Teresa Moita

BMAA in shellfish from two Portuguese transitional water bodies suggests the marine dinoflagellate Gymnodinium catenatum as a potential BMAA source

The neurotoxin β-N-methylamino-l-alanine (BMAA) and its putative role in multiple neurodegenerative diseases have been intensely studied since 2005 when the toxin was discovered to be produced by worldwide-distributed cyanobacterial species inhabiting terrestrial, marine, brackish, and freshwater ecosystems. Recently, BMAA production was also associated with one eukaryotic group, namely, diatoms, raising questions about its production by other phytoplanktonic groups. To test for BMAA bioavailability in ecosystems where abundant phytoplanktonic blooms regularly occur, samples of filter-feeding shellfish were collected in two Portuguese transitional water bodies. BMAA content in cockles (Cerastoderma edule) collected weekly between September and November 2009 from Ria de Aveiro and at least once a month from May to November from Ria Formosa, fluctuated from 0.079±0.055 to 0.354±0.066μg/g DW and from below the limit of detection to 0.434±0.110μg/g DW, respectively. Simultaneously to BMAA occurrence in cockles, paralytic shellfish toxins were detected in shellfish as a result of Gymnodinium catenatum blooms indicating a possible link between this marine dinoflagellate and BMAA production. Moreover, considerable high BMAA levels, 0.457±0.186μg/g DW, were then determined in a laboratory grown culture of G. catenatum. This work reveals for the first time the presence of BMAA in shellfish from Atlantic transitional water bodies and consubstantiate evidences of G. catenatum as one of the main sources of BMAA in these ecosystems.

Climate variability and Dinophysis acuta blooms in an upwelling system

Dinophysis acuta is a frequent seasonal lipophilic toxin producer in European Atlantic coastal waters associated with thermal stratification. In the Galician Rías, populations of D. acuta with their epicentre located off Aveiro (northern Portugal), typically co-occur with and follow those of Dinophysis acuminata during the upwelling transition (early autumn) as a result of longshore transport. During hotter than average summers, D. acuta blooms also occur in August in the Rías, when they replace D. acuminata. Here we examined a 30-year (1985-2014) time series of D. acuta from samples collected by the same method in the Galician Rías. Our main objective was to identify patterns of distribution and their relation with climate variability, and to explain the exceptional summer blooms of D. acuta in 1989-1990. A dome-shaped relationship was found between summer upwelling intensity and D. acuta blooms; cell maxima were associated with conditions where the balance between upwelling intensity and heating, leading to deepened thermoclines, combined with tidal phase (3 days after neap tides) created windows of opportunity for this species. The application of a generalized additive model based on biological (D. acuta inoculum) and environmental predictors (Cumulative June-August upwelling CUIJJA, average June-August SSTJJA and tidal range) explained more than 70% of the deviance for the exceptional summer blooms of D. acuta, through a combination of moderate (35,000-50,000m3s-1km-1) summer upwelling (CUIJJA), thermal stratification (SSTJJA>17°C) and moderate tidal range (∼2.5m), provided D. acuta cells (inoculum) were present in July. There was no evidence of increasing trends in D. acuta bloom frequency/intensity nor a clear relationship with NAO or other long-term climatic cycles. Instead, the exceptional summer blooms of 1989-1990 appeared linked to extreme hydroclimatic anomalies (high positive anomalies in SST and NAO index), which affected most of the European Atlantic coast.

A HAB warning system for shellfish harvesting in Portugal

The development of sustainable shellfish aquaculture is highly dependent on the provision of reliable monitoring and predictive information on the occurrence of harmful algal blooms (HABs). The Portuguese HAB early warning system and shellfish closures presented here is a prototype, developed in the ASIMUTH project. It relies on weekly monitoring data composed of observations of HAB species and toxin concentrations within shellfish, and ocean circulation forecasts generated by an operational oceanographic model. The shellfish harvesting areas comprise coastal areas, estuaries+rías and coastal lagoons. The weekly bulletin characterizes the current shellfish closure situation and next weeks forecasts for potentially impacted areas. The period analyzed ranged from 27 July 2013 to 17 March 2014, and describes the first skill assessment of the warning system. The forecast accuracy was evaluated, considering the number of forecasts that were verified to be correct the following week (85%) as well as the number of events not forecasted (false negatives, 12%) and those expected but did not occur (false positives, 3%). Variations were most visible in the first weeks of bulletin implementation and during autumn-winter months. The complementary use of field data, remote sensing and operational models led to more accurate predictions of blooms and range of the event.

Estimating the contribution of N-sulfocarbamoyl paralytic shellfish toxin analogs GTX6 and C3 + 4 to the toxicity of mussels (Mytilus galloprovincialis) over a bloom of Gymnodinium catenatum

Gymnodinium catenatum, a dinoflagellate species with a global distribution, is known to produce paralytic shellfish poisoning (PSP) toxins. The profile of toxins of G. catenatum is commonly dominated by sulfocarbamoyl analogs including the C3+4 and GTX6, which to date has no commercial certified reference materials necessary for their quantification via chemical methods, such as liquid chromatography. The aim of this study was to assess the presence of C3+4 and GTX6 and their contribution to shellfish toxicity. C3+4 and GTX6 were indirectly quantified via pre-column oxidation liquid chromatography with fluorescence detection after hydrolysis conversion into their carbamate analogs. Analyses were carried out in mussel samples collected over a bloom of G. catenatum (>63×103cellsl-1) in Aveiro lagoon, NW Portuguese coast. Concentration levels of sulfocarbamoyl toxin analogs were two orders of magnitude higher than decarbamoyl toxins, which were in turn one order of magnitude higher than carbamoyl toxins. Among the sulfocarbamoyl toxins, C1+2 were clearly the dominant compounds, followed by C3+4 and GTX6. The least abundant sulfocarbamoyl toxin was GTX5. The most important compounds in terms of contribution for sample toxicity were C1+2, which justified 26% of the PSP toxicity. The lesser abundant dcSTX constitutes the second most important compound with similar % of toxicity to C1+2, C3+4 and GTX6 were responsible for approximately 11% and 13%, respectively. The median of the sum of C3+4 and GTX6 was 27%. These levels reached a maximum of 60% as was determined for the sample collected closest to the G. catenatum bloom. This study highlights the importance of these low potency PSP toxin analogs to shellfish toxicity. Hydrolysis conversion of C3+4 and GTX6 is recommended for determination of PSP toxicity when LC detection methods are used for PSP testing in samples exposed to G. catenatum.

Changes in the Phytoplankton Composition in a Temperate Estuarine System (1960 to 2010)

The main aim of this study was to evaluate the temporal changes in the phytoplankton community of the Tagus Estuary and to identify the stressors involved. Environmental and phytoplankton data were gathered from several studies conducted in the 1960s, 1980s and from 1999 to 2010 (2000s). Phytoplankton data included information on the community composition through microscopy. No significant change was found between temperature and nitrogen values in the three periods. Chlorophyll a concentrations varied throughout the years, and the lowest concentrations were observed after 2004. Significant differences were also found between phytoplankton cell abundances, lower in the 1980s compared to the ones recorded in the 2000s. In the 1980s, diatoms were the most abundant group in the majority of samples and were found to be associated with nitrogen concentrations (canonical correspondence analysis), which was not observed in the 2000s. In the period 2006–2007, the importance of diatoms decreased and smaller cells became more abundant (e.g. cryptophytes, euglenophytes, prasinophytes). The ratio cryptophyta/bacillariophyta seems to yield an increase from <1 in 1980s to >1 in 2006–2007. Mesodinium rubrum and Dinophysis produce recurrent toxic blooms in the adjacent coastal area. We can speculate that the estuary can be a cryptophyte producer to sustain the cryptophytes-M. rubrum-Dinophysis trophic relationship. A top-down hypothesis (shellfish grazing) is considered to explain the change in the phytoplankton community. A quantitative tool, the Phytoplankton Community Index (PCI), yielded a significant deviation of the community, from the 1980s to the 2000s, suggesting a shift toward the dominance of small cells.

Key Questions and Recent Research Advances on Harmful Algal Blooms in Eastern Boundary Upwelling Systems

The Core Research Project (CRP) HABs in Upwelling Systems was developed as part of the implementation of the international science programme the Global Ecology and Oceanography of Harmful Algal Blooms (GEOHAB). Progress in addressing eight key questions identified as a requirement for advancing our understanding of the ecology and oceanography of HABs in upwelling systems is reviewed. The considerable diversity of HAB species found in upwelling systems is considered to reflect the mosaic of multiple and shifting sub-habitats present within upwelling systems. In developing predictive capabilities, the need to consider species-specific behaviour with reference to the environmental and ecological parameters that characterize these sub-habitats is demonstrated. However, the limited spatial resolution of many numerical models has prevented incorporation of the complexity created at the small scale by physically driven niche diversification. Observations of opportunistic exploitation of multiple seeding options rather than depending on a single seeding mode further complicate prediction. Although contrasting nutrient strategies have been demonstrated for HABs in upwelling systems, attempts to determine species-specific nutrient requirements are few. The timing of HABs is controlled by wind stress fluctuations and buoyancy inputs at the seasonal, event and interannual scales, whereas the spatial distribution of HABs is controlled by mesoscale features that interrupt typical upwelling circulation patterns leading to the identification of HAB hotspots. Here cross-shelf and alongshore currents are important in the transport, accumulation and dispersion of HAB populations. With increasing availability of long-term data sets, changing trends in HABs related to climate are emerging. Owing to the strong physical control of HABs in upwelling systems, successful prediction as a possible outcome of the integration of real-time data into model systems as a component of operational forecasting of the ocean is most likely to be achieved, thus taking a critical step towards fulfilment of GEOHAB goals.