Anadromous Atlantic salmon returning to the Connecticut River (CR) from 1996 to 1999 were assayed for variability at nine microsatellite DNA loci. Heterozygosity and allele frequencies were compared to the anadromous Atlantic salmon returning to Maine's Penobscot River from 1998 to 2000. The Penobscot River was the primary source of the salmon used to found the previously extirpated population in the Connecticut River. While there were no significant differences in heterozygosity between the source population and the Connecticut River sea-run spawners, microsatellite allele frequencies were significantly different between the populations. Two techniques of estimating effective population size (Ne) suggested a healthy level of genetic variation in the Connecticut River population of anadromous Atlantic salmon. This is significant because the sea-run population is maintained almost entirely through hatchery production. Healthy ratios of Ne to N indicate that hatchery production has not resulted in excessive inbreeding to date. ?? 2003 Elsevier B.V. All rights reserved.