Karenia Selliformis Research Articles

Allelopathic Effect of a Chilean Strain of Karenia selliformis (Gymnodiniales, Dinoflagellata) on Phytoplankton Species.

Blooms of the dinoflagellate Karenia selliformis in Chile, often associated with massive fish kills, have been noted alongside other species from the Kareniaceae family, such as Karenia spp. and Karlodinium spp. However, the potential allelopathy impact of Chilean K. selliformis on other phytoplankton species remains unexplored. Here, we assessed the allelopathic effects of cell-free exudates from a Chilean K. selliformis strain on six phytoplankton strains representing diverse microalgal groups. The findings of these experiments offer valuable insights into the varied responses of both non-toxic and toxic microalgae to allelochemicals produced by a toxic microalga, showcasing the intricate and multifaceted nature of allelopathic interactions in microalgal communities. The study revealed species-dependent effects, with variable response in cell growth, photosynthetic efficiency (i.e., Fv/Fm), and intracellular reactive oxygen species (ROS) production. While certain strains exhibited significant growth inhibition in response to the allelochemicals, others demonstrated no apparent effect on cell proliferation, indicating varying sensitivity to specific allelochemicals or potentially distinct detoxification mechanisms. Similarly, the diverse effects on Fv/Fm highlight the complexity of allelopathic interactions, with some species showing reduced efficiency without alterations in intracellular ROS production, while others displayed increased ROS production alongside impaired photosynthesis.

Bloom and phylogeny of the harmful dinoflagellate Karenia selliformis (Dinophyceae) isolated from Iran southeast coast (northern of Oman Sea)

Bloom and phylogeny of the harmful dinoflagellate Karenia selliformis (Dinophyceae) isolated from Iran southeast coast (northern of Oman Sea)

Experimental assessment of copepod survival in response to the harmful dinoflagellate <i>Karenia selliformis</i> from the southeastern coast of Hokkaido, Japan

Experimental assessment of copepod survival in response to the harmful dinoflagellate <i>Karenia selliformis</i> from the southeastern coast of Hokkaido, Japan

Five Karenia species along the Chinese coast: With the description of a new species, Karenia hui sp. nov. (Kareniaceae, Dinophyta)

Dinoflagellates within the genus Karenia are well known for their potential to cause harmful algal blooms and induce detrimental ecological consequences. In this study, five Karenia species, Karenia longicanalis, Karenia papilionacea, Karenia mikimotoi, Karenia selliformis, and a new species, Karenia hui sp. nov., were isolated from Chinese coastal waters. The new species exhibits the typical characteristics of the genus Karenia, including a linear apical groove and butanoyl-oxyfucoxanthin as the major accessory pigment. It is distinguished from the other Karenia species by a wide-open sulcal intrusion onto the epicone, a conical epicone with an apical crest formed by the rim of the apical groove, and a hunchbacked hypocone. It is most closely related to Karenia cristata, with a genetic divergence of 3.16 % (22 bp out of 883 bp of LSU rDNA). Acute toxicity tests indicated that the five Karenia species from China are all toxic to marine medaka Oryzias melastigma. Karenia selliformis and K. hui were very toxic to O. melastigma, resulting in 100 % mortality within 4 h and 24 h, respectively. Further analysis by high-performance liquid chromatography revealed that four species, K. selliformis, K. longicanalis, K. papilionacea and K. mikimotoi were capable of producing Gymnodimine-A (GYM-A). The highest GYM-A content was in K. selliformis (strain HK-43), in which the value was 889 fg/cell. No GYM-A was detected in the new species K. hui, however and its toxin remains unknown. Below we provide a comprehensive report of the morphology, phylogeny, pigment composition, and toxicity profiles of Karenia species along the Chinese coast. These findings contribute new insights for monitoring of Karenia species, with important toxicological and ecological implications.

Observations of water optical properties during red tide outbreaks off southeast Hokkaido by GCOM-C/SGLI: implications for the development of red tide algorithms

ABSTRACT A severe red tide event, caused primarily by dinoflagellate Karenia selliformis, occurred during the autumn of 2021 in the waters off southeast Hokkaido and resulted in damage of $70 million to fishery industries. There is a high demand for early warning methods based on ocean colour observations to respond to future occurrences of red tides. We therefore used a quasi-analytical algorithm to compute the total absorption coefficient (a) from the Second Generation Global Imager (SGLI) remote sensing reflectance and assessed its potential to detect blooms of K. selliformis. We discovered that, within the same range of SGLI-retrieved chlorophyll-a concentrations, the a at a wavelength of 530 nm a(530) tended to be lower during K. selliformis blooms than during diatom blooms. The a(530) observed by the SGLI therefore showed promise as an optical property for detecting K. selliformis blooms. We discuss the reasons for selection of a(530) and the limitations of the current study’s bloom detection method.

Toxicity and potential underlying mechanism of Karenia selliformis to the fish Oryzias melastigma

Karenia selliformis can produce toxins such as gymnodimines, and form microalgal blooms causing massive mortality of marine life such as fish and shellfish, and resulting in serious economic losses. However, there are a few of studies on the toxic effects of K. selliformis on marine organisms and the underlying mechanisms, and it is not clear whether the toxins produced by K. selliformis affect fish survival through the food chain. In this study, a food chain was simulated and composed by K. selliformis-brine shrimp-marine medaka to investigate the possibility of K. selliformis toxicity transmission through the food chain, in which fish behavior, histopathology and transcriptomics changes were observed after direct or indirect exposure (through the food chain) of K. selliformis. We found that both direct and indirect exposure of K. selliformis could affect the swimming behavior of medaka, manifested as decreased swimming performance and increased “frozen events”. Meanwhile, exposure to K. selliformis caused pathological damage to the intestine and liver tissues of medaka to different degree. The effect of direct exposure to K. selliformis on swimming behavior and damage to fish tissues was more severe. In addition, K. selliformis exposure induced significant changes in the expression of genes related to energy metabolism, metabolic detoxification and immune system in medaka. These results suggest that toxins produced by K. selliformis can be transferred through the food chain, and that K. selliformis can destroy the intestinal integrity of medaka and increase the absorption of toxins, leading to energy metabolism disorders in fish, affecting the metabolic detoxification capacity of the liver. Our finding provides novel insight into the toxicity of K. selliformis to marine fish.

Potential effects of climate change on the growth response of the toxic dinoflagellate Karenia selliformis from Patagonian waters of Chile

Northern Patagonia (41–44°S) is affected by climatic, hydrological and oceanographic anomalies, which in synergy with processes such as global warming and acidification of the coastal oceans may affect the frequency and intensity of harmful algal blooms (HABs). Greater frequency of HABs has been reported in the southeastern Pacific Ocean, including blooms of the toxic dinoflagellate Karenia selliformis, causing massive mortality of marine fauna in the oceanic and coastal areas of Patagonia. The objective of this study was to determine the effects of temperature and pH interaction on the growth of K. selliformis (strain CREAN_KS02), since these factors have wide seasonal fluctuations in the Patagonian fjord ecosystem. The CREAN_KS02 strain isolated from the Aysén Region (43°S) was used in a factorial experiment with five pH levels (7.0, 7.4, 7.7, 8.1 and 9.0) and two temperatures (12 and 17 °C) during a period of 18–21 days. Results indicated a significant effect of temperature and pH interaction on growth rate (range 0.22 ± 0.00 to 0.08 ± 0.01 d−1) and maximum density (range 13,710 ± 2,616 to 2,385 ± 809 cells mL−1) of K. selliformis. The highest density and growth of K. selliformis was found at 12 °C with a reduced pH (7.0–7.7). The results suggest that the current environmental conditions of coastal Patagonia, waters of low temperature and relatively low pH, may be favorable for the development of blooms of this species during autumn. We suggest that there is natural plasticity of K. selliformis in a wide pH range (7.0–8.1) but in a narrow low temperature range (10.6–12.9 °C), values that are typically recorded in the oceanic region of northern Patagonia. In contrast, in an extreme climate change scenario (ocean warming and coastal acidification) in northern Patagonia, a negative effect on the growth of K. selliformis may be expected due to amplification of the acidification effects caused by the thermal stress of high temperature water.

Karenia selliformis bloom dynamics and growth rate estimation in the Sfax harbour (Tunisia), by using automated flow cytometry equipped with image in flow, during autumn 2019

A Karenia selliformis bloom event in the Gulf of Gabès (Mediterranean Sea), was monitored over 9 days at high frequency during fall 2019, by using an automated flow cytometer (Cytosense, Cytobuoy b.v.) with an image-in-flow attachment. The instrument recorded the shape of the optical signals that lead to the resolution of six cell groups of pico-, nano- and microphytoplankton, during the Harmful Algal Bloom (HAB). K. selliformis cell dimensions derived from the hourly records, enabled to estimate the daily division rate over the bloom period. Results revealed that K. selliformis was the only bloom-forming species and it reached its highest mean abundance the fourth day of the survey. A shift in the nutrient composition occurred with a potential P limitation during the bloom growth and N limitation during the bloom collapse. The co-inertia analysis revealed opposite patterns for K. selliformis and heterotrophic prokaryotes suggesting trophic interactions and possible mixotrophic behaviour of K. selliformis at the end of the bloom. K. selliformis exhibited low growth rates generally < 1 division day−1, which could not explain the observed high abundance. The tide played a crucial role in the dynamics of K. selliformis at a semi-diurnal scale and at spring-neap tide scale and was probably enhancing K. selliformis accumulation.

Stability and Chemical Conversion of the Purified Reference Material of Gymnodimine-A under Different Temperature and pH Conditions.

Gymnodimines (GYMs) are a group of fast-acting phycotoxins and their toxicological effects on human beings are still unclear due to the lack of sufficiently well-characterized large quantities of purified toxins for toxicology studies. In this study, a certified reference material (CRM) of GYM-A was prepared from the dinoflagellate Karenia selliformis, followed by multi-step chromatography separation and purification. Subsequently, the stability of GYM-A in methanolic media was evaluated at different temperature (-20, 4, and 20 °C) and pH (3, 5, and 7) conditions for 8 months, and the conversion products of GYM-A were explored by liquid chromatography-high resolution mass spectrometry (LC-HRMS). The results show that the stability of GYM-A decreased with increasing temperature and pH values. The GYM-A was stable during storage at -20 °C regardless of pH, but it decreased rapidly (81.8% ± 9.3%) at 20 °C in pH 7 solution after 8 months. Moreover, the concentrations of GYM-A did not significantly change at all temperatures in solutions with pH 3 (p > 0.05). It is recommended that GYM-A should be stored at low temperature (≤-20 °C) and pH (≤3) conditions for long-term storage in aqueous methanolic media. In addition, two conversion products of GYM-A, tentatively named as GYM-K (m/z 540) and GYM-L (m/z 524), were identified in the samples stored at high levels of pH and temperature. Based on the LC-HRMS data, the hypothetical chemical structures of both converting derivatives were proposed. A useful strategy for long-term storage of GYM-A CRM in aqueous methanolic media was suggested and two hypothesized conversion products of GYM-A were discovered in this study.

Distribution of Harmful Algae (Karenia spp.) in October 2021 Off Southeast Hokkaido, Japan

An unprecedented large-scale outbreak of harmful algae, including Karenia selliformis and Karenia mikimotoi, was reported in mid-September 2021 in the northwest Pacific Ocean off southeastern Hokkaido, Japan. It inflicted catastrophic damage on coastal fisheries in the ensuing months. To understand the spatiotemporal distribution of Karenia spp. abundance, we conducted extensive ship-based surveys across several water masses during 4–14 October, 2021 and analyzed in-situ data in combination with Sentinel-3-derived ocean color imagery with a horizontal resolution of 300 m. High chlorophyll-a concentrations (exceeding 10 mg m–3) were identified mainly in coastal shelf–slope waters of &amp;lt;1,000-m water depth occupied by Surface Coastal Oyashio Water or Modified Soya Warm Current Water. Karenia spp. abundance was strongly correlated with chlorophyll-a concentration, which typically had a shallow vertical maximum within the surface mixed layer. Large- and small-scale distributions of Karenia spp. abundance at the ocean surface were estimated from two satellite-imagery products: maximum line height and red-band difference. Maps generated of Karenia spp. abundance revealed snapshots of dynamic Karenia bloom distributions. Specifically, the cores of Karenia blooms were located on continental shelves, sometimes locally exceeded 104 cells mL–1, and seemed to be connected intermittently to very nearshore waters. Relatively high-abundance areas (&amp;gt;103 cells mL–1) of Karenia spp. on the shelf were characterized by submesoscale (i.e., 1–10 km) patch- or streak-like distributions, or both. Within a roughly 24-h period from 12 to 13 October, Karenia-spp. abundances averaged over the shelf abruptly increased more than doubled; these abundance spikes were associated with the combined effects of physical advection and algal growth. The obtained maps and features of Karenia spp. abundance will provide basic estimates needed to understand the processes and mechanisms by which algal blooms can inflict damage on regional fisheries.

Horizontal distribution of harmful red-tide Karenia selliformis and phytoplankton community along the Pacific coast of Hokkaido in autumn 2021

Horizontal distribution of harmful red-tide <i>Karenia selliformis</i> and phytoplankton community along the Pacific coast of Hokkaido in autumn 2021

Prevalence and distribution of domoic acid and cyclic imines in bivalve mollusks from Beibu Gulf, China

Beibu Gulf is an important shellfish aquaculture area in the northwest of the South China Sea, China. In this study, the toxin profile and spatial-temporal distribution of domoic acid (DA) and 10 lipophilic phycotoxins were systematically analyzed in the bivalve mollusks collected in Beibu Gulf from October 2018 to October 2020. Neurotoxin DA was first detected in the mollusks from the investigative regions with a prevalence of 17.7%, peaking at 401 µg kg−1. Cyclic imines (CIs) including gymnodimine-A (GYM-A, 46.6%) and 13-desmethyl-spirolide-C (SPX1, 15.8%) predominated the lipophilic phycotoxins in shellfish, peaking at 10.1 µg kg−1 and 19.6 µg kg−1, respectively. Gymnodimine-A partially accompanied by SPX1 was detected in all batches of shellfish samples, suggesting that Alexandrium ostenfeldii and Karenia selliformis were possible sources of CIs-group toxins in Beibu Gulf. During the investigative period, relatively higher levels of DA occurred in shellfishes from March to August, while slightly higher contents of CIs in mollusks appeared in October and December. Spatial distribution of the targeted phycotoxins demonstrated that shellfishes tended to accumulate relatively higher contents of toxins in Lianzhou, Qinzhou and Tieshan bays.

Q-PCR-based assay for the toxic dinoflagellate Karenia selliformis monitoring along the Tunisian coasts.

Karenia selliformis is a marine dinoflagellate responsible for fish-kill events. Its presence has been reported along the Tunisian coasts (south-eastern Mediterranean Sea) since the 1990s. In the present study, a quantitative-PCR assay, based on the internal transcribed spacer (ITS) molecular marker, was developed to detect and quantify K. selliformis in environmental bivalve mollusk samples and in seawater samples. The assay was optimized, and its specificity was confirmed using cross-reactivity experiments against microalgal species commonly found on the Tunisian coasts and/or closely related to K. selliformis. Calibration curves were performed by tenfold dilutions of plasmid DNA harboring target sequence and genomic DNA, attaining a limit of detection of around 5 copies of target DNA per reaction, far below one K. selliformis cell per reaction. The field application of the developed assay showed a powerful detection capability. Thus, the designed assay could contribute to the deployment of in-field diagnostic tools for K. selliformis blooms monitoring.

Diversity and distribution of harmful microalgae in the Gulf of Thailand assessed by DNA metabarcoding

Information on the diversity and distribution of harmful microalgae in the Gulf of Thailand is very limited and mainly based on microscopic observations. Here, we collected 44 water samples from the Gulf of Thailand and its adjacent water (Perhentian Island, Malaysia) for comparison in 2018. DNA metabarcoding was performed targeting the partial large subunit ribosomal RNA gene (LSU rDNA D1–D3) and the internal transcribed spacers (ITS1 and ITS2). A total of 50 dinoflagellate genera (made up of 72 species) were identified based on the LSU rDNA dataset, while the results of ITS1 and ITS2 datasets revealed 33 and 32 dinoflagellate genera comprising 69 and 64 species, respectively. Five potentially toxic Pseudo-nitzschia (Bacillariophyceae) species were detected, with four as newly recorded species in the water (Pseudo-nitzschia americana/brasilliana, Pseudo-nitzschia simulans/delicatissima, P. galaxiae and P. multistriata). The highest relative abundances of P. galaxiae and P. multistriata were found in Trat Bay and Chumphon (accounting for 0.20% and 0.06% of total ASVs abundance, respectively). Three paralytic shellfish toxin producing dinoflagellate species were detected: Alexandrium tamiyavanichii, Alexandrium fragae, and Gymnodinium catenatum. The highest abundance of A. tamiyavanichii was found in the surface sample of Chumphon (CHO7 station), accounting for 1.95% of total ASVs abundance. Two azaspiracid producing dinoflagellate species, Azadinium poporum ribotype B, Azadinium spinosum ribotype A, and a pinnatoxin producing dinoflagellate species Vulcanodinium rugosum, with two ribotypes B and C, were revealed from the datasets although with very low abundances. Six fish killing dinoflagellate species, including Margalefidinium polykrikoides group IV, Margalefidinium fulvescens, Karenia mikimotoi, Karenia selliformis ribotype B, Karlodinium australe, and Karlodinium digitatum were detected and all representing new records in this area. The findings of numerous harmful microalgal species in the Gulf of Thailand highlight the potential risk of human intoxication and fish killing events.

Unraveling the Karenia selliformis complex with the description of a non-gymnodimine producing Patagonian phylotype

Karenia selliformis is a bloom-forming toxic dinoflagellate known for production of gymnodimines (GYMs) and causing mass mortalities of marine fauna. Blooms have been reported from coastal waters of New Zealand, Mexico, Tunisia, Kuwait, Iran, China and Chile. Based on molecular phylogeny, morphology, toxin production, pigment composition and cell growth of Chilean K. selliformis isolated in 2018 (CREAN_KS01 and CREAN_KS02), this study revealed a more complex diversity within this species than previously thought. A phylogenetic reconstruction based on the large sub-unit ribosomal nucleotide (LSU rDNA) and Internal Transcriber Spacer (ITS) sequences of 12 worldwide isolates showed that within the K. selliformis clade there are at least two different phylotypes with clear phenotypic differences. Morphological differences related to the dorsal-ventral compression, shape of the hyposome and the presence of pores on the left lobe of the hyposome. A comparison of pigment signatures among worldwide isolates revealed the existence of both acyl-oxyfucoxanthin and fucoxanthin-rich strains within the phylotypes. A LC-MS/MS screening on both Chilean 2018 K. selliformis strains showed for first time no GYMs production among cultured clones of this species. However, both CREAN_KS01 and CREAN_KS02 contained two compounds with the same mass transition as brevenal, a brevetoxin related compound. A fish gill cell-based assay showed that the CREAN_KS02 strain was highly cytotoxic but pure GYM standard did not exhibit loss of cell viability, even at cell concentrations equivalent or exceeding those reported in nature. The fatty acid profile of CREAN_KS02 included high levels of saturated (14:0; 16:0) and polyunsaturated (18:3ω6+18:5ω3; 22:6ω3) fatty acids but superoxide production in this strain was low (0.86±0.53 pmol O2− cell−1 h−1). A factorial T-S growth experiment using the CREAN_KS02 strain showed a μmax of 0.41±0.03 d−1 at high salinity and temperature, which points to its optimal environmental niche in offshore waters during the summer season. In conclusion, the present study provides evidence for significant genetic and phenotypic variability among worldwide isolates, which points to the existence of a K. selliformis “species complex”. The massive fauna mortality during K. selliformis bloom events in the Chilean coast cannot be explained by GYMs nor brevetoxins, but can to a large extent be accounted for by the high production of long-chain PUFAs and/or still uncharacterized highly toxic compounds.

Quantitative PCR assay for the simultaneous identification and enumeration of multiple Karenia species.

Quantitative PCR (qPCR) is the method of choice for specific detection and quantification of harmful algal bloom (HAB) species. Development of qPCR assay for simultaneous enumeration of species that frequently co-exist in HABs is required. A high sensitivity TaqMan qPCR assay, using probe and primers, located at ITS1-5.8S-ITS2 rDNA region, detecting, specifically, Karenia selliformis, K. bidigitata, and K. mikimotoi, was designed. ITS1-5.8S-ITS2 rDNA region copy numbers per Karenia cell genome were estimated to 217.697 ± 67.904, allowing cell quantification. An application of the designed methodology in field samples has been conducted, and it showed high sensitivity (detection of around 10-1 cell/100 mg of bivalve mollusk tissue, equivalent to about 20 copies of the target sequence). We suggest that the optimized method could contribute to early detection of three closely related Karenia species in seafood cultivating areas to promote control quality, guarantee a fast and effective intervention, and improve public health prevention.

Using a naive Bayes classifier to explore the factors driving the harmful dinoflagellate Karenia selliformis blooms in a southeastern Mediterranean lagoon

The blooms of the toxic dinoflagellate Karenia selliformis can be predicted with accuracy derived from knowledge of the main forcing variables. A naive Bayes classifier modeling framework, a member of the Bayesian network family, is used to identify the phytoplankton community and the physical and meteorological parameters involving K. selliformis blooms in the eutrophic Boughrara Lagoon (BL), Tunisia. The proposed model takes into account the physical environment parameters (salinity, water temperature, tide amplitude), meteorological constraints (evaporation, air temperature, insolation, rainfall, atmospheric pressure, and humidity), phytoplankton groups (diatoms, dinoflagellates, cyanobacteria, Euglenophyceae), and the sampling months on K. selliformis blooms. The shift to highest salinity and atmospheric pressure, associated with reduced tide, are the most favorable conditions for K. selliformis blooms in BL. The results show that K. selliformis formed nearly monospecific blooms. A shift in species composition was pointed out between the bloom and non-bloom conditions. The Euglenophycea and some dinoflagellates like Peridinium minimum, Prorocentrum minimum, Prorocentrum micans, Prorocentrum gracile, Protoperidinium minutum, and Scrippsiella trochoidea appeared during blooms, whereas diatoms, diazotrophic cyanobacteria, and dinoflagellates (Akaskiwo sanguinea, Karlodinium veneficum, Gyrodinium spirale, Oxyrrhis marina, Polykrikos kofoidii) were observed under non-bloom conditions. This study highlighted the most favorable physical and meteorological conditions for K. selliformis bloom occurrences and also pointed out species indicators for bloom establishment and others for non-bloom conditions. Monitoring the dynamics of these species with the associated physical and meteorological variability offers valuable information to plan for the best options for prediction of potentially toxic blooms of K. selliformis and associated dystrophic consequences.

Screening of Chilean fish-killing microalgae using a gill cell-based assay

Fish-killing algal species are responsible for important global economic losses to the finfish industry. Chile is the world’s second-largest salmon exporter, and fish-killing algal blooms have widely impacted its production in the last decades. The lack of standardized analytical methods to quantify and characterize the so-called “ichthyotoxins” has hindered our understanding of the underlying ichthyotoxic modes of action. The novel application of a highly sensitive and reproducible fish RTgill-W1 cell line-based assay has allowed significant progress in the field. In this study, the ichthyotoxic potency of the main microalgae species, which has been reported in fish-killing events in the historical Chilean monitoring programs, was assessed. The dinoflagellate Karenia selliformis was the most ichthyotoxic species against the RTgill-W1 (cell viability down to 8%), representing the major threat for the local salmon industry. In comparison, the raphidophyte Heterosigma akashiwo and the dinoflagellate Prorocentrum micans were the least toxic (gill cell viability down to 81 and 89%, respectively). Importantly, ichthyotoxic flagellates were more toxic to fish gill cells upon rupture. These results have important implications for the mitigation and management of algal blooms by the salmon industry.

Cyclic imines, as emerging marine toxins: Chemical properties, distribution, toxicological aspects and detection methods

Shellfish and, specifically, bivalve molluscs are a food commodity of great nterest for both commercial and public health reasons. They consume microalgae from surrounding waters, which are generally beneficial for aquaculture, but they comprise certain toxin-producing species. These species produce marine toxins which, via the filter-feeding mechanism of bivalve molluscs, accumulate in their tissues. This accumulation is more intense andmore dangerous for public health during the so-called periods of Harmful Algal Blooms (HABs) when the microalgal population grows. According to their chemical structure, marine toxins are classified into 8 groups, one of which is the cyclic imines. These lipophilic toxins were accidentally discovered during routine bioassays for the detection of other lipophilic marine toxins due to the induction of neurological symptoms and acute death in mice. They include the following subgroups: Spirolides (SPX), gymnodimines (GYM), pinnatoxins (PnTX), pteriatoxins (PtTX), prorocentrolides and spiro-prorocentrimines. The European Union (EU) is more concerned about the first three subgroups, because, in contrast with the latter three, they have already been detected in Europe or there is strong evidence supporting their presence. Spirolides are produced by the dmof\a.ge\\ate Alexandrìum ostenfeldii/peruvianum, gymnodimines by the dinoflagellate Karenia selliformis and pinnatoxins by a peridinoid dinoflagellate recently described in the new genus Vulcanodinium spp.. Although there is insufficient information regarding the geographical distribution of cyclic imines, the fact that they have been detected on multiple occasions in European waters, in combination with their aforementioned acute toxicity in mice after intraperitoneal injection, has established them, at least within the EU, as a topic of profound scientific research. In spite of their acute toxicity in mice, no incident of human intoxication has been attributed to cyclic imines. Presently, the EU has neither set any Maximum Permissible Limits for the concentration of cyclic imines in shellfish nor appointed any reference method for their detection and quantification. Currently, the methods applied are biological, biochemical and chemical. The biological method is a bioassay, which is conducted via the intraperitoneal injection of mice with an extract containing the compound under examination and it detects total toxicity. This properly is essential for the detection of unknown toxins, but the use of laboratory animals raises serious ethical concerns and animal welfare issues. The biochemical method is based on competition between cyclic imines and a fluorescently labelled compound for binding to receptors of the electric ray Torpedo marmorata. Finally, in respect of chemical methods, liquid chromatography with tandem mass spectrometry detection (LC-MS/MS) is the most significant method because it is fast, of high repeatability and specificity.

Using general linear model, Bayesian Networks and Naive Bayes classifier for prediction of Karenia selliformis occurrences and blooms

The prediction of the dinoflagellate red tide forming Karenia selliformis is a relevant task to aid optimized management decisions in marine coastal water. The objective of the present study is to compare different modeling approaches for prediction of Karenia selliformis occurrences and blooms. A set of physical parameters (salinity, temperature and tide amplitude), meteorological constraints (evaporation, air temperature, insolation, rainfall, atmospheric pressure and humidity), sampling months and sampling sites are used. The model prediction included general linear model (GLM), Bayesian Network (BN) and the simplest BN type which is, Naive Bayes classifier (NB). The results showed that three models incriminated high salinity in Karenia selliformis blooms and the sampling sites, mainly Boughrara lagoon, in the occurrences. The BN performed better than linear models (NB and GLM) for both Karenia selliformis occurrences and blooms prediction. This later is related to the facts that BN considered the inter-independency between predictive variables and that the relationships between the variables and the outcome are often non-linear such us; the transition to bloom situations appeared to be triggered by a salinity threshold. This study is useful in the management of this ecosystem so as to use the best disposal options in the early prediction of the toxic blooms.