Federal Register / Vol. 51. No. 68 / Wednesday. May 7, 1986 / Notices 
18975 
diaphragm bottles. Only needle-locking 
syringes or disposable s> rir.ge-r.eedle 
units {i.e.. needle is integral to the 
syringe) are used for the injection or 
aspiration of fluids containing organisms 
that contain recombinant DNA 
molecules. Extreme caution should be 
used when handling needles and 
syringes to avoid autoinoculation and 
the generation of aerosols during use 
and disposal. Needles should not be 
bent, sheared, replaced in the needle 
sheath or guard or removed from the 
syringe following use. The needle and 
syringe should be promptly placed in a 
puncture-resistant container and 
decontaminated, preferably by 
autoclaving, before discard or reuse. 
Appendix C-II-C-2-q. Spills end 
accidents which result in evert or 
potential exposures to organisms 
containing recombinant DN'A molecules 
are immediately reported to the 
laboratory director. Appropriate medical 
evaluation, surveillance, 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 drpendir.g on 
the agents handled or the function of the 
laboratory. 
Appendix C-Il-C-2-s. A biosafety 
manual is prepared or adopted. 
Personnel are advised of special 
hazards and are required to read 
instructions on practices and procedures 
and to follow them. 
Appendix G-lI-C-2-t. Alternative 
Selection of Containment Equipment. 
Experimental procedures involving a 
host-vector system that provides a one- 
step higher level of biological 
containment than that specified can be 
conducted in the BL3 laboratory using 
containment 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 can be 
conducted in the BL3 laboratory using 
containment equipment specified for the 
BU level of physical containment. 
Alternative combination of containment 
safeguards are shown in Table 1. 
Appendix C-/I-C-3 — Containment 
Equipment 
Appendix G-II-C-3-a. Biological 
safety cabinets (Class 1. 11. or 111) (see 
Appendix G— 111—12) or other appropriate 
combinations of personal protective or 
physical containment devices (e g . 
special 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: and harvesting 
infected tissues or fluids from 
experimental animals and embryonate 
eggs; and necropsy of experimental 
animals. 
Appendix G-lI-C—f — Laboratory 
Facilities 
.Appendix G-II-C—t-a. The laboratory 
is separated from areas which are open 
to unrestricted traffic Row 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 also be provided by a 
double-docred 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-lI-C-4-b The interior 
surfaces of walls, fioors. 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-H-C—t-c. Bench tops are 
impervious to water and resistant to 
acids, alkalis, organic solvents, and 
moderate heat. 
Appendix G-lI-C-4-d. Laboratory 
furniture is sturdy and spaces between 
benches, cabinets, and equipment are 
accessible for cleaning. 
Appendix C-II-C-4-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-Il-C-4~f. Windows in the 
laboratory are closed and sealed. 
Appendix G-U-C^t-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 C-lI-C-t~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 must verify 
that the direction of the airflow (into the 
laboratory) is proper. The exhaust air 
from the laboratory room can be 
discharged to the outside without being 
filtered or otherwise treated. 
Appendix C-II-C-t-j. The HEPA- 
filtered exhaust air from Class l 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 be recirculated 
within the laboratory if the cabinet is 
tested and certified at least every 
twelve months. If the HEPA-filtersd 
exhaust air from Class I or 11 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 
(12)) that avoids any interference with 
the air balance of the cabinets or 
building exhaust system. 
Appendix C-1I-D — Bioso r ety Level V 
(BL4). 
Appendix C-1I-D-1. Standard 
Microbiological Practices. 
Appendix G-Il-D-l-a. Work surfaces 
are decontaminated at least once a day 
and immediately after any spill of viable 
material. 
Appendix G-11-D-l-b. Only 
mechanical pipetting devices are used. 
Appendix G-II-D-l-c. Eating, 
drinking, smoking, storing food, and 
applying cosmetics are not permitted in 
the laboratory. 
Appendix G-H-D-l-d. Ail procedures 
are performed carefully to minimize the 
creation of aerosols. 
Appendix G-ll-D-2 — Special Practices 
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 are transferred to a 
nonbreakabie. sealed primary container 
and then enclosed in a nonbreakabie. 
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-ll-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 they leave the 
facility. Equipment or material which 
might be damaged by high temperatures 
or steam is decontaminated by gaseous 
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