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Antimicrobial Agents and Chemotherapy, May 2003, p. 1665-1671, Vol. 47, No. 5
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.5.1665-1671.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Wolbachia pipientis Growth Kinetics and Susceptibilities to 13 Antibiotics Determined by Immunofluorescence Staining and Real-Time PCR

Florence Fenollar, Max Maurin, and Didier Raoult*

Unité des Rickettsies, CNRS UMR 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille cedex 05, France

Received 17 May 2002/ Returned for modification 4 November 2002/ Accepted 5 February 2003

Wolbachia spp. are strict intracellular bacteria that infect a wide range of arthropods and filarial nematodes. Filarial nematodes are important causes of human diseases. There is increasing evidence that Wolbachia spp. influence important functions in the biology of the hosts, specifically, infertility. Preliminary experiments with humans and animals have suggested that antibiotics with activity against Wolbachia may help to treat filariasis. In this study, we determined using a real-time quantitative PCR assay the growth kinetics of a strain of Wolbachia pipientis from a mosquito grown in Aa23 cells. The doubling time was estimated to be 14 h. We then determined the susceptibilities of this strain to 13 antibiotics by two methods: an immunofluorescent-antibody test and a real-time quantitative PCR assay. Both techniques gave similar results. Doxycycline and rifampin were the most effective compounds, with MICs of 0.125 and 0.06 to 0.125 µg/ml, respectively. Fluoroquinolones were less effective, with MICs of 2 to 4 µg/ml for ciprofloxacin, 2 µg/ml for ofloxacin, and 1 µg/ml for levofloxacin. ß-Lactams (penicillin G, amoxicillin, ceftriaxone) were not effective at concentrations up to 128 µg/ml. The MIC of erythromycin was >32 µg/ml, whereas that of telithromycin was 8 µg/ml. Other antibiotic compounds were bacteriostatic only at high concentrations, including gentamicin, co-trimoxazole, and thiamphenicol. The real-time PCR assay was a convenient and reliable technique for determination of the antibiotic susceptibilities of Wolbachia. It may help in the future to simplify antibiotic susceptibility testing of strict intracellular pathogens.

* Corresponding author. Mailing address: Unité des Rickettsies, CNRS UMR 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 27, Boulevard Jean Moulin, 13385 Marseille cedex 05, France. Phone: (33) 04.91.38.55.17. Fax: (33) 04.91.83.03.90. E-mail: Didier.Raoult{at}medecine.univ-mrs.fr.

Antimicrobial Agents and Chemotherapy, May 2003, p. 1665-1671, Vol. 47, No. 5
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.5.1665-1671.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Makepeace, B. L., Rodgers, L., Trees, A. J. (2006). Rate of Elimination of Wolbachia pipientis by Doxycycline In Vitro Increases following Drug Withdrawal. Antimicrob. Agents Chemother. 50: 922-927 [Abstract] [Full Text]  
  • Branger, S., Rolain, J. M., Raoult, D. (2004). Evaluation of Antibiotic Susceptibilities of Ehrlichia canis, Ehrlichia chaffeensis, and Anaplasma phagocytophilum by Real-Time PCR. Antimicrob. Agents Chemother. 48: 4822-4828 [Abstract] [Full Text]  
  • Fenollar, F., La Scola, B., Inokuma, H., Dumler, J. S., Taylor, M. J., Raoult, D. (2003). Culture and Phenotypic Characterization of a Wolbachia pipientis Isolate. J. Clin. Microbiol. 41: 5434-5441 [Abstract] [Full Text]  


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