Workshop on morpho-molecular methods for the study of dinoflagellate cysts A workshop on techniques for the morphological and molecular identification of cysts from toxic HAB species, led by Drs Kenneth Neil Mertens, Kirsty Smith, Lesley Rhodes and Lincoln MacKenzie, was held recently at the Cawthron Institute in Nelson, New Zealand, 12-14 February 2018. This workshop was supported by the Catalyst:Seeding programme, financed by the Royal Society of New Zealand to establish a new research collaboration between Cawthron and IFREMER (Concarneau, France). Twenty-four researchers participated, with the majority from New Zealand and Australia, but also included people from France, Belgium, Singapore, Ireland, the UK and Canada (Fig1). Due to heavy rains on February 11th, several airplanes had trouble in landing at Nelson and experienced delays, but luckily nearly all participants arrived on time for the start of the workshop. The workshop started on 12 February with a Māori welcoming ceremony (pōwhiri), which included a speech by Cawthrons CEO Charles Eason and a response by Kenneth (IFREMER) in his Dutch native tongue. The speeches were followed by the singing of traditional Māori songs most memorable was the impressive falsetto of Dave Clark (Marine Institute, Ireland). The ceremony ended with the hongi (a traditional Māori greeting) all in all quite a spiritual experience indeed. After getting our feet back on earth, the workshop aims were introduced by Kirsty (Cawthron) and Kenneth, after which participants introduced themselves and their specific research topics, which covered several research fields. Geological applications of cysts were exemplified by Anna Pienkowski, Vera Pospelova, Erica Crouch and Lucia Roncaglia biological applications by Chris Reid, Gustaaf Hallegraeff, Janet Adamson, Aye Aye Mon, Shereen Lim, Viliami Langi and Tony Bui, and molecular tools by Dave Clarke, Rendy Ruvindy, Henna Savela, Laura Biessy and Xavier Pochon. Cawthron MSc students Enora Jaffrézic, Maïlys Picard and Julie Steyen also outlined their research projects on HAB species and cysts. After lunch, morphological techHARMFUL ALGAE NEWS NO. 60 / 2018 niques for cyst study were explained by Kenneth, and were followed by Vera Pospelova (University of Victoria, Canada) who detailed the application of sediment-traps to study cyst production. Thereafter, Lesley and Kirsty presented molecular techniques used for study of motile stages and cysts. Erica Crouch (GNS, New Zealand) outlined paleontological techniques used for late Quaternary paleocological reconstructions around New Zealand, highlighting the work of her collaborator Joe Prebble. After coffee, Lincoln shared with us how blooms of Alexandrium pacificum in New Zealand can be traced using both plankton samples as well as sediment samples. Gustaaf Hallegraeff (University of Tasmania, Australia) emphasized how selective the fossil record is, and highlighted new possibilities using modern molecular techniques. Last but not least, Susie Wood (Cawthron) presented her planned research to study the historical evolution of cyanobacterial and other microbial communities in over 300 lakes across New Zealand. The day ended with a delightful dinner at the Indian Café, where a moderate amount of craft beer was consumed. The next day included practical demonstrations of both morphological as well as molecular methods (Fig. 2). Lincoln demonstrated the use of primuline staining to enumerate Alexandrium cysts in the sediment [1]. Kenneth applied the sodium polytungstate method to isolate living cysts from the sediments for germination experiments [2] and the use of the fluorescent dye Solophenyl Flavine 7GFE500 which avoids fading of fluorescence [3]. The Cawthron laboratories were visited by the participants as well, which were guided by Kirsty and Tim Harwood everyone was stunned to see the mass production of toxins produced by Alexandrium pacificum. After lunch, Kirsty led a hands-on demonstration of three different quantitative PCR experiments that focused on Alexandrium and Azadinium the eagerly awaited results showed that everybody did a great job A for effort! The day ended with a round table discussion about morphological and molecular methods, and highlighted how both methods could be combined, as well with toxin analysis. After the workshop, several people stayed to visit the Abel Tasman National Park. A water-taxi sped us past fur seals and a rock allegedly split in two by Chuck Norris, to the start of a 12 km hike. The hike was most delightful with impressive views of Tasman Bay even at high temperatures of 27C, suppressed by tree fern shade along the way. Some pukekos and wekas could be observed, and it was easy to find sand dollars on the beach. References 1. Yamaguchi M et al 1995. Phycologia 34: 207-214 2. Bolch CJS 1997. Phycologia 36: 472-478 3. Chomérat N et al 2016. Phycologia 56: 193-203 Authors Kenneth Neil Mertens, Ifremer, LER BO, Station de Biologie Marine, Place de la Croix, BP40537, F-29185 Concarneau Cedex, France Kirsty Smith, Lesley Rhodes & Lincoln MacKenzie, Cawthron Institute, 98 Halifax Street East, Private Bag 2, Nelson 7042, New Zealand Corresponding author: kenneth.mertens@ifremer.fr Fig. 1. Group photo of participants. 17 Harmful Algae News An IOC Newsletter on Toxic Algae and Algal Blooms No. 60 - July 2018 www.ioc-unesco.org/hab Marine eukaryote and HAB monitoring in Japan with next generation technology Sequencing technologies such as Illumina MiSeq have made it possible to obtain billions of sequence reads in a Frequency distribution (%) the surface data was used in this study. Surface seawater (0.5 L) was collected weekly with a plastic bucket during the same period. For MPS-based monitoring based on amplicon-seq of 18S-rRNA gene (V79 region), PCR amplification and 454 pyrosequencing were performed accor Inland Sea, 2-17-5 Maruishi, Hatsukaichi, Hiroshima 739-045, Japan Hiroshi Shimada, Central Fisheries Research Institute of Hokkaido Research Organization, Hamanaka-cho 238, Yoichi, Hokkaido 0468555, Japan Seiji Katakura, City of Mombetsu, Kaiyo-koryukan, Kaiyo-koen, Mombetsu, Hokkaido 094-0031, Jap Quantifying dinoflagellate cysts in bottom sediments: a response to Anderson 2018 Problems associated with quantifying dinoflagellate cysts in bottom sediments featured prominently in Don Anderson s personal retrospective view of his contribution to the early days of HAB cyst research published rece viously, sample sites should be chosen to avoid such areas if possible. Clearly, there is room for questions or criticism when this dynamic system is modelled from the assumption that bottom sediments, for example in Anderson s work in The Gulf of Maine, are in a static stable condition whereby a sa Contribution of marine invertebrates to Ciguatera poisoning : the case study of French Polynesia Reported as early as the 15th century by explorers, ciguatera fish poisoning (CFP) is the most prominent non-bacterial seafood poisoning worldwide. Widespread in tropical and subtropical regions, CFP res au développement du phytoplancton ciguatérigène en Polynésie française: CARISTO-Pf n7937/MSR/REC of 4th of December 2015 and Arrêté nHC/491/ DIE/BPT of 30th March 2016). Figure 2. Comparison of Pacific ciguatoxins (P-CTXs) profiles in A) in vitro cultures of Gambierdiscus polynesiensis (TB-92 [14]) Solid Phase Adsorption Toxin Tracking (SPATT) technology for field monitoring of Gambierdiscus toxins with passive samplers Ciguatera poisoning is a seafood intoxication classically associated with the consumption of tropical coral reef fish contaminated with ciguatoxins (CTXs), although some marine Fig. 2. Summary of the results demonstrating that SPATT passive sampling could advantageously contribute to the reinforcement of ciguateric risk assessment and management programmes as a supplementary tool. or of very low densities of toxic Gambierdiscus cells. However, CTXs were detected in numero New limits of Ostreopsis distribution in the Bay of Biscay: a first report of Ostreopsis in Santander Bay, Cantabria (Northern Spain) Fig. 1. Location of sampling sites The genus Ostreopsis encompasses benthic dinoflagellates that represent one of the main microalgal threats for beach tourism in w Bloom of Ostreopsis cf. siamensis in Lisbon Bay Fig. 1. Sampling sites on Lisbon Bay Reports of benthic HAB events have increased during the last decade in temperate regions. These have been associated with proliferations of benthic toxic dinoflagellates, in particular species of Ostreopsis. The M Table1 1- List - List the macroalgaecommunity communityspecies species Table ofof the macroalgae ITSA and ITSB primers [6]. Phylogenetic analysis (not Asparagopsis armata Corallina sp. shown) revealed that all the Caulacanthus ustulatus Cystoseira sp. studied sequences grouped Ceramiacea Derbesia s A possible link between the breakdown of a fertilizer tank and a toxic Pseudonitzschia bloom fishery, leaving toxic blooms undetected in other areas. By the time of the bloom, the diatom community primarily consisted of P. seriata (Fig. 3). Identification of P. seriata as the potential culprit diat Fig. 3. Pseudo-nitzschia seriata TEM micrograph of the poroid structure arranged within the striae consisting of two outer rows of larger poroids and one or two inner rows of smaller poroids. induce DA production in P. australis [7], and the most potent DA-inducing N form has been shown to be urea. Golden alga Prymnesium parvum Carter bloom off Azhikode, southwest India In September 2009, during the southwest monsoon season, a quasi-monospecific bloom of Prymnesium parvum was collected off Azhikode (10o 11 02 N; 76o 09 22 E), on the southwest coast of India (Fig. 1). A conspicuous pale browni Regional Workshop on Monitoring and Management Strategies for Benthic HABs if possible, establish tools for standardized procedures. The exercise is open to the entire international community. The goal of achieving improved assessments of the risks associated with BHABs will help reduce the health, Workshop on morpho-molecular methods for the study of dinoflagellate cysts A workshop on techniques for the morphological and molecular identification of cysts from toxic HAB species, led by Drs Kenneth Neil Mertens, Kirsty Smith, Lesley Rhodes and Lincoln MacKenzie, was held recently at the Cawthro Cawthron Summer Scholar Explores Bloom-Forming Freshwater Cyano bacteria Globally, cyanobacteria blooms in freshwater environments are causing water quality problems and health risks with increasing frequency. New Zealand is no stranger to this, suffering from blooms of toxin-producing cyanobacteria Canadian HAB Scientists Hold Workshop to Establish National Priorities and Develop Research Network A National Harmful Algal Bloom (HAB) workshop was organized and chaired by Dr. Ian Perry at the Institute of Ocean Sciences (IOS), Fisheries and Oceans Canada (DFO), Sidney, British Columbia, July 11- Joint FAO, IAEA, IOC and WHO Technical Meeting for the development of an InterAgency Global Ciguatera Strategy Harmful Algae News has previously brought information on an initiative between the International Atomic Energy Agency (IAEA) Environment Laboratories in Monaco jointly with the Oceanographi 12th Advanced Phytoplankton Course - APC 12 Identification, Taxonomy, Systematics Roscoff Biological Station (France) - 19th May to 8th June 2019 APC12 is organized by the Station Biologique de Roscoff together with the Stazione Zoologica Anton Dohrn and the IOC Science and Communication Centre on H ICHA 2018 News: 669 abstracts evaluated ! 250 orals + 45 ignite talks selected and participants notified ICHA 2018 News: 669 abstracts evaluated ! 9 plenaries confirmed 250 orals + 45 ignite talks selected and participants notified 3 satellite sessions 9 plenaries confirmed 3 satellite sessions