This is the first study that presents concentrations of domoic acid detected in the whole shellfish tissue from breeding and harvesting areas along the Croatian coast of the Adriatic Sea during the period 2006 to 2008. Shellfish sample analyses after SAX cleaning procedures, using a UV-DAD-HPLC system, showed the presence of domoic acid in four species. The most prevalent of those species were the blue mussel (Mytilus galloprovincialis), followed by European flat oyster (Ostrea edulis), Mediterranean scallop (Pecten jacobaeus) and proteus scallop (Flexopecten proteus). Domoic acid, a potentially lethal phycotoxin that causes amnesic shellfish poisoning (ASP), was detected for the first time in January 2006 with the highest value of 6.5486 μg g^-1 in whole shellfish tissue. Pseudo-nitzschia spp. bloom events preceded these high domoic acid concentrations. According to this study, retention of domoic acid in the blue mussel M. galloprovincialis is more than 42 days. This investigation indicates the first presence of domoic acid in Croatian shellfish, but in concentrations under the regulatory limit (20 μg g^-1), therefore shellfish consumption was not found to endanger human health.

In September 2015, a massive occurrence of the Ostreopsis species was recorded in central Adriatic Kaštela Bay. In order to taxonomically identify the Ostreopsis species responsible for this event and determine their toxin profile, cells collected in seawater and from benthic macroalgae were analyzed. Conservative taxonomic methods (light microscopy and SEM) and molecular methods (PCR-based assay) allowed the identification of the species Ostreopsis cf. ovata associated with Coolia monotis. The abundance of O. cf. ovata reached 2.9 × 10⁴ cells L⁻¹ in seawater, while on macroalgae, it was estimated to be up to 2.67 × 10⁶ cells g⁻¹ of macroalgae fresh weight and 14.4 × 10⁶ cells g⁻¹ of macroalgae dry weight. An indirect sandwich immunoenzymatic assay (ELISA) and liquid chromatography–high-resolution mass spectrometry (LC-HRMS) were used to determine the toxin profile. The ELISA assay revealed the presence of 5.6 pg palytoxin (PLTX) equivalents per O. cf. ovata cell. LC-HRMS was used for further characterization of the toxin profile, which showed that there were 6.3 pg of the sum of ovatoxins (OVTXs) and isobaric PLTX per O. cf. ovata cell, with a prevalence of OVTXs (6.2 pg cell⁻¹), while the isobaric PLTX concentration was very low (0.1 pg cell⁻¹). Among OVTXs, the highest concentration was recorded for OVTX-a (3.6 pg cell⁻¹), followed by OVTX-b (1.3 pg cell⁻¹), OVTX-d (1.1 pg cell⁻¹), and OVTX-c (0.2 pg cell⁻¹).

Dinophysis spp. blooms and related shellfish toxicity events of diarrhetic shellfish poisoning (DSP) have been the most reported toxicity event through the Croatian National monitoring program. With the aim to characterize the DSP toxin profile in shellfish farmed in Croatia, for the first time a complete analysis of the toxin profile of Croatian mussels has been carried out using the LC-MS/MS technique. The obtained results showed okadaic acid (OA) as the main toxin contaminating Croatian mussels at that time. The maximum concentration of OA in shellfish tissue was recorded 12 days after the Dinophysis fortii bloom, thus suggesting that rapid growth of the toxin level in the shellfish occurred in the first week after the bloom while it was slower in the second week. Furthermore, the presence of only OA at concentrations which could endanger human health suggests D. fortii as the main organism responsible for the toxic event that occurred in Lim Bay. The presence of gymnodimine and spirolides in Croatian mussel has been detected for the first time, while the presence of yessotoxin and pectenotoxin-2 is confirmed.

The Krka River estuary is a karstic, permanently stratified estuary due to the strong freshwater inflow. It is a special environment in which to study the phytoplankton community, especially because this area is an important aquaculture site. Among other potentially toxic phytoplankton species, the diatom genus Pseudo-nitzschia occurs frequently and is a potential source of domoic acid (DA), causing shellfish toxicity and human intoxication. The main objective was to examine the dynamics of the phytoplankton community and, in particular, the genus Pseudo-nitzschia in the upper part of the Krka estuary, through monthly sampling over two years. The phytoplankton community was analysed using light microscopy and scanning electron microscopy to determine the diversity of Pseudo-nitzschia species and characterise the environmental parameters associated with a high abundance of Pseudo-nitzschia species. Seven Pseudo-nitzschia species were identified in the investigation, with higher frequencies and abundances in the less variable layer, at a 7 m depth. Blooms of Pseudo-nitzschia were noted in the late summer/early autumn, dominated by P. delicatissima/arenysensis. Winter assemblages were characterised by P. pseudodelicatissima/cuspidata, P. calliantha, and P. subfraudulenta, and were associated with domoic acid occurrence in shellfish.

With the aim of investigating whether yessotoxin (YTX) is responsible for diarrhetic shellfish poisoning (DSP) events in Croatian waters, three different methods were combined: a modified mouse bioassay (MBA) that discriminates YTX from other DSP toxins, the enzyme-linked immunosorbent assay method (ELISA) and liquid chromatography-mass spectrometry (LC-MS/MS). Among 453 samples of mussels and seawater analyzed in 2007, 10 samples were DSP positive. Results obtained by the modified MBA method revealed that most of the samples were positive for YTX, with the exception of samples from Lim Bay (LB 1) The ELISA method also identified the presence of YTX in these samples. DSP toxin profiles showed the presence of okadaic acid (OA) in three, and YTX in four out of nine samples that were analyzed by LC-MS/MS. The phytoplankton community structure pattern revealed Lingulodinium polyedrum (Stein) Dodge, which was present in the water prior to and/or during toxicity events at low concentrations (80 to 1440 cells L^-1), as a potential YTX producing species. It is proposed that L. polyedrum cells accumulated in mussels and the subsequently observed toxicity may be related to metabolism after ingestion, resulting in carboxy YTX as the major analog in the mussel.