Federal Register / Vol. 52, No. 154 / Tuesday, August 11, 1987 / Notices 
29813 
disinfectants or by heat in the liquid 
waste decontamination system by 
methods shown to be effective. The 
procedure used for heat 
decontamination of liquid wastes is to 
be monitored with a recording 
thermometer and waste is to be 
monitored for biological activity by 
introducing an appropriate indicator 
microorganism with a defined heat 
susceptibility pattern, and culturing 
samples of treated waste for presence of 
the organism. If liquid wastes from the 
shower room are decontaminated with 
chemical disinfectants, the chemical 
used is of demonstrated efficiency 
against the target or indicator 
microorganisms. Chemical disinfectants 
must be neutralized or diluted before 
release into general effluent waste 
systems." 
318. "Appendix Q-II-D-3-f. All 
equipment and floor drains will be 
equipped with deep traps (minimally 5 
inches). Floor drains will be fitted with 
isolation plugs or fitted with automatic 
water fill devices." 
319. "Appendix Q-II-D-3-g. All 
perimeter joints and openings must be 
sealed to form an insect-proof 
structure." 
320. “Appendix Q-II-D-3-h. Access 
doors to the containment area shall be 
self-closing.” 
321. "Appendix Q-Il-D-3-i. The BL4- 
N laboratory shall provide a double 
barrier to prevent the release of 
recombinant DNA containing 
microorganisms into the enviomment. 
Design of the facility will provide that, 
should the barrier of the inner facility be 
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 is 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 will be 
provided by a double-doored clothes 
change room equipped with integral 
showers and airlock.” 
322. "Appendix Q-II-D-3-j. A 
necropsy room will be provided within 
the BL4-N containment area." 
323. "Appendix Q-II-D-3-k. Each 
animal area shall contain a sink for 
handwashing. The sink shall be foot, 
elbow, or automatically operated and 
shall be located near the exit door." 
324. "Appendix Q-II-D-3-1. A ducted 
exhaust air ventilation system shall be 
provided. This system shall create 
directional airflow that draws air into 
the laboratory through the entry area. 
The exhaust air shall not be recirculated 
to any other area of the building, shall 
be discharged to the outside, and shall 
be dispersed away from the occupied 
areas and air intakes. Personnel must 
verify that the direction of the airflow 
(into the animal rooms) is proper." 
325. “Appendix Q-II-D-3-m. Exhaust 
air from BL4-N containment zone must 
be double HEPA filtered or treated by 
passing through a certified HEPA filter 
and an air incinerator before release to 
the atmosphere. Double HEPA filters are 
required in the supply air system in a 
BL4-N containment zone. Heating 
Ventiliation Air Conditioning (HVAC) 
supply and exhaust ducts and filter 
housing should comply with the NIH 
Laboratory Safety Monograph or 
supeseding volumes." 
326. “Appendix Q-II-D-3-n. 
Restraining devices for animals may be 
required to avoid damage to the 
integrity of the containment facility." 
327. “Appendix Q-D-D-3-o. All HEPA 
filters’ frames and housings must be 
certified to have nodetectable smoke 
[dioctylphthalate (DOP)] leaks when the 
exit face (direction of flow) of the filler 
is scanned above 0.01 percent when 
measured by a linear or logarithmic 
photometer. The instrument shall have a 
threshold sensitivity of at least 1 X10~* 
micrograms per liter for 0.3 micrometer 
diameter DOP particles and a challenge 
concentration of 80-120 micrograms per 
liter. The air sampling rate should be at 
least 1 cfm (28.3 liters per minute)." 
328. "Appendix Q-B-D-3-p. If an air 
incinerator is used in lieu of the second 
HEPA filter(s), it must be biologically 
challenged to prove all viable test 
agents are sterilized. The biological 
challenge must be minimally IX 10* 
organisms per cubic foot of airflow 
through the incinerator. It is universally 
accepted if bacterial spores are used to 
challenge and verify that the equipment 
is capable of sterilizing spores then 
assurance is provided that all other 
known agents will also be sterilized 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 continously monitored and 
recorded during use." 
329. "Appendix Q-II-D-3-q. The 
supply water distribution system must 
be fitted with a backflow preventer or 
break tank.” 
330. “Appendix Q-II-D-3-r. All 
utilities, liquid and gas services, are 
protected with devices that avoid 
backflow." 
331. "Appendix Q-II-D-3-s. Sewer 
and other atmospheric ventilation lines 
must be equipped with minimally a 
single HEPA filter. Condensate drains 
from these type housings must be 
appropriately connected to a 
contaminated or sanitary drain system. 
The drain position in the housing will 
dictate which system is to be used." 
332. "Appendix Q— III, Footnotes and 
References for Appendix Q. " 
333. “|1| If recombinant DNA is 
derived from a Class 2 organism 
requiring BL2 for laboratory research, 
personnel shall have specific training in 
handling pathogenic agents and are to 
be directed by knowledgeable 
scientists." 
334. “(2] Personnel who handle 
pathogenic and potentially lethal agents 
must have specific training and must be 
supervised by knowledgeable scientists 
who are experienced in working with 
these agents. BL3-N containment also 
minimizes escape of recombinant DNA- 
containing organisms from exhaust air 
or waste material from the containment 
zone." 
335. "[3] Microorganisms in Classes 4 
and/or 5 pose a high individual risk of 
life-threatening diseases to personnel 
and/or animals. Special approval must 
be obtained from USD A/ APHIS to 
Import class 5 agents." 
336. Laboratory staff have specific 
and thorough training in handling 
extremely hazardous infectious agents, 
and they understand the primary and 
secondary containment functions of the 
standard and special practices, the 
containment equipment, and the 
laboratory design characteristics. They 
are supervised by knowledgeable 
scientists who are trained and 
experienced in working with these 
agents and in the special containment 
facilities." 
337. "Within work areas of the 
facility, all activities are confined to the 
specially equipped animal rooms or 
support areas. The maximum animal 
containment area and support areas 
have special engineering and design 
features to avoid microorganisms being 
disseminated into the environment via 
exhaust air or waste disposal." 
338. “[4] Other research with 
nonlaboratory animals that may not 
appropriately be conducted under 
conditions described in Appendix Q can 
be conducted safety 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 should be provided to 
ensure that neither the organisms nor 
their gametes can escape into the supply 
or discharge system of the rearing 
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Recombinant DNA Research, Volume 13 
