The in vitro antibiotic activity of CA(1-8)M(1-18), a synthetic cecropin A-melittin hybrid peptide, was determined by broth microdilution on 20 clinical Acinetobacter baumannii isolates with different resistance profiles. The MIC(50), MIC(90) and ranges were 4 mg/l, 4 mg/l and 2-8 mg/l, respectively, and were independent of resistance pattern. Different assay parameters such as microplate plastic (polystyrene or polypropylene), addition of supplements (5-10% fetal calf serum or 5% horse blood), inoculum size (10(5), 10(6), 10(7) and 10(8) CFU/ml) or incubation period (24 or 48 h) were studied. MIC was independent of the first two parameters, although the MIC values increased both with inoculum size or incubation period. Killing curves were obtained both for a standard strain and a multiresistant isolate over a 45.7-2.8 mg/l (16-1 mM) peptide range, using an initial inoculum of 10(5)-10(6) CFU/ml and 10(9)-10(10) CFU/ml. A concentration of 45.7 mg/l was required for complete killing. Accordingly, CA(1-8)M(1-18) showed good in vitro activity against the A. baumannii strains tested irrespective of the resistance to classical antibiotics, and could be a future candidate for multiresistant A. baumannii infections, although further cytotoxicity and pharmacological studies will be needed.