(SANITIZED)UNCLASSIFIED PAPERS ON INFECTIOUS DISEASE RESEARCH(SANITIZED)

• Document Type: CREST
• Collection: General CIA Records
• Document Number (FOIA) /ESDN (CREST): CIA-RDP80T00246A003000320001-2
• Release Decision: RIPPUB
• Original Classification: C
• Document Page Count: 37
• Document Creation Date: December 21, 2016
• Document Release Date: December 9, 2008
• Sequence Number: 1
• Publication Date: February 20, 1958
• Content Type: REPORT

Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
Next 2 Page(s) In Document Denied
Q
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
FOLIA BIOLOGICA
Effect of Chlortetracycline on the Activity of a-amylase
M. BURGER, J. ROKOS and P. PROCH~ZKA
Institute of Biology of the Czechoslovak Academy of Science, Department of Microbiology, Praha
On fermentation of chlortetracycline by Actinomyces aureofaciens the fer-
mentation medium usually contains a source of starch (Van Dyck, De Sommer
1952) ~~hich splits up into lower derivatives. The extent of starch hydrolysis and the
possibility of using the sugars produced depend on the activity of amylolytic
enzymes.
It might be expected that with the accumulation of chlortetracycline in the fermen-
tation medium, particularly at higher concentrations of this antibiotic, the activity
of enzymes hydrolysing starch would be altered (Arora and Krishna Murti 1952).
Even though starchy raw materials (soya bean meal etc. ), contained in the fermen-
tation medium, probably have more functions than being the mere source of carbon,
the process must be conducted, for economic reasons, so as to make the best use of
the starch itself.
In order to study the effect of chlortetracycline on amylolytic enzymes, an enzyme
preparation was used from the mould Aspergillus oryzae: this is known, under the
experimental conditions in question, to hydrolyse starch exclusively by the action
of a-amylase (Burger and Beran 1956). Higher concentrations of chlortetracycline
were observed to have a strong inhibitory effect on this enzyme during the dextri-
nation of starch.
The purpose of the present work was to study the conditions under which inhibition
occurs and to try to find a method of preventing it, and, on the basis of the inform-
ation so obtained, to study the activity of amylase during the fermentation of
chlortetracycline by the strain of Actinomyces aureofaciens used commercially.
Analytical methods. The activity of a-amylase was determined using the author's modification of the
Wohlgemuth method (Belozersky and Proskuryakof 1951). ti'isual comparison of the starch iodine
complex with the standard solution was replaced by determination on a Pulfrich colorimeter. The red
filter S 66, through which the iodine starch complex showed the highest absorption, proved to be the
best from among the set of standard filters available. The time taken for the dextrination of starch,
required for the calculation of the Wohlgemuth amylolytic units, was determined by the extrapolation
of the final transmission values. The activity of a-amylase is expressed both in units (Belozersky and
Prokuryakof 1951) and in percentage transmission in relation to the standard solution.
Chemicals used. Soluble starch p. a. (Lachema), chlortetracycline with 50% biological activity
(Lederle Lab.) and the other chemicals analytically pure.
Method. The enzyme preparation of A. oryzae, which under the given experimental conditions
(Burger and Beran 1956), causes dextrinization of starch exclusively by the action of a-amylase, was
used for the experiments. A suspension of this preparation in physiological saline was filtered and the
filtrate was dialysed for 24 hours at room temperature against a 400 times larger volume of redistilled
water, changed three times. The amount of enzyme used was chosen so that dextrination was complete
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
in about 4i~ minutes. The final concentration of starch was 1%. If not stated otherwise, the reaction
of the mixture was maintained at pH :i.6 by 0.1 M acetate buffer. Incubation took place at 30? C in
a water bath. The concentrations of antibiotic given are in relation to the final amounts of biologicalh~
active chlortetracycline. The pH of the reaction solution was checked after each experiment by an
electron pH meter with a glass electrode.
The Effect of pH and Bu~ers on the Inhibition of Starch Dextrination
It was found that higher concentrations of chlortetracycline have an inhibiting
effect on the activity of a-amylase of the mould A. oryzae. The concentration of
chlortetracycline at which inhibition appears varies between 8 X 10-' M and
1.2 X 10-3M (fig. 1-3).
Inhibition depends on the kind of buffer used and on the pH of the solution.
In an acetate buffer at pH 6.0 (fig. 2), chlortetracycline causes inhibition at
concentrations higher than 1.2 X 10-3 M. In a phosphate buffer of the same pII,
chlortetracycline causes inhibition at a concentration as low as 8 X 10-' M.
In a phosphate buffer at pH 7.0 and with the same concentration of antibiotic,
inhibition is greater (fig. 1 and 3).
Fig. 2. Effect of Concentrations of Chlortetra-
cycline on Inhibition of A. oryzae a-amylase in
an Acetate Buffer of pH 6.0. x: time in rains.,
y: transmission in ?~o, pH of solution 6.0, buff-
ered by acetate at final concen-tration of 0.1 -I.
I: control and 400 erg., II: 600 fig., III: 800 erg.,
IV: 1000 frg. chlortetracycline in 1 ml.
Fig. 1. Effect of Concentrations of Chlortetra-
cycline on Inhibition of A. oryzae a-amylase in
a Phosphate Buffer of pH 6.0. x: time in rains.,
y: transmission in %. pH of solution 6.0, buff-
ered by phosphate at final concentration of
0.01 M. I: control, II: 400,r~g., III: 600?g., IV:
800 ?g. and 1000 ,rig. chlortetracycline in 1 ml.
The inhibiting effect of chlortetracycline on starch dextrination is present in
acetate buffer from pH 4.8 upwards (fig. 4).
The In/luence of Ca-~' on the Inhibitory Effect of Ch-lortetracycline
It is well-known that Ca `+ acts as a stabiliser for a-amylases. We have proved,
however, that under our experimental conditions, the inhibition of a-amylase is not
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
caused by the removal of Ca-~- from the solution by chlortetracycline. I~.ven a large
excess of Ca+ ~ (tab. 1) did not cause any substantial changes in the inhibiting
effect of chlortetracycline. The results and the fact that a dialysed enzyme solution
was used in all experiments, prove that the inhibitory effect of chlortetracycline is
not caused by the removal of a low molecular stabiliser.
o .~o
60 90
Fig. 3. Effect of Concentrations of Chlortetra-
cycline on Inhibition of A. oryzae a-amylase in
a Phosphate Buffer of pH 7.0. x: time in rains.,
y: transmission in %, pH of solution 7.0, buff-
ered by phosphate at final concentration of
0.01 M. I: control, II: 400 ~~g.,. III: 600 pg..
800 fig. and 1000'ig. chlortetracycline in 1 ml.
0 ~ 4,5~ 5,0 55 6,0 '
Fig. 4. Effect of pH on Inhibition of A. oryzae
n-amylase by Chlortetracycline. x: pH, y:
inhibition of A. oryzae a-amylase by chlortetra-
cycline. Buffered by acetate at final concentra-
tion of 0.1 M. Final concentration of chlortetra-
cycline 2 ~ 10-~3M.
The F,~ect of some Organic Acids on the Inhibitory Effect of Chlortetracycline
Previous experiments had already shown (fig. 1 and 2) that the inhibition of
a-amylase, caused by chlortetracycline, depends on the kind of anions present. If the
anions of some organic acids, e. g. of citric or oxalic acid in colicentrations of
2 x 10-2 M are present, inhibition disappears completely. The anions of phthalic,
benzoic, fumaric and tartaric acids partially reduced inhibition; lactic and propionic
Table 1. Effect of Cad ~ on Inhibition of a-amylase by Chlortetracycline buffered by acetate, pH 5.fi,
at final concentration 0.1 M
I
Experiment ~
Final Concentration
a-amylase
V o.
units
of Chlortetracycline
of CaC
1L
l
1.8 x
10-3 M 1.8 x LO-~
M I
0.84
2
1.8 x
10-a M ~ li x 10-3 M ~
0.74
3
1
8
10_3 M
~
~
~
.
x
i
-
0,4;;
4
-
L8 x 10_2 M '
3.30
+; ~
-
6 x 10-' M I
3.50
6
-
-
~
3..50
~I
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
Table 2. Effect of some Organic Acids on Inhibition of a-amylase by Chlortetracycline
buffered by acetate pH 5.6, final concentration 0.1 M. Final concentration of sodium salts of acids
2 X 10-2 M, of chlortetracycline 2 X 10_3 M
a-amylase units
Addition of
Sodium salt of added acid
-
Chlortetra-
Chlortetra-
0 I
Acid
I
cycline
cycline and acid
Oxalic
3.79
3.83
0.28
3.68
Citric
3.82
3.69
0.25
3.28
Phthalic.
3.83
3.83
0.26
2.71
Fumaric
3.83
3.83
0.28
1.54
Benzoic
i 3.83
3.83
0.26
1.47
Tartaric
3.79
3.67
0.28
1.44
Propionic.
3.83
3.75
0.23
0.27
Lactic
3.83
3.83
~
0.23
0.23
-
acids in acetate buffer had no effect whatsoever on inhibition caused by chlortetra-
cycline (tab. L). These findings provided a starting point for resolving other problems,
particularly as to whether inhibition of the activity of a-amylase by chlortetra-
cycline is reversible or irreversible.
The addition of citrate (acetate buffer) or acetate (phosphate buffer) to a reaction
mixture in which a-amylase has previously been inhibited by chlortetracycline,
removes the inhibition (fig. 5 and 6). The rate of starch dextrination after the
addition of these acids is the same as when chlortetracycline is not present. The
Fig. 5. Effect of Subsequent Addition of Citrate
on A. oryzae a-amylase Inhibited by Chlortetra-
cycline. x: time in mina., y: transmission in %.
pH of solution 6.0, buffered by acetate at final
concentration of 0.1 M. I: control, II: 1000 ,~~g.
chlortetracycline in 1 ml. in the 45th minute.
1 ml. of Na-citrate (final concentration 2 X
x 10-2 M), has been added. III: 1000 fig. chlor-
tetracycline in 1 ml., in the 45th minute, 1 ml.
of H2O has been added.
Fig. 6: Effect of Subsequent Addition of Acetate
on A. oryzae a-amylase Inhibited by Chlortetra-
cycline. x: time in mina., y :transmission in ?o,
pH of solution 6.0, buffered by phosphate at
final concentration of 0.01 M. I: control, II:
600'~g. chlortetracycline in 1 ml., in 45th min.
1 ml. Na?acetate (final concentration 2 X 10-1 M)
has been added, III: 600 Ng. chlortetracycline in
1 ml, in 45th min. 1 ml. of H2O has been added.
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
removal of inhibition is thus complete. The results given here prove that inhibition
of a-amylase by chlortetracycline is reversible.
In addition to studies on the bacteriostatic or bactericidal effect of antibiotics,
a number of papers have been published dealing with the question of the action of
antibiotics on separate enzyme systems both in vivo and in vitro (Agarwala et
al. 195'?, Arora et al. 1954, 1955x, b, Brody et al. 1954, Ghatak et al. 1953, Iyer et
al. 1953, Jacobson et al. 1954, Loomis 1950, Saz et al. 1954, Zimmerman et al. 1953).
It is a characteristic feature of these papers that far higher concentrations of antibio-
tics were used than those at which their bacteriostatic and bactericidal properties
are already displayed. This very fact would suggest that in these cases the effect of
antibiotics ou the enzyme system is non-specific and it is doubtful whether the
results obtained can be used to explain changes in enzymatic processes at concentra-
tions of the antibiotic which affect the multiplication of the microorganisms. ~~e also
used high concentrations of chlortetracycline when studying its effect on the activity
of a-amylase in starch dextrination. The concentrations used, however, are of
importance in the fermentation of Actinomyces aureofaciens, where the high con-
centration of the antibiotic can affect some enzyme systems.
Our experiments prove that higher concentrations of chlortetracycline inhibit
dextrination by a-amylase of A. oryzae. It is important that this inhibition can
easily be removed by some organic acids, which alone, however, do not affect the
activity of the enzyme in question. This provides the possibility of studying how
a-amylase of Actinomyces aureofaciens is affected during fermentation with the
production of chlortetracycline and, in case of inhibition, of measuring the a-amylase
content.
Van Meter and Oleson (1951) assume that chlortetracycline inhibits some reactions
in Kreb's cycle, since citric acid removed inhibition by chlortetracycline in studies
on the respiration of rat liver homogenates. The fact that citric and other acids
removed the inhibitory effect of chlortetracycline on the amylolytic enzyme system
in our experiments too, suggests that chlortetracycline may affect other systems
connected with liver respiration.
In some cases lower concentrations of antibiotics stimulate enzyme activity while
higher concentrations inhibit it (Smith et al. 1949). This may also apply for the
action of chlortetracycline on a-amylase. Arora et al. (1952) found that with lower
concentrations than those used in our experiments there is activation of the amylo-
lytic systems of malt and moulds. It must be stressed, however, that these authors
observed the activity of the amylolytic system from the point of view of increasing
the reduction rate, and did not exclude the effect of other enzymes, particularly
maltase.
.5'u~nma~?y
The inhibitory effect of chlortetracycline in concentartions of 8 x 10-' M and
higher on the dextrination activity of a-amylase of the mould Aspergillus oryzae was
demonstrated and the effect of pH, Ca- and some organic acids on this inhibition
was studied.
Inhibition of a-amylase by chlortetracycline can be removed by citrate, oxalate
and other anions of organic acids. The inhibition of a-amylase is reversible.
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
Agarvala, S. C., Krishna Murti, C. R., Shrivastava, D. L.: Studies
on Enzyme Inhibition in Relation to Drug Action. I. Effect of Certain Antibiotics on Ureas~~.
J. Sci. Industr. Res. 11B :165, 1952.
A r o r a, K. L., Ii r i s h n a M u r t i, C. R.: Studies on Enzyme Inhibition in Relation to
Drug Action, III. Action of CF~rtain Antibiotics on Amylases. J. Sci. Industr. Res. 11B :383,
195'l.
A r o r a, K. L., Krishna M u r t i, C. R.: Enzyme Inhibition Studies in Relation to
Drug Action. VI. Action of Certain Antibacterial Agents on the Succinic Oxidase System.
J. Sci. Industr. Res. 13A :482, 1954.
A r o r a, K. L., Krishna M u r t i, C. R.: Studies on Enzyme Inhibition in Relation to
Drug Action: VII. Action of Certain Antibacterial Agents on Tryptophanas~~. J. Sci. Industr.
Res. 14C : 6, 1955a.
A r o r a, K. L., Krishna M u r t i, C. R.: Enzyme Inhibition Studies in Relation to
Drug Action: VIII. Action of Certain Antibacterial Agents on the Tricarboxylic Acid Cycle
of Vibrio Cholerae. J. Sci. Industr. Res. 14C : 66, 1955b.
Brody, T. M., H u r s i t z, R., Bain, J. A.: Magnesium and the Effect of the Tetra-
cycline Antibiotics on Oxidative Processes in Mitochondria. Antibiot. Chemother. 4 : 86~i,
1954.
Burger, M., B e r a n, K.: O mechanismu ricinku maltasy plisne Aspergillus niger. I. Vliv
teploty na aktivaci hydrolysy skrobu plisirovymi enzymatrckymi preparaty. (:hem. listy
50 :133, 1956.
Van Dyck, P., D e Sommer, P.: Production and Extraction Methods of Aureomycin.
Antibiot. Chemother. 2 : 184, 1952.
G h a t a k, S., Krishna M u r t i, C. R.: Enzyme Inhibition Studies i n Relation to Drug
Action: IV. Action of Certain Antibiotics on Alkaline Phosphatase. J. Sci. Industr. Res. 1`l
1'lB :160, 1953.
1 y e r, S. N., A r o r a, K. L., Krishna 14I u r t i, C. R,: Studies on Enzyme Inhibi-
tion in Relation to Drug Action: V. Effect of Certain Antibiotics on Liver Arginase. J. Sci.
Industr. Res. 12B :536, 1953.
Jacobson, K. P? D e o d a t a d e A z e v e d o, M.: Action of Antibiotics on a Pepti-
dase and on Urease. Compt. Rend. Soc. Biol. 148:199, 1954.
Loomis, W. F.: On the Mechanism of Action of Aureomycin. Science 111 : ~i74, 1950,
Van Meter, J. C., O 1 e s o n, J. J.: Effect of Aureomycin on the Respiration of Normal
Rat Liver Homogenates. Science 113:273, 1951.
S a z, A. K., S 1 i e, R. B.: Reversal of Aureomycin Inhibition of Bacterial Cell-free Nitro
Reductase by Manganese. J. Biol. Chem. 210:407, 1954.
Smith, G. N., W o r r e 1, C. S., Swanson, A. L.: Inhibition of Bacterial Esterases
by Chloramphenicol (Chloromycetin). J. Bact. 58:803, 1949.
T i m m e r m a n n, H. J., H u m o l l e r, F. L.: Effect of Aureomycin on Choline Oxidas~~
and Other Enzyme SystE ms of Rat Liver. Am. J. Physiol. 175:468, 1953.
B e x o a e p c x x ifi, A. II., H p o c x y p >3 x o R, H. H.: HpaxTUVecxoe pyrcoso~icTlro no fN0-
xxntxn pacTexn~i. Mocxsa 1951.
BJII~IHHIIO XJIOpTOTpaLjI3I{JIIIHa Hc`I aI{TIIBHOCTb a-aMHJIa3bI
M. BYPI'EP, II. POIOC a H. HPOXA3I;A
]Iplr xblpa6oTxe xalup?rel~paurlhalulla c IIOMUII~bro 1uTamhsa :letirl~>Iiitice~ aureu-
taciens c~epMexTauKOaxax cpe~a coRep~ICIIT o6blxxo IlczouxHx xpasMa~a, I:u~~opblii
pacrnell~ReTCx B TeuexKe ~iepMe~ITauHU. 3a,xa~ie~I xacTOxnjeil pa6oThi (ibL7lrl vc?ra-
HOBYiTb B7IIIH0T JILI HaICOrl7I0HII0, a B 00060HHOCTIr 3II8i141TCJIbAble IZOH1LCN'r~)81jIIr1
YJIOpTOTpc`II~YIKJII4H8 B (~0pM0HTaIjYIOHHOI3 CpeT,(8 H8 jjeflT~JIhHOCTb ~H3I1MUI3, B613bIBa-
fOII[II% I'NJ~pO7II43 KpBXMadIa. 1~bI yCTaHOBII7III Ha7IVhll4e Cr[7bHOTU IIOj~aR.il}t~plT(CI'(r
tjONP.TBI4H XJIOpT0Tp8I~Hi{JIHHB H3 Ci-aM137I83y ASper~'LIIUS OPV'L8~ tl~rll ;ICKCTI)L1H7i[I[l'I
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
ItpaxMaala, - IIpN xoxl~ellTpaullua, >1;vletUnjux :3xa~elille il~s3 c~ej~NIel~Tai~nii a~~TII-
6uoT>2xa. IIosTOMy MbI e,iy=Ia~K ycnosKS~, npH xozopr~Ix aacTyliaeT no~yaB.nexlle.
K cnoco6, Kalc ero Ha6e>IiaTh, - u?ro6vl IIa Ix~,I3oBaxIIII IIOSI,y~e}uirlx ual~Hr~ir' upoc.~ie-
AKTb AeRTea~r xocTh a-aMr~~aatil Il~II o~paausaxRN x~opTeTpaullx~HHa r~h~~il:;rioit-
CTBeHHIxIM IiITaMMOM .~. aUP00~aCIeI1S.
~JISI OIIbITOBMhL1tOJIb30BaJIIICb 3H3~IMaTHi]eGKY1M IlPella~aTOM f~. l)Py%ae, ]:OTU~)blll
13 yCJIOBIIHY HaIIIeI'O ORbITa Q('-aXapYIBaeT ApaXMaJI I~CKJII0917TCJIbAO C IIU~fUIIl6H)
a-aMIIJIa3bI.
I~ICTYIBHOCTh a-aMIIJIa3h1 OIII)eJI,CJIHJ[acb H8MI1 ^O Me'IOAy ~~'011Ig'CIYIUth-a It HaIDeA1
BLIj[OM3M0H0HI1N; CpaBHORile KOMIIJIeHCa I30j(-I{IJaXMa~I c0 CTBHJja~TNbIM ~)BCTROhOM
npo~ssoRila~och c IIO:vIOIl~bro I.o,itopllMeTpa Pultrich-a.
MbI AoxaaanN IIa~H~IIe rlo~~as~flloiuero uei~cTS11x x~opTeTpal~IaxaIIIIIa xa ai.TE~iI-
ElocTb ocaxapNSaHhs~ n~~;i s~xxxHeM a-aMK~asbl n,aecex>~ A. oryzae IIpI9 r.~~Hl~eli-
Tpau~Il s 8.10-' M II sblme K Ilcc~e~tosa~H B.nHRrl~e pll , Cap ' II IieroTOphix
~~praxH=Ieclt~x xHC~oT xa a-r~> Ilo;~asnllxllltee ueKCTSC~e.
IloAas~ex~isr ueRTenbxocTK a-aMw~raai~ ~alopTeTpauax~~~roM Mo.IeHO Iis6e;raT~,
i II I3vICTpIIlzxxil 1J5~a). CoGpanxblil uo c~X Ilop onhlT no~~asa.rl, uTO n IIpIIpo~~e
c~13nbTpytotl~~ecs~ c~~opM>;I 6alcTepllii noaBnsiloTCII Tor~a, xor~a 6alcrepmll ;~o~Iro
~i;IIS~~T n oprax~s:~Ie II ~o.riro Ilaxo~x~rcx noA s~~Ilflxr~enl o=ieah c~o,xxUla II Tpy~Ilo
onpe~e~I~wihlY ~~c.;IOii~ni~ cpc,Ihl 3lc~rrioro oprallilaMa. I3 ycnoBlln~ onbiTa, r~anH1;IM
O~pBaOM IIpII BbIpdII~IIBc`IIIIIII B IICIiy'CCTBOHHbIS IIIITBTCZbHbIX CpBJ~IIX, OHM I303HLIIialOT
B pC3y'JIb2~T0 BJ[IIRHIIII~ ICOTOpbIO Ha3bIBaIOT }IO~OJIaTOIIpIIRTHbIMII. ~.IOJ~a OTHOCHTCII
JIOIICTBII~ aHTIIOIIOTIIICOB, COJIOYI JIIITII$, I1M\-IyIIHOI3 CbIIIOpOTICII, ~3KTepII0I~8P8,
b10XaHIdiICCKOPO pa3pV]II@~FIIIA, IICpCM~7IC8IOII~~I'OCft 3aM0(~a7iCVIBaItIIFI Ii pa3MOpa?IfI[-
BaIIIIH Ii~'JIbT~'pbI 1IJIII ~[JIIITCd'IbftOl'O IlbIpaII~HI38IlIiH R O~HOII II TOPI ?ICC IIIITaT0JI6H01I
Cj1C;IC, (O'[C~I7HIIII~CII CT8~0IOIIj130 II ~'"J1ZC pBCTId;~8IOf1~iIC^^;I bIIIIC~?0~6I, Ii T. IL J ~~)IL
O11CIIliO IICIiOTOpb75 (~)aIC1'OIJOB, KO'IOpbl(' TIpI13HaIOTCH pCIDalOll~IIMII J~dIH R03HIIliI10-
B(?HIIH (~iBIJIbTjJ~~lOII~IIZCH I~OpM ~aiiTCpIIII Iri ~'1tiP0, III CJI0,lV0'I 3a~bIBaTb, ~I'IO TfliCIIC
?fCt' yC70}III}I MOTyT C03Z~BA'CbCII II 13 OpI'tlHIi3MC, CIIO(iOGGTFIy'H TEUC)IC~ IIX BO:II(IIIC[]O-
Ii(;AIIHI ~~p~'CONAILC IIi\I3I~~IISITCT$~ ,I(?1iCTB1IC ~aIiT0pid0(~81'a~ aICI[TOL101'IIICOB II T. ll.~.
118IfOMI1II8II nCl BTOIlI, Dlbi HG O'POS]i;~~CTI3~7ACli~ ICOIIC9Il0, CpaBIII1T('JIb[IO yIIpOIIjOIlIIbIC
yC7I013IIH LIl VItPO C ~'C7IOBIIfIMIT, C ICOTOphI:bIII _VIFIIipO~bl CTBJIIiIIIIBIOTCfi B 1ICIiBOM
Opl'BTIII3M0 IiJIII B llpIlpOJjO BOO~IIj~.
Ilac~roslll~ax caoRxa ~rpe~ oT~eall,xhl~ ~py;~oB npe~cTaB~sIeT paspa6oTxy Toiz ~acTII
IIpOUJIC9IbI, B ICOTOpOII IIa OCHOBC llOJIyiI2HHIilX J1,0 CIIb IIOp J[BHHbIX IICC~CJ~OBaJILICb
BHEIIIIIIIC VC7IOBIIH, A00vY0,'jI11146IE ?~;IfI B03AIIIiHOB~HIIH (~IIIJIbTpyIOII~IIXCH (~)OpM
63ICTCpI1II, I1 BbIACI]AdIOCb~ Ii8IiI1C n10p(~)OJIOPIhI~CICII~ I33MEHCHNH IIpOHC%OJI,HT B HJIeTK0
6al:TepIIII rroR AeicTBtieM IIeIIIIrziIJIJIIIHa i2 ~py'r~~ c~IartTOpoB. B I;auecTSe Mo,~e~u
MhI i~s6pa~II nano=IIC~' 11~elir]~epa, Ilapa,Illla II callpoc~Il~ra zle~~olleha, - cx~ox-
IlocTb IcoTOpoii IC oupaao~aIIIIIO 1~ ~opM 6liI Ia ~;he xa~Il onllcalla zI c xol~opoii y xac
y~he YIIIe:aCII ~uIhIT alccllepllnzeiil~a~-~bllu-Ila~rorelleTll~e~cl~oi~ pa6uTrl (1~namiso>3II~ 1942).
B IlC~BpTO OTIEpC;[b ~[bI SO"IC~[IT II~OiI~~)L]Tb, 7I~)L~CTaI3JIgIpT JIII (~M,7IbTpyIOTI~IICCFI
(~)O~)UbT ~~IIiTE)pIIII U~fl3aTE:IbPy70 CTBJI,I110 NT OHTOI'CIi03a ~~{8JII3Ha 19j1~, ~ 9TOb1
C71~ 98C? 1IbI y[O;;Iii{IIbI C)bIJIII ObI II3 Rc`tIITII R XOjj0 pa3MH0?IiCIIIIR ~BICTCpIdII IIpII yCJIOBII$X,
I.o~ropliie nlhlc~M IIe npellszclnyloT paaI311TIIIO aTOro nporiecca.
Dlamepuarc u .rocemoaba
1, BbLiiOp 2CGriLa9L.,14,06, MrI rrpOH3BO~HJIII OIIbITbI C 3 IIIT3MMaM[I IIa~IOiIKA II~iC~c~)e 11, II30~IHpOBaII-
HbI6III B CJI~%~SaIIY BOCII?ITCH[IH OUOJIO~iC If POJIOBIIOI'0 M03Pa ~~ {(QTeIi; I~ 1I, II 2$ II I~ Z5, ~Ba I[3 HI1Y
13eeTII ceFa a sscn~,pllMexTC ~;ax b-TIIn. IIvTeM nacca~r;eH xa nlo.rroAhlx xpo.~xxax, riecoM a 480-880 r,
Blip yd'II'IITHOCTb lI TORCLIYHOCTb LIITaMMa II 23 IIOBbICN7IHCb HBCTOJIbKO, ~ITO 0,2 MJI Kp OBId 3dpa5KCH-
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
Horo NM xpo~INxa B Te=Iexxe 24 Yac, y6NBa~iN x;NBOTHOe secoM B 700 r HpN xapTllHe reMOpparN-
~ecxo# ceNTNxeetxN. I~poMe Toro MbI pa6oTa~IN c 6 NITaMMaMN, NaonNposaxxblMN Na J~blxaTeJlbxbix
IIyTe#.
2. Ilumamenbrcan cpe8a. I{ponte ~Ia6opaTOpxbix x{NBOTxbtx, B Te~Ie HoTOpbix Hano~xa II~Ie#I~epa
paaMHOx{a~acb o*Iexb cNnbHO, MbI NcnonbaoBanN B HavecTBe NNTaTenbxo# cpeubl aMHNOTN~ecxylo
H aJIJIaHTONCHyIO IIOJIOCTN 9-12-j~HOBHbIX KypNHbIX 3apOJ~biNIe#, II HOTOpbIX MbI HyJIbTNBNpOBaJIN
Ha~IO~Hy s 10 Hoc.neAosaTerlbxblx Naccaxcax.
3a ax{exxbie HcNAxocTN MbI c~NSbTposa~IN *Iepea (~NJIbTp Berkefeld-a L-3 N~IN Hce L-5. C~IeAy-
IoN~e# HNTare~Ibxo# cpeAo# 61~.n 6y~IboH Na ne=IeHN c HpN6aB~IexxeM 3,3% NnN 8,3% reMO~INaN-
OBaHHO# HpOJIN=ibe# HpOBN, e~TN KyJIbTypbI Mbi (~INJIbTpOBaJIH Ifepe3 l~1NJIbTp Se1tZ-a N NCCJIQJ~OBa7IN
~IN7IbTpaTbl Ha IIpNCyTCTBNe (~1NJIbTpyION~NXCH (~IOpM.
Pe3yJGbmambl ZLCCJGeC~08aflllll
1. Ilonbamxa norlyaenusl ~urLbmpywLLFuxcsl y6op.al ua urlg~exl~.uonxozo .,Ilamepua.~a
naeulux Kporluxoe.
Oprarlbl KporlHKa, xorx61xero xoc~Ie npxBxsxll xarloxKH II~Ieilc~epa, a xMexxo
xposb, ~IacTx ~IerKIIx, xexexx x ceneaexxx c 607IbxixM cop[epx{axxeM Mxxpo6oB,
pacTHpanllcb s cTyxxe c c~xaxo~IOrxuecKxM pacTBOpoM 1 :8. OTCTOIiBmxI3CH
3KCTpaKT (~i3JIbTpOBa7ICfr iiepe3 t~xJlbTp Seitz-a. ~HJIbTpaT ~bIJI 0603xaY0H TIxTepox A.
B3 3KCTpaKTa ~bIJIO xOJIy=I0H0 eIIje 2 (~I37IbTpaTa: xepBbIIi (B) - 3T0 hbIJIa LIaCTb
}I{xRKOCTx H8J( OCaj[KOM xOCJIe uexTpxc~yrxposaxllH xpH 1500 o6~Mxx B TegeHlte
15 Mxx. , BTOpo%i (C) - oc'raTOK xaAocaAoxxox x{xRxocTH, oTCTaxsasmeHCH B Te-
*Iexlle 24 *Iac. xpx 4? C. ~xcTpaKTbl B x C Taxx{e c~x~IbTposanHCb xepes I~x~IbTp
Seitz-a.
Bce 3 o6paaua c~xJIbTpaTOB 6bI7Ix CBOC)OJI,HbI OT 6aKTepIIH. ~II7IbTpaT A SbIJI
xOCeHH Ha xOBepXHOCTb 10% KpOBtIHOPO arapa. ~OCJIe TOPO KaK (~II7IbTpaT xpOCaiIx-
BaTICx B cpeuy, xa ee xOBepXHOCTb HaHOCxnHCb MIIKpOKOJIOHHH ?KeJITOPO CTa(~HJIO-
xoxxa. CpeRa aaKnellBaalacb R~Ix aaxpblTxH AocTyrla BosAyxa. 15-TiHeBHasl xxxy-
6auHH npx 37 ?(~ He npxxocx~Ia pocTa Konoxxil. Oxx xe HoHBnH~IIICb Rax{e TorAa,
xorRa rlosepxxocTb cpeRbl Boxpyr cTaI~H~IOxoxxa cocxa6~Ixsaaacb x xaxocx~Iacb
Ha csex{xH 10 % KpoBxxoil arap C KO7IOHHAMII cTac~xnoxoxxa.
Mo~IORoiI Kponxx, xoTOpoMy 0,15 MTI (~HJIbTpaTa A 6bIJI0 BBeRexo s Moar, a 0,9 M7I
BxyTpxsexxo, cTpaRa~I xoxocoM x xorl16 uepea 8 Rxel~. I~poMe sxTepHTa B ero
opraxax xe 6bino xaHAexo xxxaxllx xsMexexxl`I. IIa~IOVxa II~ieiic~epa xe Bbipoc~Ia.
B nerKxx 6bina xaiiRexa Escherichia coli.
TOT x{e (~I3TIbTpaT (A) 6bin xpxBIIT B aMHxOH xHTII 12-J~xeBHbIX KypHHbIX 3ap0-
J~bIIIIex HO 0,2 MJI B KaH{J~bIx. IIepBbix xaCCax{ OKa3a7ICFI CTepxJlbHbIM, OCTaJIbHble
6bIJIx 3arpA3H0HbI MIiKp06aMx aTMOC(~lepbI.
110 0,5 MJI (~IIJIbTpaTOB B K C 6bIJI0 xepeH000H0 B SKHJjKyIO CpeRy (~yJIbOH II3 He-
xexll c upH6aB~IexxeM 3,5% KpoBII). Rax{e xocne 13-RxesxoK xlrxy6aijr3x B x{HRKOK
cpeRe B xeil xe Ha6nxl~anocb xiixaxxx MHKpo6oB.
2. IZonbamxa norcy=Lenu.>z y6urlbmpy~o~.lFuxcsl y6op.n~ narlouxu lly6eu~epa nyme..u nac-
ca~ICeu rca ~cypunbcx aapoablluax.
1 MOHHHrea7lbHbIII IIITBMM xaJIO=II{H III~IeHC~epa x 2 IIITaMMa Iii JjbIXaTe7IbHbIX
xyTex ~bIJIx IIpxBIITbI B aTI7faHTOHC 9-12-JjHBBHbIX I{ypxHbIX 3apOJ~bIIIIOIi. BO BCOX
c~IyuaHx 6aI{TepHII clinbxo paapocnxcb.
I{ax{ublil xITaMM Hpoxle~t xe Mexee 10 TaKxx xaccax{eH. ?1?IiAKOCTb oT xoc~Ieuxero
xaccax{a (oxono 40 :1ial) rroc~Ie c~x~IbTpaIzIIH xepea ~ixnbTp Seitz-a aaceBanacb xa
xosepxxocTb 10% KpoBSlxoro arapa r3 B aMxlioTx=Iecxylo xo~IOCTb 11-RxesxblX
KypIIHbIx 3apOJ~bIxIex. L1epe3 3 JjHH xX aMHHOTIiLIeCKaII SKHj[KOCTb IIepeHOCx7IaCb
B aMHHOIr CBe1IfIIx 3apOj(biIIIex. T8K 6bI7I0 IIOC7IeJ(OB8T8JIbH0 IIpOII3BeJ~eHO 5-6 IIaC-
CaSKeII. HH B OJjHOM C7IyYa0 He y;[aJIOCb BbipaCTHTb xaJIOLIKy IIc~exc~epa.
~IlnbTpaT OT 10-I'O IIBCCaH{a =Iepe3 aJIJIaHTOIiCHyIO xO7IOCTb BBOJjH~ICFI B HOC
XOpbl{aM (If() 2 M.'I) IT MbiIDaM (IIO 4 I{axJII3). ~0pe3 3 JZHFI IIOCJIe IIpxBHBKH H{IIBOTHble
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
y6xBa~Il3cb H H3 MaTepga~Ia c~IIIaxcTOil Hoca, TpaxeII, a xacTblO g ~IerKHx IIpxroTOB-
BJIHTIaCb 3MyJIbCIIH C (~II3IIOJIOI'HHeCKIIM pacTBOpoM, KOTOpaH BIIpbICKIIBaJIaCb H0-
BbIM XOpbKBM HJIH MbIIIIBM B HOC B TaKI3X x{e KO7IH=IeCTBBX. 5 IIOCJIej[yIOIIjYIX IIBCCaH{eg
H8 IIpHBeJIO K IIOHBJIOHYIIO HH HOpMa7IbHbIX, HH L-(~OpM IIa7I0=IKII Hc~eHC~epa. y XOpb-
KoB B KoHue KoHI1oB axTIIBHposanacb Pasteurella.
I~Ia RpyrIIx IIITaMMOB IIano~KII H~iellc~epa, IIpoIIIeAIIInx IIaccaH{aMII xepea a~Iaax-
TOHC KypYIHOTO 3apOJjbIIIIa, (~IIJIbTpaTbI 6bITIII IIOTIy9eHbI C IIOMOII[bI0 (~HJIbTpa BOrke-
feld-a L-3 HJIII x{e L-5. OHII BIIpbICxIIBanIICb B aJIJraHTOHC KypIIHbIx aapoRblIIIeI3.
~IJIJIaHTOHCHBH 5I{HJ~KOCTb IIepeHOCIIJIaCb B a7IJIaHTOIICHble IIOJIOCTII CBe}I{HX KypFIHbIX
3apOj~bIIIIeII. HpOII3BOj[IIJIOCb OT 2 j~0 11 IIaccax{eI3. HII B OJjHOII aJInaHTOYIC?HOH
H{HJjKOCTII, 3apax{aBrueIICH 3TIIMH (~YIJIbTpaTaMII, IIaJIO~Ka II~ieHC~epa IIe paaMHOx{a-
HH B HOpMa7IbHOII, HH B L-f~OpMe.
3. IlonaamKa no.~y~cenun r~u.~bmpy>ou?uxcw g5op.-u nanouxu ll~eurfepa ua xcuBKOu
cpeaba.
2 KyJlbTypbl IIa~IOxKII Hc~eHC~epa B 6yJlboae IIS IIexeHII c IIpH6aBneHIIeM 3,3% reMO-
JIIIaHposaxxoil Kpo~III~beK KpoBII ~iHJrbTposaJlIICb -IIOCJIe 3-Axellxorl HHKy6arzHn
xepes (~HJIbTp Seitz-a. ~HJIbTpaT aacesaJlcH KaK B cse}I{yIO cpeRy Toro al;e cocTasa,
TaK Ha 10% KposaxoK arap co cTa~iIIJIOI{oKKOM. IIocJle 9-,~xesxoK IIHKy6auIIII o6e
cpeRbl ocTasaJlIICb cTepHJlbxblMH.
4. Nlopy6orcoZUS~ na.~ouxu Il~eug5epa, esapaufuaaeuceucsc ~ xypuuvlx aapol~alu~ax.
~opMa IIaJIO=IKH Hc~aHC~epa, paaMHOaKaBIIIeHCH B aJIJIaHTOHCxoH IIoJIOCTII KypII-
HOI'O 3apOJ~bIIIIa, II3y=Ia7IaCb HaMII C IfOMOH~bIO CBOTOBOPO YI 3JIeKTpOHHOI'O MIIKpO-
CKOIIOB. HaIIj[eHHble HaMH (~OpMbI HII=IeM H0 OT7fII=IaJIHCb OT (~OpM IIaJIO=IKII, BbIpaII~H-
BBBIIIeHCH Ha KpOBHHOM arape, O KOTOpbIX I'OBOpIIJIOCb B HaIIIeII IIpeAIIIeCTBOBaBIIIeH
pa6oTe (IiJIaIIIKOBII=I H I7bICTpHIjKIIYI 19556).
II.
lIOCJIe 6e3yCIleIIIHbIX IIOIIbITOK IIOJIy=IHTb [~]II7IbTpyIOIIZHeCH C~OpMbI HaJIO=IKYI
IIc~eHC~epa H3 (~IIIJIbTpaTOB CpeRbi, B KOTOpOH 6aI{TepHII pa3MII0iI{aJIYICb O=IeHb
XOpOIIIO, MbI IIbITaJIIICb BbI3BaTb 06pa30Bc`IHHe L-IIHK7Ia, OIIIIpaHCb Ha pa60TbI
Tulasne (1951), Dienes (194%), HaTO=IKH H Cyxaxosoi3 (1951), IfepMyTa H Hegaca
(1954, 19J~). UIIbITbI MbI ~jOBeJIII J~O TOH CTaj[IIH MOp(~OJIOPH=IeCI{Hx H3M0H0HHII, IIpH
KOTOpOH MbI y6eJjHJIHCb B B03HIIKHOBOHYIH T. H. L-I~YIKJIa y HaJIO=IKII llc~aHC~epa IIOJ1S
BJIHFIHIIOM pa3JIH=IHbIX (~aKTOpOB.
lllamepuarc u .nlemoava
1. Rr,16op ucma.~w.~Iloe. MbI pa6oTa~IN c 2 NITaMMaMH Haso*IKN ITc~eilc~epa Mexnxreanbxoro Hpo-
NCXO}KJ;eHNH N C 3 IIITaMMaMN, N30JINp OBaHHbIMN N3 J;bIXBTeJIbHbIX HyTe~. e~TO 6bIJIN Te }He HITaMMbI,
YTO N B HpeJ{bIJ(yH~eM OnblTe.
2, IIamamenb>~ast cpeaa, a) 17.2omrtan Cpeaa. Ilepe=IeHb HpNMeHHBIIINXCH B OHbITe RIIJ;OB HNTa-
TeTIbHO~I Cpej{bI HpNBOi~NTCH 6e3 00060~I CHeIjNI~NKaI~IIN,B paCYOTe Ha TO, =ITO OHN O~IHIelIBBOCTHbI.
e~TO T. H. IIIOKOJIa{'~HbI~I aPap, 060ran;eHHbI~ 1% proteose-pepton I~IICO ~" 3, 2O% CTCpNJIbHO~
HHaKTNBNpOBaHHO# JIOIIIaJ~xHO# CbIBOpOTHN, 1O% Apoxc}xesoro axcTpaKTa N 15% 6apaxbea xpoBN.
IlexNgN.a~tNx, BbIablBalolgNY~ L-uNx~I, MbI xaxocN.nN B H;e~IO6oK (2000 eA~MJI) x.nN Ha nosepxxocTb
arapa (150-375 ei[/M~I).
ITa~IOYKy II~IeNe~epa MbI BbIpaII~NBaJIx aapo6xo NJIN no.ayaxaapo6xo, c Ky.nbTypo~I S, mar-
cescens x,nN 6ea xee. TexxNKa NocesoB 6blBana BcerRa oAxa N Ta }Ke: MbI xaxocNJIN NccJleAyen2ylo
Ky~IbTypy NITpIIxaMN, napa.n~Ie~IbxbIMN c Hceso61soM.
b) ~ilaKa:G CpeBa, MbI HOJIbaOBaJINCb rOBH}KbNM CyJIbOHOM C HpN6aBJICHxeM 1O % Apo}Kx{eBOro
aI{CTpaKTa, 2O % JIOHIaRNHO~ CbIBOp OTKN N 2O % Kp OJIN~Ibe# Kp OBN, IC aTOI'I CpeAe MbI IlpNlrjaBJIHJIN
HOHNI~NJIJINH (I, 1O, 5O, NJIN ZOO eT'(/M.II), NJIN }KP, raMMa-PJI06ydIIiH, HOeTy=IeHHbIII N3 CbIBOpOTKN
KpO:INKOB, NMMyHN3NpOBaHHbIX Hano~xo~ IIc~e~c~epa.
e) T. x. RM-cpeaa HpeAcTaB.nHna cntecb n.noTxo~ N HcNAKOK cpe~bl B ripo6Npse (KOCO# Nloxo-
.IaAxbIl~ arap c }KHAKO~ ~Iaaol~, coT[epx{aBIHe~I 6ynbox c 10 io Apo}xxteBOro axcTpaxTa N 20 % ~IO-
IIIaJ1;NH0# CbIBOpOTI{N),
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
d) IIpexa aTr,I RnR caeTOSOro n~xxpocxoxa Bupeaam3cs xa anomxoa cpe~x no McTOAy Dienes
x Smith-a 1944) x oxpalxxsaaxcs xpacxTeneM Giemsa, passeAexablM xa 1 : 200, B Te4exxe
2 vacoH.
e) I'unepuuwyxxaR ct~eopom~:a u aa~w.wa-zao6ynutc izpxroTOB~IA~Ixca xyTeM xMMyxxaa~xx xpo-
nxH08 x KphiC xxaKTHBHpOBaxxO# HyJlbTypO# na7I0tilfx II~e#c~epa; rantMa-rso6ynxa xa rxxepxt~-
Myxxu% xpOTIHYbx% CbIBOpOTOx-IIyTOM IIpeIjxnxTaD;xx CEiBOpOTxx xpx-4? C c 25% cxxpTa
x pacTBOpexxR B c~xaxonorx*~ecxoM pacTBOpe.
Peay.~bmamsc ucc.~eaoeaxuu
1. 06paaoaarcue L-y6op.,w nar~ou~cu Ilg6eu~epa a aaeucu.atocmu om npo8o.~~cumercb-
rcocmu ~eucmeuR nercurlurercuHa.
IIpx Haxecexxx xexxuxxalr3Ha s axealo6ox B IIxTaTealbxoK cpeRe cneRyeT IIpH
OxYiCaHgx 06pa30B8HxH L-KOJIOHHx IIpxHgMaTb BO BHHMaHge (~aKTOp BpeMeHg,
YlrpalOIIjxI3 CBOIO pO7Ib IIpx IIpocaxxBaHxg axTx613oTxxa B IIgTaTealbxylo cpeRy.
Mbt gccaleAosaalx 5 1IITaMMOS xaalogxH Hc~e~I~Iepa. Bce oxx IIoR pteKCTSxeM xeHH-
uxalalHHa o6pasosaalx L-c~opMbl. LIacTggxblil axaapo6xoa ycxopseT o6paaosaHAe
xpyxHbIX KpyraIbIX Tealer~ (ISI{T), a T8M CaMbIM H L-KO7IOHxH (xCIIOnb30BaHHe
S. marcescens, aaxaleKKa ~alIIex IIeTpg).
CxcaTbli3 o6aop L-uHxala IIaalosxx IIc~eKC~epa Ha moxoalaRxoM arape c xeHxuxnalx-
HOBbIM xcealo6KOhI BblralslRxT Ha oTTi3cxax II MaSKax, oxpameHHbix IIo Giemsa,
caleuyloluxM o6paaoM:
B Tesext~e 2 *Iac. IIocale xocesa ? KyalbTypbl IIaalouxx Hc~el~c~epa Ha IIosepxxocTb
moxoalaRxoro arapa KoaloxHx xe o6paaosaalxcb. ~opMa Mxxpo60B xe McHSIalacb.
LIepea 4 oats xocale nocesa iIITpAxoBKOH IIoA alyxoi~ MecTaMx aahleTeH Ha 1aTpxxax
HaaleT MxKpo6os. Y >aTaMMa xa sepxHIIx AblxaTealbxblx IIyTer3 Mopc~oalorg~Iecxge
IIaMexexxsl MHxpo6os B cxoxnexxslx He xa6alloRalOTCtI, y Mexxxreaalbxoro mTaMMa
3aMezxbl BbIAaloluxecx xa Teals 6axzepxx oTpocTxg, yTOaIlueHi3x rI o6paaosaHxe
xpyralblx Tealeiz (Ta6. I, pxc. 1 x 2).
LIepea 6 xac. nocne xocesa s alyxy xo BceK IlosepxxocTg arapa aaNleTHbl Mealxge,
eAsa pasalx~xMble KoaloHxx. Hecxoalbxo RectlTxos xalx coTex Mgxpo6os s IIoale
apeHgx MAxpocxoxa o6paayloT Mxxpoxoaloxllx. MIIKpo6bl pacTyT s AaIr3Hy, oTalx-
xaloTCH xexpasxalbxblMx, yTOaluzexxblMx ~IacTHMx Teals x o6paayloT I~I{T. I~IHOrAa
BcTpe~aloTCSI oTRealbHble Tealbija. I{I~T 6bIBaIOT oRxopo~xble, cxxec~xoaeTOSOro
useTa (Ta6. I, pxc. 3 x Ta6. II, pxc. 4).
LIepes 8 =Iac. xocale xpHSxsxx x0 coce,~cTSy c xcealo6xoM c xexl3ur3alalxxoM xaxo-
RsITCSI MealKxe, eAsa paaalt~ut3Mble KoaloxxII, a xoRaalbiIIe oT IIeHxuIIalalr3xa oxx 6bIBaIOT
ropaaRo Kpynxee. Y mTaMMa xa sepxxi3x RblxaTealbHblx IIyTeK IfIfT o6paayloTCx
B6alxax xcealo6Ka s 607IbIIIOM Koxx~ecTSe Toalbxo Texepb, Tor~a xax y McHIIHreaalb-
Horo mTaMMa oHA yxce cc~opMxposaali~cb, x s xexoTOpblx xa xxx Ha*IxxaeTCx saxyo-
alxaanIISl.
LIepea 12 qac. xocale xocesa xa6alloRaloTCH xoaloxIIx IIepexoAxoro Txxa, a Taxxce
xOJIOHxx =IFICTbIX I{I{T B pa3alx=IHbIX CTaJ~I3fiX pa3BHTASI, - OT OJjHOpOJ~HbIX j[O BaKyO-
JIx3I3pOB8HHbIX g CerMOHTxpOBaHHbIX MHOPO=IxCJIBHHbIMI3 IIeperOpOJZK8M14. I{O7IOHII~i,
yAaaleHHble OT 5ICe7IO~ICa C xeHHuxaIaII3HOM, 6bIBaIOT Mexbme ~,1 MM (Ta6. I, pgc. 5
H Ta6. II, pxc. 6).
zIepea 24 Baca xocale xocesa xo6alxaocTH oT xexxuIIalalHxa xaxoRslTCSI optHx Toalbxo
L-xoaloxxH, cocTOSIII~xe xa L-Tealeu, - xo 6oalbiIIeK uacTx saxyoalr3ai3posaxxblx.
BpocaeTCH s ralaaa MopluHxl~cTax xosepxxocTb sTxx xoaloHt3ia s cpaBHexIIx c ralan-
xxMx cc~epr3=IecxxMH KoaloxIISlMx B~[aalexe oT xeHl3tzxalalxxa (Ta6. III, pt~c. 7).
zIepea 36 sac. nocale nocesa s6alxax xexxuxalalxxa xaxoRHTCSI, ralaBHbIM o6paaoM,
pacxastIIIIecsl L-KOnoxxA, pacxasmgecsl I?IfT x MHOxcecTSO MealxoaepHxcTOro Be-
u;eczsa Boxpyr Konoxt3~I. Hecxoalbxo RaalbiIIe oT xcealo6Ka, cpeug xopMaalbxblx
ralaAKxx xo~~IOHxH scTpeaaloTCx x Mealxi~e xoaloHHx c i3a6opoxcRexxoi~ nosepxxocTblo.
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
LIepe3 48 Sac. HocJle Hocesa pHRoM c x{eJIO6KOM nexHIIIIJIJIHIIa xacTynaeT pacrtau
L-KOTIOHI3H. FiOJIbmHHCTBO IiI~T pacuaRaeTCSi, B ApyrHX xa6JrlozjaloTCH McJlxxe
aepxxcTbie o6pa3osaxIlll HJIH KpoTKHe, HpasHnbxble KoxxoGauHJiJlHpxbie TeJlbua,
oxpamHSaIOH;HecH Kpacxoc~HOJIeTOSbIM. Ho xpoMe xxx soxpyr I~IiT H ocTaTKOs
L-xoJloxHil xaxoAHTCx Mxo~I{ecTSO 6ecc~opMexxoro aepxxcTOro BeluecTSa H xopomo
cr~iopMHposaxxbix HaJIO9eK H BoJroxox. MbI no:LaraeM, 9TO aTO MHKpo6bl, xoTOpble Boa-
xHKaIOT Ha xexoTOpbix pacnaAalolTjHxcH I~I~T B nHTaTeJlbxoil cpeRe, rye KoxueHTpauHIl
nexlluHJlJrHxa ynaJla xacTOJibI{o, =ITO yx{e xe BbI3bIBaeT o6pasosaxHx xoBbix I~IfT
H L-IjHKJIOB. B03HHKaiOIT~HC T8I{ MHKp06bI HC yCTOH9HBbI HO OTHOmexHIO I{ IICHH-
I[HJIJInHy' i3 B HpHCyTCTBHH jjOCTaT09HOr0 KOJIH9eCTBa 3TOT0 aHTH~HOTHKa HCMCjjJICHHI)
nepexoAslT B L-uHKnbl (Ta6. IV, pllc. 9).
BZjaJIII OT IieHHLlHJIJIHHa Ha6JIIOj[aIOTCH T. }I. I1CpeXOJjHble L-I{OJIOHHH (11aTO91{a
H Cyxaxosa 1951), npoxcxoH{Aexlie KoTOpbIX MbI o6T~ACxseM TaI{, 9TO y5I{e BoaxHKIIIHe
LIOpMaJIbHble KOJIOHHH IIOCTeHeHxO H3MCHfIIOTCII B L-I{OJIOHHH, Ha9HHaH C HaH60JICC
cTapblx MHKpo6os (Ta6. III, pxc. 8).
LICpC3 72 9aCa HOCJIe IIOCCBa Ha IIOBepXHOCTH CpeRbI HBXOj[HTCH 2 BH,~a KOJIOHHLI:
HOpMaJIbHble H L-ISOJIOHHH. I~OJIOHHH, BaHTax C JIIOCOPO MCCTa (BhJIH3H HJili B~aTIeKC
OT HCHHI~HJIJIHHa) H flepexeCexHaH Ha 10~~p KpOBHHOH aI'ap, j[aCT Ha9aJI0 9HCTOH
KyJIbTypC HOpMaJIbHbIX KOJIOHNH IIaJI09KH Hc~CHC~epa. BoalbmHHCTBO L-KOJIOHHH
HaXOJjHTCH B COCTOHHHH pacHaRa. BZjaJiH OT )KeJI06Ka paCHOJIO1KeHbI BIICpeMCSI{Ky
T. H. IIepeXOjjHbIC KOJIOHHH H L-KOJIOHHH.
Llepea 96 Sac. HocJle Hocesa xapTHlla TasaH x{e, KaK H 9epea 72 9aca. I~oJroxllH
HpeJjCTaBJIeHbI jtBy'MR THHaMH: L-KOJIOHHRMH H IICpeXOj[HI>IMH KOJIOHHHMLi. L-KOJIOIIHII
C COT006pa3HOII CTIJVKT~'pOH COCTOHT H3 OCTaTKOB L-TeJIeIj C MH05KCTBOM M07IK03epHH-
CTOI'O BOILjeCTBa LI C MCJIKHMH o6paaosaxliaMH B (~)OpMC naJI09eK fipKOKpaCHO-
c~HOJIeTOSOro L~BeTa (Giemsa). B MaaKax npLi yseJlH9eliHH B 1360 paa KpoMe I~I~T
HaXOJjHTCH MC7IKH0 MHKpOObI K BOJiOKHHCTbIC MHKpO~bI, a TaKSI{C ~OJIbBI08 KOJIN-
9ecTSO McJrKOaepxncTOro seH~ecTBa (Ta6. V, pxc. 10).
Llepea 120 H 144 9aca nocJle Hocesa KapTHxa KyJIbTypbl'oTJIH9aeTCH oT npeRLHe-
CTByIOLTjeH TOJIbKO TCM, 9T0, I{pOMC paCIIaJ~aI01I[HXCH COT006pa3HbIX OCTBTKOB L-KO-
JIOHHH B HCH HBXOZjHTCH H MOJIOjjble xOpMaJIbHbIC KOJIOHHn, COCTOHnjHe H3 KOpOTKHX
IIaJI09CK HJIH B07IOKHHCTbIX MHi{p060B. B CKOHJICHHHX (~HOJICTOBOrO BeLLjeCTBa
paCHBBIIIHXCH KOJIOHHH BbIJ~eJIHIOTCH MCJIKHC KpaCHO(~HOJICTOBble HaJI09KII, B03HH-
KaloruHe B peayarbTaTe T. x. xerloHxoro L-I~HI{Jra (Tulasne 1951).
:3TOT onblT, -KpoMe Ho;~TSep;ICuexHx Toro, 9TO HOJlyaxaapo6xall cpeua c S. mar-
CCSCeHS CTHMyJIHpyeT BJIHHHHe HCHHL~nJIHHa Ha B03HHI{HOBCHHO L-L~HICJIa, - IIpHHe('
eLije OjjHO OTI{pbITHe: y mTBMMOB naJI09KH Hc~eHC~epa, OUpa3y'eMbIX j(JIHHHbIMH
BOJIOKHaMH (MHOTOnJjepHble KJICTKH - Bisset 1951), L-IjHKJI IIOj( JjCYICTBHCM HCIIH-
I~HJIJIHHa H8CTy1IdCT paHbme, 9eM a I11TaMMOB B (~OpMC KOpOTI{HX IIaJI09eIf.
HpN J~OJITOBpeMCHHbIX Ha6JIIOJ1,eHHHX Hajj KyTIbTypaMH C llCHHL~HJI7IHHOM Mbl
y0ej[HTIHCb, 9T0 TaM, rj[C IIOCJIe IIOCCBa RCe MHKpOF)bI IIepeXOZ[HJIH B L-(~OIIMbI,
9epe3 4-6 JjHeH Ha9HHaJIH HOHB7IHTbCH xOpniaJibHble KOJIOHHH IIdJI09KH IIc~eHC~Cpa.
~.HeL~HaJIbHO IIOCTBBJICHHbIII OHbIT HOl{a3aJi, =ITO naJi09Ka Hc~enc~epa, HepeXOAII
B L-(~lOpMbI, CBOCO~IIa paaJIOiIiHTb 11CHIilIH7IJIHH B CBOCII OKpyrO, TaK =1T0 CIIOCOFxbIC
K pa3MHO1I{CHiIIO (~OpMbI HaJI091{H II~iCHC~epa pa3MHU?ICaIOTCH IIOTOM B 3THX MCCTaX
IIyTCM IIOIIepe9HOT0 Jje~'ICHHFI Ii ;~aloT Ha9aJI0 HOBbIM HOpMaJIbxbIM KOJIOHHHM.
2. 06pa3oeanue IfKT na~co~IKU II,gSeu~epa na. ucoKO.~aano.nti azape, na noeep.xnocml.
xomopozo 6u.~ naKanan nenu~~u.~n-un.
I~I~T 06pa3yIOTCH paBHOMepHO HO BCCH IIOBepXHOCTH CpCJ1,bI, itO~pbl3raHH011
neHHunJIJIHHOM. KyJlbTypbI HaJI09Kli H~ieLI~epa 6pa7ILiCb J~JIH HCCJIeJ~OBaHHH 9CpC3
2, 3, 4, 5, 6, 7, 8, 12, 24, 48 II 72 9aca HocJie Hocesa. LIepes 6 =Iac. 6oJrbHlnxcTBo
IIaJI09eK 06pa3VCT V?KC l{I~T, I'U1iOreHHOC COJIep51~nu
B 2-uacoBOx Ky~IbType MbI HaIIIZIH KneTKy B Haua~Ibxoli cTaRHH o6paaosaHHu
L-Te~Ibua - B c~opMe Ma~Iexbxoro ocTporo oTpocTKa B uexTpe K~IeTKH (Ta6. VI,
pnc. 1). IIpaBua, B ToK xce Ky~IbType BcTpeua~IHCb H 6o~Iee IIoaRHHe cTaAHII o6paao-
BaHHH IfIfT, TaK xaK B IIocexxaoH Ky~IbType HaxoAH~IHCb KaIeTKH paanll~HOro Bos-
pacTa, T. e. B paa~IHUHOI3 cTeneIIII pearnpylOluHe xa ReKCTBHe IIeBrIuH~I~IHBa. PHC. 2
Ha Ta6. VI npeReTaBalseT Hecxoabxo 6onee Kpynxblil Kpyr~Ibi1I oTpocTOK KaIeTKH
B 6-oNI Racy HIIxy6auHH, a pnc. 3 Ha Ta6. V I - yxce cpasxHTe~Ibxo xopomo paa-
BIITOe IfI~T, COej[IIHOHHOe C K7IeTKOH H3 2-~IaCOBOH KVJIbTypbI. C7IeJ~yIOIIZIIe CHHMKH
IIOKa3bIBaIOT B03HHKHOBOHHe I~I?T, COejjHHOHHOTO C MaTepHHCIfOII K7I0TKOII (6-uaco-
Basl Ky~IbTypa, Ta6. V I, pHC. 4 II Ta6. VII, pHC. 5, 6). B aTHx Tenbuax Moaxxo
pa37IHLIHTb McJIKyIO 3epHHCTOCTb. B 24-LIaCOBOH KyJIbTypO If ITT HBHO eIIxe CBH3aHbI
C OCTBTKaMH HJIeTOK (Ta6. VII, pHC. 7). OHH OT7IHTIaIOTCA McJIK03epHHCTOH CTpyK-
TypOH, HO He BCOrZja 6bIBaIOT OTLIeTJIHBO OrpaHHtleHbI, TiTO HaBOZ[HT Ha IIpeZ[HOJIO'
SKeHHe O~O HX pacnaAe, HO TOJIbKO CHOJIOTHiIOCKHI3 OIIbIT ObI MOP pemHTb BOIIpOC,
IIpHBOjjHT 7IH 3TOT paCIIaj[ K B03HIIKHOBeHHIO 5KH3H0CII000GHbIX McJIKHX Te7IeIj, HJIH
xce 3TII iIaCTHubI IIORBepraloTCH aBTO7IH3y H IIOrn6aIOT (Ta6. VII, pHC. 8). C~IeAyIOIuHH
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
Tab. VIII a. IIano~xa II~-e#c~epa, PacuaA xneTOx 6axTepx~ B aepxxcTOCTa. Pxc. 10-13:
ysenxYeHxe s 22.000paa, pxc. 14: yse~xeexHe B 15,000 paa.
cxxMOK 24-xacoso~i xy~bTypbt Taxxce Hao6paaxaeT ocTaTxx 6aKTepxH H pacxaAa-
ioBggecH If ITT (Ta6. VIII, pRC. 9). CpeRx HHx BxAxo MHOro Me~xHx ~acTgR, paaMe-
paMA oT 100 Ro 200 m,u. Ha6nioRaR onxcbrBaeMOe aRecb o6paaosaxAe I?I{T, MbI
B 4-gacosoK xyxbType BAAe~K cneRyioH~ee ABxeHHe: KneTxg 6aKTepHH - 6ea HaMexa
xa Ha~gxaioiReecH o6paaosaxge L-Texeu - xpespaH~anACb B rpoaRbR Mexxgx
3epHbIHIeK, IIpx6JII33YITEJIbHO OZ(YIHBKOBbIX pa3MepOB OKOJIO 1OO m~(l. HaM C~ygaxOCb
Ha6JIIOJ~aTb KJIeTKPi, coxpaHHBmxe Hepsoxa~a~bxyio c~opMy xaxo~Kx, YL oAxaxo
y Hgx xe xa6nioAanocb HHKaxoH xxeTOUxoH o6onou~fH: Bce xx coRepxcxMOe npespa-
Tgnocb B :~epxHCTOCTb (Ta6. VIIIa, pxc. 10,11, 12,13). Mbt Hoxa He MoxceM o6~,RCHHTb
3TOr0 FiBJIeHHf3. C IIOMOIIZbIO OxI3C8HHOYI xaMg MOTOJ~HKFI aTOMHbIX pexn$K Mbi
xpgrOTOBYITIH KOHTpO7IbHbIx HpeIIapaT, gCXOj[fI H3 24-iIaCOBOH KyJIbTypbl Ha arape
HOTOpaH He IIOj~BepPa7IaCb xgKaxHM BJIgRxxRM A KOTOpyIO MbI OiIHCTFIJII3 nyTeM
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
ueHTpIII~yrHposaxIItl B AecTrI~I~IITposaHxoK BoAe. Ma30K o~HUjexxoiT BasecH xa IIpeu-'
MOTHOM CTOKJIbIIIIKO IIOKpbIBaJICIi XpOMOM H CHIIMaJICH IIO7IHCTpeHOM. CHI3MKI3
aTOMHbIX peII7IHK TaHrIX Ma3KOB HCHO IIOKa3bIBaIOT BIIOJIHO COXpaHI3BIIIyIOCH ~aKTe-
pITa~Ibxylo o6o~IO*Ixy (Ta6. VIIIa pITC.14). Ha ocxosaHHII aTOro MbI cyAxM, nTO Ha6nlo-
AaBmaHCH HaMH 3epHYICTOCTb - He apTel~axT, B03HIIKHIHH B CBH3H C MOTOJ(OM
npenapoBKi3. 3auaueK RanbxeHlHHx nccneRosaxHK 6yAeT IIposepIITb, tIB~IHeTCH IIII
Ha6nIORaBiIIaxcH HaMH aepxxcTOCTb apTec~aK'roM, HJIH ~Ke aTO pacnaR IIoA AeHCTBxeM
IIOHHIjI1JIJIITHa YIJIH j[pyrOl3 B03MONCHOII IIpHLIHHbI.
(Ta6. VI, VII, VIII)
~uc~yccu w
yIay~IHS alxTepa'rypy o 61TO~IOrH~ecxoM axa~IexrlII c~r3nbTpylOluHxcx c~opM 6aKTepIIii
(B~IaIIIKOBITLI I3 Bblc'rplTuxIIT~I 1955a), MbI IIpTlcTynrI~III K IIepsoiT c~aae onblTOB c Hanon-
ISOK IIc~eHC~epa ArI>i onpeRe~IeHIIH ycnoBllil, npx Ko'ropblx BoaHnxaloT ~HIIbTpy-
IoluIIecii c~opMbT. IlpeNCRe scero Heo6xoAHMO ~bIJIO npoBepHTb, HBJIHIOTCH nII c~i3~Ib-
TpyIOIuITecH c~opMbl o6HaaTenbxoil c'rauHeiT oHTOrexeaa 6aKTepIIH (KaJIHHa 1951).
II03TOMy MbI CTpeMHJIIdCb II07IyLIIITb (~rI7lbTpyIOUZITeCH (~)OpMbI B yCJIOBIIHX, IfOTOpbIO
He HpeIIHTCTByIOT pa3MH05KOHHIO rOaKTepI3H, - IfaH H3 JIa(r)OpaTOpHbIX SKHBOTHbIX,
IIOI'II6IIIIIX OT OCTpOII COIITITKOMHLIeCKOII HHI~eKIjHII, TaK H II3 NSH~KOII CpOJ~bI HJIH K3
CpORbI pa3BIIBaIOIIZerOCH KypIIHOI'O 3apOj[bIIIIa (IIOJIOCTb aJIJIaHTOHC).
B 3TIIX yC7IOBHHX H8M He yjjaBaTIOCb IIO7IyiIIITb H3 (~HJIbTpaTOB NSYI3HOCIT0006HbI0
I~I3JIbTpyI0II~I3eCFI LIaCTIII[bI, XOTH MbI TOYHO CJIOJ~OBaJITI MOTOj[IIKe, OIIHCaHHOI3 CiyK-
HOBbIM II BOnbc~epu (1932). Mbl 3HaeM, iITO CII0006Hble K pa3MHONSOHIIIO (~lI3JIbTpy-
IOIIjIIOCH iIaCTHubI IIOJIy~IaIOTCH p0~[KO (KpaCIIJIbHHISOB 1954) II 3aBHCHT OT KOJIIILIOCTBa
HaXOjjHIuIIXCH B 3aCOBaOMOM MBTepHa7ie iIaCTI3I[ (KlleHebOP~'OP-NOb01 1951), HO,
B COOTBOTCTBIIH C IIIIIpOKO IIOCTaB7IOHHbIMH OIIbITBMH TpHKH (1955), MbI npirAepaKH-
BaeMCH TOI'O B3rdIHRa, iITO B03HIIKHOBOHIIO I~ITJIbTpyIOLuIIXCH (~OpM He MONSCT 6bITb
pOryJIHpHOII CTaJ~IIOIT B OHTOreHe3e ~aISTepIIII. II03TOMy MbI 06paTIITIHCb K OIIbITaM
II07Iy~IOHIIH L-I~IOpM IIOJ[ J~OIICTBIIOM pa3JIH~IHbIX BeII[OCTB, KOTOpbIO BCOMT[ OTHOCATCH
K f~aKTOpaM, BJIIIHIOIl[HM He6JIarOIIpHHTHO Ha IIpOIjeCCbI NSH3Hej[efiT07IbHOCTTI 6aK-
TOpUIr, BIS7fIO~IaH H pa3MHONCOHHe, - TBIS KaK y L-I~OpM ~aKTOpHIT (~HJIbTpyIOII~aHCH
CTaZ[IIII II3BOCTHa.
MbI Hcc~IeAosaalx BnHHHIIe Ha narlonKy IIc~eiic~epa neHIIuIIn~IHHa H lu)~Iy~Ii1nN
p03yJIbTBTbI, OTBOLIaI0IIjH0 TOM, KOTOpbIO TIy~JIIIISOBaJIIICb Zj0 CHX IIOp. OZ[H8K0 MbI
y~OJ[IiJIHCb, iITO J[OIICTBHe IIOHHI[HJIJIIIHa, IIpOCaLIHBaI0Iu0I'OCH CKB03b IIJIOTHyIO
CpOZ[y', pa3JIIILIHO B 3aBIICHMOCTII OT TOI'O, B KaKOII (~a30 pa3BHTIIfi HaXO~[11'1'CH CO-
061I{OCTBO MHKpOCOB. ~~ O~IOHb OOJIbIIIOH BepOHTHOCTbIO MbI C IIOMOIIjbIO MOTOJJ,a
OICpaIIIOHHbIX OTTIICKOBbIX IIpOIIapaTOB ISyJIbTyp IIaJIOLIKU 11(~leH(~Opa pa3JIIILIHOPO
Boapac'ra uoxaaa~IH Ha~IIIUHe T. H. HenonHOro L-ulTKna (Tulasne 1951). A H B-KO-
JIOHIIU L-I~OpM MbI paCCMaTpIIBaeM Ka1S p03yTibTBT pa3JIII~IHOI3 JjJIiiTCJIbHOCTIi JjeII-
CTBIIH BHOIIIHOI'O (~)aKTOpa (HaIIp., IIOHIIuIIJIJIITHa). B03MONCHO, tITO H B llpkipOZ[O
IIOZ[ J~OIICTBIIOM HOBIIOJIHO BbIHCHOHHbIX IIpHLIIIH IIpOTOICaIOT aHaJIOTII~IHble IIpOI~OCCbI.
~TIH II07IyTIOHHH L-c~opM IlaJIO~IKH IIc~eIIC~epa MbI HCIIOJIb30BaJIH TaKNCe CTapbIO
Ky7lbTypbl, Ha ~a30 KOTOpbIX B03HUKaIOT BTOpHtIHbIO L-KOJIOHIIH, a T8K)iSe j[eIICTBile
CIIOuH(~II~IOCKOI'O I'aMMa-rJIOCyJIHHa B 1ISH~KTIX H CMOIIIaHHbIX IIHTaT07IbHbIX CpOJjaX
(RM-cpeua).
B KatIOCTBO HOO~XO~[HMOPO J~OIIOJIHOHIIH K H80JIIOJ[eHIIHM B CBOTOBOM MHKpOCKOlIO
MI,I IICCJIO;(OBaJIH B03HIIKHOBOHHe H CyJ~bObI L-c~opM, a B 0006eHHOCTH I~HT, K C IIO-
MOIubIO 3JIeKTpOHHOPO MIIKpOCKOIIa, J~JIH tIOI'O MbI pa3pa60TaJIH MOTOJ~ aTOMHbIX
OTTIICKOR, COXpaHHIOII[UII ~aKTOpHaJIbHbIO K7IeTKH III S1tU. MbI II07IytIII7III Ilpej(CTaB-
TIeHHO O B03HHISHOBOHITII KKT, JjJIH ICOTOpOI'O HOT H006XOj[IIMOCTrI B ZjByX 6aKTe-
pHaJIbHbIX K7IOTKaX, I[ O COCT8B0 TOJIa ~aKTepI1H II 1~KT IIOJ~ HOIIpepbIBHbIM j1,CIICTBHCM
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
IIOHIII[gJIJIIIHa. IIpg 3TOM MbI HaIII7Ig B Te7Ie 6aHTepgg H B I{KT HpyIIgHHg J(gaMOTpOM
OHOTIO 1~0 In'GG, 3Ha~IeHge HOTOpbIX H8M IIOxa Heg3B0CTH0.
BbIIIIeH37IOHfeHble IICCJIeJ~OBaHYIFi J~OJIHfHbI IIOC7IylxgTb OCHOBaHgeM J[7IH pa6oT IIO
BOIIpOCaM CyIIjeCTBOBaHgFi H 3Ha~IeHgH [~gJIbTpyIOH;gxCH (~OpM IIaJIOLIHg rl(~ei3(~lepa.
Pe3w.~we
I~Ia ngTepaTypbl IIpeAMeTa MbI cTapangcb cocTaBHTb ce6e IIpeRcTaBneaxe o 6gonorg-
'4eCKOM 3HaLIeHIIII (~1g7IbTpyIOII~IIXCfi (~OpM 6aKTepgg. Ha MOj[e7IP[ OIIbITa C IIa7IO~IKOg
Ilc~ie~IC~epa MbI IIbITa7IIICb IIOnyuIITb I~II7IbTpyloII;geCSI ee c~lOpMbI g7lg OIIpeue~IgTb
yCJIOBI3H, IIpII xOTOpbIX OHg B03HgKaIOT. MbI y4gTbIBaJIg pe3ynbTaTbI ~gOJIUrg-
ileCxHx OIIbITOB g IIpeIIapaTbI JZ7IfI CBOTOBOI'O PI 37IeHTpOHHOPO MgxpOCHOIIOB.
B IIepBylo OLIepeAb MbI XOTenA IIpOBepgTb, HB7ISIIOTCH JtII (~II7IbTpyroIIxIIeCR (~OpMbI
06H3aTeJIbHOg CTaJ~IIeI3 OHTOreHe3a 6axTepgi3. MbI IIbITa7II3Cb II03TOMy IIO7IyLIETb
c~IIJIbTpylongIIeCx I~OpMbI IIaJIOYHII Ilc~ieg~Iepa g3 Hy7lbTyp McHI3HreaJIbHbI% IIITaM-
MOB (3) 13 IIITaMMOB g3 j~bIXaTeJIbHbIX IIyTe~I (6). MbI (~Ii7IbTpOBaJII3 xepe3 (~g7IbTp
'Seitz-a gnII Berkefeld-a L-3 g L-5 c~IeRyIOIr~IIK MaTepga~I:
a) co~Ieso~I axcTpaxT opraxoB IIorH6IIrIIx xpongxoB, HoTOpbIM 6bI7I IIpgBHT McHgH-
reanbHblil IIITaMM IIa7IO~IxIi Ilc~eilc~epa,
6) xynbTypbl IItTaMMOB pas~Ir~~HOro IIpogcxoxtRexIISI, BblpaII;eHHble s xct3RHZ3x
nIITaTe~Ibxblx cpeRax II IIpoIIreRIItIIe He Mexee, xeM 10 IIaccaxcaMg B IIonocTg aJI7IaH-
Tonc xypIIHOro aapoRblIIla.
~IITIbTpaTbI 3aCOBa7fIICb IIa COOTBOTCTBeHHyIO IIJIOTHyIO IIIIT3TOJIbHyIO cpeAy
(10% xpoBxHOi~ arap c HonoHIISIMII xcenTOro cTac~g~IOxoxxa), Ha xcIIRxylo cpeAy
{6y7IbOH s3 IIeueax C IIpm6aBneHgeM 3,5% HpOBII) II IIpgBgBaJII3Cb BOCIIpI3I3M~IIIBbIM
9xIIBOTHbIM (xpO7Igx, MbIIIIb, XOpeK) I3 xypIIHbIM 3M6pIIOHaM.
CyII~eCTBOBaHxe (~HJIbTpyIOII~IIXCIi (~OpM, HoTOpbre B03BpaTg7lgCb 6bI H gCROJ~HOK
c~opMe 6axTepllH, xe 6bIJI0 HaMg Roxaaaao. He ~ObI7III xai~ReHbI YI c~YI7IbTpy~II~Aecsl
(~OpMbI, HaTIFI~IIIe xOTOpbIX MOHfHO 6bIJI0 6bi OTMOTIITb C IIOMOIIjbIO MOTOTja ?KOpMI3JIKg?.
MbI He RyMaeM, iITO (~YIJIbTpyIOII~I3eCSi c~iOpMbI AB7IHIOTCFI CTaRIIeII OHTOreHe3a 6aH-
TepIIH, paaMHOxcaIOll~xxcx B 6naroupl~xTHblx ycnoBIISIx.
e~TII oTpII>~aze~IbHble peaynbTazbI HanpaBgnII HaIIIe BHIIMaHge Ha BoaHgxHOSeHxe
L-i;IIHnoB IIoR ReHCTBgeM paaar3~Hblx BHeniHIIx yc~IOBZ3i~.
1. MbI oIIIICa~III ~eI~cTBIIe IIeHIII~IIJIJIgIIa B II~IOTxoK IIIITaTe7IbHOi~ cpez[e Ha 5 IIITaM-
MoB IIa~IOUHII II~iegc~epa, 2 IIa xoTOpblx 6bI7IIi gao~Il3posaxbi B cny~atlx BocnaneHI3FI
MoaroBblx o6o~IOYex pe6eHHa. Bce IIITaMMbI o6paaosa~III I{I{T, a y 2 IIa Hgx IIyzeM
cl3cTeMaTIIxecxoro Ha6nIOReHIISI xy~IbTyp B 2-xacoBblx IIIITepsa~Iax c GOnbIIrOi~
BepoxTIIOCTbIO 6bI7I0 Roxaaaao IIaJIIILIIIe T. H. IIeuo7lHOro L->;gHna (Tulasne 1951).
TaH xax IIeHgr~gnngH, IIpg6aB~ISIeMbIg x cpeAe B xceno6ox i3IIg B oTBepcTHe, IIpoca~g-
BaeTCH B IIIITaTeJlbayIO cpeAy IIocTeIIesao, off oxaabIBaeT B7IgsIHge Ha paangxIIble
cTaAIIII o6paaosaHxll xonoHgK, xoTOpble, IIpogAs gepea cTauIIlo IIepexoRHblx L-xo-
7IOHgI3, xepe3 HOHOTOp08 BpeMS IIpII06peTaIOT c~OpMy TgIIgiIHbIX L-HOnOHgi3.
2. rlepBOHaLIaJIbHO HOpMaJIbHble HOJIOHgg IIa7IO~IHg II~iei~~iepa, g3MeHgBIIIPIecII
IIoR Aei~cTBgeM IIeHg>~IIII~IgHa B L-HOnoHgg, 6go~IOrII*Iecxg pasaoz;eIIHbI c ycTOguII-
BbIMg gTIII ~IaCTgLIHO CTa6g7Ig3gpOBaHHbiMII A II B-TIIIIaME L-c~-OpM.
3. IIpII L-I~gHJIe IIOJ[ BJIIIFIHgeM IIeHgL~g7I7IgHa IIaJIOLIHa Ilc~et3c~epa pa3JIaraeT 3TOT
aHTHCgOTgH H T0M CaMbIM IIpeHpaII~aeT er0 aHTII6IIOTELIeCKOe J[eIICTBI3e.
4. L-~iopMbI IIanOLIHII Ilc~ieII(~epa 6bITI$ HaMg IIO7Iy~IeHbI B CTapblx HyJIbTypax Ha
IIroxonaAHOM arape, Ha*IIIHasI c 9-ro RHSI gHxy6ai;gII. J~anee L-c~opMbl Boaxgxang
IIoR B~IxtIHgeM cne>~g~ixgecxoro roMOnorHOro raMMa-rno6y~IgHa B RM-cpeAe (cMecb
IInoTHOg T3 xcgAxol~ cpeRbl). ?l{gp[xax cpeRa yAo6Ha, xax xa}xeTCtl, Ans IIony*IeIIIISI
cTa6g~IxaosaHaol~ L-c~opMbl, Tax xax BbIablBalolgAe ee o6pasosaIIge EMIIynbcbl
J[eI3CTByIOT B TaHOI3 CpeAe J;07IbIIIe.
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
I~Ia nno~iIIslr3aHposaHHbiX Icy~IbTyp IIa~IOUKH IIc~ei~c~epa TaKxce yRanocb B TeueHIIe
3 rexepagHii IIo~IyuaTb L-c~opMbl.
OIIIICaxxbie onblTbl Ronxcxbl cnoco6cTBOSaTb BbIFicHeHHIO axaxellnx L-IzIIK~ta y 6aK-
TepHH H pemeHHlo Boupoca, KorRa B Te~exne sTOro i~IIKna BoaHHKaIOT xcHSxecnoco6Hbie
(~HJIbTpyIOHZHECH (~OpMbI ~aKTepHH. MbI XOTHM TaK C IIOMOH~bIO OHbITa IIp07IHTb CBOT
Ha IIpOI(eCCbI, IIpOTeKaIOII~IIe B OpraHII3MC X03FIIIHa.
1ITOCbI HpOBepIITb JjaHHble CBBTOBOPO MIIKpOCKOIIa IIpII yBe7IYI=IeHHH, II03BOJIIIIOIIZeM
pa3JIH=IaTb 37IeMUHTaplible =IaCTHI~bI L-(~OpM ~aKTepIIIi, MbI IIpOC7IeZ[II7II3 06pa30BaHIIe
HILT IIOj~ B7IHIIHIIUM IIBHHI~HJIJIHHa y IIa7I0=IKII II~)eIIC~epa YI B 3JIKTpOHHOI3 MIIKpO-
cxoIIHH. F'ibIJIII IIonyuexbl cneAyloH~IIe pesy~TaTbI:
MbI IIa6nloAa~III o6paaosaIIIIe HILT IIa~IOxKH II~ie~ic~iepa IIoR uei~cTBHeM IIeHHu1In-
~IIIxa. YIa yRrlIIHexxoi3 IIa~IO=IKYI BbipacTaeT oTpocTOK, KoTOpbIK ylienHUHBaeTCx
B o6'beMe II saxpyraIIeTCx. uIIaMeTp HILT 6bisaeT oxo~IO 4-5 MHKpoxa. B TexeHHe
2~f =IaC. IIC=Ie3aIOT KOHTypbI IIIIT, IfOTOpOe paCIIaj[aeTCH B 3epHHCTOCTb, 3aM0THyI0
yaKe H j[O 3TOT0 B COJjep?KIIMOM IiI~T, HJIH B 6eC(~OpMeIIHyIO MaCCy.
KpOMe O[ pa30B8HIIH KKT MbI Ha~JIIOJ~a7IH B Cpejje C IIOHHIjHJIJIIIHOM CKOII7IeHYIA
COj[ep}il'HMOPO KdIeTIfH 6aICTepHH B Oj[HOpOj[HyIO 3epHIICTUCTb, yTpaTy KJIeTO=IHOI3
o6onouKH II pacceslxIIe KpyIIIIxoK B oxpyxcaIOHlylo cpeRy BHe KJIeTKH. IIpH IIpez[-
IIIUCTBOBaBIIII2X Ha~JIIOJ[eHIIFIX C IIOMOII~bIO 3JIeKTpOHHOTO MYIKpOCKOIIa MbI TaKOPO
pacnaAa He Ha6~lioAa~1II.
~InTepaTypa
B n a m x o B n ~, J~. H Fi bI e T p n u x n i'r, B.: Mopc~o~rornH narlo~Ixu Hc~ehc~epa (Haemo-
philus influenzae Pfeiffer) B a~IexTpoxxoh Mnxpocxonnn, Fol, biol. (Praha) 1:355, 19556,
I{ a JI n H a, I', H,: ~nJIbTpyIOIi[neCx ~OpMbI 6axTepn#, TpyRbl nHCTHTyTa MHHp0~1nOJI0PItn
1:45, MOCHBa 1951.
Il p a C H JI b H n x 0 B, H, A,: O HexJIeTO=IrIbIX (~OpMaX y MHKpOOpPaHnaMOB, YCn. COBp,
bxo.aornn 37:22, 1954.
H e p M y T, D1. rI H e u a c, O.: L-c~opnibl 6axTepHti. I. I~Ianiexexnx c~opMbl Proteus vulgaris,
BbIablBaeMble nexnuHnnxxoM. iIcn. TxorlornH 3:370, 1954.
H e p M y T, M, H H e~ a c, O.: L-I~opMbl 6ax~reprr#. II. B~IHHxne axaapo6xblx yc~IOBH#
xa HanlexexnH I~opMbl Proteus vulgaris rtou T{e>;ICTBneM Hexxurrrl~rnxa. Fol. biol. (Praha) 1:257,
1955.
C y x x e B, B. B, n B o x b~ e p u> P. A,: BI;IHSSexne ~innbmpyroH;HxcH c~opM 6axTepn#
C nOMOIL{bI0 ?xOpMHJIOx? H3 6aKTepnO(~)arOBbIX (~nJlbTpaTOB n Cyn[HOCTb (~IeHOMOHa f)aKTepnO-
~IarHn. BecTxHx Mnxpo6no~IOrnH, anHA, n napaanT., 11, 1932.
Bisset, K. A.: The Morphology and Cytology of Bacteria. Ann. Rev. Microbiol. 5 : 1, 1951.
B l as k o v i ~, D, a B y s t r i c k y, V.: Biologick$~ vyznam filtrovatelnych foriem bak-
terii, Cs. biologic 4:496, 1955a
B I as k o v i ~, D.: >.7ber experimentelle Moglichkeiten~ mit dem b-Typus H. influenzae
Pfeiffer. Zbl. Bakter. I. Orig. 149:202, 1942.
Dienes, L.: Electron Micrographs Made from L-forms of Proteus and Two Human Strains
of Pleuro neumonialike Organisms. J. Batt. 66:280, 1953.
Dienes, ~., Smith, W. E.: The Significance of Pleomorphism in the Strains of Bactero-
ides. J. Batt. 48:25, 1944.
K a n d l e r, G., K a n d l e r, O.: Untersuchungen fiber die Morphologic and die Vermehrung
der Pleuropneumonieahnlichen Organismen and der L-phase der Bakterien, II. Elektronen-
mikroskopische Untersuchungen. Arch. Mikrobiol. 21:202, 1954.
K l i e n e b e r g e r- N o b e 1, E.: Filtrable Forms of Bacteria. Batt. Rev. 15:77, 1951.
P a t o~ k a, F? S u c h a n o v a, M,: Bakterialni cyklus t, zv. L-forem mikrobu. Biol.
listy 32:90, 1951.
S t e m p e n, H., Hutchinson, W. C.: The Formation and Development of Large
Bodies in Proteus vulgaris OR-19. I. Bright Phasecontrast Observation of Living Bacteria.
J. Batt. 61:321, 1951.
T r n k a, Z.: Pokusy o ziskani nebuneenych forem brucelly kultivaci na kurecim zarodku.
Kandidatska prate, Praha 1955.
T u l a s n e, R., B r i n g m a n n, G.: Donnees nouvelles apportees par le microscope
electronique sur la morphologic des formes L des batteries. Rev. Immunologic, 16 (4-5) :327,
1952.
T u 1 a s n e, R.: Les formes L des batteries. Rev, Immunologic. 15, 223, 1951_
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
By studying the literature an attempt was made to form an opinion on the bio-
logical significance of filtrable forms of bacteria. In a model experiment with Pfeiffer's
Haemophilus influenzae an attempt was made to obtain filtrable forms and, if
possible, to define the conditions under which these develop. The results were
obtained from biological experiments and from preparations studied by photo and
electron microscopy.
In the first place confirmation was sought as to whether filtrable forms develop
as an essential ontogenetic stage of bacteria. An attempt was therefore made to
obtain filtrable forms of Pfeiffer's Haemophilus influenzee, from cultures of meningeal
strains (3) and of strains from the respiratory tract (6). The following material was
filtered through a Seitz or a Berkefeld filter L 3 and L 5:
a) a saline extract of the organs of dead rabbits which had been inoculated with
a meningeal strain of Pfeiffer's Haemophilus influenzae,
b) cultures of strains of varying origin which had been cultured on fluid nutrient
media and had been passed through at least ten passages in the allantoic cavity of
the chick embryo.
The filtrates were inoculated on to suitable nutrient media (10% blood agar with
colonies of Staph. aureus), fluid media (liver broth with the addition of 3.5% blood),
on to sensitive animals (rabbits, mice, ferrets) and on to chick embryos.
The presence of filtrable forms which reverted to the original bacterial form were
not demonstrated. Nor were filtrable forms which might be demonstrated by the
phenomenon of satellitism.
We are not convinced that filtrable forms of bacteria are an ontogenetic stage of
bacteria living in conditions favourable to their multiplication.
Following these negative results attention was paid to the formation of the L-cycle
through the action of various external influences.
1. The action of penicillin in solid nutrient media on five strains of Pfeiffer's
Haemophilus two of which were isolated in children with meningitis is described.
All strains formed large round bodies (LRB) and in two strains, the cultures of
which were kept under constant observation at two-hourly intervals, a so-called
"incomplete" L-cycle was demonstrated with the greatest degree of probability
(Tulasne 1951). When penicillin placed in a cannula or in an opening in the nutrient
medium is absorbed into the medium gradually, it has an effect on the bacteria at
the various stages of development of the colonies. After a time these take on the form
of typical colonies of the L-form, via a transitional L-colony.
.`2. The originally normal colonies of Pfeiffer's Haemophilus which are changed by
the action of penicillin into L-colonies, are not equivalent biologically to stable
or partially stabilized A and B types of L-forms.
3. The L-cycle of Pfeiffer's Haemophilus produced by the action of penicillin in
turn breaks down the antibiotic and thereby destroys its antibiotic properties.
4. L-forms of Pfeiffer's Haemophilus were obtained in old cultures on chocolate
agar from the ninth day of cultivation. L-forms also developed by the action of
specific homologous gamma globulin in mixed solid and fluid media (RM environ-
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
ment). Fluid media are perhaps suitable for obtaining a stabilized L-form if the
influences evoking it have prolonged action.
L-forms in three generations were also successfully obtained from lyophilised
cultures of Pfeiffer's Haemophilus.
These experiments are intended to assist in understanding the significance of the
L-cycle of bacteria and in finding out whether viable filtrable forms of bacteria are
produced in the course of this cycle. The experimental conditions are arranged in
such a way as to approximate to the processes which take place in the host organism.
In order to 'confirm the findings obtained by photo microscopy, by the use of the
highest magnification permitting the elementary particles of the L-forms of the
bacteria to be distinguished, the formation of large round bodies of Pfeiffer's Haemo-
philus by the action of penicillin was also observed by electron microscopy. The
following results were obtained:
Using electron microscopy, the formation of large round bodies (LRB) in Pfeiffer's
Haemophilus, produced by the action of penicillin, was observed.
From longer bacilli a process grows out, which increases in size and becomes
spherical in shape. The diameter of this large, round body is about 4-5 microns.
Within 24 hours the contour of the large round body disappears and it disintegrates
into granules which were already previously visible in the contents, or into a formless
substance.
In connection with the formation of large round bodies, a grouping of the contents
of the bacterial cell into uniform granules, loss of the cell membrane and dispersal
of these granules into the medium were observed on media containing penicillin.
Such disintegration had not been found in previous observations using electron
microscopy.
(Plates I, II, III, IV, V, VI, VII, VIII)
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
F3~inrirrue nellituu~zuaa xa rla:IOVx~~ Ili~u~ii~jielri
I ~IIC. ~. ~i Tl"G`H I(f' ~i-'f~U'OBpll IIHIiVOaI~14[. IIa V;~~IiHPH HbIX LI830~f1i8S II 13p:IO1:H8A Ha'[I~HiI}pT
ou'paaosaTCft IiIiT. ~'iseamiexue a 450 paa.
I'uc. _'. li ~re~mf~n~~ 4-~~acoisoii ]IAhVval~~n~. O~ipaaosaH~rc IcIi1'. ~-s~~.ni~u~xn~~ ~; 3nn0 paa.
Ihic. :;. 1. -r~^r~~Hn~~ ft-~ru~o~;oil i~~[r.~i,ai~ui~. Ifp~~rspa.iucxiu~ trop~[ad~hHOii I.o.u>euu s L-soaot+~~IO.
1)ipasot;axirc IiliT. ~"se.~niaexut~ ~t 8511 paa.
~~III', 5. I3 Tl'41'Hlll' ~1-'f2iCUA011 11 HIZ C~i2ii~Itl(, ~_)~~?1~30i38Hlie L-IiOJIOHIIIt II hh~~~ B ];OJIOIIJIII RO.IIOIiI11I-
CI'LIS llil:f0'fPli, ~~~TR1YJf, I~pTOI)bl li IIpII U.Iii 1'OII~)fIflTH 6I_l" FCIOR4IftS [i ~':I L'PIIBiil~llll 71 iYPl'T 1~10~1 ~[V"
I:01:IfOi3, ~ 11P~[II~I(~H ITP 73 !IQQ ~)ti3,
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
i~~teet[ee neHi-~ua~s~rfixa e;~ i~aaro~tfey TTc~ei~~]~epa
l'u~+. 't. L' Te~[~~~c~~e 6-~tacosodi NHxyoageFr. ~OJI6IiILlHCTBO nano~~ex o6pasoeaato hIi'I' c ro~toreax~,inr
co,~ep:~;KMe~M. VseatHVeHxe B 3000 paa.
Puy+. G. B Te~ieKUe 12-~tacoeo~r i~xxy6aiSi-~H. ftI''P e pa:~.eF~Yxhix cTa~i-rex paneeT4~e. O;~HOpo;~~{~~c
Tesb[~a i~ eatcyo~-~[aFipoeatteE,~e ?ri~Zhi~a c rpaa3-sftt~FTeH. VeE~~tiTer-rwfa leis Apo 40, 0,95 ii oEtivsirip;~
Winkel 7eiss 9, cxirMSic 2, 4, 6 tt 9-c norto~gbio o6?~~cr,TNrsa 'Zeiss Apo 90, 1,80 i,i o~.~rzsrpa Zeiss
f[omal IV, 15, cxz3Ma; 11 - c noMOU~bio o67.e~,TNBa 7.eiss 10, 0,8 n oryns~pa ~~inkel1.eiss 9.
\Imcpo~oTOrpa~u=iecxxii annapaT Ultraphot 7eiss, paccTORxiTe naacTi?~ri~ oT ou~~,e~,~rrrsa 250 mrnr.
~huTOCx~zMKii c,~e.naxvr ;~-p MeA. YIocH~oM IIIoMO;~iT (Somogyi), PwcTOiror~~~tect;iiH u~tcTUTyT p~inu~~p-
c~tTe~ra I{o~fexci,oro, BpaTi~c:rana, as ~iTO M~,r np~aaoci~M eMy cnoro f,naro;~ap~iorri..
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
I'nc.10.,~eHCTBNC uexNuflz~Hixa flalla~iovleyII~[feH~epaHpH'i-~HeBlloii xyzrTNBaljlur. PacnaRaw-
II~HCCfI I~-ILOTOHHf[. McTIi03epHHCTOe NJIH HOHH06a1{NJIJIHpH00 COT,[epACNMOC IiIiT M3~TNBaeTCH
H olcpy.fcalou~yfo epe,*~y. YBe~THeexHe B 850 paa.
YNC, 11. 9-;SHCHHaH I.y.nbTypa na.IO~rICN II~eli~fepa Ha Iuolco~za~xoM arape. IIo HpaHnl ?cTapb}x
KO.IIOFI Ii~i? H0:3HIIfia10T rHeJJ~a BTOI1NiiHbiX fCO.TIOH}II?I~ HpeRCT1BJIHIOII(NX HepeXOT~HbI ~t TNH I.,-}CO~IOHI[#.
HanI4YNe IiI{Z' N 6a HTep Na7lbHbIX f~Op M. SrBed-I N'ieHNO B 1~0 pa3.
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
IIa~ro~[sa I~~~eif~~iepa. OopaaoRatic~e Iili'P.
Parr. 5, G: ~~ise:m~u~e>>e~ a 22.000 pas, p~ic, is ~-se~~zHe~r~te ~ 5800 paa, p~ic. 8: ~~He~ii~~t~xii~~
n 15.000 pas.
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
(RepriiNed from Nature, Vol. 179, pp. 918-919, Mrry 4, 1957)
Effect of Very Large Dases of Bacterial
Antigen on Antibody Production in New-
born Rabbits
TxE injection of large doses of antigen into very
young animals results in a weakening of the anti-
body response when a further dose of antigen is ad-
ministered to the adult animal. This phenomenon has
been described as acquired tolerances, immunological
approximations or immunological unresponsiveness'.
It is not yet clear whether the administration of large
doses of bacterial antigen during embryogenesis or
the first days of postnatal life can also inhibit
immunity responses in adult animals. Buxton found
a suppression of antibody formation to Salmonella?,
and Kerr and Robertsone to Trichomonas foetus.
Cohne and Smith and Bridges', on the other hand,
found no inhibition of antibody production after the
injection of bacterial antigens.
In the present work, five-day old rabbits were
iujocted by the intravascular or intraperitoneal routes
with a medium-sized dose (108 micro-organisms) of
heat-inactivated antigen of Salmonella paratyphi B.
Fiuther groups of animals were given excessive
amounts of the same antigen, the largest possible
dose being 4-6 x 10? organisms for five-day old
rabbits and 2 x 10? organisms for new-born rabbits.
It is evident from Table 1 that the medium-sized
dose of 108 organisms caused an antibody response
to appear, on the average, at the thirtieth day post-
natally. In both groups given excessive doses of
antigen, antibody formation was speeded up sig-
nificantly.
Bour months later, all surviving animals were re-
vaccinated with 10' organisms S. paratyphi B
(Table 2). Only where excessive doses of antigen had
been given to five-day old animals was there a
significant inhibition of antibody formation. New-
born rabbits injected with 2 x 10? organisms imme-
diately after birth showed no such inhibition on
re-vaccination.
These results show that inhibition can be achieved
with bacterial antigens, but only when administered
in amounts which are very near the lethal dose. The
negative results of Cohn? and Smithy may have been
due to the injection of insufficient amounts of bacterial
antigen.
Complete inhibition of antibody formation has
been achieved with serum albumin, and after the
first injections no antibodies were found in the serum
of infant animals?.ie, The present experiments show
that not even the injection of excessive doses of
Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
 Approved For Release 2008/12/09 :CIA-RDP80T00246A003000320001-2
;~~
Table 1. POSTNATAL DAY ON WHICH ANTIBODIES WERE FOUND FOR
'1HE FIRST TI AMOUNTSAOF ANTiBOEN (S.EParatyplTlOl )ON DIFFERENT
To 5-day-
old, 10?
micro-
organisms
(intraper-
i toneal)
No. of
animals
Median
Min.
Max.
To 5-day-
o1d,10?
intra-
vascular
29