Harmful Ostreopsis cf. ovata blooms could extend in time span with climate change in the Western Mediterranean Sea.

Fast environmental changes and high coastal human pressures and impacts threaten the Mediterranean Sea. Over the last decade, recurrent blooms of the harmful dinoflagellate Ostreopsis cf. ovata have been recorded in many Mediterranean beaches. These microalgae produce toxins that affect marine organisms and human health. Understanding the environmental conditions that influence the appearance and magnitude of O. cf. ovata blooms, as well as how climate change will modify its future distribution and dynamics, is crucial for predicting and managing their effects. This study investigates whether the spatio-temporal distribution of this microalga and the frequency of its blooms could be altered in future climate change scenarios in the Mediterranean Western basin. For the first time, an ecological habitat model (EHM) is forced by physico-chemical climate change simulations at high-resolution, under the strong greenhouse gas emission trajectory (RCP8.5). It allows to characterize how O. cf. ovata may respond to projected conditions and how its distribution could shift over a wide spatial scale, in this plausible future. Before being applied to the EHM, future climate simulations are further refined by using a statistical adaptation method (Cumulative Distribution Function transform) to improve the predictions robustness. Temperature (optimum 23-26 °C), high salinity (>38 psu) and high inorganic nutrient concentrations (nitrate >0.25 mmol N·m-3 and phosphate >0.035 mmol P·m-3) drive O. cf. ovata abundances. High spatial disparities in future abundances are observed. Namely, O. cf. ovata abundances could increase on the Mediterranean coasts of France, Spain and the Adriatic Sea while a decrease is expected in the Tyrrhenian Sea. The bloom period could be extended, starting earlier and continuing later in the year. From a methodological point of view, this study highlights best practices of EHMs in the context of climate change to identify sensitive areas for current and future harmful algal blooms.

Ecophysiological responses of Ostreopsis towards temperature: A case study of benthic HAB facing ocean warming.

Reports of the benthic dinoflagellate Ostreopsis spp. have been increasing in the last decades, especially in temperate areas. In a context of global warming, evidences of the effects of increasing sea temperatures on its physiology and its distribution are still lacking and need to be investigated. In this study, the influence of temperature on growth, ecophysiology and toxicity was assessed for several strains of O. cf. siamensis from the Bay of Biscay (NE Atlantic) and O. cf. ovata from NW Mediterranean Sea. Cultures were acclimated to temperatures ranging from 14.5 °C to 32 °C in order to study the whole range of each strain-specific thermal niche. Acclimation was successful for temperatures ranging from 14.5 °C to 25 °C for O. cf. siamensis and from 19 °C to 32 °C for O. cf. ovata, with the highest growth rates measured at 22 °C (0.54-1.06 d-1) and 28 °C (0.52-0.75 d-1), respectively. The analysis of cellular content of pigments and lipids revealed some aspects of thermal acclimation processes in Ostreopsis cells. Specific capacities of O. cf. siamensis to cope with stress of cold temperatures were linked with the activation of a xanthophyll cycle based on diadinoxanthin. Lipids (neutral reserve lipids and polar ones) also revealed species-specific variations, with increases in cellular content noted under extreme temperature conditions. Variations in toxicity were assessed through the Artemia franciscana bioassay. For both species, a decrease in toxicity was observed when temperature dropped under the optimal temperature for growth. No PLTX-like compounds were detected in O. cf. siamensis strains. Thus, the main part of the lethal effect observed on A. franciscana was dependent on currently unknown compounds. From a multiclonal approach, this work allowed for defining specificities in the thermal niche and acclimation strategies of O. cf. siamensis and O. cf. ovata towards temperature. Potential impacts of climate change on the toxic risk associated with Ostreopsis blooms in both NW Mediterranean Sea and NE Atlantic coast is further discussed, taking into account variations in the geographic distribution, growth abilities and toxicity of each species.

From micro to mesoscale: Understanding the influence of macroalgal communities on Ostreopsis Schmidt blooms.

Harmful Algal Blooms (HABs) are increasing in temperate areas, and the growth rates of benthic harmful dinoflagellates may be favoured in the context of global climate change. Benthic dinoflagellates, including species belonging to the Ostreopsis Schmidt genus, are known to develop on the surface of macroalgae and different macroalgal morphotypes and communities could host higher or lower cell abundances. The physical structure of the macroalgal substrate at the small scale (cm, microhabitat scale) and the structural complexity of the macroalgal community at the medium scale (few m, mesohabitat scale) could play a relevant role in bloom facilitation: the hypothesis that Ostreopsis species could be associated with macroalgal turfs and shrubs, structurally less complex communities than canopy-forming macroalgae, is especially under discussion and, if confirmed, could link bloom occurrence to regime shifts in temperate ecosystems. The present study, performed in two locations of the Ligurian Sea (Rochambeau, France and Vernazzola, Italy) aimed at understanding marine vegetation's role at the micro and mesohabitat scales in controlling the distribution and abundance of Ostreopsis. The abundance of the microalgal cells was quantified at different spatial scales, from cm to a few m, on different macroalgal species and communities, including artificial substrates, to tease apart the micro and mesohabitat effects. The results obtained show a high spatio-temporal variability, potentially hiding habitat-related patterns. The substrate's preferences diminish when cell abundances are very high, as in the case of Rochambeau, while in presence of moderate cell abundances as in Vernazzola or the first phases of blooms, it is possible to appreciate differences in abundances among substrates (in our study, Dictyota fasciola (Roth) Lamouroux supporting higher abundances). Our results open new research topics such as the study of blooms at a larger scale (macrohabitat) and testing different sampling methods to standardise the cells' abundances independently on the substrate.

The benthic toxic dinoflagellate Ostreopsis cf. ovata in the NW Mediterranean Sea: Relationship between sea surface temperature and bloom phenology.

Blooms of the toxic benthic dinoflagellate Ostreopsis cf. ovata can induce ecological and human health issues in certain temperate areas. In order to prevent these negative effects, long-term monitoring studies of O. cf. ovata blooms have been conducted in several impacted areas to have a comprehensive understanding of bloom dynamics and efficient tools for risk management. O. cf. ovata blooms were monitored every summer (from mid-June to the end of August) on five identified sites in Larvotto beach (Monaco, NW Mediterranean Sea), between 2007 and 2019. This time-series represents one of the largest time-series in the world describing blooms of this species. Bloom phenological features (timing, duration, maximum cell abundance and growth rate), were found to be highly variable throughout the studied period, and were analyzed as a function of different hydroclimatic parameters, including sea surface temperature (SST). The highest net growth rates were related to temperatures ranging between 21°C and 25°C, and did not coincide with maximal temperature records (27.5°C). Such results suggest that, although global warming possibly influences the expansion of O. cf. ovata from tropical to temperate waters, the definite impact of temperature on bloom dynamics might be more complex than a simple facilitation factor for algal growth, at least in NW Mediterranean waters. Furthermore, monthly SST anomalies calculated over this 13-year survey showed a strong positive correlation between spring SST positive anomalies and the bloom starting date, indicating that blooms occurred earlier in the season when spring SSTs were warmer than usual. Overall results provide tools to modelers and managers who are facing crucial challenges to predict the distribution and phenology of O. cf. ovata blooms in European coastal waters, moreover in a context of global warming.

Multiple stressors and benthic harmful algal blooms (BHABs): Potential effects of temperature rise and nutrient enrichment.

Blooms of Ostreopsis cf. ovata, causing health incidence and mass human intoxications in the Mediterranean, gained special attention over the past decades. To study the potential effects of temperature and nutrient enrichment on this benthic dinoflagellate and other associated microalgae in situ, a multifactorial experiment was set up along a temperature gradient of a heat pump system in Monaco. Microalgae were quantified in experimental units, in the natural biofilm and in the water column. No significant interaction was observed between temperature and nutrients. A species- and bloom phase-dependent effect of the increased temperature was recorded, while the nutrient enrichment had a significant effect only at the end of the experiment (when cell abundances were low). Temperature effects were also visible in the biofilm and the surrounding water. The observed assemblages were mainly driven by changes in abundances of Ostreopsis cf. ovata and Actinocyclus sp., affected in different ways.

The toxic benthic dinoflagellate Ostreopsis ovata: quantification of proliferation along the coastline of Genoa, Italy.

Toxic harmful algal bloom (HAB) occurrence is becoming more frequent and problematic in highly urbanized coastal zones. In summer 2005 along the urbanized Genoa coastline (Ligurian Sea, North western Mediterranean Sea), local first aid stations treated about 200 people, who all showed similar symptoms following exposure to marine aerosols. The link with proliferation of Ostreopsis ovata was made, and it highlighted for the first time, the risks that benthic HABs may represent in highly urbanised temperate areas. Subsequently, a specific monitoring plan was designed and implemented in the same area in July 2006, before the first signs of Ostreopsis proliferation were detected. Here we report on this quantification of an Ostreopsis ovata bloom in the Ligurian Sea. Cells were quantified both in the water column and in the epiphytic community on macrophytes. Our results suggest a role of sea water temperature and weather conditions in favouring bloom development.

Strong, weak, and missing links in a microbial community of the N.W. Mediterranean Sea.

Planktonic microbial communities often appear stable over periods of days and thus tight links are assumed to exist between different functional groups (i.e. producers and consumers). We examined these links by characterizing short-term temporal correspondences in the concentrations and activities of microbial groups sampled from 1 m depth, at a coastal site of the N.W. Mediterranean Sea, in September 2001 every 3 h for 3 days. We estimated the abundance and activity rates of the autotrophic prokaryote Synechococcus, heterotrophic bacteria, viruses, heterotrophic nanoflagellates, as well as dissolved organic carbon concentrations. We found that Synechococcus, heterotrophic bacteria, and viruses displayed distinct patterns. Synechococcus abundance was greatest at midnight and lowest at 21:00 and showed the common pattern of an early evening maximum in dividing cells. In contrast, viral concentrations were minimal at midnight and maximal at 18:00. Viral infection of heterotrophic bacteria was rare (0.5-2.5%) and appeared to peak at 03:00. Heterotrophic bacteria, as % eubacteria-positive cells, peaked at midday, appearing loosely related to relative changes in dissolved organic carbon concentration. Bacterial production as assessed by leucine incorporation showed no consistent temporal pattern but could be related to shifts in the grazing rates of heterotrophic nanoflagellates and viral infection rates. Estimates of virus-induced mortality of heterotrophic bacteria, based on infection frequencies, were only about 10% of cell production. Overall, the dynamics of viruses appeared more closely related to Synechococcus than to heterotrophic bacteria. Thus, we found weak links between dissolved organic carbon concentration, or grazing, and bacterial activity, a possibly strong link between Synechococcus and viruses, and a missing link between light and viruses.

Evaluation of the toxicological risk to humans of caulerpenyne using human hematopoietic progenitors, melanocytes, and keratinocytes in culture.

The extensive growth of Caulerpa taxifolia in the Mediterranean sea produces important quantities of bioactive secondary metabolites unable to enter the food chain. The cytotoxic effects of caulerpenyne, the major secondary metabolite from C. taxifolia, was studied in different in vitro models: skin cells, primary cultures of melanocytes and keratinocytes, immortalized keratinocytes (HaCaT and HESV), and bone marrow cells (hematopoietic progenitors CFU-GM). Typical dose-response curves from neutral red uptake and MTT assays were recorded in all models with IC50 ranging from 6 to 24 microM. Hematopoietic progenitors were more sensitive to caulerpenyne than melanocyte and keratinocyte cell lines, which could be due to their higher proliferative rate. The distribution of aggregates in colonies, macroclusters, and microclusters of hematopoietic progenitors was also altered in the presence of caulerpenyne. From our evaluation of the caulerpenyne concentrations required to result in cellular toxicity, the risks of cutaneous and/or food intoxication to humans may be considered minimal.

Cell growth inhibitory effects of caulerpenyne, a sesquiterpenoid from the marine algae Caulerpa taxifolia.

Caulerpa taxifolia (Vahl) C. Agardh (Ulvophyceae, Caulerpales) is an algae of tropical origin that was accidentally introduced into the Mediterranean in 1984. Caulerpenyne (Cau) is the major metabolite present in Caulerpa taxifolia. This metabolite has previously been shown to be cytotoxic against cell lines in culture as in KB cells and fibroblasts from hamsters. Cau along with 6 other drugs representative of the major classes of anticancer products was tested against 8 cancer cell lines of human origin. Cau demonstrated growth-inhibitory effects in all cases with some variability between cell lines; this inter-cell variability was, however, less marked than that observed with the anticancer drug tested. Cells of colorectal cancer origin were the most sensitive to the presence of Cau with IC50 values of 6.1 and 7.7 microM. Increasing the duration of contact between Cau and the cells from 75 min to 29.5 hr did not improve the cytotoxic efficacy of this compound. When Cau was pre-incubated in the culture medium for from 7 to 83 min before being exposed to CAL 27 cells (head and neck cancer origin), there was a constant loss of cytostatic action of Cau as a function of Cau pre-incubation time. As the bovine serum albumin concentration increased in the culture medium, the concentration-response curves showed a constant shift towards the right, indicating a loss of cytostatic activity of Cau. In the presence of Cau, cells in culture clearly exhibited an early and marked shift into S phase followed by a blockade into the premitotic G2 M phase. Possible targets for CAU remain to be identified. Cau needs to be tested on tumor bearing animals to confirm this promising antiproliferative activity.

Involvement of an N-Acetylglucosaminidase in Autolysis of Propionibacterium freudenreichii CNRZ 725.

Propionibacterium freudenreichii plays an important role in Swiss cheese ripening (it produces propionic acid, acetic acid, and CO(2)). Moreover, autolysis of this organism certainly contributes to proteolysis and lipolysis of the curd because intracellular enzymes are released. By varying external factors, we determined the following conditions which promoted autolysis of both whole cells and isolated cell walls of P. freudenreichii CNRZ 725: (i) 0.1 M potassium phosphate buffer (pH 5.8) at 40 degrees C and (ii) 0.05 to 0.1 M KCl at 40 degrees C. We found that early-exponential-phase cells possessed the highest autolytic activity. It should be emphasized that the pH of Swiss cheese curd (pH 5.5 to 5.7) is near the optimal pH which we determined. Ultrastructural observations by electron microscopy revealed a 16-nm-thick homogeneous cell wall, as well as degradation of the cell wall that occurred concomitantly with cell autolysis. In the presence of 0.05 M potassium chloride, there was a great deal of isolated cell wall autolysis (the optical density at 650 nm decreased 77.5% +/- 7.3% in 3 h), and one-half of the peptidoglycan material was released. Finally, the main autolytic activity was due to an N-acetylglucosaminidase activity.