In vitro susceptibilities of Ehrlichia risticii to eight antibiotics

Inhibition of the proliferation of Ehrlichia risticii cultured in murine macrophage P388D1 cells by eight antibiotics was evaluated by indirect fluorescent-antibody staining with an antiserum specific to E. risticii. There was a negative correlation between the percentage of infected cells and the log10 of the concentrations of all antibiotics examined. The ranks of the antibiotics in the order of 50% inhibitory concentrations (on a microgram-per-milliliter basis) after 48 h of exposure were as follows: demeclocycline, doxycycline, and oxytetracycline less than minocycline less than rifampin less than tetracycline less than erythromycin and nalidixic acid. When the antibiotics were removed after 48 h of incubation, continuous inhibition of proliferation was evident at 72 h. At 96 h regrowth of the organisms occurred in most of the cultures. The rate of regrowth was the highest with nalidixic acid, followed by erythromycin, at all concentrations of the antibiotic tested. Regrowth was observed with less than 0.1 microgram of minocycline per ml and less than 0.01 microgram of oxytetracycline, tetracycline, and doxycycline per ml. With more than 0.01 microgram of demeclocycline per ml, however, the inhibition persisted for up to 72 h after removal of the antibiotic. These results indicate that demeclocycline was slightly more effective than doxycycline, oxytetracycline, and minocycline in eliminating E. risticii in macrophages in vitro, whereas tetracycline and rifampin were less effective. Nalidixic acid and erythromycin were ineffective.