In the present paper we report of the results of an immunohistochemical investigation to assess tissue tropism and viral replication in developing chicken, turkey, Muscovy duck and mallard duck embryos, of Italian H7N1 isolates belonging to different genetic lineages. LPAI isolates were chosen on the basis of the location in the phylogenetic tree: a progenitor strain, A/ty/Italy/977/V99, (exhibiting no additional glycosylation site, nAGS), strain A/ty/Italy/2379/V99 (AGS in position 123) and strain A/ty/Italy/3675/V99 (AGS in position 149) were selected. The latter two strains belonged to distinct lineages originating from the pool of progenitor strains. HPAI isolate A/ty/Italy/4580/V99 was also included in the study. All the embryos tested supported the growth of HPAI. The LPAI isolates replicated readily in the allantoic layer of the CAM of all the species tested, and did not grow in the developing chicken, turkey and Muscovy duck embryos. In contrast, they replicated to different extents in the respiratory tract of the developing mallard embryo, which also presented lower mortality rates than the other species. We conclude from these findings that the pathogenesis of LPAI infections in mallard embryos is different to that observed in other species, and should be investigated further.