IV. INVESTIGATION OF C. DIPHTHEHIAE
111
cultures (Pope Linggood cystinated broth) of each strain to be tested are
spotted by means of a Pasteur pipette on to the surface of brain heart agar,
usually eight per plate. After 48 h incubation, at 37"C, growth develops
from each drop. These are killed by chloroform vapour for 2 h. Then a
drop of a 23-3 h liquid culture of the sensitive strain is applied to the edges
of each patch of growth.
After 18 h incubation at 37"C, the production of bacteriocins is expressed
by inhibition of the sensitive test cultures as presented in Fig. 21.
Storing of bacteriocinogenic strains. Bacteriocinogenic strains are cultivated
and stored either on Loeffler slants for 2-3 months in the dark at room
temperature or in the lyophilised state. These bacteriocins have not shown
up in electron microscopy. Bacteriocinogenic strains are available from the
Diphtheria Laboratory of the Cantacuzino Institute, Bucharest, Romania.
(c) Technique of bacteriocin typing of C . diphtheriae. Heavy, overnight
Loeffler cultures from each of the bacteriocinogenic strains is plated along
a 1 cm wide diametric band across each plate. After 48 h at 3 7 T , the
T A B L E I11
Characters of bacteriocinogenic strains used for C. diphtheriae
bacteriocin typing
Registered number
of bacteriocinogenic
Corynebacterium
Toxigenic
strains
species
character
Origin
153
C. hofmannii
-
Carrier
155
C. hofmannii
-
Carrier
198
C . hofmannii
-
Carrier
730
C. diphtheriae intermedius
toxCarrier
760
C. ulcerans
Di-toxCarrier
828
C . diphtheriae intermedius
toxCarrier
866
C . diphtheriae mitis
toxf
Case
868
C . diphtheriae mitis
toxf
Case
871
C. diphtheriae mitis
tox+
Case
872
C. diphtheriae mitis
toxf
Case
873
C . diphtheriae mitis
toxf
Case
876
C. diphtheriae intermedius
toxf
Case
877
C. diphtheriae mitis
tox+
Case
878
C. aiphtheriae mitis
tox+
Case
882
C . diphtheriae mitis
toxf
Case
898
C . diphtheriae mitis
toxf
Case
2378
C. diphtheriae intermedius
toxCarrier
2380
C. diphtheriae intermedius
toxCarrier
Hof.
C. hofmannii
-
-
Xer.
C. xerosis
-
-
111
cultures (Pope Linggood cystinated broth) of each strain to be tested are
spotted by means of a Pasteur pipette on to the surface of brain heart agar,
usually eight per plate. After 48 h incubation, at 37"C, growth develops
from each drop. These are killed by chloroform vapour for 2 h. Then a
drop of a 23-3 h liquid culture of the sensitive strain is applied to the edges
of each patch of growth.
After 18 h incubation at 37"C, the production of bacteriocins is expressed
by inhibition of the sensitive test cultures as presented in Fig. 21.
Storing of bacteriocinogenic strains. Bacteriocinogenic strains are cultivated
and stored either on Loeffler slants for 2-3 months in the dark at room
temperature or in the lyophilised state. These bacteriocins have not shown
up in electron microscopy. Bacteriocinogenic strains are available from the
Diphtheria Laboratory of the Cantacuzino Institute, Bucharest, Romania.
(c) Technique of bacteriocin typing of C . diphtheriae. Heavy, overnight
Loeffler cultures from each of the bacteriocinogenic strains is plated along
a 1 cm wide diametric band across each plate. After 48 h at 3 7 T , the
T A B L E I11
Characters of bacteriocinogenic strains used for C. diphtheriae
bacteriocin typing
Registered number
of bacteriocinogenic
Corynebacterium
Toxigenic
strains
species
character
Origin
153
C. hofmannii
-
Carrier
155
C. hofmannii
-
Carrier
198
C . hofmannii
-
Carrier
730
C. diphtheriae intermedius
toxCarrier
760
C. ulcerans
Di-toxCarrier
828
C . diphtheriae intermedius
toxCarrier
866
C . diphtheriae mitis
toxf
Case
868
C . diphtheriae mitis
toxf
Case
871
C. diphtheriae mitis
tox+
Case
872
C. diphtheriae mitis
toxf
Case
873
C . diphtheriae mitis
toxf
Case
876
C. diphtheriae intermedius
toxf
Case
877
C. diphtheriae mitis
tox+
Case
878
C. aiphtheriae mitis
tox+
Case
882
C . diphtheriae mitis
toxf
Case
898
C . diphtheriae mitis
toxf
Case
2378
C. diphtheriae intermedius
toxCarrier
2380
C. diphtheriae intermedius
toxCarrier
Hof.
C. hofmannii
-
-
Xer.
C. xerosis
-
-