Coxiella burnetii is a strictly intracellular bacterium. Bacteriostatic effects have been described previously on a few isolates in embryonated eggs (A. J. Spicer, M. G. Peacock, and J. C. Williams, p. 375-383, in W. Burgdorfer and R. L. Anacker, ed., Rickettsiae and rickettsial diseases, 1981). We used the shell-vial technique (D. Raoult, G. Vestris, and M. Enea, J. Clin. Microbiol. 28:2482Microbiol. 28: -2484Microbiol. 28: , 1990) to determine the susceptibility of C. burnetii to amoxicillin, amikacin, erythromycin, co-trimoxazole, pefloxacin, ofloxacin, ciprofloxacin, chloramphenicol, tetraycline, doxycycline, minocycline, and rifampin antibiotics at a single dilution. Human embryonic lung fibroblast monolayers in shell vials were seeded with 13 different C. burnetii isolates, including 3 reference strains (Nine Mile, Q212, and Priscilla) and 10 new isolates, in order to obtain 30% infected cells 6 days later. After inoculation, antibiotics were added, shell vials were incubated for 7 days, and immunofluorescence was revealed and compared with that of the positive controls. Strain Nine Mile was more susceptible than strains Q212 and Priscilla were. The heterogeneity of susceptibility to fluoroquinolones, chloramphenicol, and erythromycin was noted among the strains; all were resistant to amoxicillin and amikacin, and all were susceptible to rifampin, co-trimoxazole, tetracycline, and tetracycline analogs.Q fever is a widespread disease caused by Coxiella burnetii. Two major forms of the disease are known: acute and chronic, which includes endocarditis. The clinical form of the disease is related both to host factors (14) and to strain specificity (5). It has been reported that strains isolated from patients with endocarditis are different from those isolated from patients with the acute form of disease (22,23). Acute Q fever is a self-limiting disease, and bacteriostatic antibiotic therapy is sufficient in these cases in order to shorten the duration of disease. Chronic Q fever is a severe, frequently lethal disease in which C. burnetii survives and the disease can relapse despite years of antibiotic treatment (3, 6-8, 10, 13, 16-18, 26-31). In these cases, a bacteriocidal activity is also required, but such activity has not been shown by the use of single-drug therapy in vitro (14). For antibiotic susceptibility testing, embryonated eggs (2, 9, 24, 32) and cell cultures (34) have been used. These techniques are timeconsuming and conveniently allow the testing of only a few antibiotics with a few isolates. We describe here a new antibiotic testing method which determines the bacteriostatic activity of antibiotic compounds by the newly described shell-vial technique (19). By this method, we compared the bacteriostatic effects of 12 antibiotic compounds on 3 reference isolates and 10 new human isolates of C. burnetii.
MATERIALS AND METHODSC. burnetii isolates. The Nine Mile isolate, which is the reference isolate for acute infection (5), was obtained from 0. Baca (University of New Mexico, Albuquerque). The Q2...