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Federal Register / Vol. 59, No. 127 / Tuesday, July 5, 1994 / Notices 
of the secondary container shall be 
decontaminated. Decontamination may 
be accomplished by passage through a 
chemical disinfectant or fumigation 
chamber or by an alternative procedure 
that has demonstrated effective 
inactivation of the e.xperimental 
organism. 
Appendi.x P-ll-C-l-h. Greenhouse 
Practices Manual (BL.l-P) 
Appendix P-II-C-l-h-( l). A 
greenhouse practices manual shall be 
prepared or adopted. This manual shall; 
(i) Advise personnel of the potential 
(Xjnsequences if such practices are not 
followed, and (ii) outline contingency 
plans to be implemented in the event of 
the unintentional release of organisms 
with recognized potential for serious 
detrimental impact. 
Appendix P-II-C-l-i. Protective Clothing 
(BL3-P) 
Appendix P-Il-C-l-i-fl)- Disposable 
clothing (e.g., solid front or wrap- 
around gowns, scrub suits, or other 
appropriate clothing) shall be worn in 
the greenhouse if deemed necessary by 
the Greenliouse Director because of 
potential dissemination of the 
experimental microorganisms. 
Appendix P-II-C-l-i-(2). Protective 
clothing shall be removed before exiting 
the greenhou.se and decontaminated 
prior to laundering or disposal. 
Appendix P-II-C-l-j. Other (BL3-P) 
Appendix P-II-C-l-j-(l). Personnel are 
required to thoroughly wash their hands 
upon exiting the greenhouse. 
Appendix P-II-C-l-j-{2). All 
procedures shall be performed carefully 
to minimize the creation of aerosols and 
excessive splashing of potting material/ 
soil during watering, transplanting, and 
all experimental manipulations. 
Appendix P-II-C-2. Facilities (BL3-P) 
Appendix P-II-C-2-a. Definitions (BL.3- 
P) 
Appendix P-lI-C-2-a-(l). The term 
'greenhouse' refers to a structure with 
walls, roof, and floor designed and used 
principally for growing plants in a 
controlled and protected environment. 
The w'alls and roof are usually 
constructed of transparent or 
translucent material to allow pas.sage of 
sunlight for plant growth. 
Appendix P-lI-C-2-a-(2). The term 
'greenhouse facility’ includes the actual 
greenhouse rooms or compartments for 
growing plants, including all 
immediately contiguous hallways and 
head-house areas, and is considered part 
of the confinement area. The need to 
maintain negative pressure should be 
considered when con.stnicting or 
renovating the greenhouse. 
Appendix P-II-C-2-b. Greenhou.se 
Design (BL3-P) 
Appendix P-II-C-2-b-(l). The 
greenhou.se floor shall be composed of 
concrete or other impervious material 
with provision for collection and 
decontamination of liquid run-off. 
Appendix P-[l-C-2-b-(2). Windows 
shall be closed and sealed. All glazing 
shall he resi.stant to breakage (e.g., 
double-pane tempered glass or 
equivalent). 
Appendix P-II-C-2-b-(3). The 
greenhou.se shall be a closed self- 
contained structure with a continuous 
covering that is separated from areas 
that are open to unrestricted traffic flow. 
The minimum requirement for 
greenhouse entry shall be [.wssage 
through two sets of self-closing locking 
doors. 
Appendix P-II-C-2-b-(4). The 
greenhouse facility shall be surrounded 
by a security fence or protected by 
equivalent security measures. 
Appendix P-lI-C-2-b-(5). Internal 
walls, ceilings, and floors shall be 
resistant to penetration 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-C-2-b-(6). Bench tops 
and other work surfaces should have 
seamless surfaces that are impervious to 
water and resistant to acids, alkalis, 
organic solvents, and moderate heat. 
Appendix P-II-C-2-b-(7). The 
greenhouse contains a foot, elbow, or 
automatically operated sink, which is 
located near the exit door for hand 
washing. 
Appendix P-II-C-2-c. Autoclaves (BL3- 
P) 
Appendix P-II-C-2-c-(l). An autoclave 
shall be available for decontaminating 
materials within the greenhouse facility. 
A double-door autoclave is 
recommended (not required) for the 
decontamination of materials passing 
out of the greenhouse facility. 
Appendix P-II-C-2-d. Supply and 
Exhaust Air Ventilation ^sterns (BL.3- 
P) 
Appendix P-II-C-2-d-(l). An 
individual supply and exhaust air 
ventilation system shall be provided. 
The system maintains pressure 
differentials and directional airflow, as 
required, to assure inward (or zero) 
airflow from areas outside of the 
gn^enhouse. 
Appendix P-Il-C-2-d-(2). The exhau.st 
air from the greenhouse facility shall be 
filtered through high efficiency 
particulate air-HEPA filters and 
discharged to the outside. The filter 
chambers shall be designed to allow in 
decontamination before filters are 
removed and to facilitate certification 
testing after they are replaced. Air fillers 
shall he 80-85% average efficiency by 
the American Society of Heating, 
Refrigerating, and Air Conditioning 
Engineers (ASHRAE) Standard 52- 68 
test method using atmosphere dust. Air 
supply fans shall be equipped with a 
back-flow damper that closes when the 
air supply fan is off. Alternatively, a 
HEPA filter may be used on the air 
supply system instead of the filters and 
damper. The supply and exhaust airflow 
shall be interlocked to assure inward (or 
zero) airflow at all times. 
Appendix P-II— C-2-e. Other (BL3— P) 
Appendix P-II-C-2 -e-(l). BL3-P 
greenhou.se containment requirements 
may be satisfied using a growth chamber 
or growth room within a building 
provided that the location, access, 
airflow patterns, and provisions for 
decontamination of experimental 
materials and supplies meet the intent 
of the foregoing clauses. 
Appendix P-lI-C-2-e-(2). Vacuum 
lines shall be protected with high 
efficiency particulate air/HEPA or 
equivalent filters and liquid disinfectant 
traps. 
Appendix P-ll-D. Biosafety bevel 4 — 
Plants (BL4-P) 
Appendix P-II-D-1. Standard Practices 
(BL4-P) 
Appendix P-II-D— 1-a. Greenhou.se 
Access (BL4-P) 
Appendi.x P-II-D-l-a— (1). 
Authorized entry into the greenhou.se 
shall be restricted to individuals who 
are required for program or support 
purposes. The Greenhou.se Director 
shall be responsible for assessing each 
circumstance and determining those 
individuals who are authorized to enter 
the greenhouse facility or work in the 
greenhouse during experiments. 
Appendix P-II-D-1— a-(2). Access 
shall be managed by the Greenhouse 
Director. Biological Safety Officer, or 
other individual responsible for 
physical security of the greenhouse 
facility: and access limited by means of 
secure, locked doors. 
Apjjendix P-II— D-1— a— (3). Prior to 
entering, individuals shall be advised of 
the potential environmental hazards and 
instructed on appropriate safeguards for 
ensuring environmental safety. 
Individuals authorized to enter the 
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