Ceftiofur (XNL) and its primary metabolite, desfuroylceftiofur (DXNL), were evaluated for in vitro activity against 539 isolates from veterinary sources. Actinobacillus pleuropneumoniae, Pasteurella spp., Haemophilus somnus, Salmonella spp., Escherichia coli, staphylococci, and streptococci were tested. Overall, XNL and DXNL were equivalent in activity against the gram-negative organisms with all minimum inhibitory concentrations (MICs) within 1 serial dilution. Against the staphylococci, MIC difference of 2-3 serial dilutions were detected with an MIC90 for XNL and DXNL of 1.0 and 4.0-8.0 micrograms/ml, respectively. Although the MIC90 obtained for Streptococcus suis for each compound was within 1 dilution, the MIC values against individual strains were 2-3 dilutions greater for DXNL than for XNL. The MICs obtained with the bovine and equine streptococci for DXNL (MIC90 = 0.03 microgram/ml) were 5 serial dilutions higher than those obtained for XNL (MIC90 < or = 0.0019). Although DXNL was less active than XNL against the streptococci, these differences were not clinically important because both XNL and DXNL were highly active for these bacteria. Although these differences are of little importance with the streptococci, they may have important implications for susceptibility testing of the staphylococci. In conclusion, with the exception of the staphylococci, both XNL and DXNL were highly active against the organisms tested, with MICs for both compounds several fold lower than plasma levels achieved during dosing of XNL.