Host-Associated Differentiation (HAD) is the formation of genetically divergent hostassociated lineages maintained by ecological isolation. HAD is potentially an important route to ecological speciation in parasites including many insects. While HAD case studies are accumulating, there is a dearth of negative results in the literature making it difficult to know how common the phenomenon really is or whether there are specific traits of parasites which promote HAD. To address these two problems, studies are needed which both publish negative results (i.e., parasites not showing HAD) and test for HAD in multiple parasite species on the same pair of host species (i.e., control for host plant effects). In this study, HAD was tested in three species of herbivorous insects and one parasitoid species on the same two host tree species: pecan and water hickory. The insects were selected based on the presence or absence of two traits, parthenogenesis and endophagy. A test for HAD was considered “positive” when population substructure was explained by host-association. To test for the presence of HAD, insects were sampled sympatrically to eliminate geographical isolation as a confounding factor, sampling was replicated spatially to assure that HAD persisted, and multiple loci were sampled from each individual. Genetic data was analyzed using cluster analyses. HAD was found in both pecan leaf phylloxera and yellow pecan aphid but not in pecan bud moth or in the parasitoid of the yellow pecan aphid, Aphelinus perpallidus. Interestingly, both taxa showing HAD are parthenogenetic and both taxa not showing HAD reproduce sexually. Species showing HAD were tested for the presence of a pre-mating reproductive isolating mechanism (RIM) which could be maintaining HAD despite the potential for gene flow. Selection against migrants to the alternative host was tested in yellow pecan aphid using a no-choice fitness experiment. The overall contribution of this RIM to total isolation was positive and ranged from 0.614 to 0.850. The RIM of “habitat preference” was tested in pecan leaf phylloxera using a dual-choice preference experiment. In this species, preference was only detected for phylloxera originating from water hickory suggesting that host discrimination ability may be a less important factor promoting differentiation in phylloxera.