In vitro activity of ceftiofur and six other antimicrobial agents was assessed for 516 Streptococcus equi subsp zooepidemicus isolates collected from horses with lower respiratory tract infections in North America in 2007 and 2008 and 239 equine S. equi subsp zooepidemicus isolates received from US and Canadian veterinary diagnostic laboratories between 1989 and 2007. The lowest concentration of ceftiofur inhibiting the growth of 90% of the isolates (MIC90) was 0.12 microg/ml for both groups of isolates. The Clinical and Laboratory Standards Institute susceptible breakpoint set for ceftiofur against this organism is a minimal inhibitory concentration value of ≤ 0.25 microg/ml. The MIC90 values remained consistent for isolates collected over 19 years.