Federal Re^ster L V'ol. ^ No. 127 / Tuesdiay, fuly Sy 1994 I Niotices 
34527 
Cateoon GSne Appefxlix Kr-VI-S)i 
GkSP 
BLI-tS 
B112-LS 
Bt^S 
& Medical surveillance _ 
NR 
MR 
llt-INf-A 
IC-V-A 
7. Viable organtsms s&ouid be handled- in a system- that physically 
NR 
Kr-IIP-B 
K-lV-8 
separates the process from the external environment (dosed 
system or other primary containmentj. 
8. Culture- flukfe not removed from a system until organlsros are in- 
;nr 
,K-III-C 
K-4V^ 
Kr-V-C 
activated. 
9l Inactivaliorv at waste soludons and materials with respect to- their 
'K-ll-e 
'K-tlMD 
K-V-C 
IC-V-C 
biohazard potentiaL 
TO. Control- at aaosols by en^neering pr procedural' coobols. to> 
Minimize 
' Minimize 
' Ptevent 
Prevent 1 
prevent or minimize release erf organisms during sampling from ai 
1 Procedure 
Engmeer 
1 Engmeer 
Engineer ^ 
system, addition ot materials, to, a system, transfer of ctiltiva^edi 
ilt(-H-f 
!K-W-B 
lK-V-0 
ceJISs and reroovai ot materialv products, and effluent from a 
tem. 
1 1. Treatment of exhaust gases from a dosed system to minimize 
K-Mt-D 
; K-IV-0; 
K-V-O 
INR 
; Minimize 
, Prevent 
Pcevent 
or prevent refease of viable organisms. 
'K-m-E 
, K-IV-E 
iKr-V-E 
T2. Closed system- that has contafnerf viable organisms not R) be 
jNR 
K-m-F 
K-IV-F 
;K-V-E 
opened- urrtit sterilized by a vaRdhtied procedure. 
Closed system *> be maitrtained at as a low pressure as pos- 
NR 
NR 
NR 
' K^V-G i 
sible ta maiotaia integrity of containment featuresv 
14. Rotating seals and other penetratlans into dosed system de- 
signed: to- prevent oT' minimize; leakage. 
iNR 
INR 
1 Prevent 
K-IV-G 
' Prevent 
1 5. Closed, system, shall iiteaqx>rat 0 . monitering or sensing devices 
iNR 
;NR 
K-W-H 
; K-v-t 
to monitor the Integrity of containment 
1 6. Validated ihfegrity testing oL dosed containment system 
NR 
,MR 
,Kr+V-i 
■Kr-V-d \ 
1 7. Closed system to be permanently identified for record keeping, 
,NR 
NR 
, K-lV-d- 
.Kr-V-K 1 
purposes. 
1 8. Uruversal biosafety sign ta be posted orr each dosed system ... 
NR- 
!nr 
K-IV^ 
Kr-V-L 
19. Emergency ptens required for handfirig large losses ot cottures 
'K-ir-G 
K-IV-L 
K-V-M- 
20. Access to the- worif pfaee- 
:nr 
; G-IT-A-1^ 
G-tt-B-l-a- 
K-V-fi 
21. Requi«nents lor conttolted access areai 
>NR 
Inr 
'MR 
1K-V-M40 
NR=nof reqMired. 
Appendix K-VI. Footnotes of Appendix 
K 
Appendix K-Vl-A. This table is 
derived from the text la Appendices G 
and K. and is not ta be used ia lieu oL 
Appeadiees-Gand 1C 
Appendix K-VI-B. The criteria in this 
grid address only the hinlng^al hazards 
associated with organisms containing 
recombioani EINA. Other hazards 
accompanying, the large scale 
cultivation of such organisms Ce.g^ toxic 
properties oI products^ physical, 
mechanical, and chemical aspects ol 
downstream processing) are not 
addressed and shall be considered 
separately, albeit in coa)uncJfoa with, 
this grid 
Appendix K-VTF. E)efimtiom to 
Acc o mpan y Contomm e rtf Gtid and 
Appendix K 
Appendix IC-VH-A. Acckleatal 
Releiaee. An accidental release is the 
unintentional dkschaxgs of a 
microhlorogical agenl Ci-e., 
microorganism or virus) or eickaryotic 
cell due to a tailure in the containment 
system. 
AppeiuCx FC-VU-B. Biological Barrier. 
A bloXogical barrier is- an. impediment 
fnaturafly occurring or introduced) to 
the bkfiecllvity aad/ot survival of a 
mlcsobiological agpnt or eukaryotic ceil 
once U haabeen. released into the 
onvinanmeot. 
Appendix K-VTI-C Closed Sysfemr. A 
closed system is one in which by Its 
design and proper operation, prevents 
release of a microbiorogical agent or 
eukaryotic cell contained therein. 
Appendix K-VR-D. Cootaininent. 
Containment is the confinement of a 
microbiologicat' agent or eukaryotic cell 
that is being cultured^ stored, 
manipulated, transpocted. or destroyed 
in order to prevent or limit its contact 
with people and/or the environmenL 
Methods used to achieveithis include; 
physical and biological barriers and 
inactivation using physical or chemical 
means. 
Appendix K-VH-EL De minimis. 
Refease. De miaimi's release i& the 
release oL (i) viable microbiological 
agents or eukaryotic eeJils. that does not 
result in the establishoient of disease in 
healthy people, plants, or animals; or 
(ii)> in ancontrolledi proliferation of any 
oRcrobiofogicaP agents or eukaryotic 
cells. 
Append™ FT-VW-F. Disfirfcctfon. 
Disinfection is a proc es s by whtctr 
viable mictobiological agents or 
eukaryotic cells ace reduced ta a level 
unJdk^y to produce disease in healthy 
people, plants, or anzmals. 
Appendix Kr-VIl-G> Good Large Scale 
Practice Organism. For an organism ta 
qualify for Good Large Scale Practice; 
consideratian. U must meet the 
following cdOana IRefecencc: ' 
Organization for Economic Cooperation 
andCtevelopment, Recombinant DNA 
Safety Considerations, 1987, p. 84-85): 
(i) the host orgamsm should be Bcm- 
pathogenic. sbouM not craTtatn 
adventitious agents and should have an 
extended history of safe large scale use 
or have bufll-fn' environfmentat 
ISmitations that permit optunum growth 
in. thekege scalb setting but Brafted 
survival without adverse- consequences 
in the emfronment; (irl tfw recombinant 
DNA-engmeered OTEanlsm shtJtrM be 
non-pathogenic, shouMbe aa sale in the 
large scale setting ae Ae host oiganisniv 
and* without adVerse consequences in 
the environment; and ^rj the vector/ 
insert shonlti be weJI characterized and 
free from known harmfiii sequences; 
shouM be- hmited is size as raodb as 
possible to th« DMA required to pedbrm 
the intended funetkax; shooM not 
increase the stability of the construct in 
the. environment, unless, that is. a 
requirement of the intended function: 
should be poorly mebiHzable; and 
should not transfer any resistance 
markets to. mjrr r»rypg»nicTT^q unknown to 
acquire them natunlly if such 
acquisitioa could compeomiso the use of 
a drug to control drseaMb agpota in 
human os veterinary Biadicinaoc 
agriculture. 
Appendix K— VII-PL htactiwetton.. 
Inactilvatioti Is any prdeesa tfiat dtestroys 
[ 728 ] 
Recombinant DNA Research, Volume 19 
