Stefano Accoroni

Trends in Ostreopsis proliferation along the Northern Mediterranean coasts

Harmful benthic microalgae blooms represent an emergent phenomenon in temperate zones, causing health, ecological and economic concern. The main goal of this work was to compile records of Ostreopsis at large temporal and spatial scales, in order to study the relationship between cell abundances, the periodicity and intensity of the blooms and the role of sea water temperature in 14 Spanish, French, Monegasque and Italian sites located along the northern limits of the Mediterranean Sea. General trends were observed in the two considered basins: the north-western Mediterranean Sea, in which higher cell abundances were mostly recorded in mid-summer (end of July), and the northern Adriatic Sea where they occur in early fall (end of September). The sea-water temperature does not seem to be a primary driver, and the maximal abundance periods were site and year specific. Such results represent an important step in the understanding of harmful benthic microalgae blooms in temperate areas, and provide a good base for policy makers and managers in the attempt to monitor and forecast benthic harmful microalgae blooms.

Ostreopsis cf. ovata bloom in the northern Adriatic Sea during summer 2009: Ecology, molecular characterization and toxin profile

Intense blooms of the benthic dinoflagellate Ostreopsis cf. ovata have occurred in the northern Adriatic Sea since 2006. These blooms are associated with noxious effects on human health and with the mortality of benthic organisms because of the production of palytoxin-like compounds. The O. cf. ovata bloom and its relationships with nutrient concentrations at two stations on the Conero Riviera (northern Adriatic Sea) were investigated in the summer of 2009. O. cf. ovata developed from August to November, with the highest abundances in September (1.3×10(6) cells g(-1) fw corresponding to 63.8×10(3) cells cm(-2)). The presence of the single O. cf. ovata genotype was confirmed by a PCR assay. Bloom developed when the seawater temperature was decreasing. Nutrient concentrations did not seem to affect bloom dynamics. Toxin analysis performed by high resolution liquid chromatography-mass spectrometry revealed a high total toxin content (up to 75 pg cell(-1)), including putative palytoxin and all the ovatoxins known so far.

New approach using the real-time PCR method for estimation of the toxic marine dinoflagellate Ostreopsis cf. ovata in marine environment.

Background We describe the development and validation of a new quantitative real time PCR (qrt-PCR) method for the enumeration of the toxic benthic dinoflagellate Ostreopsis cf. ovata in marine environment. The benthic Ostreopsis sp. has a world-wide distribution and is associated during high biomass proliferation with the production of potent palytoxin-like compounds affecting human health and environment. Species-specific identification, which is relevant for the complex of different toxins production, by traditional methods of microscopy is difficult due to the high morphological variability, and thus different morphotypes can be easily misinterpreted. Methodology/Findings The method is based on the SYBR I Green real-time PCR technology and combines the use of a plasmid standard curve with a “gold standard” created with pooled crude extracts from environmental samples collected during a bloom event of Ostreopsis cf. ovata in the Mediterranean Sea. Based on their similar PCR efficiencies (95% and 98%, respectively), the exact rDNA copy number per cell was obtained in cultured and environmental samples. Cell lysates were used as the templates to obtain total recovery of DNA. The analytical sensitivity of the PCR was set at two rDNA copy number and 8.0×10−4 cell per reaction for plasmid and gold standards, respectively; the sensitivity of the assay was of cells g−1 fw or 1−1 in macrophyte and seawater samples, respectively. The reproducibility was determined on the total linear quantification range of both curves confirming the accuracy of the technical set-up in the complete ranges of quantification over time. Conclusions/Significance We developed a qrt-PCR assay specific, robust and high sample throughput for the absolute quantification of the toxic dinoflagellate Ostreopsis cf. ovata in the environmental samples. This molecular approach may be considered alternative to traditional microscopy and applied for the monitoring of benthic toxic microalgal species in the marine ecosystems.

Monitoring toxic microalgae Ostreopsis (dinoflagellate) species in coastal waters of the Mediterranean Sea using molecular PCR-based assay combined with light microscopy.

A molecular PCR-based assay was developed and applied to macrophyte and seawater samples containing mixed microphytobenthic and phytoplanktonic assemblages, respectively, in order to detect toxic Ostreopsis species in Mediterranean Sea. The specificity and sensitivity of the molecular PCR assay were assessed with both plasmidic and genomic DNA of the target genus or species using taxon-specific primers in the presence of background macrophyte DNA. The PCR molecular technique allowed rapid detection of the Ostreopsis cells, even at abundances undetectable within the resolution limit of the microscopy technique. Species-specific identification of Ostreopsis was determined only by PCR-based assay, due to the inherent difficulty of morphological identification in field samples. In the monitoring of the toxic Ostreopsis blooms PCR-based methods proved to be effective tools complementary to microscopy for rapid and specific detection of Ostreopsis and other toxic dinoflagellates in marine coastal environments.

Genetic diversity of the genus Ostreopsis Schmidt: Phylogeographical considerations and molecular methodology applications for field detection in the Mediterranean Sea

Abstract This study reports some recent phylogeographical considerations on the genus Ostreopsis distribution worldwide, with particular attention to the Mediterranean Sea, and new recent advances on the quali-quantitiative detection of Ostreopsis species along coastal areas of the Mediterranean Sea based on the PCR and quantitative real time PCR (qrt-PCR) assays. It was found that O. cf. ovata is widely dispersed throughout tropical and warm temperate coastal areas. In the Atlantic/Mediterranean region it represents a panmictic population that is highly divergent from Indo-Pacific populations. Furthermore, we demonstrated that the developed qrt-PCR assay is specific, robust and high sample throughput for the quantification of the toxic O. cf. ovata in the environmental samples. This molecular approach may be considered alternative to traditional methods of microscopy and applied for the survey of benthic toxic microalgal species in marine ecosystems.

Ecology of Ostreopsis cf. ovata Blooms in the Northwestern Adriatic Sea

Abstract The ecology of Ostreopsis cf. ovata blooms was investigated to evaluate the role of environmental factors (temperature, hydrodynamism, nutrient concentrations, depth and substratum) on the bloom dynamics. This paper reports the present knowledge on O. cf. ovata blooms along the Conero Riviera (NW Adriatic Sea), on the basis of samplings carried out from 2006 to 2010. The annual maximum of benthic cell abundance was always observed in late-summer, reaching the order of magnitude of 106 cells g-1 fw (corresponding to 107 cells g-1 dw and 104 cells cm-2) on macrophyte samples. Comparing the mean abundances settled on seaweeds with those growing on hard substrata, significantly higher abundances were observed on the latter. Hydrodynamism plays a major role in Ostreopsis blooms, as significantly higher abundances were observed in sheltered sites compared with exposed ones. The abundances of O. cf. ovata showed a marked decrease with depth. Temperature and nutrients do not seem to play an important effect on the O. cf. ovata blooms. High levels of ovatoxins were recorded in natural samples; episodes of death of both benthic invertebrates (limpets, sea urchins and mussels) and macroalgae were commonly observed during O. cf. ovata blooms.

Ostreopsis fattorussoi sp. nov. (Dinophyceae), a new benthic toxic Ostreopsis species from the eastern Mediterranean Sea.

The new benthic toxic dinoflagellate, Ostreopsis fattorussoi sp. nov., is described from the Eastern Mediterranean Sea, Lebanon and Cyprus coasts, and is supported by morphological and molecular data. The plate formula, Po, 3′, 7″, 6c, 7s, 5‴, 2′′′′, is typical for the Ostreopsis genus. It differs from all other Ostreopsis species in that (i) the curved suture between plates 1′ and 3′ makes them approximately hexagonal, (ii) the 1′ plate lies in the left half of the epitheca and is obliquely orientated leading to a characteristic shape of plate 6″. The round thecal pores are bigger than the other two Mediterranean species (O. cf. ovata and O. cf. siamensis). O. fattorussoi is among the smallest species of the genus (DV: 60.07 ± 5.63 μm, AP: 25.66 ± 2.97 μm, W: 39.81 ± 5.05 μm) along with O. ovata. Phylogenetic analyses based on the LSU and internal transcribed spacer rDNA shows that O. fattorussoi belongs to the Atlantic/Mediterranean Ostreopsis spp. clade separated from the other Ostreopsis species. Ostreopsis fattorussoi produces OVTX‐a and structural isomers OVTX‐d and ‐e, O. cf. ovata is the only other species of this genus known to produce these toxins. The Lebanese O. fattorussoi did not produce the new palytoxin‐like compounds (ovatoxin‐i, ovatoxin‐j1, ovatoxin‐j2, and ovatoxin‐k) that were previously found in O. fattorussoi from Cyprus. The toxin content was in the range of 0.28–0.94 pg · cell−1. On the Lebanon coast, O. fattorussoi was recorded throughout the year 2015 (temperature range 18°C–31.5°C), with peaks in June and August.

Effects of the bloom of harmful benthic dinoflagellate Ostreopsis cf. ovata on the microphytobenthos community in the northern Adriatic Sea

Composition and temporal variation of the microphytobenthos communities of the Conero Riviera (northern Adriatic Sea) were investigated in the course of an annual cycle, focusing on their relationships with blooms of the benthic dinoflagellate Ostreopsis cf. ovata. Sampling was carried out from March 2009 to March 2010 on undisturbed benthic substrata (macroalgae and pebbles). Samples for the study of microphytobenthos were collected with a monthly frequency, while those for the study of Ostreopsis bloom weekly. Benthic diatoms dominated the microphytobenthos communities for most of the annual cycle (except the summer), both in terms of abundance and biomass. In summer, cyanobacteria were dominant (54.04±9.18 and 24.29±11.11% of total abundance and biomass, respectively), while benthic dinoflagellates were an important component of the community in terms of biomass only at the peak of the Ostreopsis bloom in late summer (up to 91% of the total biomass). Among diatoms, the most abundant forms throughout the year were motile species (77.5±3.71% of the population), while erect diatoms formed the majority of the biomass in winter and spring (48.66±16.66 and 48.05±5.56% of total population, respectively). Diatoms were mainly affected by DIN availability, while the patterns of biomass of O. cf. ovata and cyanobacteria were related to salinity and temperature. The biomass of Ostreopsis was also affected by the availability of phosphorus. The results of this study suggest that the proliferation of Ostreopsis affected the structure of the benthic diatom community: motile diatoms were significantly more abundant during the Ostreopsis bloom peak than during the rest of summer, probably because they benefited from the abundant mucilaginous mat covering the benthic substrata. In the course of the O. cf. ovata bloom the diversity of the microphytobenthos was significantly lower than during the rest of the year, suggesting an influence of both the shading produced by the mucous mat and allelopathic compounds possibly produced by O. cf. ovata.

Consequences of Stinging Plankton Blooms on Finfish Mariculture in the Mediterranean Sea

In recent years, caged finfish mariculture across European seas suffered production losses by severe fish mortality, following episodic outbreaks of invertebrate cnidarian stingers. Due to their stinging cells and injectable venoms, medusozoan jellyfish or drifting propagules of polyp colonies at high density may impair caged fish health through toxic effects on vulnerable tissues of gills and skin, and related secondary bacterial infections. Gill disorders in European sea bass (Dicentrarchus labrax) fish farms along the Spanish Mediterranean coast are commonly reported, but regular monitoring of the frequency of cnidarian outbreaks and their potential impacts on caged fish is still poorly enforced. In this study, two sea bass mariculture farms in Southern Spain (Malaga; Almeria) were monitored biweekly for zooplankton, phytoplankton and fish gills condition, over 13 or 30 months for the Malaga and Almeria facilities respectively, within the period 2012 – 2014. Significant, direct correlations were found among low water temperature, recorded fish mortalities, and high abundances of planktonic cnidarians, particularly of the hydrozoan siphonophores Muggiaea atlantica and M. kochii, and the larval stage of Ectopleura larynx, a common member of cage biofouling communities. A significant relationship between cnidarian densities and the quantitative scoring of gill pathology was also observed. In addition, high densities of long-bristled planktonic diatoms (Chaetoceros spp.) coincided with a major fish mortality event (April 2012, Almeria farm). Standardised monitoring of plankton dynamics and composition may help in promoting response capacities of Mediterranean mariculture managers to fish health challenges (such as stinging plankton blooms) by a) improving diagnostic tools and preventative countermeasures and b) supporting the development of science-based spatial planning and sustainable growth of coastal mariculture.

Plankton dynamics across the freshwater, transitional and marine research sites of the LTER-Italy Network: patterns, fluctuations, drivers

A first synoptic and trans-domain overview of plankton dynamics was conducted across the aquatic sites belonging to the Italian Long-Term Ecological Research Network (LTER-Italy). Based on published studies, checked and complemented with unpublished information, we investigated phytoplankton and zooplankton annual dynamics and long-term changes across domains: from the large subalpine lakes to mountain lakes and artificial lakes, from lagoons to marine coastal ecosystems. This study permitted identifying common and unique environmental drivers and ecological functional processes controlling seasonal and long-term temporal course. The most relevant patterns of plankton seasonal succession were revealed, showing that the driving factors were nutrient availability, stratification regime, and freshwater inflow. Phytoplankton and mesozooplankton displayed a wide interannual variability at most sites. Unidirectional or linear long-term trends were rarely detected but all sites were impacted across the years by at least one, but in many case several major stressor(s): nutrient inputs, meteo-climatic variability at the local and regional scale, and direct human activities at specific sites. Different climatic and anthropic forcings frequently co-occurred, whereby the responses of plankton communities were the result of this environmental complexity. Overall, the LTER investigations are providing an unparalleled framework of knowledge to evaluate changes in the aquatic pelagic systems and management options.