Dinoflagellate orders with toxic/harmful representatives O. Prorocentrales O. Dinophysiales O. Gymnodiniales K. Tangen Prorocentrum Dinophysis Karenia O. Peridinales O. Noctilucales O. Dinamoebiales Pfisteria Protoceratium Noctiluca

Toxins in dinoflagellatesthat accumulate in mussels, fish, birds and mammals, and are toxic to humans (or mice) Based on chemistry Saxitoxin group (PSP) Okadaic acid group (DSP) Pectenotoxin group Yessotoxin group Brevetoxin group (NSP) Azaspiracid group (AZP) Cyclic imine group Based on symptoms PSP=Paralytic shellfish poisoning DSP=Diarrheic shellfish poisoning NSP=Neurotoxic shellfish poisoning CFP=Ciguatera fish poisoning

PSP - Paralytic shellfish poisoning Acute symptoms: Diarrhoea, nausea, vomiting, tingling sensation, respiratory and muscular paralysis Mechanisms: Na+ Ca2 channel blocker Duration: Days Incubation time: 5-30 min Poisoning: About 2500 reported cases before 1993, 1-14% death rate. Some lighter cases in Norway, no mortalities.

DSP -Diarrheic shellfish poisoning Incubation: 30 min-12h Acute symptoms: Gastrointestinal symptoms (diarrhoea, nausea, vomiting, abdominal pain), headache. Chronic: tumour promotion? Mechanisms: Phosphatase inhibitor Duration: Days Poisoning: Many thousands of cases in Europe, Asia and Australia/New Zealand. Similar symptoms to bacterial infection. No mortalities.

NSP- Neurotoxic shellfish poisoning Toxins may accumulate in clams and cause massive death of fish, birds, calms and may harm mammals. Aerosols occur. Symptomer: Similar to PSP symptoms Mechanisms: Neurotoxic, Na+-channel activator (Åpner Na+-kanaler og depolariserer hvilepotensialet i nerveceller). Aerosol-exposure may cause respiratory problems Distribution: In the Gulf of Mexico, Florida and North Carolina. Detected in New Zealand.

Ciguatera fish poisoning Related to consumption of tropical and subtropical marine fish (e.g. barracuda, red snapper, grouper) from some coral reef areas Incubation: > 24h Symptoms: gastrointestinal symptoms, neurotoxic, reversal of hot and cold sensation Mechanism: Na+ and Ca2+ channel activators Poisoning: More than 50 000 persons estimated to be poisoned every year. 0.1-12% death rate, normally not deadly, cause long term ill effects

Symptoms and history of illness Tropical distribution, especially common in French Polynesia, increasingly common in the Gulf of Mexico Symptoms and history of illness Acute phase: diarrhoea, nausea, vomiting, weekness, low heart rate and blood pressure Post-acute phase: muscle pain, tired, headache, reversal of temperature sensation Chronic phase: Ataxia (balance), problems with focussing, anguish, depression, sensitivity to wine and acetic acid

Toxin production in dinoflagellates PSP (saxitoxins) paralytic toxins Alexandrium DSP (ocadaic acid) diarrhoetic toxins Dinophysis, Prorocentrum lima NSP (brevetoxins) neuro toxins Karenia brevis PTX pectenotoxins Dinophysis YTX yessotoxins Protoceratium reticulatum, Lingulodinium polyedrum spirolids (cyclic imine) Alexandrium ostenfeldii AZP azaspiracid DSP symptomes not Protoperidinium crassipes , small thecate ichthyotoxins Karenia, Karlodinium CFP ciguatera fish poisioning Gambierdiscus toxicus

Orden Gymnodiniales - “naked” Karenia mikimotoi (Miyake & Kominami ex Oda) Hansen & Moestrup syn. Gymnodinium mikimotoi syn. Gymnodinium nagasakiense syn Gyrodinium cf. aureolum i europeiske farvann LM-graph produce ichthyotoxins SEM-graph

Karenia brevis (Davis) G.Hansen & Moestrup Syn. Gymnodinium breve Syn. Ptychodiscus brevis Produce NSP-toxins

Karlodinium veneficum Syn. Karlodinium micrum, Gymnodinium galatheanum produce ichthyotoxins

orden Prorocentrales Prorocentrum lima DSP toxin

orden Dinophysiales Dinophysis spp. Okadaic acid Pectenotoxins Venstre sulcuslist Dinophysis rotundata Dinophysis acuta

Dinophysis - diversity Okadaic acid – cause DSP Pectenotoxins – do not cause DSP in humans

Dinophysis-bloom, Southern coast of Norway

Plate formulas Kofoidian system for plate numbering Plate formulas Essential plates Practical application

Plate formulas the number of plates of each type is genus specific Po, x - at apex ’ – apical plates touch apex a – intercalar plates of epitheca ’’ – precingular plates touch cingulum c – cingulum plates s – sulcus plates ’’’ – postcingular plates touch cingulum p – intercalar plates of hypotheca ’’’’ – antapical plates at antapex epitheca hypotheca

Alexandrium-arter Plate formula; Po, 4’, 0a, 6’’, 6c, 9(10), 5’’’,2’’’’

Order Peridiniales Alexandrium Plateformula Po, 4’, 0a, 6’’, 6c, 9(10)s, 5’’’, 2’’’’ Essential plates; 1 apical, 6 pre-cingular

Alexandrium minutum

Alexandrium minutum PSP-toxins (saxitoxins)

stained with calcofluor white, observed in fluorescence microscope Produce spirolids (cyclic imine group), no evidence for effects on humans

Alexandrium tamarense

PSP-toxines (Saxitoxins)

Po, 4’, 0a, 6’’, 6c, 9(10), 5’’’,2’’’’ Po, 3’, 3a, 6’’, 6c, 7s, 6’’’, 2’’’’

Lingulodinium polyedrum (Stein) Dodge Syn. Gonyaulax polyedra Po, 3’, 3a, 6’’, 6c, 7s, 6’’’, 2’’’’ cyste – hvilespore YTX-toxins (yessotoxin) – (do not cause DSP in humans) cyst SEM-graphs E. Beckstrøm LM.graph – B. Dale UiO

Protoceratium reticulatum Plate formula Protoceratium; Po, 3’, 0a, 6’’, 6c, 6s, 6’’’, 2’’’’ Protoceratium reticulatum (Claparède & Lachmann) Bütschli Syn. Gonyaulax grindleyi YTX-toxins (yessotoxin) – (do not cause DSP in humans) Large and very reticulated plates

AZP (azaspirazid) – symptoms similar to DSP Protoperidinium crassipes (Kofoid) Balech syn Peridinium crassipes Kofoid Not the causative organism New small thecate that is in the process of being formally described has been shown to produce AZP AZP (azaspirazid) – symptoms similar to DSP

orden Noctilucales Toxic? Noctiluca scintillans (Macartney) Kofoid & Swezy Acumulates ammonium and may harm fish cellevakuole med stort ammoniuminnhold gir flyteevne

Noctiluca scintillans

orden Dinamoebales Genus with toxic species Pfiesteria Steidinger & Burkholder Pfiesteria piscicida Steidinger & Burkholder Pfiesteria shumwayae Glasgow & Burkholder Both species known from sediments in Norway (brackish water, Sandvikselva, Oslofjorden)

Pfiesteria piscicida toxic ”flesh eating” Peduncle for feeding Plate sutures after staining with calcofluor white with Fluorescence microscopy

Plateformula P. piscicida Po, cp, X, 4’, 1a, 5’’, 6c, 4s, 5’’’, 2’’’’ P. shumwayae Po, cp, X, 6’, 1a, 5’’, 6c, 4s, 5’’’, 2’’’’ triangulær rektangulær

Pfiesteria piscicida Pfiesteria shumwayae etter HAB 2003

foreslått livssyklus hos P. piscicida planozygote zoospore TOX A zoospore TOX B hypnozygote