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Antimicrobial Agents and Chemotherapy, December 2009, p. 5155-5162, Vol. 53, No. 12
0066-4804/09/$08.00+0 doi:10.1128/AAC.00532-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Effects of Subinhibitory Concentrations of Antibiotics on Colonization Factor Expression by Moxifloxacin-Susceptible and Moxifloxacin-Resistant Clostridium difficile Strains
Cécile Denève,1
Sylvie Bouttier,1
Bruno Dupuy,2
Frédéric Barbut,3
Anne Collignon,1,4* and
Claire Janoir1
EA 4043, Université Paris-Sud 11, Faculté de Pharmacie, 5 Rue Jean Baptiste Clément, 92296 Châtenay-Malabry, France,1 Unité des Toxines et Pathogénie Bactérienne, Institut Pasteur, 25 Rue du Dr. Roux, 75724 Paris Cedex 15, France,2 EA 2392, Université Pierre et Marie Curie, Paris 6, Hôpital St. Antoine, Assistance Publique-Hôpitaux de Paris, Paris, France,3 Service de Microbiologie, Hôpital Jean Verdier, Assistance Publique-Hôpitaux de Paris, Bondy, France4
Received 21 April 2009/ Returned for modification 20 June 2009/ Accepted 24 September 2009
Recent outbreaks of Clostridium difficile infection have been related to the emergence of the NAP1/027 epidemic strain. This strain demonstrates increased virulence and resistance to the C-8-methoxyfluoroquinolones gatifloxacin and moxifloxacin. These antibiotics have been implicated as major C. difficile infection-inducing agents. We investigated by real-time reverse transcription-PCR the impact of subinhibitory concentrations of ampicillin, clindamycin, ofloxacin, and moxifloxacin on the expression of genes encoding three colonization factors, the protease Cwp84, the high-molecular-weight S-layer protein, and the fibronectin-binding protein Fbp68. We have previously shown in six non-NAP1/027 moxifloxacin-susceptible strains that the presence of ampicillin or clindamycin induced an upregulation of these genes, whereas the presence of fluoroquinolones did not. The objective of this study was to analyze the expression of these genes under the same conditions in four NAP1/027 strains, one moxifloxacin susceptible and three moxifloxacin resistant. Two in vitro-selected moxifloxacin-resistant mutants were also analyzed. Moxifloxacin resistance was associated with the Thr82
Ile substitution in GyrA in all but one of the moxifloxacin-resistant strains. The expression of cwp84 and slpA was strongly increased after culture with ampicillin or clindamycin in NAP1/027 strains. Interestingly, after culture with fluoroquinolones, the expression of cwp84 and slpA was only increased in four moxifloxacin-resistant strains, including the NAP1/027 strains and one of the in vitro-selected mutants. The overexpression of cwp84 was correlated with increased production of the protease Cwp84. The historical NAP1/027 moxifloxacin-susceptible strain and its mutant appear to be differently regulated by fluoroquinolones. Overall, fluoroquinolones appear to favor the expression of some colonization factor-encoding genes in resistant C. difficile strains. The fluoroquinolone resistance of the NAP1/027 epidemic strains could be considered an ecological advantage. This could also increase their colonization fitness and promote the infection.
* Corresponding author. Mailing address: Université Paris-Sud 11, Faculté de Pharmacie, Département de Microbiologie, 5 Rue Jean Baptiste Clément, 92296 Châtenay-Malabry Cedex, France. Phone: 33 1 46 83 56 34. Fax: 33 1 46 83 55 37. E-mail: anne.collignon{at}u-psud.fr
Published ahead of print on 5 October 2009.
Antimicrobial Agents and Chemotherapy, December 2009, p. 5155-5162, Vol. 53, No. 12
0066-4804/09/$08.00+0 doi:10.1128/AAC.00532-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
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