Arjun Verma

Characterization of Gambierdiscus lapillus sp. nov. (Gonyaulacales, Dinophyceae): a new toxic dinoflagellate from the Great Barrier Reef (Australia)

Gambierdiscus is a genus of benthic dinoflagellates found worldwide. Some species produce neurotoxins (maitotoxins and ciguatoxins) that bioaccumulate and cause ciguatera fish poisoning (CFP), a potentially fatal food‐borne illness that is common worldwide in tropical regions. The investigation of toxigenic species of Gambierdiscus in CFP endemic regions in Australia is necessary as a first step to determine which species of Gambierdiscus are related to CFP cases occurring in this region. In this study, we characterized five strains of Gambierdiscus collected from Heron Island, Australia, a region in which ciguatera is endemic. Clonal cultures were assessed using (i) light microscopy; (ii) scanning electron microscopy; (iii) DNA sequencing based on the nuclear encoded ribosomal 18S and D8‐D10 28S regions; (iv) toxicity via mouse bioassay; and (v) toxin profile as determined by Liquid Chromatography‐Mass Spectrometry. Both the morphological and phylogenetic data indicated that these strains represent a new species of Gambierdiscus, G. lapillus sp. nov. (plate formula Po, 3′, 0a, 7″, 6c, 7‐8s, 5‴, 0p, 2″″ and distinctive by size and hatchet‐shaped 2′ plate). Culture extracts were found to be toxic using the mouse bioassay. Using chemical analysis, it was determined that they did not contain maitotoxin (MTX1) or known algal‐derived ciguatoxin analogs (CTX3B, 3C, CTX4A, 4B), but that they contained putative MTX3, and likely other unknown compounds.

The dinoflagellate genera Gambierdiscus and Ostreopsis from subtropical Raoul Island and North Meyer Island, Kermadec Islands

ABSTRACT An expedition to North Meyer Island, Kermadec Islands, in November 2015, resulted in the isolation of two Gambierdiscus species, G. australes and a previously unknown Gambierdiscus species maintained in the Cawthron Institute Culture Collection of Micro-algae as CAWD242. Identifications were based on morphology and DNA sequence data analysis. Nine isolates of G. australes produced maitotoxin-1 (MTX-1) ranging from present (detectable but below the confidence level) to 36.6 pg per cell. One further isolate did not produce MTX-1, but all G. australes isolates produced putative MTX-3. Isolate CAWD242 was negative for MTX-1 but produced MTX-3. Ostreopsis sp. 3 isolated from samples collected during an earlier expedition in 2013 produced low concentrations of palytoxin (PLTX)-like compounds (0.013 pg per cell), but an isolate of the same species from samples collected in 2015 was non-toxic. Other dinoflagellates isolated and identified were Coolia malayensis, Amphidinium carterae and Prorocentrum hoffmannianum.

Molecular phylogeny, morphology and toxigenicity of Ostreopsis cf. siamensis (Dinophyceae) from temperate south-east Australia

Ostreopsis is a genus of dinoflagellates that includes species producing palytoxin and structurally related compounds. The distribution of Ostreopsis species in Australia is largely unknown, but they have been reported from north Queensland (18° S) to Tasmania (41–43° S). Ostreopsis spp. have been recurrently reported from estuaries around New South Wales, with persistent occurrences in Merimbula Lake inlet throughout the year. We isolated and characterized a strain of Ostreopsis cf. siamensis using light and scanning electron microscopy as well as molecular sequences of small subunit (SSU), large subunit (LSU) and ITS regions of ribosomal DNA. The strain grew significantly faster in low nutrient concentrations. Palytoxin‐like compounds were produced by the strain, as determined by chemical analysis, and the LD 50 of the cell extract by intraperitoneal injection in mice was 25.1 mg kg−1. This is the first comprehensive molecular, morphological and toxicological study of an Ostreopsis species from Australian waters. Increasing reports of Ostreopsis from temperate waters suggest an empirical need to expand the knowledge of their diversity and distribution to aid aquaculture monitoring in Australian estuaries.

A new diatom species P. hallegraeffii sp. nov. belonging to the toxic genus Pseudo-nitzschia (Bacillariophyceae) from the East Australian Current

A new species belonging to the toxin producing diatom genus Pseudo-nitzschia, P. hallegraeffii sp. nov., is delineated and described from the East Australian Current (EAC). Clonal cultures were established by single cell isolation from phytoplankton net hauls collected as part of a research expedition in the EAC region in 2016 on the RV Investigator. Cultures were assessed for their morphological and genetic characteristics, their sexual compatibility with other Pseudo-nitzschia species, and their ability to produce domoic acid. Light and transmission electron microscopy revealed cells which differed from their closest relatives by their cell width, rows of poroids, girdle band structure and density of band straie. Phylogenetic analyses based on sequencing of nuclear-encoded ribosomal deoxyribonucleic acid (rDNA) regions showed this novel genotype clustered within the P. delicatissima complex, but formed a discrete clade from its closest relatives P. dolorosa, P. simulans, P. micropora and P. delicatissima. Complementary base changes (CBCs) were observed in the secondary structure of the 3’ nuclear ribosomal transcribed spacer sequence region (ITS2) between P. hallegraeffii sp. nov. and its closest related taxa, P. simulans and P. dolorosa. Under laboratory conditions, and in the absence of any zooplankton cues, strains of P. hallegraeffii sp. nov. did not produce domoic acid (DA) and were not sexually compatible with any other Pseudo-nitzschia clones tested. A total of 18 Pseudo-nitzschia species, including three confirmed toxigenic species (P. cuspidata, P. multistriata and P. australis) have now been unequivocally confirmed from eastern Australia.