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•Federal Register / Vol. 59, No. ).T7 / Tuesday, July 5, 1994 / Notices 
facility shall be such that if the barrier _ 
of the inner facility is breached, the 
outer barrier will prevent release into 
the environment. The animal area shall 
be separated from all other areas. 
Passage through two sets of doors shall 
be the basic requirement for entry into 
the animal area from access corridors or 
other contiguous areas. Physical 
separation of the animal containment 
area from access corridors or other 
laboratories or activities shall be 
provided by a double-door clothes 
change room equipped with integral 
show’ers and airlock. 
Appendix Q-II-D-2-h. A necropsy 
room shall be provided within the BL4- 
N containment area. 
Appendix Q-II-D-2-i. Liquid effluent 
from containment equipment, sinks, 
biological safety cabinets, animal rooms, 
primary barriers, floor drains, and 
sterilizers shall be decontaminated by 
heat treatment before being released into 
the sanitary system. Liquid wastes from 
shower rooms and toilets shall be 
decontaminated with chemical 
disinfectants or heat by methods 
demonstrated to be effective. The 
procedure used for heat 
decontamination of liquid wastes shall 
be monitored with a recording 
thermometer. The effectiveness of the 
heat decontamination process system 
shall be revalidated every 30 days with 
an indicator organism. Liquid wastes 
from the shower shall be chemically 
decontaminated using an Environmental 
Protection Agency-approved germicide. 
The efficacy of the chemical treatment 
process shall be validated with an 
indicator organism. Chemical 
disinfectants shall be neutralized or 
diluted before release into general 
effluent waste systems. 
Appendix Q-II-D-2-j. A ducted 
exhaust air ventilation system shall be 
provided that creates directional airflow 
that draws air into the laboratory 
through the entry area. The exhaust air, 
which is not recirculated to any other 
area of the building, shall be discharged 
to the outside and dispersed away from 
the occupied areas and air intakes. 
Personnel shall verify that the direction 
of the airflow (into the animal room) is 
proper. , 
Appendix Q-II-D-2-k. Exhaust air 
from BL4-N containment area shall be 
double high efficiency particulate air/ 
HEPA filtered or treated by passing 
through a certified HEPA filter and an 
air incinerator, before release to the 
atmosphere. Double HEPA filters shall 
be required for the supply air system in 
a BL4^ containment area. 
Appendix Q-U-D-2-1. All high 
efficiency particulate air/HEPA filters’ 
frames and housings shall be certified to 
have no detectable smoke 
(dioctylphthalatel leaks when the exit 
face (direction of flow) of the filter is 
scanned above 0.01 percent when 
measured by a linear or logarithmic 
photometer. The instrument must 
demonstrate a threshold sensitivity of at 
least 1 x 10 “3 micrograms per liter for 0.3 
micrometer diameter dioctylphthalate 
particles and a challenge concentration 
of 80-120 micrograms per liter. The air 
sampling rate should be at least 1 cfrn 
(28.3 liters per minute). 
Appendix Q-II-D-2-m. If an air 
incinerator is used in lieu of the second 
high efficiency particulate air/HEPA 
filter, it shall be biologically challenged 
to prove all viable test agents are 
sterilired. The biological challenge must 
be minimally 1x10® organisms per cubic 
foot of airflow through the incinerator. 
It is imiversally accepted if bacterial 
spores are used to challenge and verify 
that the equipment is capable of killing 
spores, then assurance is provided that 
all other knowm agents are inactivated 
by the parameters established to operate 
the equipment. Test spores meeting this 
criterion are Bacillus subtilis var. niger 
or Bacillus stearothermophilis. The 
operating temperature of the incinerator 
shall be continuously monitored and 
recorded during use. 
Appendix Q-II-D-2-n. All equipment 
and floor drains shall be equipped with 
deep traps (minimally 5 inches). Floor 
drains shall be fitted with isolation 
plugs or fitted with automatic water fill 
devices. 
Appendix Q-II-D-2-o. Each animal 
area shall contain a foot, elbow, or 
automatically operated sink for hand 
washing. The sink shall be located near 
the exit door. 
Appendix Q-II-D-2-p. Restraining 
devices for animals may be required to 
avoid damage to the integrity of the 
containment animal facility. 
Appendix Q-II-^D-2-q. The supply 
water distribution system shall be fitted 
with a back-flow preventer or break 
tank.' 
Appendix Q-II-E>-2^. All utilities, 
liquid and gas services, shall be 
protected with devices that avoid back- 
flow. 
Appendix Q-II-D-2-s. Sewer and 
other atmospheric ventilation lines shall 
- be equipped minimally with a single 
high efficiency particidate/HEPA filter. 
Condensate drains from these type 
housings shall be appropriately 
connected to a contaminated or sanitary 
drain system. The drain position in the 
housing dictates the appropriate system 
to be used. 
Appendix Q-lII. Footnotes and 
References for Appendix Q 
Appendix Q-llI-A. If recombinant 
DMA is derived from a Class 2 organism 
requiring BL2 containment, personnel 
shall be required to have specific 
training in handling pathogenic agents 
and directed by knowledgeable 
scientists. 
Appendix Q-IIl-B. Personnel who 
handle pathogenic and potentially lethal 
agents shall be required to have specific 
training and be supervised by 
knowledgeable scientists who are 
experienced in working with these 
agents. BL3-N contaimnent also 
minimizes escape of recombinant DNA- 
containing organisms from exhaust air 
or waste material from the containment 
area. 
Appendix Q-IIl-C. Classes 4 and 5 
microorganisms pose a high level of 
individual risk for acquiring Ufe- 
threatening diseases to personnel and/or 
animals. To import Class 5 agents, 
special approval must be obtained from 
U.S. Department of Agriculture, Animal 
and Plant Health Inspection Service, 
Import-Export Products, Room 756, 
Federal Building, 6505 Belcrest Road, 
Hyattsville, Maryland 20782. 
Laboratory staff shall be required to 
have specific and thorough training in 
handling extremely hazardous 
infectious agents, primary and 
secondary containment, standard and 
special practices, and laboratory design 
characteristics. The laboratory staff shall 
be supervised by knowledgeable 
scientists who are trained and 
experienced in working with these 
agents and in the special containment 
facilities. 
Within work areas of the animal 
facility, ail activities shall be confined 
to the specially equipped animal rooms 
or support areas. The maximum animal 
containment area and support areas 
shall have special engineering and 
design features to prevent the 
dissemination of microorganisms into 
the environment via exhaust air or 
waste disposal. 
Appendix Q-III-D. Other research 
with non-laboratory animals, which 
may not appropriately be conducted 
imder conditions described in 
Appendix Q, may be conducted safely 
by applying practices routinely used for 
controlled culture of these biota. In 
aquatic systems, for example, BLl 
equivalent conditions could be met by 
utilizing growth tanks that provide 
adequate physical means to avoid the 
escape of the aquatic species, its 
gametes, and introduced exogenous 
genetic material. A mechanism shall be 
provided to ensure that neither the 
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