icb194426a camp test original paper

8/10/2019 Icb194426a Camp Test Original Paper

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A NOTE ON A LYTIC PHENOMENON SHOWN BY GROUP B

STREPTOCOCCI

by K. CHRISTIE, N. E. ATKINS

AND E. MU NC H- PE TE HS EN

(B'rom the Commonwealth Serum Laboratories, I'niversity Bacteriology Depart-

ment and C.S.I.R. Animal Health Researeh Laboratx)ry. Melbourne).

Accepted for publication 11th July, 1944.)

Tn a recent outbreak of Scarlet Fever in a country disti'iet it was suspected

that the milk was tbe vehicle of infection. Five samples of the milk supplied to the

area were submitted to examination for haomolytie streptococci. After centrifuga-

tion, tlie mixed cream and sediment were stroked out on the surface of sheep blood

afjar plates which were inenbated over-nijclit at 37 C. From three of the samples

.streptoeocci were obtainetl whieh were apparently haemolytie ou tbe iii-iniary plates

but uon-haemolytic on sub-eultnre.

Examination of the primary plates revealed tbe presence of many .staphylo-

eocei of tbeafitype, that is their colonies were in tbe centre of a clear zone, which

in tnrn was Murronnded by a large darkened zone, wbere the stapbylocoeeal

i

toxin

had altered, bni not iysed, the sheep red cells. Wherever tliere were colonies of the

streptococcus within these zones of darkening, the colonies were surrounded iiy an

area of complete haemolysis, whilst elsewhere on the ])late.s they produeed no distinct

haemolysis

To confirm tliia ohserviitioii, a colony of

tlieae stui>liy (>t;occi wna grown by spot-

inoeuliitioii on the centre of

a

slieep blood

jigar plate. After L4 hours' incubation tho

atroptoeoeci were inofiilatod on to the darl;-

enod zone. Within two hours hiii.'nio].vsl3 was

visible aroinid the Rtrt'jitoeoccEiI growth.

Ill the ease of

n

st.iphyiococ


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198 R. CH RIS TIE , N. E. ATK INS

AXD

E . MtTNCH-PETERSEN

"i iioii-haeniolytie anaero bic stvaina of human o rigin (reeoivi'd th rough tlie courtesy of Miss

IL Btitler), none gavt> the roaetion when grown with the stapliylococcus, although aomo showed

;i narrow :nT i of iiicoHii)l('te lysiy. Fo rty -th roe stn iiiia from eases of siispofted and nini't('i.'n

from cast's of di fiiiitc liovinc mastiti s were tested . Of these, ali }mt three giivc tlie niai-tioii.

Scro logieal eX iimliKition tlicii revealed th at of all Ihe sti-aiiiN tested only those which belonged io

(rroiip B ]jaiH'e ielil gave the reaction, an d none belniigiiig to (irtui]> B failed io give H. Five

(jrou]:i B strains of liunian origin (tdiiaiis, vagina and skin lesion) also gave the positive reai'tioii,

Tliroc rep resen tative sti-iiins of each of the (iroiiis A,

V

and (i were teste


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GROUPB STREPTOCOCCI 199

incubation

for one

liour

at

37

C, the

cells were found

to be

lysed

in the

tube containing

the

undiluted filtriite,

bnt the

power

of the

filtrate

to

induce lysis

was

quickly lost

on

dilution.

An

undiluted sample which

had

been h eated

at 56 C. for 30

minutes eaused only

a

trace

of

lysis,

but

a sample lieated

at 50 C. for 30

minntes

and

then maintained

at

100

0. for

five minutes,

had a

iytic ])otency similartothatof theunheated filtrate. Asamjile heated forfive minutesat 100"C.

was asx>otent as the raw filtrate (seeTable 1). When an undiluted samjile of the filtratewas

allowed to act on the redcellsfor30 minutesat 37 C, lysis occurred immediately onadditionof

the

fi

toxin.

TABLE 1.

Results

of

haemolysis titrations, using streptoeoccal filtrate, staphylocoecal

i

toxin

and

sheep

red cells.

SeriesI. Ten

drops

of

each diluted preparation

of

streptoeoccal filtrate were added

to the

corresponding tube.

Ten

drops

of 3

toxin

(raw,

filtered toxin diluted

1: 10

with normal saline)

were added toeach tube. Two drops of a 10ji.c. suspension in salineof washed sheep redcells

were then added

and the

tubes inculiated

at

37

0. for 1

honr.

ScrictII. As above, exeept tha t the filtratewasfirst heatedto5(1 C.for 30minutes.

Sfries

III.

As in

series

I,

except that

the

filtrate

was

first heated

to 50 C. for 30

minutes,

iind then to100 C.forfive niiiiutes.

SeriesIV. As inseries^,except that the filtratewasfirst heated to10(1C .forfive minutes.

Scrien

V.

As in

series

I, but

witli saline substituted

for the /i

toxin.

Scries

VI.

As in

scries

I, bnt

with saline substituted

for the

streptoeoccal filtrate.

Initial dilutionof filtrate.

10/10 8/10 6/10 4/10 2/10

Series I

Series II

Series

III

Series IV

Series V _ .

Series

VI _ _ _ _ _

- --{-+ + Indicates comjilote lysis. -\-Indicates 25p.c. lysis.

Indicatesnolysis.

The qnantityof theagent present in the filtrate wasnotfound to beincreasedby incubation

for longer tiiau

i8

hours.

Th< staphylocot-ei most commonly found

in

animals

are

predominantly

(3

toxin-produeera

(Minett,

1936;

Cowan, 1938).

It is

only with these staphylococci that Gronp

B

strepto(;occi

produce the lysis described above. Not infreqnently staphylococci andGroupBstreptococciare

fonnd together

in the

bovine u dder; there

may be

some significant connection lietween th is

occurrence

and the

fact tha t toge ther they have

a

Iytic power which neither

has

independently.

The results obtained with

the

Group

B

streptococci

of

human origin give further evidence

of

the elose relationship of theanimal andhuman strains of this group, in spite (if the differences

in serological tyj)eand insome other characteristics establishedbySimmonsandKeogh (194 0).

The nejrative results ol>taiiied when horse blood was substituted for sheep

blood siijiuest that in routine tests for haemolytic streptococci, horse blood only

should be used, ijarticnlarly where Iitoxin-jn oducin^ stapliylococei are to be ex-

pected, as in milk samples.

The nuniber (tf members of the various rroups tested was not larfje, but the

resnlts obtained so far indicate that the phenomenon may provide a relatively

simple confirmatory or substitute test for Group B streptococci.

SUMMABY.

Strains of Group B streptococci, of aniinal and hnman orijzin. produce an

agent which will lyse sheep and ox. but not hnman, borse. rabbit or f^iinea-pij; red

cells, wh(?n these cells have been altered by staphyloeoccal itoxin.

The a^ent is extracellular, tlltrable, and thermostable.

Streptococci of liiiman and animal ori


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200 R. CPIRTSTIE, N. E . ATKTNS .\NDE . J ir 'N C H -PETER SEN

AcTcnn ivlctJgme nt. i. We are indebted to Professo r WoodrufF. of the rn iv e rs it r Itac-terio

logieal D epa rtmen t, for a(iviee and (riti( i.sni, to M iss M. Monsbourgh jiiid M r. H. A . Keddoine

for teclniU'al aasiatance, to Miss M. (iarhind for milk samples, aiul to Mr. R. T. Simmons for

providing strains reprcsentative of various groups of streptoeycei.

RE FE RE NCE S.

Christie, R. and Grnydon, J . J . (1SI41): Austr al. ,T. exp. Bio ., 19, p 9.

Cowaii, 8. T. (1938) : J . P ath . Bac t., 4(i, p. 31 .

Landsteiner, K. and von Raiiohenlnehler, R. (1909) : Z. Inimun. Porsch., 1, p. 4S9

Minett, F. C. (193{i) : J. Path. Bact., 42, p. 247.

Orcutt, M. L. and Howe, P. E. (1922) : J. exp. Med., 35, p. 409.

Simmons, R. T. and Keogh, E. V. (1940) : Austral. J. exp. Biol., 18, p. 1.^1.


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icb194426a camp test original paper

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