Dr. William J. Gartland, Jr. 
September 14, 1983 
Page 2 
Attachment II - Page 2 
1. Oral doses of 1 billion to 10 billion E. col i -K 12 cells given to 
human volunteers did not produce adverse symptoms and were 
cleared from the intestinal tract in 4-6 days (2). 
2. Less expensive but highly effective filters could be used to 
minimize losses of bacteria in the exhaust gases of fermenters 
in place of the HEPA quality filters currently employed: 
The reduction in entrained, aerosolized E. coli exiting recombinant fermentors 
via the exhaust air stream caused by leaving a mechanical coalescer (cyclone) 
and a coalescing filter (Balston) in place was examined. The accompanying 
data was included in our original submission to the RAC entitled, "Request for 
Lowering of Containment Level and Exception to the Ten-Liter Volume Limit for 
Culture of a Recombinant Organism". These documents were Lilly Protocols 24, 
25, and 25A, also called ORDA 1001, 1005 and 1007, respectively, and submitted 
from June, 1979 through April, 1980. 
In all experiments reported, air samples were taken for 10 minutes at air flow 
of approximately 0.44 SCFA per minute. 
Mechanical Coalescer (Cyclone) 
Experiment 
I 
Before 
5.00 
X 
10 s 
org/SCFA 
After 
5.50 
X 
10 4 
org/SCFA 
Remova 1 
4.45 
X 
10 s 
org/SCFA 
(89% removal) 
Coalescing 
Filter (Balston) 
Experiment 
IIA 
Before 
5.68 
X 
10 3 
org/SCFA 
After 
7.50 
X 
10° 
org/SCFA 
Remova 1 
5.67 
X 
10 3 
org/SCFA 
(99.87% remova 1 ) 
Experiment 
IIB 
Before 
5.90 
X 
10 4 
org/SCFA 
After 
0 
org/SCFA 
Remova 1 
>5.90 
X 
10 4 
org/SCFA 
( 1 00% remova 1 ) 
Experiment 
I IC 
Be f o re 
1 . 75 
X 
10 4 
org/SCFA 
After 
0 
org/SCFA 
Remova 1 
>1 . 75 
X 
10 4 
org/SCFA 
( 1 00% remova 1 ) 
[ 354 ] 
