Dinophysis Nguyen Van Nguyen RESEARCH INSTITUTE FOR MARINE FISHERIES, VIETNAM

WESTPAC workshop on the development of a research strategy for Harmful Algal Blooms Toward the future development of HAB science in the Western Pacific What we know, and what we do not know on HABs Dinophysis Nguyen Van Nguyen RESEARCH INSTITUTE FOR MARINE FISHERIES, VIETNAM Institute of Oceanography, Nha Trang, Vietnam, 19-21 December 2016

1-1. List of known toxic species (in the world) 1-2. List of toxic species in the region, and distribution Species Distribute in the region Toxins 1 Dinophysis acuminata Claparède & Lachmann, Japan, New-zealand DTX1, PTX2 2 Dinophysis acuta Ehrenberg, 39 New-zealand OA, DTX1, DTX2, PTX2, PTX11, PTX12 3 Dinophysis caudata Saville-Kent, 81 Widely ditributed OA, DTX1, PTX2 4 Dinophysis fortii Pavillard, 13 Japan OA, DTX1, PTX2 5 Dinophysis infundibulum J. Schiller, 18 Japan PTX2 6 Dinophysis miles Cleve, 1900 Tropical waters DTX-1 7 Dinophysis sacculus Stein, 83 Europe OA 8 Dinophysis tripos Gourret, 83 Japan DTX1, PTX2, 9 Dinophysis ovum Schütt, 95 Europe, US OA 10 Phalacroma mitra F.Schütt, 95 Tropical DTX1 11 Dinophysis rotundata Claparède & Lachmann, Worldwide OA, DTXs, PTXs Other more than 200 species Not yet analized There are over 200 species described, with few species appeared in high abundance. So far 11 species are known to produce toxins. The rest are unknown in terms of toxin production due to difficulty in having significant numbers of cell because most of them are scare in density and culturing techniques are unavailable yet.

1-3 Morphological characters for species identification o Conventional classification is based on cell size and shape, structure of sulcal lists and presence of chloroplast. o These parameters, especially cell size and shape, show wide range of variation within certain species, and the range may overlap among species. o Species boundary is not clear. o Classification needs re-evaluation. 3

Polymorphism More and more evidence of species go on morphological deformation process which distorts cell form and leads to interpretation as different species and make classification of boundaries complicated. Ornithocercus Metadinophysis Histioneis Dinophysis 4

The need to revise classification of Dinophysis Genus level: Phalacroma vs Dinophysis Species level: Separation

Separation of species needed Difference in DNA sequences usually implicates difference in morphology. Thus, DNA sequence is an useful criterion. But this criterion should be applied with caution on the underestimation of the difference. 6

At genus level Phalacroma and Dinophysis: merge or separate? 93 54 22 6 91 34 37 25 53 94 13 41 70 Dinophysis caudata Catba-VN 031106.5 Dinophysis caudata Okirai-JP 211206.15 Dinophysis caudata Okirai-JP 211206.9 Dinophysis caudata Omura Bay-JP 31.3 Dinophysis caudata Okirai-JP 211206.4 Dinophysis caudata MR.5 Dinophysis caudata Catba-VN 031106.4 Dinophysis caudata Catba-VN 031106.6 Dinophysis caudata Iki Bay-JP Dinophysis caudata MR38.1 Dinophysis caudata Catba-VN 031106.7 Dinophysis caudata Okirai-JP 211206.33 Dinophysis tripos Miyako-JP Dinophysis tripos French AF3238 Dinophysis tripos Okirai-JP Dinophysis miles NT-VN Dinophysis infundibulus s. Abe AB3746 Dinophysis fortii short MR40.4 Dinophysis fortii short MR40. Dinophysis schroederii MR5.4 Dinophysis fortii short MR40.19 Dinophysis fortii short MR40.5 Dinophysis fortii French AF3236 Dinophysis norvegica Okirai-JP 211206.27 Dinophysis acuminata Tokyo-JP 061106.1 Dinophysis acuminata French AF32 Dinophysis hastata f.e 200807.1 Dinophysis hastata f.c MR24.2 Dinophysis hastata f.c MR.1 Dinophysis whittingae MR40.14 Dinophysis sp MR39.1 Dinophysis argus MR40.17 Dinophysis expulsa MR10.1 Dinophysis infudibulus s.taylor MR17.4 Dinophysis infudibulus s.taylor MR20.4 Dinophysis similis MR17.5 Citharistes regius MR20.1 Citharistes regius MR30.1 Citharistes apsteinii 17.1 Ornithocercus heteroporus Ornithocercus galea MR42.5 Ornithocercus splendidus MR32.4 Ornithocercus magnificus MR14-4 Ornithocercus magnificus MR14-3 Ornithocercus magnificus MR-5 Dinophysis hastata f.a MR7.2 Dinophysis hastata f.b MR14.1 Dinophysis hastata f.a MR30.9 Ornithocercus thumii MR19.1 Ornithocercus quadratus MR7.6 Ornithocercus thumi MR22.1 Ornithocercus thumii MR5.2 Ornithocercus thumii MR22.2 Ornithocercus thumii MR14.2 Ornithocercus thumii MR22.2(2) Ornithocercus quadratus MR7.4 84 24 97 47 44 Histioneis megalocopa MR40.1 Histioneis megalocopa MR34.1 42 96 Histioneis highleyi 9.1 Histionesis isseli MR.3 Histionesis sp 40.15 Histioneis dentata MR34.3 Histioneis crateriformis MR25.8 Histioneis crateriformis MR34.8 Dinophysis amandula MR15.1 Dinophysis amandula MR5.3 Dinophysis parvula MR40.12 Dinophysis rapa MR17.11 Dinophysis sp MR25.5 Oxyphysis oxytoxoides EF6133 Dinophysis cuenus MR33.1 Dinophysis sp1 MR17.9 Dinophysis favus MR20.2 Dinophysis doryphora MR17.8 Dinophysis rotundata French AF3235 Dinophysis rotundata Tokyo-JP 6.11.06.3 75 71 68 81 79 67 Amphisolenia bidentata MR19.3 Amphisolenia bidentata MR19.4 Amphisolenia bidentata MR38.10 Amphisolenia thrinax MR.1 Amphisolenia palaeotheroides MR32.1 Amphisolenia schauinlandii MR38.4 Amphisolenia clavispes MR14.6 Amphisolenia clavispes MR38.8 Metaphalacroma skogsbergii MR31.1 Prorocentrum lima DQ3360 Heterocapsa triquetra AF0401 Alexandrium tamarense AF200668 Dinophysis sensu lato Oxyphysis and former Phalacroma 7

A. schauinslandii A. clavipes A. papaeotheroides A. bidentata A. thrinax O. splendidus O. heteroporus O. biclavatus O. magnificus O. magnificus O. steinii O. thumii O. skogsbergii O. quadratus O. galea D. parvula D. rotundata D. elongata D. amandula D. sp. D. argus D. apicata D. cuneus D. favus D. mitra D. rapa D. hindmarchii D. doryphora D. acutissima D. expulsa D. hastata D. sphaerica D. infundibulus D. norvegica D. acuminata D. fortii D. schroederi D. caudata D. tripos D. miles H.megalocopa, H. dolon, H. helenae, H. hippoperoides H. highleyi H. biremis H. crateriformis H. sp. H. isselii H. dentata C. regius C. apsteinii M. skogsbergii P. nasutum P. sp. separate separate, but further data needed merge, but further data needed M. sp. M. sinensis O. oxytoxoides 8

Future evolutionary study

93 54 22 6 91 34 37 25 53 94 13 41 70 Dinophysis caudata Catba-VN 031106.5 Dinophysis caudata Okirai-JP 211206.15 Dinophysis caudata Okirai-JP 211206.9 Dinophysis caudata Omura Bay-JP 31.3 Dinophysis caudata Okirai-JP 211206.4 Dinophysis caudata MR.5 Dinophysis caudata Catba-VN 031106.4 Dinophysis caudata Catba-VN 031106.6 Dinophysis caudata Iki Bay-JP Dinophysis caudata MR38.1 Dinophysis caudata Catba-VN 031106.7 Dinophysis caudata Okirai-JP 211206.33 Dinophysis tripos Miyako-JP Dinophysis tripos French AF3238 Dinophysis tripos Okirai-JP Dinophysis miles NT-VN Dinophysis infundibulus s. Abe AB3746 Dinophysis fortii short MR40.4 Dinophysis fortii short MR40. Dinophysis schroederii MR5.4 Dinophysis fortii short MR40.19 Dinophysis fortii short MR40.5 Dinophysis fortii French AF3236 Dinophysis norvegica Okirai-JP 211206.27 Dinophysis acuminata Tokyo-JP 061106.1 Dinophysis acuminata French AF32 Dinophysis hastata f.e 200807.1 Dinophysis hastata f.c MR24.2 Dinophysis hastata f.c MR.1 Dinophysis whittingae MR40.14 Dinophysis sp MR39.1 Dinophysis argus MR40.17 Dinophysis expulsa MR10.1 Dinophysis infudibulus s.taylor MR17.4 Dinophysis infudibulus s.taylor MR20.4 Dinophysis similis MR17.5 Citharistes regius MR20.1 Citharistes regius MR30.1 Citharistes apsteinii 17.1 Ornithocercus heteroporus Ornithocercus galea MR42.5 Ornithocercus splendidus MR32.4 Ornithocercus magnificus MR14-4 Ornithocercus magnificus MR14-3 Ornithocercus magnificus MR-5 Dinophysis hastata f.a MR7.2 Dinophysis hastata f.b MR14.1 Dinophysis hastata f.a MR30.9 Ornithocercus thumii MR19.1 Ornithocercus quadratus MR7.6 Ornithocercus thumi MR22.1 Ornithocercus thumii MR5.2 Ornithocercus thumii MR22.2 Ornithocercus thumii MR14.2 Ornithocercus thumii MR22.2(2) Ornithocercus quadratus MR7.4 84 24 97 47 44 Histioneis megalocopa MR40.1 Histioneis megalocopa MR34.1 42 Histioneis highleyi 9.1 Histionesis isseli MR.3 Histionesis sp 40.15 Histioneis dentata MR34.3 Histioneis crateriformis MR25.8 Histioneis crateriformis MR34.8 Dinophysis amandula MR15.1 Dinophysis amandula MR5.3 Dinophysis parvula MR40.12 Dinophysis rapa MR17.11 Dinophysis sp MR25.5 Oxyphysis oxytoxoides EF6133 Dinophysis cuenus MR33.1 Dinophysis sp1 MR17.9 Dinophysis favus MR20.2 Dinophysis doryphora MR17.8 Dinophysis rotundata French AF3235 Dinophysis rotundata Tokyo-JP 6.11.06.3 96 75 68 81 71 79 67 Amphisolenia bidentata MR19.3 Amphisolenia bidentata MR19.4 Amphisolenia bidentata MR38.10 Amphisolenia thrinax MR.1 Amphisolenia palaeotheroides MR32.1 Amphisolenia schauinlandii MR38.4 Amphisolenia clavispes MR14.6 Amphisolenia clavispes MR38.8 Metaphalacroma skogsbergii MR31.1 Prorocentrum lima DQ3360 Heterocapsa triquetra AF0401 Alexandrium tamarense AF200668 Metaphalacroma 10

(Abe, 1967) (Taylor, 10) (Hallegraeff et Lucas 18) Integrated phylogeny with reference of DNA sequence

Life cycle: unclear Proposed life cycle of Dinophysis by Regurea (2001) 19

Nutrient and difficulty in study on ecophysiology Dinophysis includes heterotrophic and photosynthetic species, but all feeds on other plankton (ciliate, other phytoplankton). Only photosynthetic species (mixotrophic) have been linked to DSP. Culture success with some Dinophysis Scarcity of sample (most are unable to culture) is the difficulty for taxonomy, ecology and toxins study D. acutisima D. elongata

Identified scientific problems/questions/gaps in the region (as well as in the world) Ecology (bloom dynamic) Life cycle Toxin production Roles of suspected species in DSP outbreak Taxonomy at species level and genus level (separation of species, merging, synonym)