AbstractMolecular tools increasingly refine and improve the identification of zooplankton organisms based on phenotypic features, providing a more robust and comprehensive species description. Integration of data helps revealing the hidden diversity of zooplankton and facilitating the detection of rare and non‐indigenous species. This approach, merging morphological characters and a diagnostic marker for specific identification, such as the mitochondrial cytochrome c oxidase I (COI), is here used to characterize key taxa from the zooplankton assemblage of the Gulf of Naples at the Long‐Term Ecological Research site MareChiara (LTER‐MC) (Central Tyrrhenian Sea, Western Mediterranean Sea). Zooplankton biodiversity assessment using integrated taxonomy was focused on selected crustacean groups: cyclopod copepods (Agetus typicus, Oithona plumifera, Oncaea mediterranea, Oncaea scottodicarloi); newly records of cladocerans (Evadne nordmanni), euphausiids (Euphausia krohnii, Nematoscelis megalops, Nyctiphanes spp.) and sergestids (Lucifer typus), with the aim to boost the knowledge of real zooplankton biodiversity. The results of our investigation provide new high‐quality molecular references of the analysed taxa and contribute to unveiling the genetic diversification of zooplankton species and their relevant ecological significance for Mediterranean coastal waters.