Two types of vaccine-derived polioviruses have been recently designated to emphasize the different origins of the evolved viruses: circulating vaccine-derived polioviruses (cVDPV) associated with outbreaks of paralytic disease and strains isolated from chronically infected immunodeficient individuals (iVDPV). We describe here a type 3 VDPV (PV3/EST/02/E252; later E252) isolated from sewage collected in Tallinn Polioviruses, members of the Enterovirus genus in the Picornaviridae family, are important human pathogens causing the acute paralytic disease poliomyelitis. The worldwide program for eradication of wild-type poliovirus is coordinated by the World Health Organization (WHO). The program includes two simultaneous approaches: intensive immunizations mainly with the trivalent oral poliovirus vaccine (OPV) (35) and systematic surveillance of the remaining cases. The surveillance includes, most importantly, epidemiological and virological investigation of acute flaccid paralytic (AFP) cases. The AFP surveillance is supplemented in some countries by analysis of poliovirus circulation in human populations through investigation of wastewater specimens contaminated with human fecal materials (environmental surveillance) and/or by scrutinizing the results of routine virus diagnostics (enterovirus surveillance).The evolution rate of polioviruses is very high, partly due to the high error frequency in RNA synthesis: roughly 10 Ϫ4 per base pair per replication cycle. For wild polioviruses circulating in human populations, nucleotide substitutions accumulate at a rate of approximately 1% per year and consist primarily of changes at synonymous codon positions thus not leading to amino acid substitutions (22). The genetic diversity of poliovirus strains is exploited in molecular epidemiology, a key component of poliovirus surveillance (33), currently based on sequence analysis of the VP1 coding region of the genome. The