for publication,. July 9, 1984). A pteridine compound,. 2,4-diamino-6,7-diisopropyl-pteridine. (O/129) is known to be a vibriostatic agent. (4), and susceptibility.
Microbiol. Vol.
Transferable
Resistance
to
the
Vibriostatic
6,7-Diisopropyl-Pteridine
Shigeru
Department
Yasuo
of Microbiology,
of Public
Health,
to
be
pteridine a
compound,
vibriostatic Vibrio
(1, 6). O-1 and
Sundaram non O-1
strains
during
V.
cholerae
isolates
munication, V.
1,323
for
six
(Tc),
their
The
1983,
1982,
31
Kenya seas
from
1978,
to
and
1980,
of
from
1982.
our
58 1973
6
The
All
the
the
(Km),
ampicillin
1983,
India
in
345
obtained
in
in
the
36
in
short
of
coli
trimethoprim from from
in
1978 1973
1977,
and
those
in
over
Tokyo
Non O-1 period
to
40
from in
eltor. the
U.S.A.)
tetracycline
occurred
in
were
Mo.,
period
include
biotype
travellers
and
non O-1,
and
Nigeria
of com
manner.
Philippines
which
were
this
Louis,
the
resist
features
(Cm),
in Japan
overseas
166
in
cholerae
Escherichia
(Ap),
outbreaks tested
from
Vibrio such
In
St.
Japan
1981,
domestic strains
laboratory
and Co.,
chloramphenicol
cultures
in
of O-1
in
of
biological
to
Chemical
obtained
value themselves
a plasmid-mediated
agents,
to
in
isolates
transmissible
1,157
great
among
(2).
in
of
is known
encountered and
words,
(Sigma
is or
have
countries
is
other
antimicrobial
include
agent
human also
other
cholerae,
kanamycin
1983. in
V.
this
producibility
in
to O/129
patients
and
obtained
160
bacilli
We
resistance or
1979,
.
and
their
Thailand
1980
Japan
(Sm),
in
Laboratory
(O/129) to
enterotoxin
of O-1
Malaysia
travellers
were
that
used
in
in
in
strains
strains
641
on
sensitivity
streptomycin
,4-Diamino-
9, 1984)
described (5)
, 1984
OHASHI
Tokyo
Gram-negative
however, agent
conjugation,
commonly
(Tm). to
of
total
examined and
by
Research
July
susceptibility other
the
studies
report
strains
Makoto
Shirjuku-ku,
publication,
and
from
obtained
we
cholerae A
our
for
(4),
species
and Murthy, resistant to
ant
Metropolitan
Immunol.
1159-1162
cholerae•ôNS•ô
2,4-diamino-6,7-diisopropyl-pteridine
agent
differentiating
Tokyo
and
(10),
2
in •ôNH•ôVibrio
KUDOH,
Hyakunin-cho,
(Accepted
A
Agent
(O/129)
MATSUSHITA,*
28
in
strains
from
1977
to
1982. Sensitivity was
tested
agents Society Broth One
starting 20hr,
all
of
to
of on
test
strains
the
twofold 100ƒÊg/ml.
were
agar
examined
strains
which
serially After for
were
contained
diluted
growth
of of
the 1159
the test
described
by in
U.S.A.)
at
of the
these Japan
Trypticase 37C
Soy
overnight.
106cells/ml Detroit, of
inoculated strains.
agents
(MICs)
approximately
Laboratories, concentrations
incubation
antimicrobial
precultured Md.,
(Difco
other
method
Cockeysville,
Agar
the
concentrations
dilution
Test
Systems, cultures
and
inhibitory
standard
Mueller-Hinton
containing
to O/129
minimal
(3).
Microbiology
microliter
they
the
Chemotherapy
(BBL
with
the
determining
according
inoculated plates
of by
Mich.,
antimicrobial plates
The
was U.S.A.)
MIC
at
agents 37C
was
for defined
18
to as
1160
S. MATSUSHITA
ET AL
NOTES
the
lowest
growth
concentration
ƒÊ
occurred.
and
Tm,
g/ml
or more Thirty-three
to
Strains
100ƒÊg/ml
some
were to
of
these
be
detect strains,
resistant
mutants,
One-half
were
in
35C
for
plates
a
of
CSH-2
each
of
18hr
growth
of
per
ml
Km,
of
each
W1895
C)
All
of
case
of
one
The
exceptional
the
to
quencies
of
drug
marker
and
the
plates.
the
recipient
E.
cholerae
secondary
TS1-4
with
mind
when
other
Gram-negative may
Experiments
indicate
that
to
Tm
an O/129
be
in
due
of
transfer
to
their
of
agar in
the Sm,
resistance
using
three
transfer
of out
all
was purified
resistance
with
E.
coli
except
in
the
V.
E.
or
coli
test among
common
is is
to
was
plasmid for
plasmid
or
C)
in
be
cross
the
on
more
an
selec
different 100ƒÊg/ml
by
the
resist
acquired transferable
should Vibrio resistance
diamino-pyrimidine progress.
cholerae in
as
about
resistance One
differentiating
structure,
fre
by in
the
Rfr.
the O/129
The
are
used
of
to
encoded.
used
was
times
found
themselves.
V.
experimentally
(Hfr of
and
significantly
10
trans The
respectively,
of O/129
resistance
most
chemical the
change
W1895
probably cholerae
markers
Tm,
Sm,
coli
donor,
dissociated.
and
and
E.
per
further
strains
Sm,
Tc,
recipient
not
The
donor
to
the
never
Tc,
Cm,
concentration
increased
the
markers
Cm, of
drug
did
resistance
Tm
10-2-10-4
available
strains.
sensitivity
characterizing
to
and
the
that
bacilli
for
markers
of
experiments
results
to O/129,
was
donor
from
at
5.0ƒÊg/ml;
carried
with O/129
resistant
inhibitory
strain the
Nxr
transfer
These
original
coil
were
and
incubated
agents
transfer
Nxr
37C
Penassay
Tc,
secondary
recipient
at
on
by
(Nx)-
recipients.
the
antimicrobial
on
resistance
10-4-10-5
frequency
from
of
was
5.0ƒÊg/ml;
and
resistance
of Each
The
acquired
the
the O/129
35C.
1,
acid as
either
selected
the
checked
TS1-4
Table
these O/129-
Nxr
mixture
When
1). O/129
strain,
order
of
was
jointly
(Table
only of
at
cholerae
transferred
strain
the
of
50ƒÊg/ml.
in
Laboratories)
The
markers
latter
sensitive
recipient.
markers
transfer in
V.
as
non O-1
cross
selective
used
recipients
were
overnight
by
Rfr
non O-1
the
Ap,
resistance
the
nalidixic
TS1-4
4.5ml
Cm,
of
in
using
(Difco
Experiments
acquired (Hfr
ance
of
transconjugant.
experimentally
strains
and
segregation
colonies
ferred
20ƒÊg/ml;
100ƒÊg/ml;
demonstrated,
and
one
12.5
resistant
strains
shown
Tc,
and
be
two
the
plasmids
by
were of
to
and of
Cm,
Ap,
found
As
out,
mixed.
for
of
resistance.
cholerae, O-1,
Broth
and
for
former
R
Transconjugants Nx
concentrations: O/129,
100ƒÊg/ml;
V
were
more
None
Tm
strains
phase,
or
to O/129.
carried
and
Penassay
shaking.
25ƒÊg
the
transferable
resistant in
of
inhibition
more
were
time.
with
were
the
or
strains
16
same
resistance
Nxr
cultivated
with
and
conjugally
complete
6.25ƒÊg/ml
resistant. non O-1
the
which
50ƒÊg/ml
associated
experiments coli
logarithmic
following
O/129
presence
were
containing
ciated
always
of
which
at
of
Km,
tested,
showed
was
E.
milliliter
strains,
V.
agents
at
MIC
and
to O/129
mating
an
Sm
considered 20 of the
agents
the
agent
gave
for were and
resistant
resistance
resistant
tive
more
antimicrobial
To
which
or
antimicrobial
to
the O/129
antimicrobial
for O/129 of the O-1
the
found
in
of
1161
closely bear
asso this
species to
in
from Tm
and
moiety.
1162
S. MATSUSHITA
ET AL
The authors wish to thank Dr. Ratanasda Phan-Urai, Department of Medical Sciences, Thai land, Dr. Hiroshi Ogonuki, Liaison Officer in Manila for the Japan International Cooperation Agency, Dr. M. Jegathesan, Institute for Medical Research, Malaysia, Mr. D.C. Metha, State Public Health Laboratory, Pune, India, and Dr. Masaaki Iwanaga, Institute for Tropical Medicine, Nagasaki University, for their generous supply of isolates. REFERENCES 1) 2)
3)
4) 5) 6)
Davis, G.H.G., and Park, R.W.A. 1962. A taxonomic study of certain bacteria currently classi fied as Vibrio species. J. Gen. Microbiol. 27: 101-119. Kudoh, Y., Matsushita, S., Yamada, S., Tsuno, M., Ohta, K., Sakai, S., and Ohashi, M. 1981. Enterotoxin producibility and some biological features of O-1 and non O-1 Vibrio cholerae isolates, p. 214-224. In Kuwahara, S., and Zinnaka, Y. (eds), Symposium on cholera, Gifu, 1980, Toho University, Tokyo. Japan Society of Chemotherapy. 1981. Method for minimal inhibitory concentration (MIC) determination of antimicrobial agents by the agar dilution technique (Revised). Chemotherapy 29: 76-79 (text in Japanese). Shewan, J.M., Hodgkiss, W., and Liston, J. 1954. A method for the rapid differentiation of cer tain non-pathogenic, asporogenous bacilli. Nature 173: 208-209. Sundaram, Sp., and Murthy, K.V. 1983. Occurrence of 2,4-diamino-6,7-diisopropyl-pteridine (O/129) resistance in human isolates of Vibrio cholerae. FEMS Microbiology Letters 19: 115-117. WHO Scientific Working Group. 1980. Cholera and other vibrio-associated diarrhoeas. Bull. WHO 58: 353-374. (Received for publication,
April 6, 1984)
