Federal Register / Vol. 59, No. 127 / Tuesday, July 5, 1994 / Notices 
34539 
shall M constructed to form a sealed 
internal shell that facilitates fumigation 
and is animal and arthropod-proof. 
These internal surfaces shall be resistant 
to penetration and degradation by 
liquids and chemicals to facilitate 
cleaning and decontamination of the 
area. All penetrations into these 
structures and surfaces (e.g., plumbing 
and utilities) shall be sealed. 
Appendix P-II-D-2-a-{7). Bench tops 
and other work surfaces shall have 
seamless surfaces impervious to water 
and resistant to acids, alkalis, organic 
solvents, and moderate heat. 
Appendix P-n-D-2-a-{8). A double- 
door autoclave, fumigation chamber, or 
ventilated airlock shall be provided for 
passage of all materials, supplies, or 
equipment that are not brought into the 
greenhouse facility through the change 
room. 
Appendix P-ll-D-2-b. Autoclaves 
(BL4-P) 
Appendix P-ll-D-2-b-(l). A double- 
door autoclave shall be provided for the 
decontamination of materials removed 
from the greenhouse facility. The 
autoclave door, which opens to the area 
external to the greenhouse facility, shall 
be sealed to the outer wall and 
automatically controlled so that it can 
only be opened upon completion of the 
sterilization cycle. 
Appendix P-Il-D-2-c. Supply and 
Exhaust Air Ventilation Systems (BL4- 
P) 
Appendix P-lI-D-2-c-(l). An 
individual supply and exhaust air 
ventilation system shall be provided. 
The system shall maintain pressure 
differentials and directional airflow as 
required to assure inward (or zero) 
airflow from areas outside of the 
greenhouse. Differential pressure 
transducers shall be used to sense 
pressure levels. If a system 
malfunctions, the transducers shall 
sound an alarm. A backup source of 
power should be considered. The 
supply and exhaust airflow shall be 
interlocked to assure inward (or zero) 
airflow at all times. The integrity of the 
greenhouse shall have an air leak rate 
(decay rate) not to exceed 7 percent per 
minute (logarithm of pressure against 
time) over a 20-minute period at 2 
inches of water gauge pressure. 
Nominally, this is 0.05 inches of water 
gauge pressure loss in 1 minute at 2 
inches water gauge pressure. 
Appendix P-II-I>-2-c-(2). Exhaust air 
from the greenhouse facility shall be 
filtered through higli efficiency 
particulate air/HEPA filters and 
discharged to the outside and dispersed 
away from occupied buildings and air 
intakes. Filter chambers shall be 
designed to allow in situ 
decontamination before fillers are 
removed and to facilitate certification 
testing after they are replaced. HEPA 
filters shall he provided to treat air 
supplied to the greenhouse facility. 
H^A filters shall be certified annually. 
Appendix P-II-D-2-d. Other (BL4-P) 
Appendix P-Il-D-2-d-(l). Sewer 
vents and other ventilation lines contain 
high efficiency particulate air/HEPA 
filters. HEPA filters shall be certified 
annually, 
Appendix P-lI-D-2-dH2). A pass- 
through dunk tank, fumigation chamber, 
or an equivalent method of 
decontamination shall be provided to 
ensure decontamination of materials 
and equipment that cannot be 
decontaminated in the autoclave. 
Appendix P-II-D-2-d— (3). Liquid 
effluent from sinks, floors, and 
autoclave chambers shall be 
decontaminated by heat or chemical 
treatment before being released from the 
maximum containment greenhouse 
facility. Liquid wastes from shower 
rooms and toilets may be 
decontaminated by heat or chemical 
treatment. Autoclave and chemical 
decontamination of liquid wastes shall 
be evaluated by appropriate standard 
procedures for autoclaved wastes. 
Decontamination shall be evaluated 
mechanically and biologically using a 
recording thermometer and an indicator 
microorganism with a defined heat 
susceptibility pattern. If liquid wastes 
are decontaminated with chemical 
disinfectants, the chemicals used must 
have demonstrated efficacy against the 
target or indicator microorganisms. 
Appendix P-lI-D-2-d-(4). If there is 
a central vacuum system, it shall not 
serve areas outside of the greenhouse 
facility. In-line high efficiency 
particulate air/HEPA filters shall be 
placed as near as practicable to each use 
point or vacuum service cock. Other 
liquid and gas services to the 
greenhouse facility shall be protected by 
devices that prevent back-flow. HEPA 
filters shall be certified annually. 
Appendix P-III. Biological Containment 
Practices 
Appropriate selection oJ the following 
biological containment practices may be 
used to meet the contaimnent 
requirements for a given organism. The 
present list Is not exhaustive; there may 
be other ways of preventing effective 
dissemination that could possibly lead 
to the establishment of the organism or 
its genetic material in the environment 
resulting in deleterious consequences to 
managed or natural ecosystems. 
Appendix P-lII-A. Biological 
Containment Practices (Plants) 
Appendix P-IIl-A-1. Effective 
dissemination of plants by pollen or , 
seed can be prevented by one or more 
of the following procedures: (i) Cover 
the reproductive structures to prevent 
pollen dissemination at flowering and 
seed dissemination at maturity; (ii) 
remove reproductive structures by 
employing male sterile strains, or 
harvest the plant material prior to the 
reproductive stage; (iii) ensure that 
experimental plants flower at a time of 
year when cross-fertile plants are not 
flowering within the normal pollen 
dispersal range of the experimental 
plant; or (iv) ensure that cross-fertile 
plants are not growing within the 
known pollen dispersal range of the 
experimental plant. 
Appendix P-III-B. Biological 
Containment Practices (Microorganisms) 
Appendix P-III-B-1. Effective 
dissemination of microorganisms 
beyond the confines of the greenhouse 
can be prevented by one or more of the 
following procedures: (i) Confine all 
operations to injections of 
microorganisms or other biological 
procedures (including genetic 
manipulation) that limit replication or 
reproduction of vimses and 
microorganisms or sequences derived 
from microorganisms, and confine these 
injections to internal plant parts or 
adherent plant surfaces; (ii) ensure that 
organisms, which can serve as hosts or 
promote the transmission of the virus or 
microorganism, are not present within 
the farthest distance that the airborne 
virus or microorganism may be expected 
to be effectively. disseminated; (hi) 
conduct experiments at a time of year 
when plants that can serve as hosts are 
either not growing or are not susceptible 
to productive infection; (iv) use viruses 
and other microorganisms or their 
genomes that have known arthropod or 
animal vectors, in the absence of such 
vectors; (v) use microorganisms that 
have an obligate association with the 
plant; or (vi) use microorganism^ that 
are genetically disabled to minimize 
survival outside of the research facility 
and whose natural mode of transmission 
requires injury of the target organism, or 
assures that inadvertent release is 
unlikely to initiate productive infection 
of organisms outside of the 
experimental facility. 
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