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CONFIDENTIAL
INTERIM REPORT 100
PRINCIPLES AND PRACTICE OF BW DEC CN T AMIN ATI CN
22* Evaluation of Corps of togineers Mobile Mater Purification
' Ihit for the Removal of 13 globieii Spores from Cold Mater
By
Bernard F-, Surkiewiez
Isaac J. Fish, Jr.
and
Saul Keye
This is a report of record
and does not necessarily
reflect the doctrine of the
Army BW Program
Work completed February 1955
Authority to Reproduce
Granted to ASl ft-DSC
Per
;.c. <■
■■ ii. ' '
This Document or any portion thereof may not be
reproduced without specific authorisation by the
Assistant Chief Chemical Officer for BW, Camp Detrick
Physical Defense Division
Camp Detrick lhiB material contains information
Frederick, Maryland affecting the national defense of
the United States within the m a yng
of Espioift"-’ Laws. Title is, t.
secs. 79." 5 V- l 1 h’* lvati.-ir ‘:"on
or revete i . n;* '*on!cnls in any „
Published August 1955 manrn: a ; : hurLed person is
prohibited ‘r* taw.
55 A 4 4005a
FIDENTIAL
CONFIDENTIAL
INTERIM REPORT 100
PRINCIPLES AND PRACTICE OF BW DECONTAMINATION
22. Evaluation of Corps of Engineers Mobile Water Purification
Unit for the Removal of B globlgii Spores from Cold Water
APPROVAL RECOMMENDED:
APPROVED:
Chief, Physical Defense Division Director of Research
Camp Detrick
Frederick, Maryland
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INTERIM REPOUT 100
PRINCIPLES AND PRACTICE OF BW DECONTAMINATION
22. Evaluation of Corps of Engineers Mobile Water Purification
Unit for the Removal of B globigii Spores from Cold Water
CONTENTS
■ JBtgS.
Acknowledgment s . iii
Summary . . iv
I. INTRODUCTION
A. Authorization ............... 1
B. Purpose . 1
C. Description of the Water Purification Unit . 2
D. Preliminary Tests . 5
E. Test Agent . 6
F. Test Site . 6
III. TEST PROCEDURE
A. Tests Performed . 6
B. Sampling . 8
III. -INDIVIDUAL TESTS
A. Test No. 1 . 10
B. Test No. 2 . 11
C. Test No. 3 . • . 12
D. Test No. 4 . 13
E. Test No. 5 . 14
F. Test No. 6 . 15
IT.. CONCLUSIONS . 16
Bibliography . 17
Appendix . 19
i
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CONTENTS (Ccntd)
FIGURES Pag e.
1. Water Purification Uhit, Mobile Electrified, 1500 gph.
Project 8-75-05-012 3
2. Cross-Section of Diatamite Filter and ERDLator Showing
Points of Sampling . 4
TABLES
I. Chemical and Analytical Data, Test No. 1 . 21
II. Viable Bacteria Count, Test No. 1 . . . 22
III. Chemical and Analytical Data, Test No. 2 . 23
XV. Viable Bacteria Count, Test No. 2 . 24
V. Chemical and Analytical Data, Test No. 3 . 25
VI. Viable Bacteria Count, Test No. 3 . 26
VII. Chemical and Analytical Data, Test No. 4 . 27
VIII. Viable Bacteria Count, Test No. 4 . 28
U, Chemical and Analytical Data, Test No. 5 . 29
Z. Viable Bacteria Count, Test No. 5 ......... 30
H. Chemical and Analytical Data, Test No. 6 . 31
HI. Viable Bacteria Count, Test No. 6 . 32
ii
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ACKwnBT.TOfanprrs
Personnel of the Sanitary Engineering Branch, Corps of Engineers,
Engineer Research and Development Laboratories, Fort Belvoir, Virginia,
under the direction of Mr. Don C. Lindsten, Chief, Waste Disposal
Section, operated the ERDL Mobile Water Purification Unit, provided
laboratory facilities, and exhibited unlimited cooperation in all
matters pertaining to the BW phase of this field test. Portions of
the descriptive material and the two text figures were furnished by
theao Corps of Engineers personnel.
iii
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SUMMARY
A series of tests was performed to evaluate the use of the Corps
of Engineers Research and Development Laboratories Mobile Water
Purification Unit on cold water containing approximately 10^ spores
of B globigil per ml. The coagulation and filtration processes
involved in the normal use of the unit did not produce safe water
under these circumstances, nor did the normal chlorination procedure
of 1 ppm improve the quality of the affluent water.
The recommended method of obtaining safe water when the water
source is cold and contains a high concentration of resistant
spores is to employ superchlorinaticn to 100 ppm residtal available
chlorine at a pH lower than 7 (6.6) for 45 minutes. Alter this, the
water is dechlorinated by addition of 600 ppm activated carbon,
treated in the ERDL unit with coagulants, and filtered clear.
This method appears to be practical and to produce potable water
with safety.
Heating the water to 100°P before superchlorinaticn, rather than
reducing the pH, resulted in safe water but produced a contaminated
sludge and, in addition, required special equipment.
iv
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INTERIM REPORT 100
PRINCIPLES AND PRACTICE OF BW DECONTAMINATION
22. Evaluation of Corps of Engineers Mobile Water Purification
Unit for the Removal of B globigli Spores from Cold Water
£, INTRODUCTION
A. AUTHORIZATION
Letter TECRD MS 8-75-07-214 (8-75-05-014), dated 19 November 1954,
subject "Field Test, Evaluation of Corps of Engineers Mobile Water
Purification Unit for Removing Chemical and Simulated Biological
Warfare Agents from Water," from Chief, Military Engineering Department,
Corps of Engineers, US Army, Engineer Research and Development Labora¬
tories, Fort Belvoir, Virginia, to Chief, Biological Laboratories,
Camp Detrick, Frederick, Maryland, requested active participation
in subject test in the form of two bacteriologists, a supply of B
globigii spores, and the equipment and materials necessary for the
bacteriologists to assay all biological samples. First Indorsement
thereto, CMLCD-10-FD, dated 7 January 1955 > from Assistant Chief
Chemical Officer for BW, Camp Detrick, Frederick, Maryland, to Commanding
Officer, Engineer Research and Development Laboratories, Fort Belvoir,
Virginia, concurred with this request.
B. PURPOSE
A previous evaluation of the ERDL Mobile Water Purification Uhit
has been completed and presented in Camp Detrick Interim Report 66 (l)f.
Briefly, the conclusions were as follows*
"1. Water contaminated with lcA to. 10^ vegetative cells of S mar-
cescens per ml was sterilized when the unit was operated in the standard
manner (coagulation, disinfection with 1 ppm available chlorine residual,
and filtration).
* See Bibliography
I
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"2. Water contaminated with 10 B globigii spores per ml was not
sterilized when the unit was operated in the standard manner even with
chlorination up to 7.5 ppm.
"3. A definite and increasing BW hazard existed in sludge disposal
when B globigii was the agent, even when the unit was operated with
chlorinaticn. When Stfarcescens was the agent used, a sludge hazard
existed for at least 1 hour, even when the unit was operated with
chlorination.
"4. A definite respiratory BW hazard existed within the unit.”
As a result of the above findings, the Corps of Engineers decided
to conduct further field tests in an effort to rid water of g globigii
spores under cold-weather conditions by various pretreatment processes
which consist of "superchlorination" and subsequent dechlorination,
followed by treatment with the ERDL Mobile Water Purification Unit.
C. DESCRIPTICN OF THE WATER PURIFICATION UNIT
A cut-away drawing of the unit is presented as Figure 1, and a
complete description is contained in Reference (l). The unit (also
called anvERDLator) is a solids contact clarifier arranged for con¬
tinuous coagulation. A sketch of the unit (not to scale) is included
as Figure 2. In operation, raw or pretreated water is admitted to
the mixing zone, where it is thoroughly intermingled with the primary
coagulant (ferric chloride) fed by a solution feeder, the coagulant
aid (powdered limestone) fed by a slurry feeder, and, generally,
with a disinfectant (calcium hypochlorite) fed by a solution feeder.
This intermingling is accomplished by a series of belt-driven rotating
discs. The intermingled water along with the developing ferric hydroxide
floe passes upward through the multiple vanes into the clArification
zone. As the water retches the level above the slurry or sludge
blanket, which is maintained by a continuous withdrawal of the excess
slurry into the sludge concentrator, the upper effluent is relatively
clear. The concentrated sludge of the concentrator is vented off to
waste. The clear water of the separator zone is carried over to a
holding tank or wet well along with the top clear portion of the con¬
centration. From the wet well the effluent is pumped into a diatomite
filter for further clarification.
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The effluent of the ERDLator Is pumped into the interior of the
diatomite filter, having picked up a predetermined amount of filter
aid which is deposited on the precoated diatomaceous earth cake held
on the wire elements. Water passing through the cake to the interior
of the elements is carried over to a clear well for distribution.
The continuous addition of filter aid to the cake results in a slow
pressure rise on the outside, and eventually the cake must be removed
and replaced. The time for backwashing is determined by the differential
of internal and external pressures on the elements, which indicate the
density of cake impeding the output of filtered effluent. Removal of
the cake is accomplished by using the impounded air in the upper portion
of the filter on the air pump principle (i.e, a quick reduction of the
external pressure) to literally blast the cake from the face of the
elements and wash the expended filter aid to waste. By a reversal of
the process, a new cake is developed by a precoat and a new cycle of
filtering is begun.
D. PRELIMINARY TESTS
Upon being informed by the Corps of Engineers that pretreatment
consisting of "superchlorinstion." of the contaminated water for 45
minutes would be employed in these trials, preliminary tests were
conducted at Camp Detrick to determine the titratable residual
chlorine required in the field.
A general formula for the sterilization of distilled water con¬
taining anthrax spores by hypochlorites has been derived by Fair and
co-workers (2). The formula is;
„ , oo1.073(25-T) &
t3/2
Here, R
N
T
t
K
(HjO+)
required titratable residual chlorine in ppm
number of spores present per ml
temperature of water °C
time of contact, minutes
ionization constant of H0C1 (varies with T; 2.0 x 10”® at 0°C)
hydrogen ion concentration
Since B globlgii spores are as resistant as B anthracis. the above
formula ttss used to calculate the amount of residual chlorine required
to destroy 1 x ICk b globigil spores at pH 7 within 45 minutes at 0°C.
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Laboratory tests were then performed and the applicability of the
formula was confirmed. Between 75 and 100 ppra titratable residual
chlorine would be required under the specified conditions.
E. TEST AGENT
A paste containing 2200 grams of B globigii spores (Camp Detrick
Production Lot No. 112-BG-204) was suspended in 18 liters of sterile
distilled water and the suspension was homogenized under aseptic
conditions. The final bacteriological count was 2 x 10^® spores
per ml. The spore suspension was kept at approximately 40°P by
immersing the 5-P'^lon container of spores in a spring at the test site*
F. TEST SITE
The field trials were conducted at the edge of Miller's Pond,
Camp A. P. Hill, Bowling Green, Virginia. All the raw water used
during the trials was pumped from Miller's Pond.
. II. TEST PROCEDURE
A. TESTS PERFORMED
Each of the following six tests was performed under conditions
determined by Corps of Engineers personnel during the actual trials.
After each test, the ERDL Mobile Water Purification Chit was decontaminated
by pumping water containing 100 ppm chlorine and adjusted to pH 4
through the entire unit for at least 1 hour, followed by a thorough
rinse with pond water.
1. Test No. 1
The effect of superchlorination (100 ppm titratable residual
chlorine) on spores in cold water at three different pH values.
2. Teat No. 2
Superchlorination (100 ppm) in cold water at a lowered pH,
followed w:
a. Dechlorination with 600 ppm activated carbon.
b. Treatment with the ERDL Mobile Water Purification Unit
operating under the following conditions1
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(1) Coagulation, A coagulant bed was preformed by
operating the unit for approximately 4 hours with pond water.
(2) No chlorination.
(3) Filtration. Standard "precoat" method, i.e, adding
a^diatomaceous earth slurry (0.1 lb/sq ft of filter area) directly
to the filter elements.
3. Test No. 3
No superchlorinaticn. Treatment with the ERDL Mobile Water
Purification Unit operating under the following conditions :
a. Coagulation with no preformed coagulant bed.
b. Chlorination to approximately 1 ppm titratable residual
chlorine .
c. Filtration. Standard precoat method.
4. Test No. 4
No superchlorinaticn. Treatment with the ERDL Mobile Water
Purification Unit operating under the following conditions:
a. Coagulation. A coagulant bed was preformed by operating
the unit for approximately 4 hours with pond water.
b. No chlorination.
c. Filtration. Standard precoat method for the first
2^ hours of the runj "body-feed" method used for the last 3i hours
of the run. In the body-feed method, approximately 20 ppm of diatcmaceous
earth were added directly to the water entering the filter unit.
5. Test No. 5
No superchlorinaticn. Treatment with the ERDL Mobile Water
Purification Unit operating under the following conditions:
a. No coagulation.
b. Chlorination to approximately 1 ppm titratable residual
chlorine.
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c. Filtration. Standard precoat method for the first 3 hours
of the run; a combinaticn of the precoat and body-feed methods during
the last 3 hours of the run.
6. Test No. 6
Superchlorination (100 ppm) with no adjustment of pH, but with
the water heated to 100°F, followed by:
a. Dechlorination with 600 ppm activated carbon.
b. Treatment with the ERDL Mobile Water Purification Unit
operating under the following conditions:
(1) Coagulation with no preformed coagulant bed.
(2) No chlorination.
(3) Filtration. A combination of the precoat and body-
feed methods during the entire run.
B. SAMPLING
1* Chemical Samples
Samples of water for chemical analysis were collected in
bottles other than those used for collecting the bacteriological
samples. All chemical analyses were performed by Corps of Engineers
personnel.
2. Bacteriological Samples
a. Samples for viable bacteria assay, when the ERDL Mobile
Wfcter Purification Unit was employed, were taken at the following
five points (see Figure 2):
(1) Raw contaminated, at feed water rotameter inlet.
(2) Coagulated, at effluent trough overflow.
(3) Filtered, at hose discharging filtered water.
(4) Sludge, at sludge concentration waste outlet or from
near the bottom of the coagulator.
(5) Filter bed backwash, at waste line.
Samples from the tanks where superchlorination was employed were taken
at various intervals after the microorganisms and disinfectants were
added.
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b. All the sample bottles were sterile and contained 1 ml
of sterile, aqueous 18 percent sodium thiosulfate to neutralize chlorine
when present (l ml of 18 percent sodium thiosulfate neutralizes more
than 250 ml of H^O containing 100 ppm C^). Since the volume of every
sample was at least 200 ml, the resulting concentration of sodium thio¬
sulfate (less than 0.09 percent) is one known not to inhibit the growth
of the test agent.
c. Samples of the filtered water and of water from tanks where
superchlorination was employed were assayed by both the membrane
(mill! pore) filter and pour plate methods. These samples were taken in
sterile 250-ml ground-glass-top graduates. All other samples were
assayed only by the pour plate method, and these samples were taken in
sterile 8-oz screwcapped prescription bottles.
(l) Membrane Filter Method of Assay
Ringed Lovell Millipore Filters*, sterilized by exposure
to the vapors of ethylene oxide, were used. Water samples of 50-,
60-, or 100-ml portions (limited to less than 100 ml in cases where
unfiltered water samples, which clogged the MFs, ware used) were
filtered through each of two MFs held in the MF holders designed at
Camp Detrick. After each filtration, the MFs held in the holders
were rinsed once with 10 ml of sterile distilled water. MFs were
placed in glass dishes an blotters containing 2 ml of trypticase soy
broth**. The dishes were incubated at 37*PC for 15 hours. The number
of colonies appearing on the MFs was counted with a 91 stereoscopic
microscope.
(2) Pour Plate Method of Assay
Five-ml pipettes were used to deliver 5»0 ml of the
water samples to petri dishes. Che-ml pipettes were used to make
decimal serial dilutions in 9-ml sterile distilled water blanks and
to deliver 1.0 ml of the dilutions in duplicate petri dishes. Nutrient
agar*** was poured into the dishes and incubated at 37°C for 40 hours.
At the request of the Corps of Ehgineers, all samples were collected
by Corps of Engineers personnel. Accordingly, instructions issued to
these personnel emphasized the necessity of maintaining aseptic techniques
while taking the samples for bacteriological assay.
* Lovell Chemical Company, Watertown, Massachusetts.
** Baltimore Biological Laboratories, Baltimore, Maryland.
*** Camp Detrick Media Catalog Type 103.
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III. INDIVIDUAL TESTS
A. TEST NO. 1
1. General
This test was designed to determine the jffect of 100 ppm
tit rat able residual chlorine ("superchlorination" ) on the spores of
B globigii in cold water at three different pH's. The ERDL Mobile
Water Purification Unit was not used in thiB test. Since it had been
determined in previous Corps of Engineers trials that a concentration
of 100 ppm chlorine is effective in the decontamination of water con¬
taining certain CW agents, this concentration was used for all super¬
chlorination processes employed during this field trial.
2. Test Procedure
Each of three collapsible GRS-coated nylon water tanks (tanks
No. 1, 2, and 3) was filled with 3*000 gallons of water from Miller's
Pond, and 115 ml of the stock B globigii suspension of 2 x 10^° spores
per ml were added to each tank. The organisms were mixed in the water
for 5 minutes by means of canoe paddles and a pump which circulated
water continuously at a rate of 55 gallons per minute. At this point
samples were taken from each tank to determine the extent of spore con¬
tamination. A slurry of 3.6 lb "HTH" (70 percent calcium hypochlorite)
was added to each tank, along with 2,200 ml of concentrated HC1 in
tank No. 1 and 1,350 ml of concentrated HC1 in tank No. 2. No acid
was added to the water in tank No. 3« The HTH and acid were mixed
with the water for 5 minutes in the same manner in which the micro¬
organisms were mixed. The water-circulating pumps were operated at
each tank throughout the duration of the test. Samples of the water
from each tank were taken at 0 time (after the 5-minute mixing period)
and at the indicated intervals after 0 time.
3. Results
Chemical and bacteriological analyses of the water samples
are presented in Tables I and II in the Appendix.
4. Conclusions
It is evident that a pH of 7 or lower is necessary to attain
complete kill within 45 minutes of 1 to 2 x 105 spores per ml in cold
water containing 100 ppm chlorine.
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B. TEST NO. 2
1. General
This test was designed to determine the effect of superchlorination
(100 ppn for 45 minutes at a low pH), dechlorinaticn (with 600 ppm activated
carbon), and treatment with the ERDL Mobile Water Purification Unit (oper¬
ating with coagulation and a preformed bed, no chlorination, and filtration
with i the standard precoat method) on the spores in cold water.
2. Test Procedure
a. Superchlorination
Water tanks 2 and 4 were each filled with 3 >000 gallons
of water, and 115 ml of the stock spore suspension were added to each
tank. After mixing in the manner described in Test 1, samples were
taken from each tank. HTH (3.6 lb) and 2,000 ml of concentrated HC1
were added to each tank and mixed in the manner previously described.
Samples were taken at 0 time (after the 5-minute mixing period) and 0
plus 45 minutes.
b. Dechlorination
At the end of the 45-minute superchlorination period, the
contents of tank No. 2 were pumped to tank No. 3, and the contents of
tank No. 4 were pimped to tank No. 1. Water tanks 1 and 3 were empty
except for 15 pounds of activated carbon. A water circulating pump
was operating at each tank during the entire dechlorinaticn process.
Aa soon as tanks 1 and 3 were filled ( approximately 1 hour), samples
were taken.
c. Treatment with the EkDL Mobile Water Purification Unit
At the end of the dechlorination time (1 hour), the water
was led from tanks 1 and 3 to the ERDL Mobile Water Purification Unit.
Coagulated, filtered, and sludge water samples were taken after 1, l£,
2i, and 3 hours' operation of the unit. The sludge samples were taken
at the waste outlet of the sludge concentrator.
3. Results
Chemical and bacteriological analyses of the water samples
are presented in Tables III and 17 in the Appendix,
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4. Conclusions
Water containing up to 2 x 10^ B. globigii spores per ml can
be sterilized under the conditions of this test. It is of interest
to note that the sludge samples contained many organisms other than
B glofeigii. Since a bacteriological assay had shown that approximately
150 organisms (not g. globigii) per ml were present in the water from
Miller's Pond, and since no chlorine was used in the ERDL MSbile Water
Purification Unit, it may be concluded that the sludge contained
organisms as a result of the 4 hours' operation of the unit with the
pond water to preform the coagulant hed. It is felt that the presence
of a few B^ globigii colonies on the MFs or pour plates may result
from chance contamination, rfcther than the actual presence of the
spores in the water. Such contamination could occur in the laboratory
where traffic was necessarily heavy, or in the field where the sample
bottles were handled by personnel who worked near the contaminated
water and where both the personnel and the sample bottles were exposed
to any bacterial aerosol or spray resulting frcm the constant pumping,
mixing, and treating of the test waters.
C. TEST NO. 3
1. General
This test was designed to determine the extent of removal of
spores from cold water by means of the ERDL Mobile Water Purification
Unit operating in the standard manner (coagulation with no preformed
bed, chlorination to approximately 1 ppm, and filtration with the
standard precoat method).
2. Test Procedure
A stock spore suspension of 19 ml was added to 500 gallons
of pond water contained in a small tank. A water-circulating pump
was operating at the small tank during this entire run. Pond water
was pumped into the tank at the rate of 25 gallons per minute and
water from the same tank was pumped simultaneously at the same rate
into the ERDL Mobile Water Purification Unit. At the same time, a
spore suspension, consisting of 345 ml of the stock suspension mixed
with 30 gallons of water, was bled into the small tank at the rate
of 5 gallons per hour by means of a Signamotor. Thus a constant flow
of water, contaminated to approximately 1 x 10^ spores per ml, was
fed into the ERDL Mobile Water Purification Unit for 6 hours. Raw
contaminated, coagulated, and filtered water samples were taken after
1, lj, 2,. 3, 4, 5 and 6 hours' operation of the unit. No samples
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were taken from the sludge concentrator, but at the end of 6 hours'
operation a sample of sludge released from a waste line near the bottom
of the coagulator was taken and assayed.
3. Results
Chemical and bacteriological analyses of the water samples
are presented in Tables V and 71 in the Appendix.
4. Conclusions
Complete sterilization of spore- contaminated water was not
achieved under the conditions of this test. Significant reductions
resulted: Coagulation alone caused a spore reduction of 27 to 97
percent, depending on time, while the combination of coagulation and
filtration caused a reduction of 97.6 tc 99.998 percent. Upon
assay of the sludge sample removed from near the bottom of the coagulant
bed, it was apparent that the low concentration of chlorine used in
this test had no appreciable effect upon the spores and that a BW
hazard exists in the waste products of tie unit.
D. TEST NO. 4
1, General
This test was designed to determine the extent of mechanical
removal of spores from cold water by means of the ERDL Mobile Water
Purification Unit operating without chlorination (coagulation with a
preformed bed and filtration with the precoat method fcr the first 2$
hours, and the body-feed method for the last 3i hours of operation).
2. Test Procedure
Water was contaminated and led into the unit in the same
manner as described in Test 3. Raw contaminated, coagulated, filtered
and sludge water samples were taken at the indicated intervals during
the operation of the unit. The sludge samples were siphoned from
nefr the bottom of the coagulator with a long length of tygon tubing.
The tubing was flushed for at least 30 seconds gust before taking
each sludge sample. This method was used, rather than drawing the
sludge from the bottom of the coagulator through the waste line, to
prevent "breaking" the coagulant bed. In the course of operating
the unit, it was necessary to backwash the filter bed (diatomaceous
earth and floe) from the filter elements when the filter clogged to
the point where the proper flow of water was inhibited. After 5 hours'
operation in this test, a sample of the filter bed backwash was taken
at the waste line.
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3. Results
Chemical and bacteriological analyses of the water samples
are presented in Tables VII and VIII in the Appendix.
4. Conclusions
It is evident that the spores were not removed couplet ely
under the conditions of this test. Significant reduction resulted:
Coagulation caused a bacterial reduction of 94 to 98 percent, vhile
the combination of coagulation and filtration caused a reduction of
98 to 99.999 percent. Analysis of the sludge samples and the filter
bed backwash sample showed that a BW hazard exists in the waste
products of the unit.
E. TEST NO. 5
1. General
This test was designed to determine the effect on spores in
cold water when treated with the ERDL Mobile Water Purification Unit
employing the method used to treat non-turbid waters (no coagulation,
chlorination to approximately 1 ppm, and filtration with the precoat
method for the first 3 hours, and a combination of the precoat and
body-feed methods for the last 3 hours of operation).
2. Test Procedure
Water was contaminated and led into the unit in the same
manner as in Tests 3 and 4. Raw contaminated, coagulated, and filtered
water samples were taken at the indicated intervals. Since there was
no coagulation process in this run, no sludge samples were taken. Water
samples were taken at point (2) (see page 8) to determine the effect of
the chlorine (minus the effect of filtration) on the spores. Filter
bed backwash samples were taken after 2^, 5 and 6 hours’ operation of
the unit.
3. Results
Chemical and bacteriological analyses are presented in Tables
IX and X in the Appendix.
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1$
4. Conclusions
Little, if any, reduction in spore count resulted under the
conditions of this test. It is evident that the low concentration of
chlorine had no effect on the test agent in cold water. The filter
bed backwash samples indicated that spores were entrapped and might
constitute a BW hazard in the waste products of the unit.
F. TEST NO. 6
1. General
This test was designed to determine the effect of superchlori-
nation in water at 100°F with no adjustment of pH (100 ppm chlorine
for 45 minutes), dechlorination (with 600 ppm activated carbon), and
treatment with the ERDL Mobile Water Purification Unit (operating
with coagulation and no preformed bed, no chlorination, and filtration
with a combination of the precoat and body-feed methods) on the spores
in water. This method was evaluated because of a possible future
need for the Corps of Engineers to develop means of supplying warm
water in the field.
2. Test Procedure
a . Supe rchlorinati on
Water tanks 1 and 4 were each filled with 3,000 gallons
of water heated to approximately 100°F by means of a "Cyclothera"
heater. After adding and mixing 115 ml of the stock sp>ore suspension
to each tank in the manner previously described, samples were taken
from each tank. Added and mixed in each tank were 3.6 lb of HTH
(no HC1 was added), and samples were taken at 0 time (after the
5-minute mixing period) and at the indicated intervals after 0 time*
b. Dechlorination
Dechlorination with 600 ppm activated carbon proceeded
in tanks 2 and 3 in the manner previously described (the contents of
tank 1 were pumped into tank 2, and the contents of tank 4 were pumped
into tank 3). Samples of ohe dechlorinated water in each tank were
taken just prior to being led into the ERDL Mobile Water Purification
Unit.
CONFIDENTIAL
u
CONFIDENTIAL
c. Treatment with the ERDL Mobile Water Purification Unit
It the end of the dechlorination time (approximately
1 hour), the water was led from tanks 2 and 3 to the unit. Coagulated,
filtered, and sludge water samples were taken after 3/4, 1 3/4, 2 3/4,
and 3 3/4 hours' operation of the unit. The sludge samples were siphoned
from near the bottom of the coagulator in the manner previously described.
Filter bed backwash samples were taken after 1 3/4 and 3 3/4 hours'
operation of the unit.
3. Results
Chemical and bacteriological analyses are presented in Tables
XT and XII in the Appendix.
4. Conclusions
It is possible to sterilize spore-contaminated water under the
conditions of this test. As indicated by the sludge and filter-bed
backwash samples, superchlorination alone for 45 minutes with
100 ppm Cl2 at 100°F with no adjustment of pH does not destroy all
of the 2 x Hr spores per ml in water; therefore, coagulation
and filtration are necessary to remove mechanically the relatively
few remaining viable spores. However, under these circumstances
a BW hazard exists in the waste products of the unit.
IV* CONCLUSIONS
Replicate tests were not performed in this series, so that the
absolute significance of some of the data cannot be estimated.
For this same reason, and because operating conditions were changed
frequently within a single test, it is not possible to compare the
efficiency of various combinations of operating procedures nor to
recommend a single best method of operating the ERDL unit without
superchlorination. It may, however, be concluded that superchlori¬
nation (to 100 ppn) of water at 0°C for 45 minutes is ineffective
in removing B globigii spores (10? per ml) from water if the pH
is not adjusted. However, if the pH is adjusted to 6.6 or lower and
the ERDL unit is used to remove the carbon needed for dechlorination,
the water is rendered safe for drinking. Heating the water to
100°F and treating it with 100 ppm available chlorine for. 45 minutes
without pH adjustment was found to give as good an effluent as
water superchlorinated at 0°C at a pH of 6.6. However, the sludge
was found to be highly contaminated, indicating a lesser effect
of chlorine. In view of the greater simplicity and economy involved
CONFIDENTIAL
CONFIDENTIAL
in employing a small amount of acid rather than a large piece
of water-heating equipment, the pH adjustment method is certainly
to be preferred. When no superchlorination was used, the various
stages of the ERDL Mobile Water Purification Unit could be evaluated.
Coagulation reduced the concentration from 2 x 10-* to approximately
lCr; this process alone, therefore, is useless. When both coagulation
and filtration were employed, the reduction was much greater.
Since tests showed that filtration alone was ineffective, it is
evident that the filter removes organisms ’which are physically
trapped in the floe formed by the coagulants!. However, the water
resulting from the combination of coagulation and filtration processes
was variable in quality and had an average of 10^ spores per ml,
which is considered unsafe, and the sludge and filter cake were
highly contaminated. As might be expected, a residual available
chlorine concentration of 1 ppm had no effect on the spores and
did not affect the filterability of the ccagulum.
It is concluded that for cold water containing approximately
105 spores per ml, the following prodedure should be employed!
Superchlorination at 100 ppm residual available chlorine at a pH
lower than 7 for 45 minutes, followed by dechlorination ( by addition
of: 600 ppm activated carbon), and coagulation and filtration in
the ERDL Mobile Water Purification Unit.
BIBLIOGRAPHY
1, Camp Detrick Interim Report 66, hBW Evaluation of the ERDL
Mobile Water Purification Unit," O'Leary, Francis M. , dated
21 July 1954, SECRET.
2, Final Report, Contract QHfcmr-251, Harvard College and OSRD,
"Disinfection of Water and Related Substances," Fair, G. and
co-workers, p. 272, December 1945*
CONFIDENTIAL
CONFIDENTIAL
APPENDIX
Tables of Data
u
CONFIDENTIAL
CONFIDENTIAL
CONFIDENTIAL
Phenoiphthalein
Methyl orange
CONFIDENTIAL
22
TABLE XI. VIABLE BACTERIA COUNT, TEST NO. 1
Sample
Pond H£0
Tank 1,
after
contamination
Tank 1,
tt
tt
4 HTH t
acid,
0
Tank 1,
fi
It
tt
tt
0
Tank 1,
tt
.If
N
tr:
0
Tank 1,
n
tt
tt
R
0
Tank 2,
after
contamination
Tank 2,
w
II
+ HTH +
acid 0
Tank 2,
rt
tt
n •
tt
0
Tank 2,
ft
tt
a
If
0
Tank 2,
tt
ft
R
II
0
Tank 2,
w
tt
It
",
0
Tank 3,
after
contamination
Tank 3,
*
.*
+ HTH
0
Tank 3,
n
tt
n
0
Ta* k 3,
H
*
4
0
Tank 3,
H
tt
N
0
Tank 3,
tt
ft
R
0
Tank 3,
If
It
It
0
Tank 3*
tt
tt
R
0
MF» PP**
Bg/ml Be/ml
-
0(150 microorgr
other than Bg)
2 x 105
time
120
3.4 x 103
t 15 min
■ 0.15
0
+ 30 min
0.14
0,1
+ 45 min
0.14
0.1
1.4 x 105
time
268
540
+ 15 min
4.7
4.4
■f 30 min
0.85
0.4
*■ 45 min
0.3
0.1
1 hr
0.1
0.1
2 x 105
time
tntc***
1.9 x 105c
4- 15 min
INTO
1.7 x 10|
30 min
TNTC
1.9 x 10?
+ 45 min
TNTC
1.4 x 10?
♦ L-hr
me
1.2 x 10?
* l£ hr
TNTC
9x104
t 2 hra
TNTC
9 x 104
* Millipore filter
** Pour plate
Too numerous to count
CONFIDENTIAL
CONFIDENTIAL
23
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CONFIDENTIAL
•r
id i ty
CONFIDENTIAL
2k
TABLE IV. VIABLE BACTERIA COUNT, TEST NO. 2
Sample
MF*
Bg/ml
PP»*
Bg/ml
Tank 2, after
contamination
-
1.8 x 10^
Tank 2. after
contain. + HTH + acid, 0 + 45 niin
0.66
0
Tank 3, after dechl. of contents
of tank 2
—
0.2
Tank 4. after
contamination
—
2 x 105
Tank 4, after contain. + HTH + acid, 0 + 45 oin
0.14
0.2
Tank 1, after dechl. of contents
of tank 2
—
0.2
Time
.fad.
Coagulated
i
-
0
Sludge
i
-
0 (600 microorg.
other than Bg)
Filtered '
i
0.01
0
Coagulated
ii
-
0
Sludge
1*
-
$600 microorg.
other than Bg)
Filtered
0.1
0
Coagulated
2i
-
0.2
Sludge
2*
-
0 (200 microorg.
other than Bg)
Filtered
2i
0.5
0.4
Coagulated
3
-
0
Sludge
3
-
0 (150 microorg.
other than Bg)
Filtered
3
0.08
0
* Millipore filter
Pour Plate
CONFIDENTIAL
CONFIDENTIAL
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CONFIDENTIAL
CONFIDENTIAL
26
TABLE VI. VIABLE BACTERIA COUNT, TEST NO. 3
Water Sample
Time
(hr)
MF*
Bg/ml
PP**
Bg/ml
Raw contaminated
—
1.5 x lol
1.1 x 105
Coagulated
*
-
Raw contaminated
1
1.6 x 105
Coagulated
1
-
7.5 x 104
Filtered
1
7
14
Raw contaminated
1J
1.6 x 10?
2.2 x 104
Coagulated
1|
-
Filtered
ll
3
10
Raw contaminated
2
1.5 x 10?
Coagulated
2
-
1.2 x 104
Filtered
2
3
3
Raw contaminated
2i
—
1.5 x 10?
Coagulated
29
2 x 103
1.6 x 104
Filtered
25
3.7 x 103
Raw contaminated
3
1.5 x 10?
1.2 x 104
Coagulated
3
-
Filtered
3
Approx 100
80
Raw contaminated
4
1.1 x 10?
7.3 x 104
Coagulated
4
-
Filtered
4
Approx 200
160
Raw contaminated
5
—
1.3 x 10?
Coagulated
5
-
4.6 x 103
Filtered
5
4
4
Raw contaminated
6
1.7 x 10?
4.7 x 103
Coagulated
6
-
Filtered
6
8
16 6
Sludge
6
-
5 x 106
* Millipore filter
** pour plate
CONFIDENTIAL
TABlE VII. CHEMICAL AND ANALYTICAL DATA, TEST NO.
CONFIDENTIAL
27
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CONFIDENTIAL
Methyl orange
CONFIDENTIAL
TABLE VIII. VIABLE BACTERIA COUNT, IEST NO. 4
Water Sample
Raw Contaminated
Coagulated
Filtered
Raw Contaminated
Coagulated
Fi ltered
Raw Contaminated
Coagulated
Sludge
F i 1 tered
Raw Contaminated
Coagulated
S I udge
F i I tered
Raw Contaminated
Coagulated
S 1 udge
Filtered
Raw Contaminated
Coagulated
Sludge
Filtered
Raw Contaminated
Coagu 1 ated
Sludge
Fi ltered
Raw Contaminated
Coagulated
Sludge
F j ie i. * backwash
:: ' : i? re '
Raw «, ated
Coag u i u hud
Sludge
Filtered
“Mill ipore f i Iter
**Pour plate
"“Too numerous to count
Time
(hr)
hr
hi
• i
2
2
2
2
H
1;
2;;
2i
3
3
3
3
4
4
4
4
5
5
5
5
5
6
6
6
6
MF*
fifl/mt
TNTC*
TNTC
Approx 30
TNTC
TNTC
Approx 9
Approx 60
pp**
fig/ ml
2.2 x 10®
I x lg
3.5 x I02
2.1 x I05
9 x IO3
3.25 x I03
2.3 x PO5
8.8 x IO3
2.75 x 10®
67
1.8 x I05
9 x IO3
4.6 x I06
1.42 x I03
1.7 x 10®
9.9 x I03
5.6 x iO®
1.18 x IO3
1.4 x 10®
6 x IO3
6.7 x IO6
17
1.5 x iO®
4.6 x IO3
7 x 10®
1.7 x IO2
1.4 x 10®
3.2 x IO3
7 x 10®
2.7 x 10®
2
1.4 x 10®
3.7 x IO3
8 x 10®
II
CONFIDENTIAL
i
TABLE IX. CHEMICAL AND ANALYTICAL DATA, TEST NO.
CONFIDENTIAL
29
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<0 <0
o
o
o —
o —
o —
o
o —
o —
3 —
O —
3fc co
CL
o
o u.
o u.
ou.
o
u.
O U.
o u-
o u.
o u.
# #
#
CONFIDENTIAL
Phenol phthate in
Methyl orange
30
CONFIDENTIAL
TABLE X. VIABLE BACTERIA COUNT, TEST NO. 5
Water Sample
Time
(hr)
MF*
Bg/ml
PP»*
Bg/al
Raw contaminated
Coagulated
i
-
2.1 x 105
2 x 105
Raw contaminated
i
213 x 105
Coagulated
i
-
1.8 x 105
Filtered
i
TNTC***
1.6 x 105
Raw contaminated
l£
1.8 x 105
Coagulated
li
1.8 x 105
Filtered
lj
TNTC
Approx 1 x 105
Raw contaminated
2
1.7 x 105
Coagulated
2
-
1.8 x 105
Filtered
2
TNTC
Approx 1 x 10*
Raw contaminated
2i
1.5 x 105
Coagulated
Filter bed backwash
24
1.7 x 10j
3 x 106
Filtered
3
TNTC
Approx 1 x 10^
Raw contaminated
3
1.8 x 105
Coagulated
3
-
1.5 x 105 „
Filtered
3
TNTC
Approx 1 x 10*
Raw contaminated
4
1.5 x lOf
Coagulated
4
-
1.4 x 105
Filtered
4
TNTC
Approx 1 x 103
Raw contaminated
5
1.5 x 105
Coagulated
5
-
1.2 x 103
Filter bed backwash
5
-
9.7 x 105
Filtered
5
TNTC
Approx 1 x lO^
Raw contaminated
6
1.5 x lof
Coagulated
6
-
1.3 x 105
Filter bed backwash
6
-
3.7 x 105
Filtered
* u-nunrtT.*
6
TNTC
Approx 7 x 10^
** pour plat#
*#*Too numerous tc count
CONFIDENTIAL
s
CONFIDENTIAL
<0
co
£
O
g
<
8
««
-4
3
*
X
2
AS
<0 c*-*
2"l
o
0
1 1 C.7
116.0
106.1
0
d(J>0
• fa
ONgiflO
OoOOOOOOC
Color
(PP«)
o
=1*
O O O O |
sf =e s* sr
S3S3 '
• o
• 10 10 10 10 10 |0
10 > ' ✓ V v v V
i?
o
•
10
oo oo
• • • • 1
10 10 10 10
o © o o
• • • • |
10 10 10 10
0-000-
1 a a a * a •
• OIOOnO
K
V
Q.
CO
<o
h* co & 3* eg
■ ■ • ■ 0
lewacoN
10 10
d- d d-WO
• ■ a a •
N O) O) 0> N
a- eo0) — 5 — cm
• ••*••*
=*■ (000 N* N- C*. N.
10
in in in
8#-
o
10
CM
h* <0—i0
n i Wtf d-
St <o OO
d 1 d' 1 d-
.00 00 1
( St 1 1 St 9 St
cn
CO
fe i i fe “J
8
5 ,8 .8
• 8 i i S i S
Iss
x at-
o
■
2
o
2*r = S
2S-22
d O d*
1 1 2 1 ft 1 2
§»
u co
o
■
00
O
Q.(0 oo eg
2 1 "Z 1 eg « co
•
£ ^
o
•
o
O
• O' O oo
ar — Sd-OCN
Ssisss
O eg — (0 (0 (o k
co co co co <n co
z'fi*
<0 a#
i w S
-* £
o
© 3- =f co ©
eg eg 10
O (0 O d O
— iR eg
ooooooooo
g!0£
3 + -*
© o © +
g 10 10
'+» + +
© o © —
0)
Is.
X g-»
£c 4-
6
X X
X fe •
+4
4-
t*
•• a
4!
• l
g*
40
e
-c «e
• a
O
k
as
i—
a
as
o
g-» +* *
gj
gj gg t.
X
II
S£
i
ss
8
o u
0*0
o
y u
X
k k
ka k.
w
W L.
k.
0 0
gJ V
g- 4- r
«0 0
V 0
g~» g-»
4- 4-
0 (0
43
«*-
40
X]£'
g- <4-
(0 «
0
gj
g-
©
I
&
^ *■ co — — — — eg
-K JC J* JL JL JC Jt Jt J* Jt
sssss sssss
1
3
S 1
■U *0 g-* *0
0 « © 40 0
w *— C — k.
© 3 © 3 ©
*?=?*
u- o u. o u.
-• &
IS
I55
15
li
31
CONFIDENTIAL
52
CONFIDENTIAL
TABLE XII. VIABLE BACTERIA COUIT, TEST 10. 6
Hatar Staple
PP**
||Al
Pond
•
0.1 (27G aioroorg.
other than B|)
Tank 4, aft ar oontaalnation
m
1.97 x 10®
Tank 4, aftar o on tan. ♦ HTH,
0 tina
..TVTC***
1.66 x 10®
Tank 4, " " "
0 a 16 ain
Approx 64
1.17 x 10®
Tank 4, " " "
0 4 50 ain
2.5
*5
Tank 4, * " *
0 4 45 ain
TVTC
1.17 x 10®
Tank 5, aftar daohlor. of tank 4
-
1
Tank 1, aftar oontaalnation
•
2 x 10®
Tank 1, aftar oontaa. 4 HTH,
0 tiaa
TVTC
2.06 x 10®
Tank 1, " " "
0 4 16 ain
TVTC
1.96 x 10*
Tank 1, " " "
0 4 50 ain
TVTC
1.71 x 10*
Tank 1, " * "
0 4 45 ain
5.0
5
Tank 2, aftar daohlor. of tank 1
-
0.6
i
* Tiaa
Coagulated
w-
m
1.6
Sludge
3/4
-
5
Coagulated
1 5/4
-
0
Sludge
1 »A
-
80 5
Pi 1 tar bad baokaaah
1 5/4
-
1.06 x 10®
PI 1 tar ad
3 ’ (4
0.2
0.5
Coagulatad
2 */4
-
0.1
Sludga
2 3/4
•
15
Piltarad
2 5/4
0.16
0.2
Coagulatad
5 5/4
-
0
81udga
5 5/4
-
46 *
Piltar bad baokvaoh
5 5/4
-
4.9 x 10*
Piltarad
5 5/4
0.35
« 0.2
* Mlllipora Piltar
** Pour plata
*** Too nunerous to oount
CONFIDENTIAL
i
CONFIDENTIAL
33
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1
* 2
3
4
5
6
7
8
9
10
11
12
13, 14
15
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17-22
23 - 25
26 - 31
32 - 43
49
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INTERIM REPORT 100
Distribution List
Addressee
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Technical Records
Director of Research
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USDA Liaison Office
Commanding Officer, Naval Unit
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Dugway Proving Ground, Utah
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Pins Bluff Arsenal, Arkansas
CONFIDENTIAL
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CONFIDENTIAL
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51
52 - 54
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56
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ATTN: Research and Development Division
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ATTN: PTfcl Division (for G-2 and CIA)
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Army Chemical Center, Maryland
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Fort McClellan, Alabama
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kray Chemical Canter, Maryland
ATTN: Technical Library
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Surgeon General, USA
Department of the Army
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Office of the Secretary of Defense
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ATTN: Librarian
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CONFIDENTIAL
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Distribution List (Cont'd)
C6py No.
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75
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79-83
84
85
Addressee
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Building No. 1
Amy Chemical Center. Maryland
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Building No. 1
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Johns Hopkins University
6410 Connecticut Avenue
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Room 3E 1025* The Pentagon
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CONFIDENTIAL
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INTERIM REPQFT 100
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CONFIDENTIAL
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