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Antimicrobial Agents and Chemotherapy, March 2001, p. 990-991, Vol. 45, No. 3
0066-4804/01/$04.00+0 DOI: 10.1128/AAC.45.3.990-991.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
LETTERS TO THE EDITOR
Quinolone Resistance-Determining Regions of gyrA and
parC in Pasteurella multocida Strains with
Different Levels of Nalidixic Acid Resistance
 |
LETTER |
Pasteurella multocida causes sporadic or epidemic
diseases among different animal species, including focal and systemic
infections in humans. Penicillin is the drug of choice for treatment of
Pasteurella infections, but third-generation cephalosporins
and fluoroquinolones are a good alternative for
beta-lactamase-producing strains or for allergic patients (1,
10).
Quinolone resistance in gram-negative bacteria is increasing, with
different mutations occurring in the quinolone
resistance-determining regions (QRDR) of the gyrA and
parC genes, one of the main causes of resistance (3,
5-9). In an attempt to determine if this mechanism also
occurs in P. multocida, both QRDR were isolated and sequenced from six isolates from animal (PM25 and its
derivative PM1024) and human clinical (16Q, 14Q, and 15Q) origins
(Table 1). All strains were identified
by standard methods (4), MICs were determined by the
E-test method (AB Biodisk), and the epidemiological relationship of
strains was corroborated by pulsed-field gel electrophoresis (data
not shown). Strains PM25 and 16Q were fully susceptible to all
quinolones assayed, while PM1024, 14Q, and 15Q presented different
levels of nalidixic acid resistance (Table 1).
PCR amplification with degenerate oligonucleotide primers, described
below for Haemophilus influenzae, was used to amplify the
QRDR of the gyrA and parC genes of strain PM25,
which were further sequenced (3). The nucleotide sequences
of both QRDR (accession numbers AF173979 and AF173980 of GenBank for gyrA and parC, respectively) were compared
with data from The Institute for Genomic Research
(http://www.tigr.org). Identities found were 98 and 96% with
P. multocida PM70, 83 and 82% with Actinobacillus actinomycetemcomitans, 81 and 85% with
H. influenzae, and 78 and 78% with Escherichia
coli for the QRDR of the gyrA and parC
genes, respectively. PM25 nucleotide sequences obtained were used to
design specific primers to amplify both QRDR of the other
strains (gyrA, 5'-GATGCACGAAGGCGGGAATGCC-3'
and 5'-CCGGTATTGCCGTCGGTATGG-3', and
parC, 5'-GAACTTGGTTTAAATGCCGCC-3' and
5'-CTCGACTGCCGCATATTT-3', at amplicon positions in
E. coli of 153 to 535 and 151 to 493, respectively). A
comparison of the deduced amino acid sequences of both QRDR with
E. coli revealed two changes in the GyrA subunit: a Ser-to-Ile mutation (AGC
ATC) in PM1024 and an
Asp-to-Gly mutation (GACGGC) in 14Q and 15Q at positions
exactly analogous to Ser-83 and Asp-87 of E. coli,
respectively (Table 1). These mutations may be responsible for the
different levels of nalidixic acid resistance and for the decreased
susceptibilities to fluoroquinolones that these strains exhibit.
However, other mechanisms could be involved in the MIC increase,
because after four subcultures of strains 14Q, 15Q, and PM1024 in 5%
blood agar (Oxoid) without quinolones, nalidixic acid MICs were 2.0, 2.0, and 48 µg/ml, respectively. Therefore, mutations found could be
the starting point for additional changes that, in conjunction with
other mechanisms, could lead to a high level of fluoroquinolone
resistance in P. multocida.
| |
ACKNOWLEDGMENTS |
This research was supported by grants PM97-0170 (Dirección
General de Enseñanza Superior), 98/1293 (Fondo de Investigaciones Sanitarias de la Seguridad Social), and 1999SGR-106 (Comissionat per a
Universitats i Recerca de la Generalitat de Catalunya) of Spain.
We acknowledge the technical assistance of Joan Ruiz and Mar
López.
| |
FOOTNOTES |
*
Phone: 34-93-5812615 Fax: 34-93-5812387 E-mail: mllagostera{at}einstein.uab.es
| |
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Maribel Cárdenas
Jordi Barbé
Montserrat Llagostera*
Departament de Genètica i
Microbiologia
Facultat de Ciències
Edifici
Cn
Universitat Autònoma de Barcelona
08193
Bellaterra (Barcelona), Spain
|
| | | | |
Elisenda Miró
Ferran Navarro
Beatriz Mirelis
Guillem Prats
Departament de Genètica i de
Microbiologia
Hospital de Sant Pau
Facultat de
Medicina
Universitat Autònoma de
Barcelona
Barcelona, Spain
|
| | | | |
Ignasi Badiola
Unitat de Sanitat Animal
Institut de Recerca
i Tecnologia Agroalimentària
Barcelona, Spain
|
Antimicrobial Agents and Chemotherapy, March 2001, p. 990-991, Vol. 45, No. 3
0066-4804/01/$04.00+0 DOI: 10.1128/AAC.45.3.990-991.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
