An important epidemiological tool in the control of epidemics of Foot-and-mouth disease (FMD) is genetic tracing using complete virus genome sequence data. However to interpret these genetic data, it is important to quantify underlying variation present in FMDV populations from individual tissue samples. Cloned complete capsid sequences from two virus populations from epithelium from a cow (n=26) and from a sheep (n=15) infected during the UK 2001 outbreaks were generated. Genetic diversity of the two virus populations differed significantly, with sequences representing virus from the cow having a mutation frequency of 2.79 x 10(-4) mutations per nucleotide sequenced (mpns) and those from the sheep having 3.94 x 10(-4) mpns (chi(2)=8.24, P=0.004). The dN/dS ratio of sequences from the cow was higher (1.228) than that from the sheep (0.187) although not significantly so. The sequences from the cow epithelium exhibited significantly higher than expected number of changes within neutralising antigenic sites (P=0.0007). The performance of two different reverse transcriptase enzymes was found not to differ with respect to the frequency (P=0.559, chi(2)=0.341) or dN/dS ratio (P=0.863, chi(2)=0.03) of the mutations observed. These data provide insight into the population diversity that exists within a single lesion and help toward understanding the mechanisms that underpin sequence evolution of FMDV.