24572 
Federal Register / Vol. 48, No. 106 / Wednesday, June 1, 1983 / Notices 
contaminated materials such as 
glassware, animal cages, laboratory 
equipment, and radioactive wastes shall 
be decontaminated by a method 
demonstrated to be effective before 
washing, reuse, or disposal. 
Appendix G-11-D-l-d. Mechanical 
pipetting devices shall be used; pipetting 
by mouth is prohibited. 
Appendix G-If-D-l-e. Eating, 
drinking, smoking, and storage of food 
are not permitted in the P4 facility. 
Appendix G-II-D-l-f. Persons shall 
wash their hands after handling 
organisms containing recombinant DNA 
molecules and when they leave the 
laboratory. 
Appendix G-II-D-l-g. Care shall be 
exercised to minimize the creation of 
aerosols. For example, manipulations 
such as inserting a hot inoculating loop 
or needle into a culture, flaming an 
inoculation loop or needle so that it 
splatters, and forceful ejection of fluids 
from pipettes or syringes shall be 
avoided. 
Appendix G-II-D-l-h. Biological 
materials to be removed from the P4 
facility in a viable or intact state shall 
be transferred to a nonbreakable sealed 
container, which is then removed from 
the P4 facility through a passthrough 
disinfectant dunk tank or fumigation 
chamber. 
Appendix G-11-D-l-i. No materials, 
except for biological materials that are 
to remain in a viable or intact state, 
shall be removed from the P4 facility 
unless they have been steam-sterilized 
(autoclaved) or decontaminated by a 
means demonstrated to be effective as 
they pass out of the P4 facility. All 
wastes and other materials as well as 
equipment not damaged by high 
temperature or steam shall be steam 
sterilized in the double-door autoclave 
of the P4 facility. Other materials which 
may be damaged by temperature or 
steam shall be removed from the P4 
facility through a passthrough 
fumigation chamber. 
Appendix G— II-D-l-j. Materials 
within the Class III cabinets shall be 
removed from the cabinet system only 
after being steam-sterilized in an 
attached double-door autoclave or after 
being contained in a nonbreakable 
sealed container, which is then passed 
through a disinfectant dunk tank or a 
fumigation chamber. 
Appendix G-II-D-l-k. Only persons 
whose entry into the P4 facility is 
required to meet program or support 
needs shall be authorized to enter. 
Before entering, such persons shall be 
advised of the nature of the research 
being conducted and shall be instructed 
as to the appropriate safeguards to 
ensure their safety. They shall comply 
with instructions and all other required 
procedures. 
Appendix G — II— D— 1—1. Persons under 
18 years of age shall not enter the P4 
facility. 
Appendix G-II-D-l-m. Personnel 
shall enter into and exit from the P4 
facility only through the clothing change 
and shower rooms. Personnel shall 
shower at each egress from the P4 
facility. Air locks shall not be used for 
personnel entry or exit except for 
emergencies. 
Appendix G-II-D-l-n. Street clothing 
shall be removed in the outer side of the 
clothing-change area and kept there. 
Complete laboratory clothing, including 
undergarments, head cover, shoes, and 
either pants and shirts or jumpsuits, 
shall be used by all persons who enter 
the P4 facility. Upon exit, personnel 
shall store this clothing in lockers 
provided for this purpose or discard it 
into collection hampers before entering 
the shower area. 
Appendix G-II-D-l-o. The universal 
biohazard sign is required on the P4 
facility access doors and on all interior 
doors to individual laboratory rooms 
where experiments are conducted. The 
sign shall also be posted on freezers, 
refrigerators, or other units used to store 
organisms containing recombinant DNA 
molecules. 
Appendix G-II-D-l-p. An insect and 
rodent control program shall be . 
instituted. 
Appendix G-II-D-l-q. Animals and 
plants not related to the experiment 
shall not be permitted in the laboratory 
in which the experiment is being 
conducted. 
Appendix G-II-D-l-r. Vacuum outlets 
shall be protected by filter and liquid 
disinfectant traps. 
Appendix G-II-D-l-s. Use of the 
hypodermic needle and syringe shall be 
avoided when alternate methods are 
available. 
Appendix G — 1 1— D— 1— t. The laboratory 
shall be kept neat and clean. 
Appendix G-II-D-l-u. If experiments 
involving other organisms which require 
lower levels of containment are to be 
conducted in the P4 facility concurrently 
with experiments requiring P4-level 
containment, they shall be conducted in 
accordance with all P4-level laboratory 
practices specified in this section. 
Appendix G-II-D-2. Containment 
Equipment. 
Appendix G-II-D-2-a. Experimental 
procedures involving organisms that 
require P4-level physical containment 
shall be conducted either in: (i) A Class 
III cabinet system or in (ii) Class I or 
Class II cabinets that are located in a 
specially designed area in which all 
personnel are required to wear one- 
piece positive-pressure isolation suits. 
Appendix G-II-D-2-b. Laboratory 
animals involved in experiments 
requiring P4-level physical containment 
shall be housed either in cages 
contained in Class III cabinets or in 
partial containment caging systems 
(such as Horsfall units [11], open cages 
placed in ventilated enclosures, or solid- 
wall and -bottom cages covered by filter 
bonnets, or solid-wall and -bottom cages 
placed on holding racks equipped with 
ultraviolet irradiation lamps and 
reflectors) that are located in a specially 
designed area in which all personnel are 
required to wear one-piece positive- 
pressure suits. 
Appendix G-II-D-2-c. 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 P4 facility using 
containment equipment requirements 
specified for the P3 level of physical 
containment. Alternative combinations 
of containment safeguards are shown in 
Table II. 
Table II.— Possible Combinations of Containment Safeguards 
Classification of experiment 
Alternate combinations of physical and biological containment 
Physical containment 
Biological 1 
containment 
Physical containment 
Laboratory design 
specified for 
Laboratory 
practices 
specified for 
Containment 
equipment 
specified for 
Biological 
containment 
P4 
HV1 
P4 
P4 
P4 
HV1 
HV2 
P4 
HV1 
P4 
P4 ■ 
P3 
1 see Append** I for description of biological containment. 
■In this case gloves shall be worn, in addition to the clothing requirements specified m Appendix G-11-D-l-n. 
Appendix G-II-D-3. Special 
Laboratory Design. 
Appendix G-II-D-3-a. The laboratory 
shall be located in a restricted-access 
facility which is either a separate 
building or a clearly demarcated and 
isolated zone within a building. 
Clothing-change areas and shower 
rooms shall be provided for personnel 
entry and egress. These rooms shall be 
ri.52] 
