Two decades of data on the neurotoxic Gymnodinium catenatum dynamics and paralytic shellfish toxins contamination of molluscs in the southwestern Mediterranean: What have we learned?

Harmful algal blooms have been documented in the Moroccan Western Mediterranean region since 1993, primarily associated with the presence of paralytic shellfish toxins (PSTs) produced by the dinoflagellate Gymnodinium catenatum. The proliferation of this neurotoxic species has led to recurring bans on the harvesting of molluscs, resulting in significant socio-economic repercussions and threats to human health. In the present study, we examine the dynamics of G. catenatum and mollusc PST contamination patterns over a 20-year period (2002-2021) in two distinct marine ecosystems: M'diq Bay and the Oued Laou Estuary. For the PST contamination, we considered two commercially important shellfish species: the smooth clam, Callista Chione, and the cockle, Acanthocardia tuberculata. The highest G. catenatum abundances were consistently observed from November to February in both sites. Our data revealed inter-annual variations in G. catenatum abundance, peaking at 91,840 cells.L-1 in November 2011. PST contamination levels in A. tuberculata were significantly higher than those observed in C. chione. Furthermore, we identified a significant correlation (Pearson, P-value <0.05) between PST contamination of smooth clams and the abundance of G. catenatum. The contamination of A. tuberculata by PSTs reached very high levels, with up to 13,500 µg STX di-HCl eq. kg-1 of shellfish meat, exceeding the established safety thresholds by 16-fold. Additionally, there has been an increase in the prevalence and incidence of PSTs over the years. Notably, we observed a substantial increase in G. catenatum blooms and PST events in the Western Mediterranean during the last decade (2010-2021). The examined data suggest that rainfall could play a pivotal role in G. catenatum bloom dynamics by enriching marine waters with nutrients. The statistical model selection approaches indicated that nutrient concentrations (i.e., nitrate and phosphorus) were the most significant parameters for G. catenatum blooms in the studied area.

New insights into the dynamics of causative dinoflagellates and the related contamination of molluscs by paralytic toxins in the southwestern Mediterranean coastal waters of Morocco.

The distribution of the two potentially toxic dinoflagellates Gymnodinium catenatum and Alexandrium spp. was investigated in the Mediterranean Moroccan Sea from March 2018 to March 2019. The cockle Acanthocardia tuberculata and the smooth clam Callista chione were collected at four stations, and their toxin levels were assessed using the mouse bioassay. The toxin profile was analysed by LC-MS/MS in G. catenatum and in the bivalves harvested in M'diq and Djawn. The species G. catenatum was present throughout the year, whereas Alexandrium spp. was less abundant. The paralytic shellfish toxin (PST) level in cockles was, on average, six times above the sanitary threshold; GTX5 was the major contributor to the total PST level, followed by dc-STX and STX. The toxin level of the smooth clam was considerably lower than that of the cockle; GTX5 and C-toxins were the dominating analogues. Our results suggest the responsibility of G. catenatum for the recurrent PST contamination in the Moroccan Mediterranean Sea, with a west-east gradient.

Development of harmful algal blooms species responsible for lipophilic and amnesic shellfish poisoning intoxications in southwestern Mediterranean coastal waters.

Mediterranean waters have undergone environmental changes during the last decades leading to various modifications of the structure of phytoplankton populations, especially Harmful Algal Blooms (HABs) species. Monitoring of the potentially toxic phytoplankton species was carried out biweekly in the western Mediterranean coast of Morocco from March 2018 to March 2019. Lipophilic Shellfish Toxins (LSTs) using LC-MS/MS and Domoic Acid (DA) using HPLC-UV were measured in the exploited mollusks, the cockle Acanthocardia tuberculata and the smooth clam Callista chione. We also determined the prevailing environmental factors in four surveyed sites (M'diq bay, Martil, Kaa Asras, and Djawn) selected to cover a variety of coastal ecosystems. Results showed that Pseudo-nitzschia spp. a DA producer species, was abundant with a pick of 50 × 103 cells l-1 on October 2018 in Djawn. Dinophysis caudata was the dominate Dinophysis species and showed a maximum density of 2200 cells l-1 on July in Djawn. Prorocentrum lima, an epibenthic dinoflagellate, appeared rarely in the water column with densities <80 cells l-1. Gonyaulax spinifera and Protoceratium reticulatum were found occasionally with a maximum density of 160 cells l-1. Karenia selliformis was detected only five times (<80 cells l-1) throughout the survey period. LC-MS/MS analyses revealed the presence of OA/DTX3, PTX-2, PTX-2 sa, and PTX-2 sa epi in the cockle at concentrations of up to 44.81 (OA/DTX-3+PTXs) ng g-1 meat. GYM-A was detected in the clam at concentrations of up to 4.22 ng g-1 meat. For the first time, AZAs and YTXs were detected in the southwestern Mediterranean with maximum values of 2.49 and 10.93 ng g-1 meat of cockle, respectively. DA was detected in moderate concentrations not exceeding 5.65 µg g-1 in both mollusks. Results showed that the observed toxic algae in the water column were responsible from the analysed toxins in the mollusks. It is likely that the southwestern Mediterranean waters could see the development of emergent species producing potent toxins (YTXs, AZAs, GYM-A). These dinoflagellates have to be isolated, ribotyped, and their toxin profiles determined.