CONTINUATION OF MKULTRA, SUBPROJECT 133 (W/ATTACHMENTS)

• Document Type: SPECIALCOLLECTION
• Collection: General CIA Records
• Document Number (FOIA) /ESDN (CREST): 00693632
• Release Decision: RIPPUB
• Original Classification: U
• Document Page Count: 11
• Document Creation Date: January 23, 2025
• Document Release Date: February 12, 2002
• Publication Date: July 24, 1962

(111[31-
DRAFT
MEMORANDUM FOR: THE RECORD
JUL1962
SUBJECT : Continuation of MKULTRA, Subproject 133
1. The purpose of MKULTRA, Subproject 133 is to enable TSD/BB
to utilize the services ofillMTEMprAs80ciatetErofes-s-oz1tifaili0
Bit>16'gy at-Ren t3selaerfetilyte-chnislinstitute0.1TroymNeweYtork14)
� 2. During the first year of the program as made
significant contributions to the all too scanty knowledge of the mechanisms
of mineral transformations. A technical discussion of these accomplishments
is attached hereto with an outline of proposed work for the coming year.
It is possible that these investigations may well lead to new approaches
for energy transfer systems (bio-batteries) and deterioration, of metals.
� un.ctioned as GoVet'4'
and ellout during the first year of this project.
This service will heretofore
-be furnished by th Geildhitkter or:TMedic-altRese'axtbso The cost of -
_
this program for�the second year Will be $9 000.00 to which must be added
$360.00 which represents a 4% service charge to be retained by the sutou
The total cost of the program, therefore, will not exceed $9,360.00. Charges
should be made against Allotment No. 3125-1390-3902.
4. It is not anticipated that permanent equipment other than that
listed in the budget will be required for this program. Title to the
att:. A
Tot
APPROVED FOR RELEASE
DATE: JAN 2002.
n . r
equipment listed will be retained by thegbrstitute-in. lieu of higher
overhead rates.
5. Documentation and accounting for travel expenses which
are reimbursable by
the accepted practice of that organization.
6. 411111thas been cleared4O1
,AA)
Ali conform to
Si
TLY and is unwitting
and will remain Unwitting of the true nature of the s
APPROVED FOR OBLIGATION OF FUNDS:
DATE:
`.ACITSD"-- � -
Attached:
Budget
Project Summary and Proposal
Distribution:
Original only
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APPROVED FOR RELEASE
DATE: JAN 2002
Research.
The Geschicter Fund
9) 1834, Connecticut Avenue
Washington, D. C.
Attention:'
Subject: Proposal entitled .
Please find enclosed one co of the subject pro-
posal �submitted on behalf of
The proposal is not being submitted elsewhere for
possible support.
Your consideration .of our proposal will be a.ppre-
ciated and we look forward to hearing from you.
Very tru
APPROVED FOR RELEASE
DATE:JAN 2002
I 63i-35
Pr9posal entitled
Submitted on behalf of
June 1962
APPROVED FOR RELEASE
DATE:JAN 2002
DEPARTMENT OF BIOLOGY
June 14 1962
Purpose of Study:
The purpose .of this proposal:is a-request for financial support
to continue, an investigation of microbial action On marine manganese
nodules and terrigenouSmineral.sulfideS, which the principal investi-
gator has been pursuing since 1958. Very intensive work on these
materials is being carried on by him, with fruitful results during
the current year, 1961-62 umder a grant4-fraM the
4111111111111111111FtiStanfordUniVettitYWalifornia* Since relatively
little is known aboutacrobiallminetalftransforciation and in view of
.current academic and_practical interest ofmicrabiologistt
mining engineers, soil scientists, oceanographers .etc., in .the subject,
this research should make a valuable Contribution to:science.
Summary of Past Work:
a. Bacteriology of mineral sulfides.
geologists,
� Attempts vete,made ta:evaluatetWmictobial_floia'itolable from
unsterilized; crushed sulfide minerals by enrithment-in mineral. solution.
The following minerals were studied: alabandite, artenopyrite, bornite,
Chalcocite, chalcopyrite, cinnabar, cobaltite, covellite, enargite galena,
marcasite, orpiment, pyrite; pytrhotite realgar, and.sphalerite. Of these
minerals, only cobaltite,"enargite, galena, pyrite, pyrrhotite, realgar,
and sphalerite yielded microorganisms. For the most part these organisms
were heterotrophic and probably represented contaminants. However,
Hymhomicrobium, isolated from realgar, a pink yeast repeaSE81041EILEDA1IEAIN
DATEAAN2M12
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from sphalerite, and Arthrobacter, -isolated from cobalt,ite galena
pyrrhptite,. realgar, and sphalerite may constitute part of 4 normal
� flora. The action:ofany of these' organisms with respect to the mineral
with which they were found associated remains to be established.
After surface-sterilization some of the above:mineral:sulfides,
when enriched in mineral solution, have yielded iron-oxidizing auto-
trophs. These minerals- include arsenopyrite, pyrite pyrrhotite chal
copyrite, enargite, galena Marcasite and sphalerite. At least some of
the isolated bacterial strains are not restricted to a diet of iron for
'energy, but can use sulfur or, probably, some other oxidizable metals.
The ability to grow on any-of-I:the above sulfide minerals was tested
by toCulatingsurfaCe74terile samples in Oxidiiing.coluMns.with'Ferta-
_ _ _
,bacillus fetrooxidans and attempting to recover the organism Irdia.effluent
feeding solution over a period of tWo Months or more. So far, positive
results; have been obtained-Witharsenopyrite, enargite, chalcopyrite,
�
� � �
marcasite, galena pyrite, pyrrhotite, and sphalerite. Negative results �
. �
have been.obtained-with-alabandite,i bornite, cobaltite, covellite
chalco-
. cite, and one sample of galena.. Cinnabar, orpiment and realgar are.being
currently, investigated
In addition tatha4oregoing.qualitatiye work, quantitative studies,
4
on the rates of oxidation of' synthetic Cu2S and natural arsenopyrite are
presently being undertaken. From these studies it has became clear that
synthetic Cu2S can be oxidized at least 4x as fast by bacteria than by
autoxidation, and that arsenopyrite can be more rapidly oxidized by
bacteria than by autoxidation, Results with the latter material are not
yet sufficient to establish an exact rate comparison. The precise mach-
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DATE: JAN 2002
anitia of bacterial oxidation remains to be established. The work with
:synthetic Cu2S proves, what some other workers seem to doubt, that F.
--�
ferrooxidanaCan oxidize metals other than iron.
�
� b.. Manganese Nodules
-
:Oceanographers have felt pretty strongly in the past that the origin
. and. development of manganese nodules in the oceans is attributable to
purely. physicochemical
.on finding organic nitrogen in nodules,
� concluded that biological agentsWere.invOlved in nodule genesis. At his
:tuggestiOnt the principal investigator attempted to find out if bacteria
might play a.role. in this. He found that bacteria were indeed present
4n_thenodular sUbStince.after'sUrface-sterilization (4 rough estimate. at
� � � � �
present is 104. per gram). They included Achromobacteri. Arthrobacter,
:
. Bacillus, BrevibacteriuM 'Staphylococcus, Vibrio an unidentified rod and
an unidentified coccus. The principal investigator showed in quantitative
experiments. that nodular'substance:can adsorb manganous ion from sea water,
and that adsorption is accelerated by bacteria that grow from the
nodular material. The acceleration of manganous Ion adsorption is eXplain-.
� . :
- �
- able on the basis that the bacteria oxidize the adsorbed manganese, which
_ . � .
� � 7
facilitates further adsorption otmanganous manganese. The acceleration
� requires the presence of peptone to permit bacterial development. If
peptone and glucose are present, manganese is released from the nodular
substance rather than adsorbed .at least in a net effect. Since some
nodules were apparently initiated around shark's teeth, ear bones of whales,
pumice, etc., in the sea, attempts were made to see if oyster shells can
adsorb manganous manganese and thus serve as possible foci of nodules.
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It was found that they do adsorb it and that peptone did not stimulate this
adsorption (no bacteria were present!). As far as a survey of the lterature
has gone, these observations with respect to manganese nodules have not
been reported before.
Pertinent-literature:
The early literature dealing with microbial. action on minerals his
been covered by Alexander (1). A review by Lyalikova summarizes much of
the-past important work on Thiobacillus ferrooxidans and Ferrobacillus
ferrooxidane (2). .An intimate association of iron-oxidizing autotrophs
with natural mineral sulfides has been indicated by the work ofl
111111111d by that of Lyalikova (5). Differences of opinion exist between
Bryner and Anderson (6), MAlouf and Prater (7), and Ivanov, Nargirvyak,
and,Stepanov.on the one hand, and (8) on the, other
about amechanism of mineral sulfide oxidation of-chalcopyrite, molybdenitei,
chalcocite, and sphalerite, for instance. No previo.:_s studies on .bacteria
in manganese nodules has been reported. However, bacterial manganese oxi-
dation and reduction by soil bacteria has been known for some time. An
important quantitative study on large-scale bacterial manganese metabolism
is that of Mann. and Quastel (9). Descriptions of manganesejtodulesare-givin
by Murray (1) and Dietz (11), A.Chamical.and physical study of nodules.
was made by Buser and Gruetter (12). The finding of organic nitrogen in
nodules was first reported by.. Graham (13) and Graham and Cooper (14), who .
also suggested a biological origin of the nodules on this basis.
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References:
er. 4
1. Alexander, M. INTRODUCTION TO SOIL MICROBIOLOGY, John Wiley
& Sons Co., 1961.
2. 'Lyalikova, N. N., Mikrobiologiya 29: 773-779 (1960).
e./.49W
(1962)ili'Presa.'
5. Lyalikova, N. N. Mikrobiologiya, 30: 135-139 (1961).
6. Bryner, L. C., and R. Anderson Ind. Eng. Chem. 49: 1721-1724 (1957).
7. Malouf, E. E., and J. D.Prater J. Metals 13: 353-356 (1959).
8. Razzell, W. E., Annual Western Meeting, Vancouver, Oct. 1960
Transactions, LXV, 1962 pp. 135-136.
9. Mann, G., and J. H. Quastel, Nature 158:154-156 (1946).
10. Murray, J., VOYAGE OF H.M.S. CHALLENGER. DEEP SEA DEPOSITS.
Her. Majesty's Stationary Office. 1891.
11. Dietz, R. S., J. Calif. Mines and Geol. 51: 209-220, (1955).
12.- Buser W. and A. Gruetter, Schweiz. mineralog. petrogr. Mitt.
36: 49-62, (1956).
13. Graham, J. W., Science 129: 1428-1429 (1959).
14. Graham, J. W. and Susan Cooper Nature 183: 1050-1051.
Pertinent Publications by Principal Investigator:
1
C-
(1962) in press.
01
1'33 3.33
Proposed New Work:
Continuation of Present lines of investigation:
-
a. Mineral Sulfides: � i
1. Continuation of survey of natural sulfide minerals for
a normal- flora, with particular emphasis on large-scale
microbial action on minerals.
2. Characterization of isolates (physiological and morpho-
logical)..,
3. Examination of isolates for specific mineralizing activities.
Elucidation of biochemidal mechanisms of mineral trans-
formation.
b. Manganese Nodules:
1. Qualitative and quantitative bacteriological comparison
of manganese nodules from different oceans.
2. Study of the biochemical mechanism of manganous oxidation
- and Mn02 reduction in the various bacteria isolated.
Determination of the mechanism of iron-incorporation into
manganese nodules.
4.
The methods to be used in .these studies will'be adaptations of Standard
procedures* of bacteriology, physiology, and biochemistry.-
.
Personnel:
ASO.C7,7Troff-Siology
Principal Investigator
Technician:
Graduate Student:
Undergraduate student: -
(NSF undergraduate
research fellow)
ci
dtlY"-SUpported)w
e*(summer 1962)
(summer 1963)
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DATE:JAN 2002
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Proposed Budget:
PERSONNEL
Principal Investigator
1/8 time-academic year
1/4 time-suer months
Technician-full time
PERMANENT EQUIPMENT
Fluorimeter
..CONSUMABLE SUPPLIES
- 'Chemicals
.-Glassware
� TRAVEL
To scientific meetings
OTHER EXPENSES
Publication
Telephone
INDIRECT. COST:
@ 147. of Total Direct Cost.
$ 1,050
700
4l55
5,905
900�
"
300
340
640
250
180
20
� 200
Total Direct Cost
,Total Cost
$. 7,895
1,105
$ 9,000
APPROVED FOR RELEASE
DATE:JAN 2002