The extent to which host biology, ecology and phylogeny determine the diversity of macroparasite assemblages has been investigated in recent years in several taxa, including fish. However, consensus has not been reached probably as a result of data being collected from different sources, different temporal scales or host and parasite biogeography and phylogeny having greater influence than expected. The present study evaluates the relative importance of 27 biological, ecological and phylogenetic characteristics of 14 flatfish species on the diversity of their ecto- and endoparasite assemblages, comprising a total of 53 taxa. Redundancy analyses were applied to the mean abundance of each parasite taxa infecting each host and to the richness, taxonomic distinctness and variance in taxonomic distinctness calculated for each assemblage within each host. Only a few host characteristics contributed significantly to the observed patterns: host distribution was more important in determining the type and mean abundance of ectoparasites present in an assemblage, whereas diversity of these assemblages were mainly related to the host's maximum size. Endoparasite mean abundance and diversity were mostly influenced by the number of food items ingested and by the presence of Crustacea and Polychaeta in the diet. However, the sympatric occurrence of related hosts also played an important role in the diversity values found in macroparasite assemblages. Results showed that a host characteristic has different importance according to the host-parasite relationship being examined, suggesting an important role for host-parasite co-evolution on the diversity of extant macroparasite assemblages.