MKULTRA SUBPROJECT 143 (23 DOCUMENTS WITH PUBDATES FROM 621211 TO 6602 23)
Document Type:
Keywords:
Collection:
Document Number (FOIA) /ESDN (CREST):
00017403
Release Decision:
RIPPUB
Original Classification:
U
Document Page Count:
39
Document Creation Date:
August 23, 2024
Document Release Date:
September 8, 1976
Sequence Number:
Case Number:
Publication Date:
December 11, 1962
File:
Attachment | Size |
---|---|
MKULTRA SUBPROJECT 143 [8144510].pdf | 3.54 MB |
Body:
}am-cm exit-procleet� - -
Date of Cr:Act-Al _ PericA, T Antiga-of At
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ACCOUNTING BY INDIVIDUAL
SUBMITTED gy ,
PEMLZRA ablprAleTt i 2.43
DATE � VOUCHER NO, 2.12
FOR ADVANCE
PERI� OF ACCOUNTING
FROM
IL julanzazt:
.WOrg: Follow Inatfoctiona on 2.03
TO
3.965
I. CASH ON HAND BEGINNING Of PERIOD
_22._Coetobekr
2_39,83600001 S, EXPENSES THIS PERI00:
2. OUT5TANO9NG ADVANCES 'BEGINNING Of PERIOD
vOuCHER NUMDER
DATE
DESCRIPTION A OUNT
2. RECEIPTS THIS PERIOD,
L_EVenBee___�.�_$=.-..efthgt3AL______S
I
UNDER
DATE
OE CRIP ON
callalextIot
390836.00
TOTAL EXPENSES
n 16.00
. REFUNDED
MER
T
I ICA1311
_
j 11 1 CHIECR /NOWAY GAGA
1
1
11
7. OUTSTAND
HGl
AovApipcs
EN loll PERIOD
A f
chl fito011il 1
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A. cA5
ON NAND
IENOHOF
PER
00 ON
BALANCE
DUE APVAN E
III
I
A. I TOTAL TO ACCOUNT FOR
S j8
36.1pou, 1 7
1 7
TOT4L
ACCOUNTED
1
111
11
I
I 1 I
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$ 710.P,avoNnor
,o,�4,,�,
CERTIFY F SAEAVAILA LE
11 1 APPROVED 1
I
III
III I
/I
eer4
ly
thai
..1.
Al!f0A74flurrovl
hinted
hereon an.
09L
.
oo
f. NO.
I
CHARGE FAN ACCOUNT MO.
I I 1 , 11
I
1 11
1 1 II
$19*813640
DATE
I
� I
SIGNATURE
003YME:f
OFIAP
1
001150203
1
1
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1
r
1
A
OFF IC
1
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on
0
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of
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Farman
14 6
for
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o
Ihs
LiAfiruA
werealinmort44
ill not
ltostmra,
Seen
a efeortoFt:
for
rocelved,
offAciallpu
Ithat 'payment
and that
1
1 1
-
or
DAYL
SIGNATURE
Or AUTHORIZING
OFF
CER
CERTIFIED
FOR
PAYMENT
OR
CREDIT
s GNATUR
or
PAv�1111
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ATE
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or
CERTIFYING1
OFFICER
I
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TBD/BB
1
1
SPACE
BE.LOWHIFOR
EXCLUSIVE
USE
OF OFFICE
OF FINANCE
DESCRIPTION.ALL.
OTHER ACCOUNTS 13.33
34.39 1
.1
162.47 1
49.
SO
$1.34
H
55.66
67.70
71.60
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CODE
41
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'ACCOUNTS 03.27
1111
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606
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7
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60.70
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DATE
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CREDIT
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FORM 282 :=1:: 4.65 vioui
1,
OROUP I
"trairorirtrelYOZIMIZIP-
I I - )
,
�
This Jilt* certify-that I-have received an accounting fruit HAMM
&lb-project # 03 far the period 1 January 1963 . 4,06tOber 1905. Ite
- accounting refleatexpeasei ia the amount of439,843.29 agaiast the
paining grant balance ar139,834.00. The residia1ha3.sme_c1412.71
should be vtittea:Off as Wag inprecticle for collection or detailed
accounting.:
The acCounting statement end paymeat certification vill he retained
by TO and will be made available for review in TaiLnetessary.
I certify that satisfactory servicaa revesenteChy_the accouttIng
have been received and that-to the best of myknodledge and belief the
funds expended:were for the purposes authoriU.d by ,;he project approval
A. � , . � ,t t� � ..;'. 4 � K
. -
. �
111110221.0,
Consumable Supplies_
PICA
Telephone &ITtlegram
Tubes 6 pipettes
Misc. Supplies
E lee tr ieelfSuppi ires
ChelliC31311'
freight bills
0 9;089,17
650.00
7,710.00
400.00
1,995.00
250,00
1,458.00 -
1110-40
2160.00
200.00
_Jur
16,157.17
8.34-
5.55
575;43- -4
066.35'
115.93
176.07
15.33-
'Filter apparatus
111;16
Culture 3
5.99.71
Flasks, stoppers, syringes, etc.
357.99
Repair on Vacuum pump
8965 ___
Test
59.50
cube mixer
Service cnia-Sartorids Balance 120m
Totel:Censumehle Supplies 3003.11
Travel
1/634/6/63
/3/64-4/4/64,
0/19/64-10/24/64,-
Continued inet page.
457;04
27.9011
21.82
174.25
i61
t
-
Cost_ Account_
Object Class_
Retrairits and_References
Obligations Obligations tInliquidated
Incurred Liquidated Balance
/fij&1
LTA
070 era ex
e
cvh :
���
�
NAME OF A VA C E ,
DATE '
Am0 7.1.*L00.00
FEEHER No, (Finance one only)
REQUEST FOR ADVANCE
,30 jnnityAxy 10G4
$20..000,00
..*. � OF FUNDS �
PAYARLE TO
' taChad.-412.- �� , � '
ROOM
' ,
OUILDING
TELEPHONE EAT,
,
PURPOSE
� 4 Advance t - or funding )ULTRA SubprejeciCiA.13,
C-4--....."-=----�
invoice 0 2 bleb activity was
approved by C on 25 January
Pineal Annex attachment B.
....- .ai �_.'7..SD'
1964.
�
Accounting
� .
�
.
to be in iFeer anee with ITKULTRA
I 11! X0 1a_tes_ttlian 1.0 February 1964.
, , 1 11
[S7AirOS
�I 1,
1, ,
OF OLITSTA
I li 'II
1 III 1
IND ADVANCES
' II I I 1
,i, atrAelf At I
ko ehl �leportingipoint
Noy lunexpended
alp fully;
s
balance.
account
fl atetf
sathorixe
for ti
and Ibr thedu
deduct
advance
!dale
on
by submieelon Of woucharc;eactire frd
cReekodNolow. Init,Ie event of apifell
Fr up eale , 'to effect set dement,.
of any onexpended
re to 7. account
,iiimil
balence
rotund
...." I ATE I 1
II ' I AMOUR
uwAclaxon/pRwAce,
i
, REPORTING
POINT 11 I
11
I
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DIVISION
- H
DOUARTERS
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APPROVED
H I
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1
RECEIPT
FOR. 'FUNDS
ADVANCED':
DATE
SIGNATURE
OF
APP
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OFFICER',
under
so
Cc
whom
nouledge receipt
to he cowed
above.
of funds
for the purpose
in
the autatot]i
atited'andaccountled
elatedlhere.
for
ATE
1
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' F CERT 1 I
, I
* ORLIGATION REFRcNcE NO.
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9� 0
CHARGE ALLOTMENT NO.
4125.4390-111902
CERTIFIED
FOR PAY ENT1OR CREO
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I
ATE
1
AUTHORIZED CERTIFYING
1
OFFICER
$ GRAVURE
OF
AOVANCEE
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SPACE BELO FOR EXCLUSIVE USE OF
FINANCE DIVISION
I
� 'ARCO 111Y
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VOUCHER O.
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DESCRIPTION-ALL OTHER ACCOUNTS 1 3� 3
43.4�
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NO.
fl
GENERAL
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ALL.1 OT5.11 OR r Cosi
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- .
. ADVMAICEIACCOIUNTS
1 DESCRIPTION
- 1
19-27'
,
11
I.LIT2NUI.
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No.
STAT !ION
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60YIER0000T Tnerfinis �Tract Oyez 0-040h)
1
UnVORANDUU FOR
-
, Under the authority Id in-the m-�mcrandu dit d ',.pril-1953,--
-; from the DC! to: the DOA and the, extension-of -this autboyi.ty in fsub-
4
scqueat memoranda; SubprojeCt_143
has ''ssn approved, $20, 890 JKL:...
-of the over.--all_Project.UKUIZTRVfunds ha" 70'n cOligated--1-0-cc=v
trio
subprojecv'- r.mengt1..s and stiould-u-'-tle__r,Iirgoci-LCV CO 4125.1390=739.0.2
- Concur:
'0 pstE
Concur:
APPROVED FOR .OBL 'Da Tiff
OF FUSDS:
13 N
ifig4
Date:
Distribution:
Orig & 1 -
BB
es
GROUP 1 �
_.Exc1u-ded7frm 7 auto7
Obligationi- Obligations Unliquidated
Incurred Liquidated Balance
eDCLt�t
and
_ ensin of this-antho,rity
,tu-brre,i 3k,ct 143
I.?. 5� een. approveL, = 01 .144 _
rnernor,ind
-
IKULTR4. &rids have been obligated to cover the'
3125.1390.T.3902
. THAT-::;;NZ;,S�Ars:
'
GROUP I
Excluded from
matic_downgrading and
�aeifioation �
,
- , �
NAME OF A AN
DATE
*
.00
VOUCHER ,NO. flnence use on
FFOUNRDASDVANCE 7
REQUEST
lie
, OF
t
AYABLE
Atejlitititieblegt iftyleetnt Inst.
ROOM RCM-DING191;$
ns
TELEPHONE CRT.
L PURPOSE
� .7, Mesta. to NIA.VIA gibproject 1b3 Undo" IF X far siottrIt/ approve& by CAM) am 17 2*.iceveter 1962.
,�:t , Acootatiog for tags arroreoca it'll be in acoordusoo with atorobsetat A of NICIZMA nava Aare*.
... ,
.., �
--.
/Imam frevara tba cheek to ete.terottlefritoloarival. trench tbzwilk 11:30bet7et Crater Irs 3 aleatory 30631
-,, . --.
STATUS OF
1
OUTSTAN!ING ADVANCES
111 II i 11
. I agree that I
to "ff r�Pol'thill
ORF ROFFPORdea
will
Point
Wane.
fully
minted,
7 4402,10,1744
account for this ad by auhuieigian
and by th� due (trete checked below.
41160/44444 Ili.* Hy priari
of unuehers and refund 0/ onymonorponded
In the *rant alley faIiure to as account end
to effort sot fleapit. 11 Ir I I 1 1 11 1
beloonce
refund
�
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AMOUNT 1 1 I
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II
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FOR FUNDS ADVANCED
1
DATE
'�
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P 1
I
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SIGNATURE OF
1
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I �-
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1
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7 44knowita.244 0�444.1,44 el fund. dr, the
oo
under tor bia wed for ths prowlers iota
ea *shown *bore.
001100101t
*cloud
'stated
accounted
bore-
fo r
0 '� CERTIFY FUNDS ARE AVAILABLE .
..
52:1241M23:23LACItSki
DATE
AMOUNTd
0212,14AT ION REFERENCC NO.
...............-,
CHARGE ALLOTMENT NO...
CERTIFIED
FOR PAYMENT OR CREDIT
I
I i
DATE 4, ,
AUTHORIZED CERTIFYING OFFICE
SIGNATURE of ARTA/4st(
ATE.
SIGNATURE
1
. SPACE 0 ELOW FOR EXCLUSIVE USE OF FINANCE � ION
RED BY
�Ii .
REVIEWED BY
1
,...,:-
VOUCHER NO. T� 12
-, 0 coital. or4- ALL OTHER AC 01P4 S 13.3S
4.
43
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ir
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DESCRIPTION.
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AMC OF P A
DATE
,
*62
SIVA36600
VOUCHER NO. (F/R4Fce "IR
FOR ADVANCE
19 Det ...a. ...x. ,79
,REQUEST
'�,..: OF FUNDS *
$ g.
PAVAOL -
ROOM SU1S.ONG
T LEP 0 E EXT.
0...........,
PUNFOSIL -r
*, . , f-
. . AdraVF040 to 14103Intik titlibpro4iot 143 Uteri*. # 3. far mot%v ty approve& by' MO
frd. Accountimg for tbin edience 10133. be In accordance ifltlx ftttecbment A co? ICULTRik
,;' � ,..
. -;�kg., Please rotifer% the check to CbierirSDfritological Broth tbromA 1311D/S4get �facer
,
on 3.7 December 1962.
/local Annex. ,
by 3 January 2963/
,-
i ' STATUS O OUTSTAND
'I 1
011ii III
IN VANCSES
1 I 1 1 1
/ a�,,.. //H.*
toirmo reiirrvino
any'unoxpoododi
s fully pranunt far this
polfte prated and by the due
bottoclo, / authorise deductilon
advance
dolts
bV oobolooilOn
check.d below.
frosrevg rotary
of
ln
to effact
voucher. and re fund
the averse *fop failure
kletelewent. 11
of
ill 1
any unexpended balance
to so sweetest and refund -
1 II hill
liv DATE 1
I
IAMOUNT I
I
top
I I
I
1 REPORTING
1 p INTI 11
11 �
1 II IH
1
11
V
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1 T
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quire. .
Iteaearch Accountant
told se he-diseussed7thee
ou-this'morning and it was. a
You lioAlld again contant_th4 'contractor
re the $18,20 unexpended balance reflected in the
..acesAinting ftf�-� the period ending August 31,- 1962,
_not carried forward forward to the text grant year;
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current grant period would be extendCtd.
-tinte74nlyi.tc-30 June 1966, giving the grantee
Jopportunie.. in the field the $123.69
the January 10th, 1966
--- letter. fro the original of of this
et0-tr-it(4
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,._ oe
the February 10t
1966 letter from th which forwarded
this 'grantee's. atcoun urea �uring the period -
January ifkt-i-jpp3-4UOT-Igh�Ootiober 22nd, ,1963. . .
_ , .
s.
auks Very!, such for /our help in the first ites.
, , _ . .
MEMCAXDUKirOK : Chief, TSDAB
=MIXT :Dallnquent Accountings
0----MKULTRAi-SUbproject 143----
,
1. Eeview_of MKULTBA-p-Subproject 143 financial_records indicate
that accOuntings for grants are now past due. It is_xequested-that
your officeiinitiate action -to obtain the de3.in4pent_accounting8.
2. YOr_yow information fundi have been adVanced to the grantee
U follows
AuthOrization 1 $19,83&00
Grant period 1 Jan. - 31:Dec. 1963 !
AuthoriSation 2 i20,000.00
Grant periOd 1 Jan. - 31 Dec, I61
�-�"" 1
Distribution:
Original-fe IMdressee
- 4
�
cluidwiedgediaf
dated
_ - "rt, � , -
PROJECT CRYPTO :MKUMTRA-#143- _
GRANTEE
PRINCrPAL INNESTIGATORI:
DATE RENEWED :H=1 January 1964
During the past year, the investigators havestudied a
total of 57 compounds for_their capacity to stimulate bacteria.
These 57 compounds-consisted of 34 phenols, 14 amines, 5 alde-
hydes and ketones, 3 quaternary ammonium compounds_and 2 un-
classified chemicals. This stady has shown that_the_phenolic
chemicals are_theiibest compounds for stimulating-bacteria. Of
this group they_have-noted-that Q-phenyl--phenbl, p,itert-batyl-
phenol and nonyl-phenol are the-bett compoundsistudied-so far.
They havebeen_able to produce as much as_aJfour4old in-
crease in microbial growth using these compounds, This degree
of stimulation-is not as great as they had hoped it would be. _ _
For this reason,_during the past three weeks they_changed their
line of approachito the problem. -- -
They havepradaced_a product consisting of_phenol, casein
hydrolyzate, yeast_extract and peptone which has_a_much-greater
stimulating capacity, It appears that concentrations as small
as 50 ppm will_give them a consistant 10 fold'increase-in-grow-
th. The deterioration-of the cutting fluid-isgreater in three
days than they-have ever observed in their studies-vith these
products at anytime,In addition, as yet unpublished inform-
ation from thislaboratory indicates that this-produce-should
neutralize practically any known biocide that might be added
to cutting flaids.
It is belieVed"Nthat they have arrived at a point where
dynamic progresscan be-anticipated. In view of_the_high-pro- -
udcitivity and promise of resultS-that will be=of direct bene-
fit to the operational aspects of DDP. It willTbe the purpose
of the renewed project to combine-the two principles-of bact-
erial stimulation(the-use of subeffective concentrations of
Inhibitors and_the_use_of_non-specific protective compounds)
to determine the_ultimate stimulation to be expected-in cut-
ting fluids. More emphasis will be given to actual-degradative
process a r se ban stimulation of growth per_se. In add-
ition as agreed to investigate iiriai fouling
of die e coo ng systems in conjunction with!his-other work.
� .
: .
NAME OF
DATE '
30 Jartvz4ry 190g,
AmouNT WO .00
FENNIEN NO. (Patience Use 001Y)
REQUEST FOR ADVANCE
$20_,000 CO
OF FUNDS
PAYABLE TO �
Ir 0 0 �lea
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itunditglarMITA Subproject 143, invoice 4:# 2 mrhich activity
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SOVIEMOOKOT POINT.OtiOnt.cit 1142
;
rw.u. &January 1964
COVERT:ACTION-SUPPORT
B ! --Sabotage .
Branch B Category
IVa
roject Title _Item Cla-fication ied (Data)
_ Task No
Type of Contra. DateIrdMiateci ,21_January_1964
TT $2 CcO2M4
$20 RO 0 Completion Date
-
Purpose: The objedtive of the first year was acnieved._:An approach- o
the stimulati6h_of soluble-oil degradation vas-proved feasible. Iin-
addition a system for negating microbial inhibitors was developed.
These two accomplishments appear Wbe the firstsignificant break-
through for mitrobialidegradation of materials. It is planned-to----
study furtberthelliditatiens of-th-etia-approAches and-theit-applicatiol
to a variety7of-praCtiCal-bituations during the coming year. -
Status: Performance has been excellent. Has lrogressed beyond-expect-
ations. Current in fiscal matters, -
Requirement: �Generated by undocumented requests_from DP units and div-
isions as well As necessity for TSD-t0 maintain_a_quick delivery cap-
ability to meet anticipated future operational needs.
4. The cost of this program for the second year is
$20,000.00. To this sum must be added $800.00 representing
a four percent service-charge to -the'
The ill function as cut-out and
0-thisi-grant--The cover title of the_project_is _
. Total cost 61-the project will hotexceed-$20,800;0
Charges shOtild be made agaitst7Allotment No.--4125;=.1aga.=3902.
6.
Ill request th
to submit :a summary-accounting of mohies-receiVed and to
return any_unexpended grant funds at-the end_of tht litoject
year.
7. Title to any permanent equipment purchased by funds
granted th shall be retained by7thet-in-lieu of
higher overhead rates.
-
8. It was mutually agreed that documentation and acc-
ounting for-travel expenses which are reimbursablt-hY the
hall conform with the acceptedipractites of _
that institUtion.
Attachments:-
Pr op o_sal.ik- budget re Vint 4r:s
Distribution:
Original-only
el- a CO7-44 ;* a 11
� 4 1 y�
r-777777JW7171. T
MEMORANDUM-1'0R
THE RECORD
SUBJECT' :--MKULTRAI Subproject 143
143 is to-continue support
1. The_purpost-of subpro
of the work:Of:
on an approach-to microbial t
deteriorationof_materialS,
2. Intringthe -first year a'syStem-was.develOed whiCh
allows A 10_ftild-Stimulation of the microbial deterioration
-of solubleoilltoilants (as:4 type system) usinvextremelr7-7:7--
small qUantities-:_of.actiVe material, In additian a system -
was found:which_appartntly_serves to inactivate-most-micro--
bial inhibitors in-suc 'a product. This wasAuuch-more-of an
accomplistiment_than was-expected. For the coming year-it is
planned to-explore-the limitations of-these approaches as
_ _ _
they apply-tocutting=oils and other petroleum based products.
It is hoped that these techniques will T?e a_pplicableacross-
the-boarcLtoiall_sabotage or harass rent systems involving
microorganisms, promoting a more-deIlverable package-to_the
operator.'
3. principal investigator;_has-been-
cleared through SECRET.
_AMMIM
"
� � �
ii
:
5.u3.!�:-..4.1.a..N.:.Y73--,I Fort
SUMECT- -L-mxt.L.TRA --Subp,:ojee
. _
a uary 964
Under the : au t bori ty-- gr atdth t _cle:=2oranduridat.o.d_13:-A-nril- 1953 ..
. .
from th rcr to th-a-DO/A , and the edraenclon o.f th -aa,.1tho-r. -I vs; 1 1.1 filth- �
4.equer,t � rzi.tmoranda, ----143 hashtr
-of tne
ubproject4a e
- �
APPROVED
OF FlItelD
Date:
Pros: MKULTAT-fur 3 .7...a vs
_
ATION-
_
DistributIcn:
�rig Z1 1 -
2 - BB
GROUP_T�.. .
fn)m auto .;�7::
3 � � :ill �
72'
-
As you know, the grant :from,
one year old and itAs_tiMelt6
1
During the past year wellave_studied a-total of 57 comounda=for-their
capacity to stimnate_bacteria. These_57 compounds consiSted_or 34 �
phenols, 13 amines, 5 aldehydes and ketones, 3 quaternary-ammonium--
compounds and 2 tnclassifiediCheiicals�This:tudy_has_shown-that-the
phenolic chemicalsiare-the_hest_compounds_for_stimulatinEbetteria.- Of
this group we haveinoted-that Qphenyl-phenol, p-test-batylphenol, di-
sec-butylphenol,:dbdecyl pehnol, 214-dimethyl--6-test-butylphenoand�
nonyl phenol are-the_best coovounds_studied_so_far_._.
We have been ableAoiprodUce as-much_as_a fourfold_increase-inFMiorobial -
growth usinuthetecompounds. This degree of stimulation-ia-tiOtTas- -----
great as we had hoped-it7Wciuld-bo.__For_this_reason� during-the-past
three weeks we changed our line of_approach to the problem;
We have produced aL-product_consisting of phenol, casein, hydrolyzate,
yeast extract and_paptone_whichlhas a much_greater stimulating capacity.
It appears that eoncentrationt_as_small_as_5() ppm will giveus_a_consis-
tent 10 fold increaseAn=groWth._ The_deteriorationlof_the-cutting fluid
is greater in threedays_thanTwe have ever observed in our-studies_with- -
these products at any-time.- In addition, as yet umpublished_infermation
from this laboratory indiCates that this product should neutralize prac-
tically any knowA_bibcideLorLstat_that might,be_added_to:cutting fluids.
We feel that we how haveferritediata_point where_we can_makeisomeAYnamic
progress. During th coming year we hope to: :
1. Study additional chemicals and_media_products_for_theirstiMulatory
capacity.
2. Determine the best_ratios of_the media_components_and generally__ _
improve the fOrmulition.
3. Determine the effect of the formulation_on_other_petroleum_produots.-
4. Determine if the:formulation_has the_capacity_to_neutralizeanti-
microbial agents.
S. Begin work on how the_formulition_stimulates microorganisms.
6. Determine the effect of the formulation plus urine_on microbial-growt
I believe these points cover the main objectives for the coming-year.
Kindest personal regards.
Sincerely yours,:
PROPOSED_OPAIT_COST
Prineipalinvestigator_( time)
3990.00
Full time-research _assistants 9417.00
B� Expendal1e_eq4pment-_ata supplies 2560.00
C. Typing reports; papers, etc. 700.00
D� Overhead (20%) 3333.00
sicirAL 20,000-00
-
27 January_1964
MEMORANDUM-FOR_TRE RECORD
SUBJECT: MKULTRA_Subproject_143
. At the request of both Chief and_Deputy_Chief, TS
wai-contactedifor the purpose of funding this MKULTRA
u hrau h one of the established funding mechanisms controlled
by Due primarily to the futiaing date, it was_riot_possible -
to accomplish this at this time.
2. Arraigements illkemadewit to project
our funding requirements_fcTr MKULTR Zu� p ojects on a serni-annu-al
basis. With thisAea
of funding our total_re_quirernents.
3.
�11 meet with
funding echamsm _at_1115 this_date:
feels that they_may ca.pable
o discuss other_Subprojects_
ExeMd v!?,713111,
triax'2!..;
-
- '
. COVERT ACTION SUPPORT
Branch._ BB Category: SAbotage -
(formerly :IIIb)_
iProject Engineer
Task No
e of Contract -
----
Purpose: 'To 'investigate the poter.tial of a new approach- to- the-stiir.
ulation--Of raicrobia-l=deterioration-of petroleum 151"odutI
and its possible-application to other. degredative Crocesse
. _
_ .
Status: Contractor has-develooed-a. model system for desired system.
Will now attempt-to toiis'nftl scle uriwt
fiscalatters. -
� �
Requirement: Generated by-undocumented requests from DDP units-and
divisions as well_as.necessity for TSD to maintain a quick
delivery capability-to meet anticip-ated-future operational
needs. -
a
�
7: ' .-
BECELE'T
DRAFT__
11 December 1962
on an approach to the stimulationiotpetroleum
_product deterioration.
2. Although the-tOdel system for these studiee_vill-be_pet-
roleum productsi-thia_appreoeh to stitulation of deterioration may
have vide-application to all areas of biologicar,sabbtage-currently
, of interest ito TSD.
3. principal investigatrjM3beencl�.
through SECRET,
4. The cost of this program for the first year is $19,636.00.
To this sum-must be-tddid $793.44 representing a:rour_per cent service_
functiona!vsut out and cover f6f-this grant. The-cover-title Of the
5. Total_cost_of-the project for a period-of one-year will -tot
exceed $20,62944.' Charges should be made againstJAllotmen't No.
#
11 request th
a summary acCbunting-of monies received and to returnLany-nex-
-
-
_ _
_
;ended grant funds at the end of the project year.
Title to any permaneni:ewipment purchased by fUndalra
the hall be retained by theiln lieu ofbieler over-
head rates.
. It iossmutually agreed-that7documentation WI-accounting__
for travel expenseevbilch are reimbursable by
Cceformvith the_accepted practices of that-institution.
9. The requirement for a six months' informal accounting on_
the part of the7principal investigatbr is waived.
hall
Attachment:
Proposal &iudge t
Distribution:
Original only
� "Tr
TSD iological Branch
1,--1 c�.(117
p
arbdrg-
d can place the following equipment a th-ediposa
-Ultra-centrifuge
--*
Ratheon: 'Son i 6- Osc i I
--Mickie Disintegrator
Centrifuges-(refrigerated, Sorv� International, Ultra-centrifuge,
Si4inco, etc.)
4 _ _
_
Sharpl esSUper Centr I fuge:'
-BeckMan 0141-60 Spectrophotometers
�
Beckman 8
Incubators_
Research pH Meters
Shaking Incubators
Kjeldahl Apparatus
Colorimeters
Pilot Plant Fermentors
Chromatography Equipment'
Fraction coil ectors .
Isotope laboratory with-complete supporting�equipment
- To determine�If_the _growth of microorganisms_in petroleum
products can be markedly increased by the addition of substatic
concentrations of antimicrobial agenis._,
ITEWANKEll_IEE
as been ca_refuLLy searched b_ deterpine_what
microbial, growti
relate to thitopic,,
mshen`ei-iiii and PeroVa institute_of Microbiology, Academy o
�
_ -� - �. ��-- � �
cierice U.S:S.
ikrobiologiya', 23. 53-56, 1954) found that Cultures_
of SaccharOmyces:ceeviTiae�itr_ained to grow In of -
,
� .
.,---
small concentrations pUligC12;,orphenols have much higher _fermentation:,
,,,,�-�._%�� '-'..--;:i0.:::;:�"4 ;:'_1�:'-�: . ti:L-.. - 4,-. ,,, '-.,.-:. 1.-:-�; '''''' ..:' `. � f-r� ': : ". _ , 7 ", � .:, �:, - .":�:' _ : . , �,.. . ' - ' '
rates r.than..,culturesthat_have_not_been_trained to_grow_in_the...presence��
:As ,iittre_as_50_ ppm of HgCli accelerates 7 the_.
oworkers believe_thatthls effect is
to traces of inhiUtor_in_the environment .and_not_because_of some
change in the physiology of the
' cells
reported that 50 ppm of-2 richloro phenol CouLd_rnarkedly stimulate
the growth of bacteria in-acraision_oils. in some instances this
produceslmost a'three fold increase in the number of ceLl s
in the environment,
found that .25 to' 150 ppm of--2,4-dinitrophenol increases the oxidation
�
of
Cie), _C12, C13, C14, and c6fatty acids. The
1;�f, �
increase was asinuch-as 100% and was not associated with an 'increase in
�
_
t tnthe environment
These results imply that two types of stimulation may be
- .
_
produced by subtatic concentrations-of inhibitors. First, a
metabolic stimulation-ma/ be produced where the cells -do not increase
in number but thisimetabotc rate is�increased. Second, the number
. .
. of cells in the environment may e increased by the-compouncl. It
'-
remains to be see_Cy'l-whetlier. there I a stimulation-in-the individua
'
metabolic activity ofeach cell in this case. -.There is an increase
_
in activity, however,--ihis may only be due to the_�reater-ntimbir-of
,
_
have found that
- �
ro- Vut-y1 -acetate produce as 3 fciid increase in
duct ion oLpeniciltiifbii�ium chrysogenum.
PLAN OF ATTACK.
is well known -that the -rate of oxidation of hydrocarbons ma
be limited by the rate of_d-C.H. fi-Sion of the compound across the
- - _ � - �
� permeability barrrer It is also well known that -surface active agents
increase the permeability of--the cytoplasmic membrane. Several-years
ago
developed the idea that the use of surface active
compounds may increase the accessibility of hydrocarbon substrates
to microbial attack and thus' speed.up the oxidation of these compounds.
Due to several factors including a lack of funds, this idea has
�
never been tested to any extent. To date, we have studied 3 phenol id
compounds. Two of these compounds produced-stityulation and one did
not.
We propose_to_study the effect of substatic concentrations o
�
atty-amines t:Oh-031s, and 06 t-epill'y an nlurcornpourids on
. - �
7the oxidation of_two different synthetic coolantsi.andone petroleum:
type-prodUct..: may also use Other antimicrobi_alegents other
:than those -1is_te4==abOve.
-.
microorganisms belonging :te Genus
used in this investigation.i.- The cultures
ri samples Of used cutting fluids _obtained
MEDIA
Whenever possible, the organisms will be waratatned in cutting
�
luids In SoWinstanCeS cony ntitinar Midi-6-$uCh_aiJmitrient'brath,.
_
meat -infuiionrbicith or'en-inor-ganic-sal $ medium7may-be used
,
INCUBATION TEMPERATURE
_
All experiments wit] be done at 30�C The length of the
incubation perib0 Will vary according to the organiMend'the
particular experiment.
CHEMICALS
- -
The-best grade of chemical obtainable-will be used throughout
this investigatiOn-
GLASSWARE
All flasks, tubes and other-glasS-eOuipment will be actd cleaned
prior to each experiment.
HYDROGEN ION CORCENTRATION
_
The pH of the environment will-vary according to the _particular
experiment. In general, we will let-theFPh of the cutting Mid
'Le
determine the pH 'of the
INpcut.um
:Thercells-used_in_the experimentS will be grown on an-appropriate
_ .
medium. They will_be_harvested_after_a suitable incubation_period
_
nd wasfle('..1 thretmes1n_iso.ionia_sarine. The cells_w_1111 be
suspended in is�n!csa1ine--,
quantities undorm- lithissuspens ion
will be used 'tc:Linoctilate the tubes or. flasks
STERILITY'f
. We will employ suitable procedures for sterilizing the media
- -
Appropriate_controls Will_be included An each eXpertment. Lack
experiment will�be_done_inricate and repeated_at least once
CUTTING FLUIDS.
Three different cutting iLuids will be used in-thts-investigation
-.-
consisting of'two_chemicar coolants and one petroleum product.
.EXPERIMENTAL TECHMQUE
Each cuttinj-ftuid will beIprepared in a II =25-d1lution in tap
water., Ore huriclred_ml_othis_coolant will he placed in each 250 ml
Erlenmeyer flask. The number oflasks_used_ in each experiment will
vary from time to time. T_h_e_i_n_hibitor_will_be added to the series of
flasks in vary ing-conceramt tons. these f_Lasks_cli I lb�rLli�
autoclaving and then inoculated with the test orgartiSms. The flasks _
will be incubated for a period of time (probably two_weeks) in a
shaking incubator.
At periodic -Untervals of_about 2 days time the number of cells
per ml will be-determined for each flask. At the same-time the pH
of the flask will be_determined and other biochenlical tests_(such as
dehydrogenase' activity) may be conducted to determine the degree of
m.tAhnItn e'etti.:/itv_th_the_environment- At the _ehd-of the exoertment
'
the. miaining cuttIng_fluid will Iii�poure :into a_vOlumetrIc cyl inder
to determine- thequanti,ty of free oil in tfle- product (this test
will be used on the petroleum coolant only). The quantity of free
oil present is ars7_-Cel lent index of the degree of_attack_on_the
emulsifier.:
We will study as many compound's as -possible during the year
period of this grant. In no case will�thRibe less than twenty-
five: compounds
caST:OF�PROJEeT
-
PriAcipal Investigator $200.00/month (9 months)
Full time salary to work on project in
,
ummer_-
575imonth each
Publication Cost
_
Typing- report, papers, and -Ourchasihg-reprints , -
of publications that prise from this: work r
. .LC0 001
$. 19,836.00