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According to experts, pasturing in Europe has been decreasing for many years. Today, a lot of different pieces of regulation in Germany and in Europe exist, affecting farmers when they bring their cows to pasture. Furthermore, there are social requirements for pasturing, which need to be considered. Factors like increasing relevance of animal welfare, environmental protection, and the recreational use of the environment and esthetics of landscape come into play. Therefore, scientific research and new solutions are needed. This paper aims to give an overview across selected pasture-related regulations affecting farmers in Germany and Europe. Furthermore, aspects which are not directly regulated by the government, but are expectations of society will be highlighted. Concluding from this, technological possibilities - already on market and in the pipeline of scientific research will be described. Results show that today several different attempts are made to improve pasturing. Most of these attempts focus on improvement of management tasks. Attempts also directly focus on technological improvements. It can be concluded that new tools for pasturing can increase productivity and simultaneously decrease management tasks. Nevertheless, pricing of these tools as well es the amount of bureaucracy will be the most crucial factors for their adaptation.

A unialgal culture of aPseudo-nitzschiaspecies dominant in the plankton of Algoa Bay in the spring of 2012 was established by isolation of clonal chains of cells. Identification of the species asPseudo-nitzschia multiserieswas based on frustule morphometrics provided by light and scanning electron microscopy, and confirmed by phylogenetic analysis of the LSU rDNA gene. Cultures were shown to produce domoic acid (DA) as measured by ELISA and LC/MS-MS methods, and levels of cellular DA were ∼0.1 pg cell–1. Although it is recognised as a cosmopolitan species, these observations provide the first account of this toxic diatom in the coastal waters of South Africa. [ABSTRACT FROM AUTHOR]

Diarrhetic shellfish poisoning (DSP) poses a significant threat to the safe consumption of shellfish in the southern Benguela ecosystem. The accumulation of DSP toxins was investigated in two cultivated bivalve species, the Pacific oyster Crassostrea gigas and the mussel Choromytilus meridio-nalis, suspended from a mooring located off Lambert's Bay on the west coast of South Africa. The dinoflagellate Dinophysis acuminata, a known source of polyether toxins associated with DSP, was common through most of the study period. The toxin composition of the dinoflagellate was dominated by okadaic acid (OA) (91%), with lesser quantities of the dinophysistoxin DTX-1 (6.5%) and pecteno-toxin PTX-2 (2.4%), and traces of PTX-2sa and PTX-11. The mean cell toxin quota of D. acuminata was 7.8 pg OA cell–1. The toxin profile in shellfish was characterised by a notably higher relative content of DTX-1. The study showed the average concentration of DSP toxins in the mussels to exceed that in the oysters by approximately 20-fold. The results indicate a need to establish species-specific sampling frequencies in shellfish safety monitoring programmes. [ABSTRACT FROM PUBLISHER]

Mussels Choromytilus meridionalis and oysters Crassostrea gigas were suspended from a mooring off Lambert's Bay, South Africa, to study the kinetics of lipophilic phycotoxin accumulation and detoxification. The shellfish were subsequently harvested daily over approximately three weeks and analysed for lipophilic phycotoxins by liquid chromatography coupled with tandem mass spectrometry in the multiple reaction monitoring mode. A mass transition typical for the cyclic imine toxin gymnodimine was detected in low but consistent levels in all oyster samples throughout the sampling period, whereas this peak was detected with lesser intensity in only some mussel samples and was frequently below the limit of quantitation (0.02 μg g-1 fresh weight). Comparison of retention times and collision-induced mass spectra of a certified standard of gymnodimine and an oyster extract provided unambiguous confirmation of the identity of gymnodimine in the shellfish extracts. The absence of known producers of gymnodimine in the plankton, and the non-detection of gymnodimine in filtered water samples collected during the period of study, lead to the conclusion that the shellfish were contaminated at their site of initial collection in Saldanha Bay prior to deployment off Lambert's Bay. This finding is the first confirmed evidence of gymnodimine in the southern Benguela upwelling system. [ABSTRACT FROM AUTHOR]

Abstract Background The dinoflagellate Alexandrium minutum typically produces paralytic shellfish poisoning (PSP) toxins, which are known only from cyanobacteria and dinoflagellates. While a PSP toxin gene cluster has recently been characterized in cyanobacteria, the genetic background of PSP toxin production in dinoflagellates remains elusive. Results We constructed and analysed an expressed sequence tag (EST) library of A. minutum, which contained 15,703 read sequences yielding a total of 4,320 unique expressed clusters. Of these clusters, 72% combined the forward-and reverse reads of at least one bacterial clone. This sequence resource was then used to construct an oligonucleotide microarray. We analysed the expression of all clusters in three different strains. While the cyanobacterial PSP toxin genes were not found among the A. minutum sequences, 192 genes were differentially expressed between toxic and non-toxic strains. Conclusions Based on this study and on the lack of identified PSP synthesis genes in the two existent Alexandrium tamarense EST libraries, we propose that the PSP toxin genes in dinoflagellates might be more different from their cyanobacterial counterparts than would be expected in the case of a recent gene transfer. As a starting point to identify possible PSP toxin-associated genes in dinoflagellates without relying on a priori sequence information, the sequences only present in mRNA pools of the toxic strain can be seen as putative candidates involved in toxin synthesis and regulation, or acclimation to intracellular PSP toxins.

Azaspiracid-59 (AZA-59) was detected in plankton in coastal waters of the Pacific Northwest USA. Given that bivalves metabolize and transform accumulated phycotoxins, a strain of Azadinium poporum isolated from the coast of Washington State that is a known producer of AZA-59 was used in a controlled feeding experiment with mussels (Mytilus edulis) to assess AZA-59 accumulation rates and transformation into shellfish metabolites. Mussels started feeding immediately after the addition of A. poporum. Mussels were generally healthy during the entire experimental exposure of 18 days with prevailingly high rates of clearance (ca 100 mL per mussel and hour) and ingestion. Mussel were extracted after different exposure times and were analyzed by liquid chromatography coupled with low- and high-resolution mass spectrometry. In the course of the experiment a number of putative AZA-59 metabolites were detected including hydroxyl and carboxy analogs that corresponded with previously reported mussel metabolites of AZA-1. A significant formation of 3-OH fatty acid acyl esters relative to free AZAs was observed through the time course of the study, with numerous fatty acid ester variants of AZA-59 confirmed. These results illustrate the potential for metabolism of AZA59 in shellfish and provide important information for local AZA monitoring and toxicity testing along the Northern Pacific US coast., Competing Interests: Declaration of Competing Interest ☒ The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Harmful dinoflagellates and their resulting blooms pose a threat to marine life and human health. However, to date, global maps of marine life often overlook harmful microorganisms. As harmful algal blooms (HABs) increase in frequency, severity, and extent, understanding the distribution of harmful dinoflagellates and their drivers is crucial for their management. We used MaxEnt, random forest, and ensemble models to map the habitats of the representative HABs species in the genus Alexandrium, including A. catenella, A. minutum, and A. pacificum. Since species occurrence records used in previous studies were solely morphology-based, potentially leading to misidentifications, we corrected these species' distribution records using molecular criteria. The results showed that the key environmental drivers included the distance to the coastline, bathymetry, sea surface temperature (SST), and dissolved oxygen. Alexandrium catenella thrives in temperate to cold zones and is driven by low SST and high oxygen levels. Alexandrium pacificum mainly inhabits the Temperate Northern Pacific and prefers warmer SST and lower oxygen levels. Alexandrium minutum thrives universally and adapts widely to SST and oxygen. By analyzing the habitat suitability of locations with recorded HAB occurrences, we found that high habitat suitability could serve as a reference indicator for bloom risk. Therefore, we have proposed a qualitative method to spatially assess the harmful algae risk according to the habitat suitability. On the global risk map, coastal temperate seas, such as the Mediterranean, Northwest Pacific, and Southern Australia, faced higher risks. Although HABs currently have restricted geographic distributions, our study found these harmful algae possess high environmental tolerance and can thrive across diverse habitats. HAB impacts could increase if climate changes or ocean conditions became more favorable. Marine transportation may also spread the harmful algae to new unaffected ecosystems. This study has pioneered the assessment of harmful algal risk based on habitat suitability., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The dinoflagellate Alexandrium pseudogonyaulax, a harmful algal bloom species, is currently appearing in increasing frequency and abundance across Northern European waters, displacing other Alexandrium species. This mixotrophic alga produces goniodomins (GDs) and bioactive extracellular substances (BECs) that may pose a threat to coastal ecosystems and other marine resources. This study demonstrated the adverse effects of A. pseudogonyaulax on four marine trophic levels, including microalgae (Rhodomonas salina), microzooplankton (Polykrikos kofoidii) and mesozooplankton (Acartia tonsa), as well as fish gill cells (RTgill-W1, Oncorhynchus mykiss), ultimately leading to enhanced mortality and cell lysis. Furthermore, cell-free supernatants collected from A. pseudogonyaulax cultures caused complete loss of metabolic activity in the RTgill-W1 cell line, indicating ichthyotoxic properties, while all tested GDs were much less toxic. In addition, cell-free supernatants of A. pseudogonyaulax led to cell lysis of R. salina, while all tested GDs were non-lytic. Finally, reduced egg hatching rates of A. tonsa eggs exposed to cell-free supernatants of A. pseudogonyaulax and impaired mobility of P. kofoidii and A. tonsa exposed to A. pseudogonyaulax were also observed. Altogether, bioassay results suggest that the toxicity of A. pseudogonyaulax is mainly driven by BECs and not by GDs, although further research into factors modulating the lytic activity of Alexandrium spp. are needed., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

The production of allelochemicals by the toxigenic dinoflagellate Alexandrium catenella is one of the suggested mechanisms to facilitate its bloom formation and persistence by outcompeting other phototrophic protists and reducing grazing pressure. In Southern California, toxic events caused by A. catenella and paralytic shellfish toxins (PSTs) regularly impact coastal ecosystems; however, the trophic interactions and mechanisms promoting this species in a food web context are still not fully understood. In the present study, we combined a dynamical mathematical model with laboratory experiments to investigate potential toxic and allelochemical effects of an A. catenella strain isolated off the coast of Los Angeles, Southern California, on competitors and a common zooplankton consumer. Experiments were conducted using three toxigenic strains of A. catenella, comparing the new Californian isolate (Alex Cal) to two strains previously described from the North Sea, a lytic (Alex2) and non-lytic (Alex5) strain, testing for donor density-dependent effects on two phytoplankton species (Rhodomonas salina, Tetraselmis sp.) and on the rotifer Brachionus plicatilis. Bioassays revealed a steep decline in competitor and consumer populations with increasing Alex Cal concentrations, indicating an intermediate lytic activity compared to the North Sea strains (lytic Alex2 and non-lytic Alex5). The rotifer fed and grew well on the PST- toxic, but non-lytic Alex5 strain, while its survival significantly decreased with increasing concentrations of the two lytic strains Alex Cal and Alex 2, indicating that negative effects on the rotifer were mediated by allelochemicals rather than PST-toxins. Mixed culture experiments including both competitors and consumers demonstrated that the intensity of allelochemical effects not only depended on the A. catenella density but also on the target density. Negative effects on grazers were alleviated by co-occurring competitors with a lower sensitivity to allelochemicals, thus reducing harmful compounds and allowing grazing control on the dinoflagellate to come into effect again. Results from mixed culture experiments were supported by the mathematical approach used in this study which was calibrated with data from simple monoculture growth, pairwise competition and predator-prey experiments, demonstrating the applicability of this model approach to predict the outcome of more complex food web dynamics at the community level., Competing Interests: Declaration of competing interest I hereby declare on behalf of all authors that we have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Prevalent interactions among marine phytoplankton triggered by long-range climatic stressors are well-known environmental disturbers of community structure. Dynamic response of phytoplankton physiology is likely to come from interspecies interactions rather than direct climatic effect on single species. However, studies on enigmatic interactions among interspecies, which are induced by bioactive extracellular compounds (BECs), especially between related harmful algae sharing similar shellfish toxins, are scarce. Here, we investigated how BECs provoke the interactions between two notorious algae, Alexandrium minutum and Gymnodinium catenatum, which have similar paralytic shellfish toxin (PST) profiles. Using techniques including electron microscopy and transcriptome analysis, marked disruptions in G. catenatum intracellular microenvironment were observed under BECs pressure, encompassing thylakoid membrane deformations, pyrenoid matrix shrinkage and starch sheaths disappearance. In addition, the upregulation of gene clusters responsible for photosystem-I Lhca1/4 and Rubisco were determined, leading to weaken photon captures and CO 2 assimilation. The redistribution of lipids and proteins occurred at the subcellular level based on in situ focal plane array FTIR imaging approved the damages. Our findings illuminated an intense but underestimated interspecies interaction triggered by BECs, which is responsible for dysregulating photosynthesis and organelle function in inferior algae and may potentially account for fitness alteration in phytoplankton community., (© 2024 John Wiley & Sons Ltd.)

In the context of harmful algal blooms, fish can be exposed to the combined effects of more than one toxin. We studied the effects of consecutive exposure to Microcystin-LR (MCLR) in vivo and paralytic shellfish toxins (PST) ex vivo/in vitro (MCLR+PST) in the rainbow trout Oncorhynchus mykiss's middle intestine. We fed juvenile fish with MCLR incorporated in the feed every 12 h and euthanized them 48 h after the first feeding. Immediately, we removed the middle intestine to make ex vivo and in vitro preparations and exposed them to PST for one hour. We analyzed glutathione (GSH) and glutathione disulfide (GSSG) contents, glutathione S-transferase (GST), glutathione reductase (GR), catalase (CAT), and protein phosphatase 1 (PP1) activities in ex vivo intestinal strips; apical and basolateral ATP-biding cassette subfamily C (Abcc)-mediated transport in ex vivo everted and non- everted sacs; and reactive oxygen species (ROS) production in isolated enterocytes in vitro. MCLR+PST treatment decreased the GSH content, GSH/GSSG ratio, GST activity, and increased ROS production. GR activity remained unchanged, while CAT activity only increased in response to PST. MCLR inhibited PP1 activity and activated Abcc-mediated transport only at the basolateral side of the intestine. Our results show a combined effect of MCLR+PST on the oxidative balance in the O. mykiss middle intestine, which is not affected by the two toxins groups when applied individually. Basolateral Abcc transporters activation by MCLR treatment could lead to an increase in the absorption of toxicants (including MCLR) into the organism. Therefore, MCLR makes the O. mykiss middle intestine more sensitive to possibly co-occurring cyanotoxins like PST., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

An unusual mass mortality event (MME) of harbour seals (Phoca vitulina) and harbour porpoises (Phocoena phocoena) occurred in Denmark and Sweden in June 2007. Prior to this incident, the region had experienced two MMEs in harbour seals caused by Phocine Distemper Virus (PDV) in 1988 and 2002. Although epidemiology and symptoms of the 2007 MME resembled PDV, none of the animals examined for PDV tested positive. Thus, it has been speculated that another - yet unknown - pathogen caused the June 2007 MME. To shed new light on the likely cause of death, we combine previously unpublished veterinary examinations of harbour seals with novel analyses of algal toxins and algal monitoring data. All harbour seals subject to pathological examination showed pneumonia, but were negative for PDV, influenza and coronavirus. Histological analyses revealed septicaemia in multiple animals, and six animals tested positive for Klebsiella pneumonia. Furthermore, we detected the algal Dinophysis toxin DTX-1b (1-115 ng g -1 ) in five seals subject to toxicology, representing the first time DTX-1b has been detected in marine vertebrates. However, no animals tested positive for both Klebsiella and toxins. Thus, while our relatively small sample size prevent firm conclusions on causative agents, we speculate that the unexplained MME may have been caused by a chance incidence of multiple pathogens acting in parallel in June 2007, including Dinophysis toxin and Klebsiella. Our study illustrates the complexity of wildlife MMEs and highlights the need for thorough sampling during and after MMEs, as well as additional research on and monitoring of DTX-1b and other algal toxins in the region., Competing Interests: Declaration of competing interest The authors declare that they know of no competing financial- or personal relationships that could influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Enhancing ocean productivity by artificial upwelling is evaluated as a nature-based solution for food security and climate change mitigation. Fish production is intended through diatom-based plankton food webs as these are assumed to be short and efficient. However, our findings from mesocosm experiments on artificial upwelling in the oligotrophic ocean disagree with this classical food web model. Here, diatoms did not reduce trophic length and instead impaired the transfer of primary production to crustacean grazers and small pelagic fish. The diatom-driven decrease in trophic efficiency was likely mediated by changes in nutritional value for the copepod grazers. Whilst diatoms benefitted the availability of essential fatty acids, they also caused unfavorable elemental compositions via high carbon-to-nitrogen ratios (i.e. low protein content) to which the grazers were unable to adapt. This nutritional imbalance for grazers was most pronounced in systems optimized for CO 2 uptake through carbon-to-nitrogen ratios well beyond Redfield. A simultaneous enhancement of fisheries production and carbon sequestration via artificial upwelling may thus be difficult to achieve given their opposing stoichiometric constraints. Our study suggest that food quality can be more critical than quantity to maximize food web productivity during shorter-term fertilization of the oligotrophic ocean., (© 2024. The Author(s).)

This study describes two novel species of marine dinophytes in the genus Alexandrium. Morphological characteristics and phylogenetic analyses support the placement of the new taxa, herein designated as Alexandrium limii sp. nov. and A. ogatae sp. nov. Alexandrium limii, a species closely related to A. taylorii, is distinguished by having a shorter 2'/4' suture length, narrower plates 1' and 6'', with larger length: width ratios, and by the position of the ventral pore (Vp). Alexandrium ogatae is distinguishable with its metasert plate 1' having almost parallel lateral margins, and by lacking a Vp. Production of paralytic shellfish toxins (PSTs), cycloimines, and goniodomins (GDs) in clonal cultures of A. ogatae, A. limii, and A. taylorii were examined analytically and the results showed that all strains contained GDs, with GDA as major variants (6-14 pg cell -1 ) for all strains except the Japanese strain of A. limii, which exclusively had a desmethyl variant of GDA (1.4-7.3 pg cell -1 ). None of the strains contained detectable levels of PSTs and cycloimines., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Goniodomin A (GDA) is a polyketide macrolide produced by multiple species of the marine dinoflagellate genus Alexandrium. GDA is unusual in that it undergoes cleavage of the ester linkage under mild conditions to give mixtures of seco acids (GDA-sa). Ring-opening occurs even in pure water although the rate of cleavage accelerates with increasing pH. The seco acids exist as a dynamic mixture of structural and stereo isomers which is only partially separable by chromatography. Freshly prepared seco acids show only end absorption in the UV spectrum but a gradual bathochromic change occurs, which is consistent with formation of α,β-unsaturated ketones. Use of NMR and crystallography is precluded for structure elucidation. Nevertheless, structural assignments can be made by mass spectrometric techniques. Retro-Diels-Alder fragmentation has been of value for independently characterizing the head and tail regions of the seco acids. The chemical transformations of GDA revealed in the current studies help clarify observations made on laboratory cultures and in the natural environment. GDA has been found to reside mainly within the algal cells while the seco acids are mainly external with the transformation of GDA to the seco acids occurring largely outside the cells. This relationship, plus the fact that GDA is short-lived in growth medium whereas GDA-sa is long-lived, suggests that the toxicological properties of GDA-sa in its natural environment are more important for the survival of the Alexandrium spp. than those of GDA. The structural similarity of GDA-sa to that of monensin is noted. Monensin has strong antimicrobial properties, attributed to its ability to transport sodium ions across cell membranes. We propose that toxic properties of GDA may primarily be due to the ability of GDA-sa to mediate metal ion transport across cell membranes of predator organisms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Phytoplankton succession is related to hydroclimatic conditions. In this study we provide the first description of a toxic phytoplankton succession in the Patagonian Fjord System. The shift was modulated by atmospheric-oceanographic forcing and consisted of the replacement of the marine dinoflagellate Dinophysis acuta in a highly stratified water column during austral summer by the diatom Pseudo-nitzschia calliantha in a mixed water column during late summer and early autumn. This transition, accompanied by a change in the biotoxin profiles (from lipophilic dinophysis toxins to hydrophilic domoic acid), was induced by the arrival of an intense atmospheric river. The winds in Magdalena Sound may have been further amplified, due to its west-east orientation and its location within a tall, narrow mountain canyon. This work also documents the first known appearance of toxic P. calliantha in Northern Patagonian. The potential impacts of the biotoxins of this species on higher trophic levels are discussed., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

To date, the putative shellfish toxin azaspiracid 59 (AZA-59) produced by Azadinium poporum (Dinophyceae) has been the only AZA found in isolates from the Pacific Northwest coast of the USA (Northeast Pacific Ocean). Anecdotal reports of sporadic diarrhetic shellfish poisoning-like illness, with the absence of DSP toxin or Vibrio contamination, led to efforts to look for other potential toxins, such as AZAs, in water and shellfish from the region. A. poporum was found in Puget Sound and the outer coast of Washington State, USA, and a novel AZA (putative AZA-59) was detected in low quantities in SPATT resins and shellfish. Here, an A. poporum strain from Puget Sound was mass-cultured and AZA-59 was subsequently purified and structurally characterized. In vitro cytotoxicity of AZA-59 towards Jurkat T lymphocytes and acute intraperitoneal toxicity in mice in comparison to AZA-1 allowed the derivation of a provisional toxicity equivalency factor of 0.8 for AZA-59. Quantification of AZA-59 using ELISA and LC-MS/MS yielded reasonable quantitative results when AZA-1 was used as an external reference standard. This study assesses the toxic potency of AZA-59 and will inform guidelines for its potential monitoring in case of increasing toxin levels in edible shellfish., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

The genus Amphidinium has been the subject of recent attention due to the production of polyketide metabolites. Some of these compounds have shown significant bioactivities and could be related to species interactions in the natural benthic microenvironment. Among these compounds, amphidinols (AMs) are suspected to be related to fish kills and probably implicated in ciguatera symptoms associated with the occurrence of benthic harmful algal blooms (bHABs). Here, we present the first report of a variety of AMs produced by cultured strains from several species from the Mexican Pacific, the Gulf of California, and the Gulf of Mexico. Through ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS), ten previously known AMs (AM02, -04, -05, -06, -07, -09, -11, -14, -15, and -17), four recently reported AMs (N7, N8/N9, N12, and N13), and three new variants (U1, U2, and U3) were identified. Of the twelve analyzed Amphidinium cultures, five were not AM producers, and the cell quotas of the remaining seven strains ranged from close to nondetectable to a maximum of 1694 fg cell -1 , with many intermediate levels in between. The cultures from the Mexican North Pacific coast produced AMs in a higher quantity and variety than those from worldwide locations. This is the first study of AMs from Mexican Amphidinium strains, and our results confirm the relevance of continuing the investigation of the genus bioactive metabolites.

Dinoflagellates of the genus Karlodinium are ichthyotoxic species that produce toxins including karlotoxins and karmitoxins. Karlotoxins show hemolytic and cytotoxic activities and have been associated with fish mortality. This study evaluated the effect of toxins released into the environment of Karlodinium veneficum strain K10 (Ebro Delta, NW Mediterranean) on the early stages of Danio rerio (zebrafish). Extracts of the supernatant of K10 contained the mono-sulfated KmTx-10, KmTx-11, KmTx-12, KmTx-13, and a di-sulfated form of KmTx-10. Total egg mortality was observed for karlotoxin concentration higher than 2.69 μg L -1 . For 1.35 μg L -1 , 87% of development anomalies were evidenced (all concentrations were expressed as KmTx-2 equivalent). Larvae of 8 days postfertilization exposed to 1.35 µg L -1 presented epithelial damage with 80% of cells in the early apoptotic stage. Our results indicate that supernatants with low concentration of KmTxs produce both lethal and sublethal effects in early fish stages. Moreover, apoptosis was induced at concentrations as low as 0.01 μg L -1 . This is of great relevance since detrimental long-term effects due to exposure to low concentrations of these substances could affect wild and cultured fish., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Rationale: More than half of surveyed microalgae and over 90% of harmful algae have an obligate requirement for vitamin B 12 , but methods for directly measuring dissolved B 12 in seawater are scarce due to low concentrations and rapid light-induced hydrolysis., Methods: We present a method to detect and measure the four main congeners of vitamin B 12 dissolved in seawater. The method includes solid-phase extraction, separation by ultrahigh-performance liquid chromatography and detection by triple-quadrupole tandem mass spectrometry utilizing an electrospray ion source. This method was applied to coastal field samples collected in the German Bay, Baltic Sea and the Danish Limfjord system., Results: The total dissolved B 12 pool ranged between 0.5 and 2.1 pM. Under ambient conditions methyl-B 12 and adenosyl-B 12 were nearly fully hydrolyzed to hydroxy-B 12 in less than 1 h. Hydroxy-B 12 and a novel, corresponding isomer were the main forms of B 12 found at all field sites. This isomer eluted well after the OH-B 12 peak and was also detected in commercially available OH-B 12 . Both compounds showed very high similarity in their collision-induced dissociation spectra., Conclusions: The high instability of the biologically active forms of Me-B 12 and Ado-B 12 towards hydrolysis was shown, highlighting the importance of reducing the duration of the extraction protocol. In addition, the vitamin B 12 pool in the study area was mostly comprised of a previously undescribed isomer of OH-B 12 . Further studies into the structure of this isomer and its bioavailability are needed., (© 2022 The Authors. Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd.)

The study aims to unravel the variability of Dinophysis spp. and their alleged toxins in conjunction with environmental drivers in Ambon Bay. Phytoplankton samples, lipophilic toxins and physiochemical water properties were analysed during a 1.5-year period. Three Dinophysis species (D. miles, D. caudata, and D. acuminata) were found in plankton samples, of which D. miles was the most abundant and persistently occurring species. Pectenotoxin-2 (PTX2) and its secoacid (PTX2sa) were detected throughout, and PTX2sa levels strongly correlated with D. miles cell abundance. The toxin showed a positive correlation with temperature, which may suggest that D. miles cells contain rather constant PTX2sa during warmer months. Dissolved nitrate concentrations were found to play a major role in regulating cell abundances and toxin levels. This study adds adequate information regarding marine biotoxins and potentially toxic species for future Harmful Algal Bloom management in Ambon and Indonesia at large., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Numerous potentially toxic plankton species commonly occur in the Black Sea, and phycotoxins have been reported. However, the taxonomy, phycotoxin profiles, and distribution of harmful microalgae in the basin are still understudied. An integrated microscopic (light microscopy) and molecular (18S rRNA gene metabarcoding and qPCR) approach complemented with toxin analysis was applied at 41 stations in the northwestern part of the Black Sea for better taxonomic coverage and toxin profiling in natural populations. The combined dataset included 20 potentially toxic species, some of which ( Dinophysis acuminata , Dinophysis acuta , Gonyaulax spinifera , and Karlodinium veneficum ) were detected in over 95% of the stations. In parallel, pectenotoxins (PTX-2 as a major toxin) were registered in all samples, and yessotoxins were present at most of the sampling points. PTX-1 and PTX-13, as well as some YTX variants, were recorded for the first time in the basin. A positive correlation was found between the cell abundance of Dinophysis acuta and pectenotoxins, and between Lingulodinium polyedra and Protoceratium reticulatum and yessotoxins. Toxic microalgae and toxin variant abundance and spatial distribution was associated with environmental parameters. Despite the low levels of the identified phycotoxins and their low oral toxicity, chronic toxic exposure could represent an ecosystem and human health hazard.

Historical records (1980-2018) of potentially toxic phytoplankton and phycotoxins on the Argentine Continental Shelf (35°S-56.5°S) and adjacent ocean waters were systematically reviewed from scientific literature to assess their abundance and diversity over spatial and temporal scales. Records increased from 124 in the period 1980-1992 to 638 in 2006-2018, and the scanned area expanded from coastal to offshore waters including the shelf-break front. Alexandrium was the most reported genus (54%) during 1980-1992 and Pseudo-nitzschia (52%) during 1993-2005. By 2006-2018, a higher diversity was documented: Alexandrium (20%), Dinophysis (32%), Pseudo-nitzschia (31%), and the most recently described potentially toxic dinoflagellates of the family Amphidomataceae (8%). Likewise, a wider spectrum of phycotoxins was documented in the last decade, with lipophilic (LSTs) and paralytic shellfish toxins (PSTs) as the most recorded. Increased records are related to intensified monitoring, more detailed taxonomic analyses and more sensitive chemical techniques for marine biotoxin detection. This quantitative assessment brings light to the widespread occurrence of HABs along contrasting areas of the Patagonian Shelf and sets the basis for ecosystem risk evaluation. Moreover, comparison of toxic phytoplankton reported in the SW Atlantic with those in similar temperate seas in the North Atlantic and the Pacific Ocean, disclose ocean basin differences in strain toxicity of A. ostenfeldii, D. tripos and Azadinium species., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier B.V.)