New invasive Nemertean species (Cephalothrix Simula) in England with high levels of tetrodotoxin and a microbiome linked to toxin metabolism

The marine nemertean Cephalothrix simula originates from the Pacific Ocean but in recent years has been discovered in northern Europe. The species has been associated with high levels of the marine neurotoxin Tetrodotoxin, traditionally associated with Pufferfish Poisoning. This study reports the first discovery of two organisms of C. simula in the UK, showing the geographical extent of this species is wider than originally described. Species identification was initially conducted morphologically, with confirmation by Cox 1 DNA sequencing. 16S gene sequencing enabled the taxonomic assignment of the microbiome, showing the prevalence of a large number of bacterial genera previously associated with TTX production including Alteromonas, Vibrio and Pseudomonas. LC-MS/MS analysis of the nemertean tissue revealed the presence of multiple analogues of TTX, dominated by the parent TTX, with a total toxin concentration quantified at 54 µg TTX per g of tissue. Pseudomonas luteola isolated from C. simula, together with Vibrio alginolyticus from the native nemertean Tubulanus annulatus, were cultured at low temperature and both found to contain TTX. Overall, this paper confirms the high toxicity of a newly discovered invasive nemertean species with links to toxin-producing marine bacteria and the potential risk to human safety. Further work is required to assess the geographical extent and toxicity range of C. simula along the UK coast in order to properly gauge the potential impacts on the environment and human safety

Fatal Canine Intoxications Linked to the Presence of Saxitoxins in Stranded Marine Organisms Following Winter Storm Activity.

At the start of 2018, multiple incidents of dog illnesses were reported following consumption of marine species washed up onto the beaches of eastern England after winter storms. Over a two-week period, nine confirmed illnesses including two canine deaths were recorded. Symptoms in the affected dogs included sickness, loss of motor control, and muscle paralysis. Samples of flatfish, starfish, and crab from the beaches in the affected areas were analysed for a suite of naturally occurring marine neurotoxins of dinoflagellate origin. Toxins causing paralytic shellfish poisoning (PSP) were detected and quantified using two independent chemical testing methods in samples of all three marine types, with concentrations over 14,000 µg saxitoxin (STX) eq/kg found in one starfish sample. Further evidence for PSP intoxication of the dogs was obtained with the positive identification of PSP toxins in a vomited crab sample from one deceased dog and in gastrointestinal samples collected post mortem from a second affected dog. Together, this is the first report providing evidence of starfish being implicated in a PSP intoxication case and the first report of PSP in canines

Detection of Tetrodotoxin Shellfish Poisoning (TSP) Toxins and Causative Factors in Bivalve Molluscs from the UK

Publication history: Accepted - 28 August 2017; Published online - 30 August 2017.Tetrodotoxins (TTXs) are traditionally associated with the occurrence of tropical Pufferfish Poisoning. In recent years, however, TTXs have been identified in European bivalve mollusc shellfish, resulting in the need to assess prevalence and risk to shellfish consumers. Following the previous identification of TTXs in shellfish from southern England, this study was designed to assess the wider prevalence of TTXs in shellfish from around the coast of the UK. Samples were collected between 2014 and 2016 and subjected to analysis using HILIC-MS/MS. Results showed the continued presence of toxins in shellfish harvested along the coast of southern England, with the maximum concentration of total TTXs reaching 253 µg/kg. TTX accumulation was detected in Pacific oysters (Crassostrea gigas), native oysters (Ostrea edulis) common mussels (Mytilus edulis) and hard clams (Mercenaria mercenaria), but not found in cockles (Cerastoderma edule), razors (Ensis species) or scallops (Pecten maximus). Whilst the highest concentrations were quantified in samples harvested during the warmer summer months, TTXs were still evident during the winter. An assessment of the potential causative factors did not reveal any links with the phytoplankton species Prorocentrum cordatum, instead highlighting a greater level of risk in areas of shallow, estuarine waters with temperatures above 15 °CFunding from Cefas internal funding sources (Seedcorn, DP345B and DP402); and FS

Current Status of Forecasting Toxic Harmful Algae for the North-East Atlantic Shellfish Aquaculture Industry

Across the European Atlantic Arc (Scotland, Ireland, England, France, Spain, and Portugal) the shellfish aquaculture industry is dominated by the production of mussels, followed by oysters and clams. A range of spatially and temporally variable harmful algal bloom species (HABs) impact the industry through their production of biotoxins that accumulate and concentrate in shellfish flesh, which negatively impact the health of consumers through consumption. Regulatory monitoring of harmful cells in the water column and toxin concentrations within shellfish flesh are currently the main means of warning of elevated toxin events in bivalves, with harvesting being suspended when toxicity is elevated above EU regulatory limits. However, while such an approach is generally successful in safeguarding human health, it does not provide the early warning that is needed to support business planning and harvesting by the aquaculture industry. To address this issue, a proliferation of web portals have been developed to make monitoring data widely accessible. These systems are now transitioning from “nowcasts” to operational Early Warning Systems (EWS) to better mitigate against HAB-generated harmful effects. To achieve this, EWS are incorporating a range of environmental data parameters and developing varied forecasting approaches. For example, EWS are increasingly utilizing satellite data and the results of oceanographic modeling to identify and predict the behavior of HABs. Modeling demonstrates that some HABs can be advected significant distances before impacting aquaculture sites. Traffic light indices are being developed to provide users with an easily interpreted assessment of HAB and biotoxin risk, and expert interpretation of these multiple data streams is being used to assess risk into the future. Proof-of-concept EWS are being developed to combine model information with in situ data, in some cases using machine learning-based approaches. This article: (1) reviews HAB and biotoxin issues relevant to shellfish aquaculture in the European Atlantic Arc (Scotland, Ireland, England, France, Spain, and Portugal; (2) evaluates the current status of HAB events and EWS in the region; and (3) evaluates the potential of further improving these EWS though multi-disciplinary approaches combining heterogeneous sources of information.Versión del edito

Distribution of Tetrodotoxin in Pacific Oysters (Crassostrea gigas)

A potent and heat-stable tetrodotoxin (TTX) has been found to accumulate in various marine bivalve species, including Pacific oysters (Crassostrea gigas), raising a food safety concern. While several studies on geographical occurrence of TTX have been conducted, there is a lack of knowledge about the distribution of the toxin within and between bivalves. We, therefore, measured TTX in the whole flesh, mantle, gills, labial palps, digestive gland, adductor muscle and intravalvular fluid ofC. gigas using liquid chromatography-tandem mass spectrometry. Weekly monitoring during summer months revealed the highest TTX concentrations in the digestive gland (up to 242 µg/kg), significantly higher than in other oyster tissues. Intra-population variability of TTX, measured in the whole flesh of each of twenty animals, reached 46% and 32% in the two separate batches, respectively. In addition, an inter-population study was conducted to compare TTX levels at four locations within the oyster production area. TTX concentrations in the whole flesh varied significantly between some of these locations, which was unexplained by the differences in weight of flesh. This is the first study examining TTX distribution inC. gigas and the first confirmation of the preferential accumulation of TTX in oyster digestive gland

Analysis of Microcystins in Cyanobacterial Blooms from Freshwater Bodies in England

Cyanobacterial blooms in freshwater bodies in England are currently monitored reactively, with samples containing more than 20,000 cells/mL of potentially toxin-producing species by light microscopy resulting in action by the water body owner. Whilst significantly reducing the risk of microcystin exposure, there is little data describing the levels of these toxins present in cyanobacterial blooms. This study focused on the quantitative LC-MS/MS analysis of microcystins in freshwater samples, collected across England during 2016 and found to contain potentially toxin-producing cyanobacteria. More than 50% of samples contained quantifiable concentrations of microcystins, with approximately 13% exceeding the WHO medium health threshold of 20 μg/L. Toxic samples were confirmed over a nine-month period, with a clear increase in toxins during late summer, but with no apparent geographical patterns. No statistical relationships were found between total toxin concentrations and environmental parameters. Complex toxin profiles were determined and profile clusters were unrelated to cyanobacterial species, although a dominance of MC-RR was determined in water samples from sites associated with lower rainfall. 100% of samples with toxins above the 20 μg/L limit contained cell densities above 20,000 cells/mL or cyanobacterial scum, showing the current regime is suitable for public health. Conversely, with only 18% of cell density threshold samples having total microcystins above 20 μg/L, there is the potential for reactive water closures to unnecessarily impact upon the socio-economics of the local population. In the future, routine analysis of bloom samples by LC-MS/MS would provide a beneficial confirmatory approach to the current microscopic assessment, aiding both public health and the needs of water users and industry

The invasive sea slug Pleurobranchaea maculata is a vector of two potent neurotoxins in coasts of Argentina

Toxic exotic organisms can have profound effect as new vectors of keystone compounds in non-native areas. In recent years the invasive sea slug Pleurobranchaea maculata has been reported as thriving along the oriental coasts of South America. The same species had been previously found to contain high levels of tetrodotoxins (TTXs) in its native range. With the aim of determining toxin contents for the introduced individuals we performed mouse bioassays (MBA) and liquid chromatography tandem?mass spectrometry analyses (LC?MS/MS) in three distant populations (− 38°02′11″, − 57°31′28″; − 40°29′59″, − 60°14′09″; − 42°44′15″, − 65°01′40″) and followed the temporal variation in toxin contents in one of them, from June 2014 to January 2015. Relative low levels of TTXs were detected jointly with high levels of PSTs. This is the first identification of TTXs in the temperate coasts of the southwestern Atlantic and the first detection of PSTs in a pleurobranch, in both adults and eggs. Concentrations of PSTs and TTXs varied widely among individuals and populations, and through time. Our results provide new hints on the origin and acquisition mechanisms of these toxins in P. maculata and highlight the risk posed by the introduction of this new vector of potent neurotoxins for seafood safety and marine communities in the invaded area.Fil: Farias, Nahuel Emiliano. Universidad Nacional de Mar del Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; ArgentinaFil: Goya, Alejandra Beatriz. Centro Regional Buenos Aires Sur Senasa (servicio Nacional de Sanidad y Calidad Agroalimentaria); ArgentinaFil: Schwindt, Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Biología de Organismos Marinos; ArgentinaFil: Obenat, Sandra Mabel. Universidad Nacional de Mar del Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; ArgentinaFil: Dhanji-Rapkova, Monika. Centre For The Environment Fisheries And Aquaculture Science; Reino UnidoFil: Turner, Andrew D.. Centre For The Environment Fisheries And Aquaculture Science; Reino Unid