Federal Register / Vol. 59, No.- 127 / Tuesday, July 5, 1994 / Notices 
34517 
containing recombinant DNA molecules 
are immediately reported to the 
Biological Safety Officer, Institutional 
Biosafety Committee, and NIH/ORDA. 
Reports to NIH/ORDA shall be sent to 
the Office of Recombinant DNA 
Activities, National Institutes of Health, 
Building 31, Room 4B11, Bethesda, 
Maryland 20892, (301) 49&-9838. 
Appropriate medical evaluation, 
sur%’eillance, and treatment are provided 
and written records are maintained. 
Appendix G-II-C-2-r. Baseline serum 
samples for all laboratory and other at- 
risk personnel should be collected and 
stored. Additional serum specimens 
may be collected periodically 
depending on the agents handled or the 
function of the laboratory. 
Appendix G-II-C-2-s. A biosafety 
manual is prepared or adopted. 
Persormel are advised of special hazards 
and are required to read and follow the 
instructions on practices and 
procedures. 
Appendix G-II-C-2-t. Alternative 
Selection of Containment Equipment 
(BL3) Experimental procedures 
involving a host-vector system that 
provides a one-step higher level of 
biological contaiiunent than that 
specified may be conducted in the BL3 
laboratory using contaiiunent 
equipment specified for the BL2 level of 
physical containment. Experimental 
procedures Involving a host-vector 
system that provides a one-step lower 
level of biological containment than that 
specified may be conducted in the BL3 
laboratory using containment 
equipment specified for the BL4 level of 
physical containment. Alternative 
combination of containment safeguards 
are shown in Appendix G-Table 1. 
Appendix G-lt-C~3. Containment 
Equipment (BL3). Appendix G-II-C-3- 
a. Biological safety cabinets (Class I, II, 
or III) (see Appendix G-III-L) or other 
appropriate combinations of personal 
protective or physical containment 
devices (e.g., specid protective clothing, 
masks, gloves, respirators, centrifuge 
safety cups, sealed centrifuge rotors, and 
containment caging for animals) are 
used for all activities with organisms 
containing recombinant DNA molecules 
which pose a threat of aerosol exposure. 
These include: manipulation of cultures 
and of those clinical or environmental 
materials which may be a source of 
aerosols; the aerosol challenge of 
experimental animals; the h^esting of 
Infected tissues or fluids from 
experimental animals and embryonate 
eggs; and the necropsy of experimental 
animals. 
Appendix G-lI-C-4. Laboratory 
Facilities (BL3) Appendix C-II-C-4-a. 
The laboratory is separated from areas 
which are open to uruestricted traffic 
flow within the building. Passage 
through two sets of doors is the basic 
requirement for entry into the laboratory 
from access corridors or other 
contiguous areas. Physical separation of 
the high containment laboratory from 
access corridors or other laboratories or 
activities may be provided by a double- 
doored clothes change room (showers 
may be included), airlock, or other 
access facility which requires passage 
through two sets of doors before 
entering the laboratory. 
Appendix G-II-C— 4-b. The interior 
surfaces of walls, floors, and ceilings are 
water resistant so that they can be easily 
cleaned. Penetrations in these surfaces 
are sealed or capable of being sealed to 
facilitate decontaminating the area. 
Appendix G-II-C-4-c. Bench tops are 
impervious to water and resistant to 
acids, alkahs, organic solvents, and 
moderate heat. 
Appendix G— II-C— 4-d. Laboratory 
furniture is sturdy and spaces between 
benches, cabinets, and equipment are 
accessible for cleaning. 
Appendix G-II-C— 1 -e. Each 
laboratory contains a sink for hand 
washing. The sink is foot, elbow, or 
automatically operated and is located 
near the laboratory exit door. 
Appendix G-II-C-4-f. Windows in 
the laboratory are closed and sealed. 
Appendix G-II-C-4-g. Access doors 
to the laboratory or containment module 
are self-closing. 
Appendix G-II-C— 4^h. An autoclave 
for decontaminating laboratory wastes is 
available preferably within the 
laboratory. 
Appendix G-II-C— 4-i. A ducted 
exhaust air ventilation system is 
provided. This system creates 
directional airflow that draws air into 
the laboratory through the entry area. 
The exhaust air is not recirculated to 
any other area of the building, is 
discharged to the outside, and is 
dispersed away from the occupied areas 
and air intakes. Personnel shall verify 
that the direction of the airflow (into the 
laboratory) is proper. The exhaust air 
from the laboratory room may be 
discharged to the outside without being 
filtered or otherwise treated. 
Appendix G-II-C-4-j. The high 
efficiency particulate air/HEPA filtered 
exhaust air from Class I or Class II 
biological safety cabinets is discharged 
directly to the outside or through the 
building exhaust system. Exhaust air 
from Class I or II biological safety 
cabinets may bo recirculated within the 
laboratory if the cabinet is tested and 
certified at least every twelve months. If 
the HEPA-filtered exhaust air from Class 
I or II biological safety cabinets is to be 
discharged to the outside through the 
building exhaust air system, it is 
connected to this system in a manner 
(e.g., thimble unit connection (see 
Appendix G-III-L)) that avoids any 
interference with the air balance of the 
cabinets or building exhaust system. 
Appendix G-II-D. Biosafety Level 4 
(BL4) 
Appendix G-Il-D-1. Standard 
Microbiological Practices (BL4) 
Appendix G-II-D-l-a. Work surfaces 
are decontaminated at least once a day 
and immediately after any spill of viable 
material. 
Appendix G-II-D-l-b. Only 
mechanical pipetting devices are used. 
Appendix G-II-D-l-c. Eating, 
drinldng, smoking, storing food, and 
applying cosmetics are not permitted in 
the laboratory. 
Appendix G-II-D-l-d. All 
procedures are performed carefully to 
minimize the creation of aerosols. 
Appendix G-II-D-2. Special Practices 
(BL4). Appendix G-II-D-2-a. Biological 
materials to be removed from the Class 
III cabinets or from the maximum 
containment laboratory in a viable or 
intact state lue transferred to a non- 
breakable, sealed primary container and 
then enclosed in a non-breakable, sealed 
secondary container which is removed 
from the facility through a disinfectant 
dunk tank, fumigation chamber, or an 
airlock designed for this purpose. 
Appendix G-II-D-2-b. No materials, 
except for biological materials that are 
to remain in a viable or intact state, are 
removed from the maximum 
containment laboratory unless they have 
been autoclaved or decontaminated 
before exiting the facility. Equipment or 
material which might be damaged by 
high temperatures or steam is 
decontaminated by gaseous or vapor 
methods in an airlock or chamber 
designed for this purpose. 
Appendix G-II-D-2-c. Only persons 
whose presence in the facility or 
individual laboratory rooms is required 
for program or support purposes are 
authorized to enter. The supervisor has 
the final responsibility for assessing 
each circumstance and determining who 
may enter or work in the laboratory. 
Access to the facility is limited by 
means of secure, locked doors; 
accessibility is managed by the 
Principal Investigator, Biological Safety 
Officer, or other persons responsible for 
the physical security of the facility. 
Before entering, persons are advised of 
the potential biohazards and instructed 
as to appropriate safeguards for ensuring 
their safety. Authorized persons comply 
with the instructions and all other 
applicable entry and exit procedures. A 
logbook signed by all personnel 
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Recombinant DNA Research, Volume 19 
