Microbiol. Immunol., 42( 1), 61-63, 1998
In Vitro Susceptibility of Chlamydia pecorum Macrolides, Tetracyclines, Quinolones and
to
β-Lactam Pudjiatmoko, Hideto Fukushi*, Yoshitsugu Ochiai, Tsuyoshi Yamaguchi, and Katsuya Hirai Department ofVeterinary Microbiology,Faculty ofAgriculture, GifuUniversity, Gifu,Gifu501-11,Japan Received August7, 1997.Accepted October 20,1997 Abstract:
The
romycin),
in
two
one ƒÀ-lactam
for
and
The
results
Key
words:
susceptibility
was
to
Chlamydia
and
1.0 ƒÊg/ml
pecorum,
pecorum
to
minocycline), The
were
0.063
ciprofloxacin. doxycycline
MIC,
Macrolide,
macrolides
0.004
are
MIC the
two
Quinolone,
to
for
for
most
effective
ithromycin pon
(Dynabott,
Japan)
quinolones,
Seika,
and
Japan); and
Japan);
Japan).
The
solubilized
for
macrolides,
tetracyclines,
and
antibiotics use
according
were
was
(Dai
Ichi
supplied to the
also
used
pecorum
strains
were
cell
centrifuged
then
aspirated
at
drugs
was
After
the
cultures
including
(MIC)
was
deter-
Japan
Society
were
done
in
a 24-well
with
g for
C.
tra-
cell-culture ml
incubation
of
fixed
10
of
wells
methanol
FITC-conjugated
the
as
the
(Denka lowest
in
medium were
dilution
cell at 37
C for
and
stained
for
72
hr,
inclu-
genus-spe-
Seiken,
the
of
monolayers.
cells
Japan).
antibiotic
inclusions
ml was
Drugs
anti-Chlamydia
antibody
per
culture ml.
Each of
organism
inoculum
of
per
inoculated
with
chlamydial
The
1 ml
of
HeLa-229
units
dilutions.
onto
defined
with
0.2
1 hr.
with
serial
overlaid
The
concentration cell
culture
at were
inhibited. MICs in
of
the
Table
1.
with
an
effective with
erythromycin
Abbreviation: 61
(16),
the
cycloheximide
was
cline,
*Address correspondence to Dr . Hideto Fukushi, Department of Veterinary Microbiology, Faculty of Agriculture, Gifu University, 1-1 Yanagido, Gifu, Gifu 501-11, Japan. E-mail:
[email protected]
1969
tests
overlaid
monoclonal
most
in
of
at 900 •~
MIC
shown
and
A
isolate
inoculated
cific
The
(11),
(12),
(13).
104 inclusion-forming
two-fold
completely
1968
1954
1954
method
coverslips
was
and
with
which
powders
on
were
in an
Drug
1 ƒÊg of
tested
in in
concentration
standard
well
containing
Phar-
lamb
containing
and
lung brain
comparison.
(8).
Each
calf calf
L2/454/Bu, for
the
suspension
sions
from
from
monolayers
plate.
manufacturer's
tested
C. pecorum.
from
inhibitory
by
antibiotic, C.
isolate
Chemotherapy
instruction. Three
for
1,024 ƒÊg/ml.
against
isolate
strain,
mined
(Meiji as
an
Minimum
Japan);
ampicillin
0.008
to 0.5 ƒÊg/ml
than
isolate
an
chomatis
doxycycline
ofloxacin
and ƒÀ-lactam,
0.25 greater
an
Ov/IPA,
(Dainip-
(Lederle,
and
clarithromycin,
drugs
Bo/Shizuoka,
clar-
erythromycin
minocycline
ciprofloxacin
maceutical,
and
Japan)
Pharmaceutical,
(Pfizer,
included:
eryth-
ciprofloxacin)
Tetracycline
cultures. tested
for
was
and
and
erythromycin,
ampicillin
Bo/Maeda,
antibiotics
(ofloxacin
0.008 ƒÊg/ml
Chlamydiapecorum, a newly described chlamydial species, causes enteritis, pneumonia, encephalitis and conjunctivitis in cattle (2, 7, 10-12, 15), polyarthritis and intestinal infection in sheep (13, 15), polyarthritis, abortion and pneumoniain swine (15), and a severe reproductivetract disease in koala bears (3, 9). Data on the treatment of C. pecorum infection in animals are limited,and little is known about the susceptibilityof C. pecorum to antimicrobial agents. Some antibioticshave been used for treating chlamydiosis. Tetracyclines,macrolides,and lately,quinolones have been used for treating human chlamydial infections (5, 6). Tetracyclinesare the drug of choicefor treating avian chlamydiosis(18). In this study, we evaluated the susceptibilityof C. pecorum in vitro to macrolides, tetracyclines,quinolonesand plactam antibioticsin cell The
(clarithromycin
quinolones
to 0.125 ƒÊg/ml The
and
two
two
MICs
minocycline,
for
clarithromycin
Chlamydia
and
determined.
doxycycline
0.25
show
of
(doxycycline
(ampicillin)
to 0.031 ƒÊg/ml ofloxacin
vitro
tetracyclines
antibiotics
Clarithromycin MIC
of
antibiotics MICs
against
of with
MIC,
was
0.004
0.008 MIC
minimum
to
the
pecorum most
doxycycline
0.063
inhibitory
the and
0.031 ƒÊg/ml, of
are
effective
to 0.008 ƒÊg/ml;
were
an
C.
followed to
0.125 ƒÊg/ml,
concentration.
next
minocyby
62
PUDJIATMOKO
Table
1. Activity
and
of seven
ofloxacin
and
0.5 ƒÊg/ml
and
cillin,
at
tion.
The
MICs
The
than
drug
to be
pecorum.
has
tration,
most
been
used
for
psittacine
tested
by
LCR
or
vivo
In
and of
and
determination
crobial
activity
showed
no
against
activity.
conducted
to
advantages
had C.
in
(14). be
Therethe
drugs of
in
treatment
animals.
doxycycline,
antimi-
while
clinical
8)
ampicillin
studies
antibiotics
1) Fraschini, F., Scaglione, F., Pintucci, G., Maccarinelli, G., Dugnani, S., and Demartini, G. 1991. The diffusion of clarithromycin and roxithromycin into nasal mucosa, tonsil and lung in humans. J. Antimicrob. Chemother. 27 (Suppl. A): 61-65. 2) Fukushi, H., and Hirai, K. 1992. Proposal of Chlamydia pecorum sp. nov. for Chlamydia strains derived from ruminants. Int. J. Syst. Bacteriol. 42: 306-308. 3) Girjes, A.A., Hugall, A.F., Timms, P., and Lavin, M.F. 1988. Two distinct forms of Chlamydia psittaci associated with disease and infertility in Phascolarctos cinereus (koala). Infect. Immun. 56: 1897-1900. 4) Gylstorff, I. 1984. The treatment of chlamydiosis in psittacine birds. Isr. J. Vet. Med. 43: 11-19. 5) Hammerschlang, M.R., Hyman, C.L., and Roblin, P.M. 1992. In vitro activities of five quinolones against Chlamydia pneumoniae. Antimicrob. Agents Chemother. 36: 682683. 6) Hammerschlang, M.R., Qumei, K.K., and Roblin, P.M. 1992. In vitro activities of azithromycin, clarithromycin, 1ofloxacin, and other antibiotics against Chlamydia pneumoniae. Antimicrob. Agents Chemother. 36: 1573-1574. 7) Kawagami, Y., Omori, Y., Fukuhara, S., Tokuda,G., Ishii, S., and Matumoto, M. 1956. Studies on the disease of cattle caused by a psittacosis-lymphogranulomagroup virus (Miya-
minocy-
excellent
pecorum,
for
animals
evaluation
optimal
showed
which
chemotherapy
to
in
stain
in
Prospective
determine
shown
been
the
has of
should
of
infection
most
a variety
agent
for
clarithromycin,
next
in has
applicable
erythromycin
the
alveolar
Doxycycline
doxycycline
C. pecorum
conclusion,
cline
be
C. Clarpene-
and
was
C.
8).
intracellular
epithelium
immunoperoxidase
may
efficacy
regimens
and
to
against (6,
chlamydial
and
and
active
Doxycycline
clarithromycin
choice,
also
chlamydiosis
eradicating
similar
against
trachomatis
C. pecorum.
avian
(4).
in
is
clarithromycin
antibiotic
Doxycycline
against
birds
study,
tissue
treating
effective
pecorum
bronchial
for
ciprofloxacin,
ampicillin.
C.
is
17).
antibiotic
to
0.031 ƒÊg/ml
this
C.
References
forma-
comparable
and
effective
and
into (1,
effective
of
of
excellent
especially
macrophages
fore,
the
to
Ampi-
0.125 ƒÊg/ml
for
In
C. pneumoniae
ithromycin
were
species
0.25
inclusion
ofloxacin
Clarithromycin
psittaci,
be
for
species.
of
clarithromycin,
susceptibility
shown
inhibit
minocycline,
1,024 ƒÊg/ml
chlamydial
was
to
for
and
MICs
respectively.
trachomatis
0.25 ƒÊg/ml
greater
other
failed C.
four Chlamydia
with
1.0 ƒÊg/ml,
0.008 ƒÊg/ml
doxycycline
and
to
for
erythromycin,
against
ciprofloxacin
0.25
1,024 ƒÊg/ml,
C. pecorum: for
antibiotics
ET AL
should
offer
be
potential
animals.
9) We script.
thank This
Research Sports
Dr. study
No. and
C.-C.
Kuo
was
supported
10156763
Culture,
from
Japan.
for
the
critical by
reading
Grants-in-Aid
Ministry
of for
of Education,
the
manu-
Scientific Science,
10)
11)
gawanella). VII. Isolation of a virus belonging to this group from feces of cattle. Jpn. J. Exp. Med. 25: 51-63. Kumamoto, Y., Matsumoto, A., Nagayama, A., Soejima, R., Hirai, K., Hashizume, S., and Hagiwara, T. 1992. Method for in vitro determinationof chlamydial susceptibility (Minimum inhibitory concentration; MIC) to antimicrobial agents. Chemotherapy 40: 308-314. McColl, K.A., Martin, R.W., Gleeson, L.J., Handasyde, K.A., and Lee, A.K. 1984. Chlamydia infection and infertility in the female koala (Phascolarctos cinereus). Vet. Rec. 115: 655. McNutt, S.H., and Waller, E.F. 1940. Sporadic bovine encephalomyelitis (Buss disease). Cornell Vet. 30: 437448. Omori, T., Ishii, S., and Harada, K. 1954. Studies on the dis-
NOTES
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63
ing of the European
society for Chlamydia research, Study
group for STD and dermatological microbiology of the Austrian Society for dermatology and venerology, Vienna, Austria. 15) Perez-Martinez, J.A., and Storz, J. 1985. Antigenic diversity of Chlamydia psittaci of mammalian origin determined by microimmunofluorescence. Infect. Immun. 50: 905-910. 16) Schachter, J., and Meyer, K.F. 1969. Lymphogranuloma venerum. II. Characterization of some recently isolated strains. J. Bacteriol. 99: 636-638. 17) Schentag, J.J., and Ballow, C.H. 1991. Tissue-directed pharmacokinetics. Am. J. Med. 91 (Suppl. 3A): 5S-115. 18) Vanrompay, D., Ducatelle, R., and Haesebrouck, F. 1995. Chlamydia psittaci infection: a review with emphasis on avian chlamydiosis. Vet. Microbiol. 45: 93-119.
