Federal Register / Vol. 59. No. 127 / Tuesday, July 5. 1994 / Notices 
34487 
10 productive infection; (iv) u.se viruses 
and other microorganisms or their 
genomes that have knowm arthropod or 
animal vectors, in the absence of such 
vectors; (v) use microorganisms that 
have an obligate association with the 
plant; or (vi) use microorganisms 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. 
.Appendix P-III-C. Biological 
Containment Practices 
(Mat.roorganisms) 
Appendix P-III-C-1. Effective 
dissemination of arthropods and other 
small animals can be prevented by using 
one or more of the following 
procedures; (i) Use non-flying, flight- 
impaired. or sterile arthropods; (ii) use 
non-motile or sterile strains of small 
animals; (iii) conduct experiments at a 
time of year that precludes the survival 
of escaping organisms; (iv) use animals 
that have an obligate association with « 
plant that is not present within the 
dispersal range of the organism; or (v) 
provent the escape of organisms present 
in run-off water by chemical treatment 
or evaporation of run-off water. 
I. Addition of Appendix Q. Physical and 
Biological Containment for 
Beroinhinant DNA Research Involving 
Animals, to the NIH Guidelines 
The following new appendix. 
Appendix Q. reads as follows; 
Appendix Q specifies containment 
and confinement practices for research 
involving whole animals, both those in 
which the animal's genome has been 
altered by stable introduction of 
recombinant DNA. or DNA derived 
therefrom, into the germ-line (transgenic 
animals) and experiments involving 
viable recombinant DNA-modified 
microorganisms tested on whole 
animals. The appendix applies to 
animal research activities with the 
following modifications: 
Appendix Q shall supersede 
Appendix C w'heii research animals are 
of a size or have growth requirements 
that prcc.lude the use of containment for 
laborator)' animals. Some animals may 
require other types of containment (see 
Appendix Q-III-D). The animals covered 
in Appendix Q are those species 
normally categorized as animals 
in<. hiding but not limited to cattle, 
swine, sheep, goats, horses, and poultry. 
The Institutional Biosafety Committee 
sliall include at least one scientist with 
expertise in animal containment 
principles when experiments utilizing 
Appendix Q require Institutional 
Biosafety Committee prior approval. 
The institution shall establish and 
maintain a health surveillance program 
for personnel engaged in animal 
research involving viable recombinant 
DNA-containing microorgani.sms that 
require Biosafety Level (BL) 3 or greater 
containment in the laboratory. 
Appendix Q-I. General Considerations 
Appendix Q-I-A. Containment Levels 
The containment levels required for 
re.search involving recombinant DNA 
associated with or in animals is based 
on classification of experiments in 
Section III. For the purpose of animal 
research, four levels of containment are 
established. These are referred to as 
BLl-Animals (N). BL2-N, BL3-N, and 
BL4-N and are described in the 
following sections of Appendix Q. The 
descriptions include: (i) standard 
practices for physical and biological 
containment, and (ii) animal facilities. 
Appendix Q-l-B. Disposal of Animals 
(BLl-N through BL4-N) 
Appendix Q-l-B-1. When an animal 
covered by Appendix Q containing 
recombinant DNA or a recombinant 
DNA-derived organism is euthanized or 
dies, the carcass shall be dispo.sed of to 
avoid its use as food for human beings 
or animals unless food use is 
specifically authorized by an 
appropriate Federal agency. 
Appendix Q-l-B-2. A permanent 
record shall be maintained of the 
experimental use and disposal of each 
animal or group of animals. 
Appendix Q-11. Physical and Biological 
Containment Levels 
Appendix Q-Il-A. Biosafety Level 1 - 
Animals (BLl-N) 
Appendix Q-Il-A-1. Standard Practices 
(BLl-N) 
Appendix Q-11-A-l-n. Animal Facility 
Access (BLl-N) 
Appendix Q-Il-A-l-a (l). The 
containment area shall be locked. 
Appendix Q-lI-A-l-a-(2). Access to 
the containment area shall be limited or 
restricted when experimental animals 
are being held. 
Ajipondix Q-lI-A-l-a-(3). The 
containment area shall be patrolled or 
monitored at frequent intervals. 
Appendix Q-II-A-l-b. Other (BLl-N) 
Appendix Q-lI-A-l-b-(l). All 
gcmetically engineered neonates shall be 
permanently marked within 72 hours 
after fiirth. if their size permits. If their ' 
size does not permit marking, their 
containers should be marked. In 
addition, transgenic animals should 
contain distinct and biochemically 
assayable DNA sequences that allow 
identification of transgenic animals 
from among non- transgenic animals. 
Appendix Q-lI-A-l-b-(2) A double 
barrier .shall be provided to separate 
male and female animals unless 
reproductive studies are part of the 
experiment or other measures are taken 
to avoid reproductive transmission. 
Reproductive incapacitation may be 
used. 
Appendix Q-II-A-l-b-(3). The 
containment area shall be in accordance 
with state and Federal laws and animal 
care requirements. 
Appendix Q-Il-A-2. Animal Facilities 
(BLl-N) 
Appendix Q-lI-A-2-(a). Animals shall 
be confined to securely fenced areas or 
be in enclosed structures (animal rooms) 
to minimize the possibility of theft or 
unintentional release. 
Appendix Q-ll-B. Biosafety Level 2 - 
Animals (BL2-N) (see Appendix Q-lIl- 
A) 
Appendix Q-lI-B-1. Standard Practices 
(BL2-N) 
Appendix Q-Il-B-l-a. Animal Facility- 
Access (BL2-N) 
Appendix Q-Il-B-l-a-(l). The 
containment area shall be locked. 
Appendix Q-ll-B-l-a-(2). The 
containment area shall be patrolled or 
monitored at frequent interv'als. 
Appendix Q-Il-B-l-a-(3). The 
containment building shall be 
controlled and have a locking access. 
Appendix Q-Il-B-l-a-(4). The Animal 
Facility Director shall establish policies 
and procedures whereby only persons 
who have been advised of the potential 
hazard and who meet any specific entry 
requirements (e.g.. vaccination) may 
enter the laboratory or animal rooms. 
Appendix Q-II-B-l-a-(5). Animals of 
the same or different species, which are 
not involved in the work being 
performed, shall not be permitted in the 
animal area. 
Appendix Q-Il-B-l-b. 
Decontamination and Inactivation (BL2- 
N) 
Appendix Q-ll-B-l-b-(l). 
Contaminated materials that are 
decontaminated at a site away from the 
laboratory shall be placed in a clo.sed 
durable leak-proof container prior to 
removal from the laboratory. 
Appendix Q-II-B-l-b-(2). Needles and 
syringes shall be promptly placed in a 
puncture-resistant container and 
decontaminated, preferably by 
autoclaving, before discard or reuse. 
Recombinant DNA Research, Volume 19 
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