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VOL.
No.
LIl .. "
l,c.
~Durnal
",,\
':;
®rhjinal (tommitntcattons.
.--.
'~J~'
.
i.
:NO'l'ES ,ON SqN,NE, DYSENTERY' IN, LAHORE, ,DISTRIO,T.·
By.
MAJ OR W. T. H. ·LARGE,
. Royal Army, Medical COrp8.
, SINCE the publication of Manifold's paper-(1927) on~' :aacillary:Dy.septery
in Poona," the military laboratorjes all over India 'havebeen reporting. ona
si'milar state of affairs in other districts, i.e:, that most of the 'dysentery
amongst the troops in India, and ,~p:erefore presumably s,Inongst thegeI,leral
population also, is bacillary.in o,rigin, only a small· proportion, about 25 per
cent, being attributable to. Ent(Z'mwba., histolytica.
.,
During' Manifold's . research, he . isolated a late lactose.fermenting
organism, giving the characteristic. sugarreactiolls of B. Sonne" ,from a,
certain number of his cases. Some of. his, strains did not ferment the
lactose until the seventeenth ,day:' Little also, -working :in' l\:t:'how,
mentions the, occurrence of this organism and describes it as B. Sonne. '
~ ,Study of recent literature ,on the subject shows that B.,Sonne is: now
being recognized as a cause of dysentery in, all parts of the world, even by ,
as widely separa,ted .obser:vers. a~ .Patter.soIl ,and,! Williams,in ,Australia.
(1922), and Perry and; Beusted, in Egypt.-(1927).
"
In Lahore District ,Laboratory, fQll.owing the completion of :0. research
on ,bacillary dysentery ill' generlltl, which forms the subject ofa separate
paper, 'an inquiry into the ch,aracters of Sonne' dysentery was commenced.
In this investigation three type cultures of B. Sonne, Group Ill, were
used~, ,
'. '" :, ',' ;' , '.', (' '.,. "',, ,','
,
, ,(l)~rl?,""" Distrf,c~, ,Laborat01'y, ,.l'oona..,....This .was, o~tained about
, eighteen 'Inohtlisprevipus)y from the R.A.M. College,~on~on.. . ',:
',,;1 " ,"
,.
1.
,
,
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2
Notes on Sonne Dysentery
i1i
Lahore District
(2) From the Bombay Bacteriological.Laboratory.-Originalsource, the
National Collection of Type Cultures. '
(3) From the R.A.1l'I. Oollege, London, direct, in May, 1928;
Before describing the results obtained 'with the lactose-fermenting
dysentery organisms locally isolated, a few of the points noted in the
behaviour of the three types on the media. in this laboratory are mentioned
here, with which the behaviour of the locally-isolated organisms will be
compared .
.Plate Oultures.-On lactose litmus agar the colonies showed differences
.in the three types, as follows ;(1) B. Sonne obtained from Poona formed discrete,. slightly crenated,
rounded colonies of 1'5 to 2 millimetres in dIameter, which do not differ
markedly from Flexner colonies .except in their crenation (hereafter noted
as first type).
.
(2) B. Sonne obtained from Bombay showed discrete, distinctly
crenated, irregularly circular colonies, about 2 to 2'5 millimetres in
diameter (second type).
(3) B. Sonne obtained Jrom R.A.M. College, London, showed spreading,
flat, crenated.colonies, quite different from those of B. Flexner; average size
4 to 5 millimetres (t4ird type).
.
The types are thus arranged in the order in which they most nearly
resemble Flexner .colonies. .
,
Broth Culture8.~Judged by the amount of deposit at the bottom·of the
broth tube the types follow a similar order. The first type deposits slightly~
the second type more so, while the third type deposits markedly.
. Lactose Broth Oultures.-All three types developed acid· in- lactose after
a. few days' incubation, the .period varying between three and fi~e- days.
No difference, between the types . was noted 'in this respect. Andrade's
fuchsin indica.tor was used, and the media were of standard reaction.
Glucose and Mannite Broth.~Showed 'acidity immediately the growth
"
of the organism became apparent.
Peptone Water.'---Indol was not formed.
Serum Tests.~High titre serum prodttced frqm rabbits in the. usual
wa.y agglutinated the'· homoiogous organism to 1-2,500 on. most occasions.
Dreyer's method of performing the agglutination test was used. A higher
titre was never obtained, and with some rabbits a titre of 1-1,250 only could
be produced .. A point common to all these, sera was that low dilutions of
1'-25 and, 1-50 were strongly positive, those of 1 in 125 and 1 in 250 very
weakly positiveaIid sometimes completely negative, while from 250 upwards
.
to 1 in 2;500, distinct agglutination was again Iioted.
'LAOTOSE-FERMENTING DYSENTERY ORGANISMS OF LAHORE DISTRICT;
During the course of eighteen months' work, twenty-four different
strains were examined, forming about i2per cent of' the :total
number of organisms isolated from bacillary dysentery cases.- All were
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W. T. H. Large
3
isolated from mild cases of dysentery, often in almost pure culture. Such
cases.wer,e considered clinically, and were at first reported bacteriologically,
on the biochemical findings, to be cause,d by B. dysenterice Flexner, so
common in this part of the world. No clinical difference between them
and the ordinary mild Flexner infections was noted by the physician in
charge of them, a~d all recovered in a few days under the ordinary saline
treatment.
0
When tested after not more than ten to twelve subcultures, the
organisms can be grouped in two types depending on their cultural and
serological reactions. (On first isolation from the patient, it is difficult to
detect any distinction between the colonies of the two types, unless an
almost pure culture is obtained, in which the colonies have space to grow
to their full size.)
A. The less common type resembles B. Sonne (third type) in all
particulars, i.e., its colonies on litmus lactose agar platea are crenated and
irregularly circular. It deposits in broth. It agglutinates to full titre with
B. Sonne high titre serum and, shows similarly a weakening of agglutination in dilutions of moderate amount, i.e., 1-50 and 1 in 125. Seven out of
twenty-four belonged to this type (local Sonne type A).
B. The commoner type appears at this early stage of its existence as a
circular colony with smooth edges, closely resembling a Flexner colony
and, because of its smooth circular outline, it is distinguishable from any
of the types of B. Sonne as observed in this laboratory. It resembles the
first type Sonne, however, more than the second and third. It does il0t
agglutinate, however, with Sonne serum. It does not deposit in broth
(local Sonne, type B).
Neitber type agglutinates to any appreciable extent with Flexner V, W,
X,Y or Z serum.
Carbohydrate reactions of 'the two types are similar, with the exception
of that in lactose broth. Both develop acidity, but while the A type
usually produces acid in carefully prepared broth on the third day; the
B type seldom turns the indicator until the fifth day. The period before
development of acidity was variable however in both types, and unless the
media were very carefully prepared, acidity might not develop until several
days later.
Peptone Water.-Indol was not formed.
Serological Tests.-High titre sera prepared from two of the locally
isolated B type organisms at about the twelfth subculture, and within about
a month 'of their isolation from the patient, agglutinated the homologous
organism to a titre of 1 in 500. The type of agglutination was that found
with dysentery organisms generally, i.e., fine and granular. There was no
hint of an inhibition zone (a negative phase); as in the case of the Sonne
organism.
Of the twenty~four strains isolated, ten were agglutinated to full titre
with each Of tbese'sera, but were completely negative to Sonne serum.
'
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4
Notes on Sonne Dysentery in Lahore District
.Tests of these type B sera against the stock Sonne organisms and the
locally isolat.ed A type organisms produced a variable and somewhat
peculiar result. In a few cases no agglutination occurred. The majority
of the organisms, however, showed a light flocculation up to full titre
resembling in appearance the agglutination of typhoid organisms, and
differing from the hard granular agglutination of the type B organisms ..
Serologically, therefore, as well as culturally, there appeared to be two
types of lactose-fermenting dysentery bacilli in the Lahore district: Type A,
which apparently was identical with B. Sonne, and type B, the commoner
of the two, which differed culturally from B. Sonne and was not agglutinated
by Sonne serum.
Animal Tests.-Rabbits inoculated with t cubic centimetre living
broth culture of two type B organisms died in twenty-four hours with
symptoms of enterocolitis and bloody diarrhrea. '1'he organisms were
recovered from the intestinal contents of the animals and from the blood,
heart, spleen, liver, &c.
Important work of another nature prevented the development of this
research until June, 1928, i.e., when the majority of the strains were about
a year old.
A new set of sera was prepared from B. Sonne (three types) a,nd from
several of the local type B organIsms.
The results were at first extremely puzzling, for while the Sonne serum
acted in identically the same manner as before with the type A organism,
it agglutinated to full titre two of the type B organisms with which
previously it. had been completely negative. The local type B serum, newly
prepared, agglutinated all type B organisms as before to full titre, but it
also. agglutinat~d B Sonne and the type A organism to full titre in the
normal way, as if it had been a Sonne serum.
On plating the orgl;tnism .from which the· type B serum had just been
prepared, it was discovered that there were now two distinct types of
colonies, the f?mooth circular type referred to as type B and crenated
irregular colonies referred to .above as type A, which differed in no way
from the colonies of the B. Son ne .(third type) recently obtained from
London.
All the laGtose-fermenting organisms of type B were now plated, and in
everyone two types of colonies were found. Some showed a vast majority
of type A colonies, others of type B, and some showed a mixture of about
equal number of the two types. It is to be remembered that the colony
originally isolated was in every case of the smooth circular type B.
No reason could be discovered for the preponderance on the plate culture
of one or other type of colony. Some of the oldest and most frequelltly
subcultured organisms showed so marked. a preponderance of type B
colonies that it was with difficulty that a single A colony could be found,
while. one or two. more recently isolated organisms (derived originally
from a type B colony and subcultured about twelve times only) showed a
preponderance of type A colonies.
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W. T. B. Large
5
The reverse however was not thl3 case, with one possible e~ception to be
referred to later. The three stock cultures' of B. Sonne were again plated
to see if any type B colonies could be found, but with negative results. The
seven local Sonne organisms were also plated out, and all showed complete
absence of type B colonies except one, "Cobbold," in which a single B
colony was found. The explanation for this exception in to be found in the
following test .. All the type A colonies and type B colonies were separated
into broth cultures and again replated separately. The type A colonies
bred true type A colonies without exception, but on one out of ten plates of
type B colonies an occasional type A could be found, even on this single
subculture from a B type colony. In addition, in one or two colonies,
a transitional stage was observed in which one side of the colony showed
the bold circular outline of a type B colony, while the other side sloped off,
and spread out into the fiat, irregular shape of the type A colony.
In the case of "Cobbold" noted above, the change from type B to type
A colony had progressed so far, during the ten subcultures made before
agglutination tests were performed, that practically only the type A colony
now remained.
Thjotta (1918) notes, in his description of the Sonne organism, that the
colonies may be of different types. One appears to be identical with the
type A noted above. The other, he says, is "small and dense, at other
times large and dense, with an edge growing out like a bunch of hair.
Both these colonies show irregularities of agglutination, either being inagglutinable or showing spontaneous agglutination in saline solution. If
cultivated for some generations on agar, these colonies will. as It rule,
develop colonies of the ordinary type both as regards agglutination and
growth."
I do not think the description" small and dense" applies to the colonies
described above as type B, which are clear, circular, sharp colonies, indistinguishable in appearance from those of Flexner's bacillus, and forming
possibly still another variation of the Sonne colony.
This distinction between the colonies of the Sonne bacillus is of great
importance for several reasons:(1) While the type B colony resembles those of other pathogenic
organisms, e.g., B. Flexner, and therefore is not likely to be missed,the
type A colony, from its peculiarity in size, and irregular al!pearance, may
easily be considered of no pathogenic importance, and be therefore disregarded. This may be the explanation for failure to find the causative
organism in many cases diagnosed on microscopical examination of the
exudate as bacillary dysentery.
(2) The serological reactions of the two types of colonies are different.
Type A colonies are identical with those of Sonne, and go to full titre with
Sonne serum. Type B colonies do not agglutinate at all with Sonne serum.
hut agglutinate to full titre with a serum prepared from one of themselves.
No data ar~ yet available as to the length of time necessary for the develop-
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6
Notes on SonneD,yseutery in Lahore District
ment from type B to type A, but considerably more than palf of the strains
of 'type B colonies, maintained in this laboratory for over a year, are still
inagglutinable with Sonne serum, although, on plating them out, a few type
A or Sonne colonies may be found. Presumably, in time they will follow
the example of the others and develop so many of the A type colonies that
they will become agglutinable with Sonne serum, but in the meantime
their serological diagnosis can be completed by the use of a type B serum.
Fyffe (1927) described a crenated and a smooth type of colony. His
crenated type appears to be identical with the true B Sqnne, and the A
type of colony described above. His smooth type differs from the B type
described above in the following part~culars: (a) The B type colony is
seldom larger than 2 millimetres. (b) The B colony never fermented lactose
in less than three days, and generally took five days at least to produce acid.
Presuming that his smooth type is identical with the B type described
above, the fact that it required several subcultures before it agglutinated
with Sonne serum is explained by the- observations above on the type B
colony, that is, that it really is an early type of the true B, Sonne.
CONCLUSIONS.
Two types of Sonne organisms are found in the Lahore district.
(1) Type A, crenated, irregular, and varying in size from 1 millimetre
(when the irregularity of the colony is not easily observed) to, perhaps,
6 millimetres! when the colony is markedly indented and irregular in shape.
'rhi~ type agglutinates with Sonne serum.
(2) Type B, smooth circular, and about 1'5 millimetres in diameter.
This type does not agglutinate with Sonne serum, but with a type B serum.
From it, at a varying period of time, are developed type A colonies which
h,ave the same characters as the typical B. Son ne, Group +II.
(3) As the type B colony is the more common, and as it may not give
rise tothe type A colony and agglutinate with Sonne serum for as long as
a year after isolation, it would appear that a type B serum is necessary for
the diagnosis of Sonne dysentery, in addition to the type A or true Sonne
serum.
REFERENCES.
,SONNE, CARL (1914). Journ. Bact., iv, 356.
. ,THJOTTA, TH. (1918). Ibid.
PATTERSON,~S. W., and WILLIAMS, F .. E. (1922), JOU1"n. Path. and Bact., xxv, 393.
SMITH, J. (1924). Journ. Hygiene, xxiii, 94.
FYFFE, G. (1927). Ibid., August.
PERRY, H. M., and BENSTED, H. G. (1928). Tmns. Roy. Soc. Trop. Med. and Hyg., xxi,
No. 5, February.
MANIFOLD, J. A. (1926). JOURNAL OF THE ROYAL ARMY MEDICAL CORPS, February.
MANIFOLD, J. A. (1928). Ind. Journ. Med. Research, xv, No. 3, January.
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Notes on Sonne Dysentery
in Lahore District
W. T. H. Large
J R Army Med Corps 1929 52: 1-6
doi: 10.1136/jramc-52-01-01
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