Abstract A Murray Valley encephalitis virus (MVE) field isolate of high neuroinvasiveness (BH3479) and a neutralization escape variant of low neuroinvasiveness (BHv1) selected from BH3479 (which differ by a single amino acid at residue 277 in the envelope glycoprotein) were examined for their distribution in the tissues of weanling Swiss mice at various times after footpad inoculation. BH3479 was first detected in lymph nodes draining the inoculated limb at 24 hr postinoculation (pi) and was found in serum between 36 and 72 hr pi. BH3479 was first detected in the central nervous system (CNS) at 4 days pi and reached maximum CNS titers (>10 9PFU/g) between 6 and 9 days pi. All BH3479-infected mice developed encephalitis and died before 10 days pi. In contrast, BHv1 was not detected in lymph nodes draining the footpad at any time after inoculation; BHv1 was first detected in the serum between 60 and 72 hr pi—24 hr later, and at a 20-fold lower titer than for BH3479. BHv1 was first detected in the CNS at 7 days pi 3 days later and at a 300-fold lower titer than for BH3479. After 10 days pi, BHv1 could not be isolated from the CNS or from other host tissues. Most BHv1-infected mice experienced a subclinical infection; the mortality rate from BHv1 infection was less than 1%. Both viruses appeared to enter the CNS via the olfactory lobes. BH3479 spread throughout the CNS in a rostral to caudal direction over 3–4 days. In contrast, BHv1 infection in the CNS was restricted to the olfactory lobes and adjacent structures of the forebrain.