Federal Register / Vol. 59. No. 127 / Tuesday. July 5. 1994 / Notices 
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1 through Biosafety Level 4. For large 
scale (over 10 liters) research or 
production. Appendi.x K supersedes 
Appendix G. Appendix K defines Good 
L.arge Scale Practice through Biosafety 
Level 3 — Large Scale. For certain work 
with plants. Appendix P supersedes 
Appendix G. Appendix P defines 
Biosafety Levels 1 through 4 — Plants. 
For certain work with animals. 
Appendix Q supersedes Appendix G. 
.Appendix Q defines Biosafety Levels 1 
through 4 — Animals. 
Appendix G-I. Standard Practices and 
Training 
The first principle of containment is 
strict adherence to good microbiological 
practices (see Appendices G— III-A 
through Consequently, all 
personnel directly or indirectly 
involved in experiments using 
recombinant DNA shall receive 
adequate instruction (see Sections I\'- 
B-l-e and IV-B— 4-d). At a minimum, 
these instructions include training in 
aseptic techniques and in the biology of 
the organisms used in tlie experiments 
so tliat the potential biohazards can be 
understood and appreciated. 
Any research group working with 
agents that are known or potential 
biohazards shall have an emergency 
plan that describes the procedures to be 
followed if an accident contaminates 
personnel or the environment. The 
Principal Investigator shall ensure that 
everyone in the laboratory is familiar 
with both the potential hazards of the 
work and the emergency plan (see 
Sections IV-B— 4-d and IV-B— 4-e). If a 
research group is working with a known 
pathogen for which there is an effective 
vaccine, the vaccine should be made 
available to all workers. Serological 
monitoring, when clearly appropriate, 
will be provided (see Section lV-B-1- 
n 
The Laboratory Safety Monograph 
(see Appendix G-Ill-O) and Biosafety in 
Microbiological and Biomedical 
l.aboratories (see Appendix G-lll-B) 
describe practices, equipment, and 
facilities in detail. 
// Addition of Appendix P. Physical 
and Biological Containment for 
fU conihinant DNA Bcsearch Invoicing 
Plants, to the NIH Cuidclines 
The following new appendix, 
Appendix P, reads as follows: 
Appendix P specifies physical and 
biological containment conditions and 
practices suitable to the greenhouse 
conduct of experiments involving 
recombinant DNA-contnining plants, 
plant -assfx.iated microorganisms, and 
small animals All provisions of the Nil I 
Guidelines apply to plant research 
activities with the following 
modifications; 
Appendix P shall supersede 
Appendix G when the research plants 
are of a size, number, or have grow'th 
requirements that preclude the use of 
containment conditions described in 
Appendix G. The plants covered in 
Appendix P include but are not limited 
to mos.ses. liverworts, macroscopic 
algae, and vascular plants including 
terrestrial crops, forest, and orna.mental 
species. 
Plant-associated microorganisms 
include viroids. virusoids, viruses, 
bacteria, fungi, protozoans, certain small 
algae, and microorganisms that have a 
benign or beneficial association with 
plants, such as certain Rhizobium 
species and microorganisms known to 
cau.se plant diseases. The appendix 
applies to microorganisms which are 
being modified with the objective of 
fostering an association with plants, 
Plant-as.sociated small animals 
include those arthropods that: (i) Are in 
obligate association with plants, (ii) are 
plant pests, (iii) are plant pollinators, or 
(iv) transmit plant disease agents, as 
well as other small animals such as 
nematodes for which tests of biological 
properties necessitate the use of plants. 
Microorganisms associated with such 
small animals (e.g., pathogens or 
symbionts) are included. 
The Institutional Biosafety Committee 
shall include at least one individual 
with expertise in plant, plant pathogen, 
or plant pest containment principles 
when experiments utilizing Appendix P 
require prior approval by the 
Institutional Biosafety Committee. 
Appendix P-1. General Plant Biosafety 
Levels 
Appendix P-l-A. The principal 
purpose of plant containment is to avoid 
the unintentional transmission of a 
recombinant DNA-containing plant 
genome, including nuclear or organelle 
hereditary material or release of 
recombinant DNA-derived organisms 
associated with plants. 
Appendix P-l-B. The containment 
principles are based on the recognition 
that the organisms that are used pose no 
health throat to humans or higher 
aiiiinals (unless deliberately modified 
for that purpose), and that the 
( orilainment conditions minimize the 
possibility of an unanticipated 
deleterious effect on organisms and 
ecosystems outside of the experimental 
facility, e g., the inadvertent spread of a 
serious pathogen from a greenhouse to 
a local agricultural crop or the 
unintentional introduction and 
establishment of an organism in a new 
(•( osystern. 
Appendix P-I-C. Four biosafety 
levels, referred to as Biosafety Level 
(BL) 1— Plants (P). BL2-P. BL3-P, and 
BL4-P, are established in Section II. The 
selection of containment levels required 
for research involving recombinant DNA 
molecules in plants or associated with 
plants is specified in Section III. The.se 
biosafety levels are described in 
Appendix P-II. This appendix describes 
greenhouse practices and special 
greenhouse facilities for physical 
containment. 
Appendix P-l-D. BLl-P through 
BL4-P are designed to provide 
differential levels of biosafety for plants 
in the absence or presence of other 
experimental organisms that contain 
recombinant DNA. These biosafety 
levels, in conjunction with biological 
containment conditions described in 
Appendix P-III, provide flexible 
approaches to ensure the safe conduct of 
research. 
Appendix P-l-E. For experiments in 
which plants are growm at the BLl 
through BL4 laboratory settings, 
containment practices shall be followed 
as described in Appendix G. These 
containment practices include the use of 
plant tissue culture rooms, growlh 
chambers within laboratory facilities, or 
experiments performed on open 
benches. Additional biological 
containment practices should be added 
by the Greenhouse Director or 
Institutional Biosafety Committee as 
neces.sary (see Appendix P-Ill), if 
botanical reproductive structures are 
produced that have the potential of 
being released. 
Appendix P-11. Physical Containment 
Levels 
Appendix P-II-A. Bio.safety Level 1 — 
Plants (BLl-P) 
Appendix P-II-A-1. Standard 
Practices (BLl-P) 
Appendix P-Il-A-l-a. Greenhouse 
Access (BLl-P) 
Appendix P-Il-A-l-a-(l). Access to 
the greenhouse shall be limited or 
restricted, at the discretion of the 
Greenhouse Director, w'hen experiments 
are in progress. 
Appendix P-II-A-l-a-(2). Prior to 
entering the greenhouse, personnel shall 
be required to read and follow 
instructions on BLl-P greenhouse 
practices and procedures. All 
procedures shall be performed in 
accordance with accepted greenhouse 
practices that are appropriate to the 
exireriinenlal organism. 
Appendix P-lI-A-l-b. Records (lU.l-P) 
Appendix P-lI-A-l-b-(l). A record 
shall Ire kept of experiments currently in 
progress in the greenhouse facility. 
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Recombinant DNA Research, Volume 19 
