not be permitted to 
enter the controlled area. 
Appendix K-IV-N-5. The universal 
biohazard sign shall be posted on entry 
doors to the controlled area and all 
internal doors when any work involving 
the organism is in progress. This 
includes periods when decontamination 
procedures are in progress. The sign 
posted on the entry doors to the 
controlled area shall include a statement 
of agents in use and personnel 
authorized to enter the controlled area. 
Appendix K-IV-N-6. The controlled 
area shall be kept neat and clean. 
Appendix K-IV-N-7. Eating, drinking, 
smoking, and storage of food are 
prohibited in the controlled area. 
Appendix K-IV-N-8. Animals and 
plants shall be excluded from.the 
controlled area. 
Appendix K-IV-N-9. An effective 
insect and rodent control program shall 
be maintained. 
Appendix K-IV-N-10. Access doors 
to the controlled area shall be kept 
closed, except as necessary for access, 
while work is in progress. Serve doors 
leading directly outdoors shall be sealed 
and locked while work is in progress. 
Appendix K-IV-N-1 J. Persons shall 
wash their hands when leaving the 
controlled area. 
Appendix K-IV-N-12. Persons 
working in the controlled area shall be 
trained in emergency procedures. 
Appendix K-IV-N-13. Equipment and 
materials required for the management 
of accidents involving viable organisms 
containing recombinant DNA molecules 
shall be available in the controlled area. 
Appendix K-IV-N-14. The controlled 
area shall be decontaminated in 
accordance with established procedures 
following spills or other accidental 
release of viable organisms containing 
recombinant DNA molecules. 
Appendix L — Release Into the 
Environment of Certain Plants 
Appendix L-I — General Information 
Appendix L specifies conditions under 
which certain plants as specified below, 
may be approved for release into the 
environment. Experiments in this 
category cannot be initiated without 
submission of relevant information on 
the proposed experiment to NIH, review 
by the RAC Plant Working Group, and 
specific approval by NIH. Such 
experiments also require the approval of 
the IBC before initiation. Information on 
specific experiments which have been 
approved will be available in ORDA 
and will be listed in Appendix L— III 
when the Guidelines are republished. 
Experiments which do not meet the 
specifications of Appendix L-II fall 
under Section III-A and require RAC 
review and NIH and IBC approval 
before initiation. 
Appendix L-II — Criteria Allowing 
Review by the RAC Plant Working 
Group Without the Requirement for Full 
RAC Review 
Approval may be granted by ORDA in 
consultation with the Plant Working 
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Recombinant DNA Research, Volume 11 
