Federal Register / Vol. 59, No. 127 / Tuesday, July 5, 1994 / Notices 
34537 
Appendix P-II— C-l-i. Protective 
Clothing (BL3-P) 
Appendix P— II— C— l-i-{l). Disposable 
clothing (e.g., solid front or uTap- 
around gowns, scrub suits, or other 
appropriate clothing) shall be worn in 
the greenhouse if deemed necessary by 
the Greenhouse Director because of 
potential dissemination of the 
experimental microorganisms. 
Appendix P-II-C-l-i-{2). Protective 
clothing shall be removed before exiting 
the greenhouse 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-Il-C-l-j— (2). All 
procedures shall be performfed carefully 
to minimize the creation of aerosols and 
excessive splashing of potting material/ 
soil during watering, transplanting, and 
ail e.xperimental manipulations. 
Appendix P-II-C-2. Facilities (BL3-P) 
Appendix P-II-C-2-a. Definitions 
(BL3-P) 
Appendix P-II-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 walls and roof are usually 
constructed of transparent or 
translucent material to allow passage of 
sunlight for plant growth. 
Appendix P-II-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 constructing 
or renovating the greenhouse. 
Appendix P-II-C-2-b. Greenhouse 
Design (BI.3-P) 
Appendix P-II-C-2-b-(l). The 
greenhouse floor shall be composed of 
concrete or other impervious material 
with provision for collection and 
decontamination of liquid run-off. 
Appendix P-II-C-2-b-{2). Windows 
shall be closed and sealed. All glazing 
shall be resistant to breakage (e.g., 
double-pane tempered glass or 
equivalent). 
Appendix P-II-C-2-b-(3). The 
greenhouse 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 passage 
through two sets of self-closing locking 
doors. 
Appendix P-Il-C-2-b-(4). The 
greenhouse facility shall be surrounded 
by a security fence or protected by 
equivalent security measures. 
Appendi.x P-Il-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-l>-(6). Bench tops 
and other work surfaces should have 
seamless surfaces that are impervious to 
water and resistant to adds, alkalis, 
organic solvents, and moderate heat. 
Appendix P-II-C-2-b-(7). The 
greenhouse contains a foot, elbow, or 
automatically operated sink, w'hich is 
located near Uie 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-<l. Supply and 
Ej^aust Air Ventilation Systems (BL3- 
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 
greenhouse. 
Appendix P-II-C-2-d-(2). The 
exhaust 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 
situ decontamination before filters are 
removed and to facilitate certification 
testing after they are replaced. Air filters 
shall be 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 
HEP A 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. 
Appendi.x P-lI-C-2-e. Other (BL3-P) 
Appendix P-II-C-2-e-(l). BL3-P 
greenhouse 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-II-C-2-e-(2). Vacuum 
lines shall be protected with high 
efficiency particulate air/HEPA or 
equivalent filters and liquid disinfectant 
traps. 
Appendix P-Il-D. Biosafety Level 4 — 
Plants (BL4-P) 
Appendix P-Il-D-1. Standard Practices 
(BL4-P) 
Appendix P— II-D-l-a. Greenhouse 
Access (BL4-P) 
Appendix P-II-D-l-a-(l). 
Authorized entry into the greenhouse 
shall be restricted to individuals w’ho 
are required for program or support 
purposes. The Greenhouse 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-E>-l-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. 
Appendix P-II-D-l-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 
greenhouse facility shall comply with 
the instructions and all other applicable 
entry/exit procedures. 
Appendix P-II-D-l-a-(4). Personnel 
shall enter and exit the greenhouse 
facility only through the clothing 
change and shower rooms and shall 
shower each time they exit the 
greenhouse facility. Personnel shall use 
the airlocks to enter or exit tlie 
laboratory only in an emergency. In the 
event of an emergency, every reasonable 
effort should be made to prevent the 
possible transport of viable propagulos 
from containment. 
Appendix P-Il-D-l-a-(5). Prior to 
entering the greenhouse, personnel shall 
be required to read and follow 
instructions on BL4-P practices and 
procedures. 
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Recombinant DNA Research, Volume 19 
