24970 
Federal Register / Vol. 45, No. 72 / Friday, April 11, 1980 / Notices 
VII-D-2. Culture fluids (except as 
allowed in VII-D-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 
organisms that will serve as the host for 
propagating the recombinant DNA 
molecules. 
VII-D-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. 
VII-D-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. 
VII-D-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 organisms that will 
serve as the host for propagating the 
recombinant DNA molecules. 
VII-D-6. A closed system used for the 
propagation and growth of viable 
organisms containing recombinant DNA 
molecules shall be bperated so that the 
space above the culture level will be 
maintained at or slightly below 
atmospheric pressure. 
VII-D-7. Rotating seals and other 
mechanical devices directly associated 
with a closed system used to contain 
viable organisms containing 
recombinant DNA molecules shall be 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. 
VU-D-8. 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. 
VII-D-9. 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 host for 
propagating the 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 containmentifeatures. 
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. 
VII-D-10. 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 or production 
activities involving viable organisms 
containing recombinant DNA molecules. 
VII-D-11. 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. < 
VH-D-12. Emergency plans required 
by Section IV-D-3-d shall include 
methods and procedures for handling 
large losses of culture on an emergency 
basis. 
VH-D-13. 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 requirements: 
VH-D-13-a. 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. 
VII-D-13-b. The surfaces of walls, 
ceilings, and floors in the controlled 
area shall be such as to permit ready 
cleaning and decontamination. 
VII-D-13-c. Penetrations into the 
controlled area shall be sealed to permit 
liquid or vapor phase space 
decontamination. 
Vfl-D-13-d. All utilities and service or 
process piping and wiring entering the 
controlled area shall be protected 
against contamination. 
VD-D-13-e. Handwashing facilities 
equipped with foot-, 6lboW-, or 
automatically-operated valves shall be 
located at each major work area and 
near each primary exit. 
VII-D-13-f. A shower facility shall be 
provided. This facility shall be located 
in close proximity to the controlled area. 
VU-D-13-g. 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. 
VII-D-13-h. The controlled area shall 
have a ventilation system that is 
capable 6f controlling air movement. 
The movement of air shall be from areas 
of lower contamination potential to 
areas 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 of occupied 
buildings and air intakes. 
Vfi-D-14. The following personnel 
and operational practices shall be 
required: 
VH-D-14-a. Personnel entry into the 
controlled area shall be through the 
entry area specified in section VII-D- 
13-a. 
VII-D-14-b. 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. 
VII— D — 14 — c. 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 jail persons 
shall be informed of the operating 
practices, emergency procedures, and 
the nature of the work conducted. 
VII-D-14-d. Persons under 18 years of 
age shall not be permitted to enter the 
controlled area. 
VII-D-14-e. The universal biohazard 
sign shall be posted onentry doors to 
the controlled area and all internal 
doors when any work involving the 
organism is in progress. This includes 
periods when decontamination 
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