The third serotype of human rotavirus shares neutralization specificity with rotaviruses derived from monkeys, dogs, cats, horses, rabbits, and mice. In addition, viruses of this serotype include strains of all known subgroup specificities (i.e., I, II, neither I nor II, both I and II). The relationships of the major neutralization protein, VP7 of the various serotype 3 strains were analyzed by testing their reactivity with VP7 monoclonal antibodies (mAbs) and by comparing nucleotide and amino acid sequence. This analysis was performed in an attempt to clarify antigenic relationships and to better understand to evolution of serotype 3 rotavirus stains which have established an extraordinarily broad host range.