ABSTRAGT An existing microsatellite genotype database has been used for several years in population genetic assignment analyses of boll weevils, Anthononiu.'í grandis grandis-Boheman (GOleoptera: Gurculionidae), captured in eradication zones. It is important to update it in case of changes in genotype frequency at any of the locations over time. Such changes at neutral loci could be caused by drift, immigration, or population bottlenecks. We examined alíele frequency distribution for 10 microsatellite loci to determine genetic differentiation among 10 boll weevil populations sampled from Texas and Mexico in 2009. In addition, temporal changes in genetic composition were examined in the eigbt populations for which samples were available from previous years. Svibstantial levels of spatial genetic structvu'e were observed, witb the 10 populations clustering as four major gi-oups. Pairwise F^^estimates in 2009 samples ranged from 0.001 (College Station-Cameron) to 0.492 (College Station-Ojinaga). There was little change in genetic profiles over time at four of the eigbt locations. Thus, for those four locations, genotype and alíele fi'equency data can be pooled over the two sample dates, which will provide greater statistical power in future population assignment tests. However, genetic profiles changed substantially at Ojinaga, and to a lesser extent at Uvalde, Cameron, and Rosales, so tlie 2009 genotype data should be substituted in futvn-e analyses. Finally, populations from two new locations, Brownsville and Lockhart, TX, were sampled, genotyped, and added to the database. The addition of Lockhart is particularly important given its suiprisingly high diffeientiation from the relatively neai'by populations of Cameron and Uvalde.