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Federal Register / Vol. 49, No. 227 / Friday. November 23, 1984 / Notices 
handled or the function of the 
laboratory. 
"Appendix G-II-C-2-8. 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-II-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 
BL4 level of physical containment. 
Alternative combination of containment 
safeguards are shown in Table 1. 
“Appendix G-II-C-3. Containment 
Equipment. 
"Appendix G-II-C-3-a. Biological 
safety cabinets (Class I, II, or III) (see 
Appendix G — III— 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-II-C— 4. Laboratory 
Facilities. 
"Appendix G-II-C-4-a. The 
laboratory is separated from areas 
which are open to unrestricted 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 also 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, alkalis, 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 4 - c . 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 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 G-II-C-4-j. The 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 be 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 
[12]) that avoids any interference with 
the air balance of the cabinets or 
building exhaust system.” 
i. The following language is 
substituted for Appendix G-II-D, P4 
Level. 
"Appendix G-II-D. Biosafety Level 4 
(BL4). 
"Appendix G-II-D-1. Standard 
Microbiological Practices. 
“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, 
drinking, 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. 
“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 
nonbreakable, sealed primary container 
and then enclosed in a nonbreakable, 
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-U-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 
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 
laboratory director, biohazards control 
officer, or other person 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, indicates the date and time of 
each entry and exit. Practical and 
effective protocols for emergency 
situations are established. 
"Appendix G-II-D-2-d. Personnel 
enter and leave the facility only through 
the clothing change and shower rooms. 
Personnel shower each time they leave 
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