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1. Genome wide conditional mouse knockout resources

Author

Kaloff, C, Anastassiadis, K, Ayadi, A, Baldock, R, Beig, J, Birling, M-C, Bradley, A, Brown, SDM, Bürger, A, Bushell, W, Chiani, F, Collins, FS, Doe, B, Eppig, JT, Finnell, RH, Fletcher, C, Flicek, P, Fray, M, Friedel, RH, Gambadoro, A, Gates, H, Hansen, J, Herault, Y, Hicks, GG, Hörlein, A, de Angelis, M Hrabé, Iyer, V, de Jong, PJ, Koscielny, G, Kühn, R, Liu, P, Lloyd, KCK, Lopez, RG, Marschall, S, Martínez, S, McKerlie, C, Meehan, T, von Melchner, H, Moore, M, Murray, SA, Nagy, A, Nutter, LMJ, Pavlovic, G, Pombero, A, Prosser, H, Ramirez-Solis, R, Ringwald, M, Rosen, B, Rosenthal, N, Rossant, J, Noppinger, P Ruiz, Ryder, E, Skarnes, WC, Schick, J, Schnütgen, F, Schofield, P, Seisenberger, C, Selloum, M, Smedley, D, Simpson, EM, Stewart, AF, Teboul, L, Valentini, GP Tocchini, Valenzuela, D, West, AP, and Wurst, W

Subjects
Pharmacology and Pharmaceutical Sciences, Biomedical and Clinical Sciences, Biotechnology, Genetics, Stem Cell Research - Embryonic - Non-Human, Stem Cell Research, Human Genome, 2.6 Resources and infrastructure (aetiology), Generic health relevance, Neurosciences, Neurology & Neurosurgery, Pharmacology and pharmaceutical sciences
Abstract

The International Knockout Mouse Consortium (IKMC) developed high throughput gene trapping and gene targeting pipelines that produced mostly conditional mutations of more than 18,500 genes in C57BL/6N mouse embryonic stem (ES) cells which have been archived and are freely available to the research community as a frozen resource. From this unprecedented resource more than 6,000 mutant mouse strains have been produced by the IKMC and mostly the International Mouse Phenotyping Consortium (IMPC). In addition, a cre-driver resource was established including 250 inducible cre-driver mouse strains in a C57BL/6 background. Complementing the cre-driver resource, a collection of comprising 27 cre-driver rAAVs has also been produced. The resources can be easily accessed at the IKMC/IMPC web portal (www.mousephenotype.org). The IKMC/IMPC resource is a standardized reference library of mouse models with defined genetic backgrounds that enables the analysis of gene-disease associations in mice of different genetic makeup and should therefore have a major impact on biomedical research.

Published
2016

2. Response of planktonic microbial assemblages to disturbance in an urban sub-tropical estuary.

Author

Ajani, PA, Savela, H, Kahlke, T, Harrison, D, Jeffries, T, Kohli, GS, Verma, A, Laczka, O, Doblin, MA, Seymour, JR, Larsson, ME, Potts, J, Scanes, P, Gribben, PE, Harrison, L, Murray, SA, Ajani, PA, Savela, H, Kahlke, T, Harrison, D, Jeffries, T, Kohli, GS, Verma, A, Laczka, O, Doblin, MA, Seymour, JR, Larsson, ME, Potts, J, Scanes, P, Gribben, PE, Harrison, L, and Murray, SA

Abstract

Microbes are sensitive indicators of estuarine processes because they respond rapidly to dynamic disturbance events. As most of the world's population lives in urban areas and climate change-related disturbance events are becoming more frequent, estuaries bounded by cities are experiencing increasing stressors, at the same time that their ecosystem services are required more than ever. Here, using a multidisciplinary approach, we determined the response of planktonic microbial assemblages in response to seasonality and a rainfall disturbance in an urban estuary bounded by Australia's largest city, Sydney. We used molecular barcoding (16S, 18S V4 rRNA) and microscopy-based identification to compare microbial assemblages at locations with differing characteristics and urbanisation histories. Across 142 samples, we identified 8,496 unique free-living bacterial zOTUs, 8,175 unique particle associated bacterial zOTUs, and 1,920 unique microbial eukaryotic zOTUs. Using microscopy, we identified only the top <10% abundant, larger eukaryotic taxa (>10 µm), however quantification was possible. The site with the greater history of anthropogenic impact showed a more even community of associated bacteria and eukaryotes, and a significant increase in dissolved inorganic nitrogen following rainfall, when compared to the more buffered site. This coincided with a reduced proportional abundance of Actinomarina and Synechococcus spp., a change in SAR 11 clades, and an increase in the eukaryotic microbial groups Dinophyceae, Mediophyceae and Bathyoccocaceae, including a temporary dominance of the harmful algal bloom dinoflagellate Prorocentrum cordatum (syn. P. minimum). Finally, a validated hydrodynamic model of the estuary supported these results, showing that the more highly urbanised and upstream location consistently experienced a higher magnitude of salinity reduction in response to rainfall events during the study period. The best abiotic variables to explain community dissimil

Published
2023

3. Abiotic and biotic factors controlling sexual reproduction in populations of Pseudo-nitzschia pungens (Bacillariophyceae).

Author

Kim, JH, Ajani, PA, Murray, SA, Kang, S-M, Kim, S-H, Lim, HC, Teng, ST, Lim, PT, Park, BS, Kim, JH, Ajani, PA, Murray, SA, Kang, S-M, Kim, S-H, Lim, HC, Teng, ST, Lim, PT, and Park, BS

Abstract

Pseudo-nitzschia pungens is a widely distributed marine pennate diatom. Hybrid zones, regions in which two different genotypes may interbreed, are important areas for speciation and ecology, and have been reported across the globe for this species. However, sexual reproduction between differing clades in the natural environment is yet to be observed and is difficult to predict. Here we carried out experiments using two mono-clonal cultures of P. pungens from different genotypes to measure the frequency and timing of sexual reproduction across varying biotic (growth phases and cell activity potential) and abiotic conditions (nutrients, light, turbulence). We found the mating rates and number of zygotes gradually decreased from exponential to late stationary growth phases. The maximum zygote abundance observed was 1,390 cells mL-1 and the maximum mating rate was 7.1%, both which occurred during the exponential growth phase. Conversely, only 9 cells mL-1 and a maximum mating rate of 0.1% was observed during the late stationary phase. We also found the higher the relative potential cell activity (rPCA) in parent cells, as determined by the concentration of chlorophyll a per cell and the ratio of colony formation during parent cultivations, revealed higher mating rates. Furthermore, sexual events were reduced under nutrient enrichment conditions, and mating pairs and zygotes were not formed under aphotic (dark) or shaking culture conditions (150 rpm). In order to understand the sexual reproduction of Pseudo-nitzschia in the natural environment, our results highlight that it is most likely the combination of both biotic (growth phase, Chl. a content) and abiotic factors (nutrients, light, turbulence) that will determine the successful union of intraspecific populations of P. pungens in any given region.

Published
2023

4. Genomic copy number variability at the genus, species and population levels impacts in situ ecological analyses of dinoflagellates and harmful algal blooms.

Author

Ruvindy, R, Barua, A, Bolch, CJS, Sarowar, C, Savela, H, Murray, SA, Ruvindy, R, Barua, A, Bolch, CJS, Sarowar, C, Savela, H, and Murray, SA

Abstract

The application of meta-barcoding, qPCR, and metagenomics to aquatic eukaryotic microbial communities requires knowledge of genomic copy number variability (CNV). CNV may be particularly relevant to functional genes, impacting dosage and expression, yet little is known of the scale and role of CNV in microbial eukaryotes. Here, we quantify CNV of rRNA and a gene involved in Paralytic Shellfish Toxin (PST) synthesis (sxtA4), in 51 strains of 4 Alexandrium (Dinophyceae) species. Genomes varied up to threefold within species and ~7-fold amongst species, with the largest (A. pacificum, 130 ± 1.3 pg cell-1 /~127 Gbp) in the largest size category of any eukaryote. Genomic copy numbers (GCN) of rRNA varied by 6 orders of magnitude amongst Alexandrium (102- 108 copies cell-1) and were significantly related to genome size. Within the population CNV of rRNA was 2 orders of magnitude (105 - 107 cell-1) in 15 isolates from one population, demonstrating that quantitative data based on rRNA genes needs considerable caution in interpretation, even if validated against locally isolated strains. Despite up to 30 years in laboratory culture, rRNA CNV and genome size variability were not correlated with time in culture. Cell volume was only weakly associated with rRNA GCN (20-22% variance explained across dinoflagellates, 4% in Gonyaulacales). GCN of sxtA4 varied from 0-102 copies cell-1, was significantly related to PSTs (ng cell-1), displaying a gene dosage effect modulating PST production. Our data indicate that in dinoflagellates, a major marine eukaryotic group, low-copy functional genes are more reliable and informative targets for quantification of ecological processes than unstable rRNA genes.

Published
2023

5. Gamay (Botany Bay, Australia): What we know and still need to know about this highly modified coastal waterway?

Author

Stelling-Wood, TP, Gribben, PE, Birch, G, Bishop, MJ, Blount, C, Booth, DJ, Brown, C, Bruce, E, Bugnot, AB, Byrne, M, Creese, RG, Dafforn, KA, Dahlenburg, J, Doblin, MA, Fellowes, TE, Fowler, AM, Gibbs, MC, Glamore, W, Glasby, TM, Hay, AC, Kelaher, B, Knott, NA, Larkum, AWD, Parker, LM, Marzinelli, EM, Mayer-Pinto, M, Morgan, B, Murray, SA, Rees, MJ, Ross, PM, Roughan, M, Saintilan, N, Scanes, E, Seymour, JR, Schaefer, N, Suthers, IM, Taylor, MD, Williamson, JE, Concejo, AV, Whittington, RJ, Figueira, WF, Stelling-Wood, TP, Gribben, PE, Birch, G, Bishop, MJ, Blount, C, Booth, DJ, Brown, C, Bruce, E, Bugnot, AB, Byrne, M, Creese, RG, Dafforn, KA, Dahlenburg, J, Doblin, MA, Fellowes, TE, Fowler, AM, Gibbs, MC, Glamore, W, Glasby, TM, Hay, AC, Kelaher, B, Knott, NA, Larkum, AWD, Parker, LM, Marzinelli, EM, Mayer-Pinto, M, Morgan, B, Murray, SA, Rees, MJ, Ross, PM, Roughan, M, Saintilan, N, Scanes, E, Seymour, JR, Schaefer, N, Suthers, IM, Taylor, MD, Williamson, JE, Concejo, AV, Whittington, RJ, and Figueira, WF

Published
2023

6. Developing model systems for dinoflagellates in the post-genomic era.

Author

Ishida, H, John, U, Murray, SA, Bhattacharya, D, Chan, CX, Ishida, H, John, U, Murray, SA, Bhattacharya, D, and Chan, CX

Abstract

Dinoflagellates are a diverse group of eukaryotic microbes that are ubiquitous in aquatic environments. Largely photosynthetic, they encompass symbiotic, parasitic, and free-living lineages with a broad spectrum of trophism. Many free-living taxa can produce bioactive secondary metabolites such as biotoxins, some of which cause harmful algal blooms. In contrast, most symbiotic species are crucial for sustaining coral reef health. The year 2023 marked a decade since the first genome data of dinoflagellates became available. The growing genome-scale resources for these taxa are highlighting their remarkable evolutionary and genomic complexities. Here, we discuss the prospect of developing dinoflagellate models using the criteria of accessibility, tractability, resources, research support, and promise. Moving forward in the post-genomic era, we argue for the development of fit-to-purpose models that tailor to specific biological contexts, and that a one-size-fits-all model is inadequate for encapsulating the complex biology, ecology, and evolutionary history of dinoflagellates.

Published
2023

8. Geographical distribution, molecular and toxin diversity of the dinoflagellate species Gambierdiscus honu in the Pacific region.

Author

Stuart, J, Smith, KF, Rhodes, L, Murray, JS, Viallon, J, Henry, K, Darius, HT, Murray, SA, De Azevedo, CD, Argyle, P, Chinain, M, Stuart, J, Smith, KF, Rhodes, L, Murray, JS, Viallon, J, Henry, K, Darius, HT, Murray, SA, De Azevedo, CD, Argyle, P, and Chinain, M

Abstract

An increase in cases of ciguatera poisoning (CP) and expansion of the causative species in the South Pacific region highlight the need for baseline data on toxic microalgal species to help identify new areas of risk and manage known hot spots. Gambierdiscus honu is a toxin producing and potential CP causing dinoflagellate species, first described in 2017. Currently no high-resolution geographical distribution, intraspecific genetic variation or toxin production diversity data is available for G. honu. This research aimed to further characterize G. honu by investigating its distribution using species-specific real-time polymerase chain reaction assays at 25 sites in an area spanning ∼8000 km of the Coral Sea/Pacific Ocean, and assessing intraspecific genetic variation, toxicity and toxin production of isolated strains. Assessment of genetic variation of the partial rRNA operon of isolates demonstrated no significant intraspecific population structure, in addition to a lack of adherence to isolation by distance (IBD) model of evolution. The detected distribution of G. honu in the Pacific region was within the expected tropical to temperate latitudinal ranges of 10° to -30° and extended from Australia to French Polynesia. In the lipophilic fractions, the neuroblastoma cell-based assay (CBA-N2a) showed no ciguatoxin (CTX)-like activity for nine of the 10 isolates, and an atypical pattern for CAWD233 isolate which showed cytotoxic activity in OV- and OV+ conditions. In the same way, liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis confirmed no Pacific-CTXs (CTX-3B, CTX-3C, CTX-4A, CTX-4B) were produced by the ten strains. The CBA-N2a assessment of the hydrophilic fractions showed moderate to high cytotoxicity in both OV- and OV+ condition for all the strains showing a cytotoxic profile similar to that of gambierone. Indeed, this study is the first to show the cytotoxic activity of gambierone on mouse neuroblastoma cells while no cytotoxicity was observe

Published
2022

10. Temperature variability interacts with mean temperature to influence the predictability of microbial phenotypes.

Author

Fu, F-X, Tschitschko, B, Hutchins, DA, Larsson, ME, Baker, KG, McInnes, A, Kahlke, T, Verma, A, Murray, SA, Doblin, MA, Fu, F-X, Tschitschko, B, Hutchins, DA, Larsson, ME, Baker, KG, McInnes, A, Kahlke, T, Verma, A, Murray, SA, and Doblin, MA

Abstract

Despite their relatively high thermal optima (Topt ), tropical taxa may be particularly vulnerable to a rising baseline and increased temperature variation because they live in relatively stable temperatures closer to their Topt . We examined how microbial eukaryotes with differing thermal histories responded to temperature fluctuations of different amplitudes (0 control, ±2, ±4°C) around mean temperatures below or above their Topt . Cosmopolitan dinoflagellates were selected based on their distinct thermal traits and included two species of the same genus (tropical and temperate Coolia spp.), and two strains of the same species maintained at different temperatures for >500 generations (tropical Amphidinium massartii control temperature and high temperature, CT and HT, respectively). There was a universal decline in population growth rate under temperature fluctuations, but strains with narrower thermal niche breadth (temperate Coolia and HT) showed ~10% greater reduction in growth. At suboptimal mean temperatures, cells in the cool phase of the fluctuation stopped dividing, fixed less carbon (C) and had enlarged cell volumes that scaled positively with elemental C, N, and P and C:Chlorophyll-a. However, at a supra-optimal mean temperature, fixed C was directed away from cell division and novel trait combinations developed, leading to greater phenotypic diversity. At the molecular level, heat-shock proteins, and chaperones, in addition to transcripts involving genome rearrangements, were upregulated in CT and HT during the warm phase of the supra-optimal fluctuation (30 ± 4°C), a stress response indicating protection. In contrast, the tropical Coolia species upregulated major energy pathways in the warm phase of its supra-optimal fluctuation (25 ± 4°C), indicating a broadscale shift in metabolism. Our results demonstrate divergent effects between taxa and that temporal variability in environmental conditions interacts with changes in the thermal mean to mediate micr

Published
2022

11. The extracellular matrix gene, Svep1, orchestrates airway patterning and the transition from lung branching morphogenesis to alveolar maturation in the mouse

Author

Foxworth, N, primary, Wells, J, additional, Ocaña-Lopez, S, additional, Muller, S, additional, Denegre, J, additional, Palmer, K, additional, McGee, T, additional, Memishian, W, additional, Murray, SA, additional, Donahoe, PK, additional, Bult, CJ, additional, and Loscertales, M, additional

Published
2021
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12. The NIH Somatic Cell Genome Editing program.

Author

Saha, K, Sontheimer, EJ, Brooks, PJ, Dwinell, MR, Gersbach, CA, Liu, DR, Murray, SA, Tsai, SQ, Wilson, RC, Anderson, DG, Asokan, A, Banfield, JF, Bankiewicz, KS, Bao, G, Bulte, JWM, Bursac, N, Campbell, JM, Carlson, DF, Chaikof, EL, Chen, Z-Y, Cheng, RH, Clark, KJ, Curiel, DT, Dahlman, JE, Deverman, BE, Dickinson, ME, Doudna, JA, Ekker, SC, Emborg, ME, Feng, G, Freedman, BS, Gamm, DM, Gao, G, Ghiran, IC, Glazer, PM, Gong, S, Heaney, JD, Hennebold, JD, Hinson, JT, Khvorova, A, Kiani, S, Lagor, WR, Lam, KS, Leong, KW, Levine, JE, Lewis, JA, Lutz, CM, Ly, DH, Maragh, S, McCray, PB, McDevitt, TC, Mirochnitchenko, O, Morizane, R, Murthy, N, Prather, RS, Ronald, JA, Roy, S, Sabbisetti, V, Saltzman, WM, Santangelo, PJ, Segal, DJ, Shimoyama, M, Skala, MC, Tarantal, AF, Tilton, JC, Truskey, GA, Vandsburger, M, Watts, JK, Wells, KD, Wolfe, SA, Xu, Q, Xue, W, Yi, G, Zhou, J, SCGE Consortium, Saha, K, Sontheimer, EJ, Brooks, PJ, Dwinell, MR, Gersbach, CA, Liu, DR, Murray, SA, Tsai, SQ, Wilson, RC, Anderson, DG, Asokan, A, Banfield, JF, Bankiewicz, KS, Bao, G, Bulte, JWM, Bursac, N, Campbell, JM, Carlson, DF, Chaikof, EL, Chen, Z-Y, Cheng, RH, Clark, KJ, Curiel, DT, Dahlman, JE, Deverman, BE, Dickinson, ME, Doudna, JA, Ekker, SC, Emborg, ME, Feng, G, Freedman, BS, Gamm, DM, Gao, G, Ghiran, IC, Glazer, PM, Gong, S, Heaney, JD, Hennebold, JD, Hinson, JT, Khvorova, A, Kiani, S, Lagor, WR, Lam, KS, Leong, KW, Levine, JE, Lewis, JA, Lutz, CM, Ly, DH, Maragh, S, McCray, PB, McDevitt, TC, Mirochnitchenko, O, Morizane, R, Murthy, N, Prather, RS, Ronald, JA, Roy, S, Sabbisetti, V, Saltzman, WM, Santangelo, PJ, Segal, DJ, Shimoyama, M, Skala, MC, Tarantal, AF, Tilton, JC, Truskey, GA, Vandsburger, M, Watts, JK, Wells, KD, Wolfe, SA, Xu, Q, Xue, W, Yi, G, Zhou, J, and SCGE Consortium

Abstract

The move from reading to writing the human genome offers new opportunities to improve human health. The United States National Institutes of Health (NIH) Somatic Cell Genome Editing (SCGE) Consortium aims to accelerate the development of safer and more-effective methods to edit the genomes of disease-relevant somatic cells in patients, even in tissues that are difficult to reach. Here we discuss the consortium's plans to develop and benchmark approaches to induce and measure genome modifications, and to define downstream functional consequences of genome editing within human cells. Central to this effort is a rigorous and innovative approach that requires validation of the technology through third-party testing in small and large animals. New genome editors, delivery technologies and methods for tracking edited cells in vivo, as well as newly developed animal models and human biological systems, will be assembled-along with validated datasets-into an SCGE Toolkit, which will be disseminated widely to the biomedical research community. We visualize this toolkit-and the knowledge generated by its applications-as a means to accelerate the clinical development of new therapies for a wide range of conditions.

Published
2021

14. Using qPCR and high-resolution sensor data to model a multi-species Pseudo-nitzschia (Bacillariophyceae) bloom in southeastern Australia.

Author

Ajani, PA, Verma, A, Kim, JH, Woodcock, S, Nishimura, T, Farrell, H, Zammit, A, Brett, S, Murray, SA, Ajani, PA, Verma, A, Kim, JH, Woodcock, S, Nishimura, T, Farrell, H, Zammit, A, Brett, S, and Murray, SA

Abstract

Harmful algal blooms, including those caused by the toxic diatom Pseudo-nitzschia, can have significant impacts on human health, ecosystem functioning and ultimately food security. In the current study we characterized a bloom of species of Pseudo-nitzschia that occurred in a south-eastern Australian oyster-growing estuary in 2019. Using light microscopy, combined with molecular (ITS/5.8S and LSU D1-D3 rDNA regions) and toxicological evidence, we observed the bloom to consist of multiple species of Pseudo-nitzschia including P. cf. cuspidata, P. hasleana, P. fraudulenta and P. multiseries, with P. cf. cuspidata being the only species that produced domoic acid (3.1 pg DA per cell). As several species of Pseudo-nitzschia co-occurred, only one of which produced DA, we developed a rapid, sensitive and efficient quantitative real-time polymerase chain reaction (qPCR) assay to detect only species belonging to the P. pseudodelicatissima complex Clade I, to which P. cf. cuspidata belongs, and this indicated that P. cuspidata or closely related strains may have dominated the Pseudo-nitzschia community at this time. Finally, using high resolution water temperature and salinity sensor data, we modeled the relationship between light microscopy determined abundance of P. delicatissima group and environmental variables (temperature, salinity, rainfall) at two sites within the estuary. A total of eight General Linear Models (GLMs) explaining between 9 and 54% of the deviance suggested that the temperature (increasing) and/or salinity (decreasing) data were generally more predictive of high cell concentrations than the rainfall data at both sites, and that overall, cell concentrations were more predictive at the more oceanic site than the more upstream site, using this method. We conclude that the combination of rapid molecular methods such as qPCR and real-time sensor data modeling, can provide a more rapid and effective early warning of harmful algal blooms of species of Pseudo-n

Published
2021

15. Impacts of harmful algal blooms on marine aquaculture in a low-carbon future.

Author

Lenzen, M, Li, M, Murray, SA, Lenzen, M, Li, M, and Murray, SA

Abstract

The IPCC Special Report on Global Warming of 1.5 °C highlights the potential for dietary shifts to reduce greenhouse gas emissions from livestock. Reductions in the consumption of terrestrial animal protein require increases in the consumption of other food categories, to maintain food security, balanced dietary patterns, and protein intake. Aquaculture has long been suggested as one way to meet future food security needs, and marine and estuarine aquaculture in particular is associated with comparatively low greenhouse gas emissions. However, marine and freshwater aquaculture is affected by factors including harmful algal blooms (HABs), which have been increasingly documented around the world, correlated to increases in worldwide aquaculture. In this study, we applied a global multi-region input-output model to capture the direct effects as well as the indirect and induced effects HABs might pose to a global dietary transition from terrestrial livestock to increased seafood consumption from marine and estuarine aquaculture sources. We found that marine and estuarine aquaculture has a substantial potential to replace meat consumption from terrestrial livestock sources, as increases in CO2 emissions from aquaculture were more than offset by reductions in emissions from mainly cattle grazing and associated land clearing. HABs were found to have a minor monetary impact, but the impact on protein supply was found to be potentially sizeable. For example, in a future setting where 40% of terrestrial protein sources were replaced by aquaculture, a HAB-caused global loss of 5% would set in motion numerous supply-chain cascades, affecting industries auxiliary to aquaculture, indirectly and ultimately reducing protein intake by 10-20%. Such reductions have the potential for pushing parts of Sub-Saharan populations into protein-energy malnutrition. Nevertheless, there remains a significant potential for a dietary transition to increased aquaculture seafood to contribute to red

Published
2021

16. A Comparative Analysis of Methods (LC-MS/MS, LC-MS and Rapid Test Kits) for the Determination of Diarrhetic Shellfish Toxins in Oysters, Mussels and Pipis

Author

Ajani, PA, Sarowar, C, Turnbull, A, Farrell, H, Zammit, A, Helleren, S, Hallegraeff, G, Murray, SA, Ajani, PA, Sarowar, C, Turnbull, A, Farrell, H, Zammit, A, Helleren, S, Hallegraeff, G, and Murray, SA

Abstract

Rapid methods for the detection of biotoxins in shellfish can assist the seafood industry and safeguard public health. Diarrhetic Shellfish Toxins (DSTs) are produced by species of the dinoflagellate genus Dinophysis, yet the comparative efficacy of their detection methods has not been systematically determined. Here, we examined DSTs in spiked and naturally contaminated shellfish–Sydney Rock Oysters (Saccostrea glomerata), Pacific Oysters (Magallana gigas/Crassostrea gigas), Blue Mussels (Mytilus galloprovincialis) and Pipis (Plebidonax deltoides/Donax deltoides), using LC-MS/MS and LC-MS in 4 laboratories, and 5 rapid test kits (quantitative Enzyme-Linked Immunosorbent Assay (ELISA) and Protein Phosphatase Inhibition Assay (PP2A), and qualitative Lateral Flow Assay (LFA)). We found all toxins in all species could be recovered by all laboratories using LC-MS/MS (Liquid Chromatography—tandem Mass Spectrometry) and LC-MS (Liquid Chromatography—Mass Spectrometry); however, DST recovery at low and mid-level concentrations (<0.1 mg/kg) was variable (0–150%), while recovery at high-level concentrations (>0.86 mg/kg) was higher (60–262%). While no clear differences were observed between shellfish, all kits delivered an unacceptably high level (25–100%) of falsely compliant results for spiked samples. The LFA and the PP2A kits performed satisfactorily for naturally contaminated pipis (0%, 5% falsely compliant, respectively). There were correlations between spiked DSTs and quantitative methods was highest for LC-MS (r2 = 0.86) and the PP2A kit (r2 = 0.72). Overall, our results do not support the use of any DST rapid test kit as a stand-alone quality assurance measure at this time

Published
2021

17. Gambierdiscus, the cause of ciguatera fish poisoning: an increased human health threat influenced by climate change

Author

Farrell, H, Kohli, GS, and Murray, SA

Abstract

New species of Gambieriscus are continually being described, with evidence showing that each species might have its own characteristic toxin profile [10, 12, 13].As in the case of other dinoflagellate genera such as AlexandriumorKarenia, the pro-duction or not of certain toxin groups appears to generally vary at the species level, rather than being consistent within the genus [16]. For this reason, species of harmful algal bloom (HAB)-forming taxa are monitored, acting as early-warning systems for shellfish and seafood safety. This review highlights the significant advances in the study of Gambierdiscus and Fukuyoa. We provide a summary of the morphology and phylogenetics of species of Gambierdiscus and Fukuyoa, their toxicology, distribution, chemistry and methods for the detection of CTXs and MTXs in seafood. While, in re-cent years, advances have been made in our understanding of these genera, this re-view further outlines the major gaps in our current understanding of Gambierdiscus and Fukuyoa and outlines goals for future research in this field

Published
2020

18. Phenotypic trait variability as an indication of adaptive capacity in a cosmopolitan marine diatom

Author

Ajani PA, Petrou K, Larsson ME, Nielsen DA, Burke J, and Murray SA

Subjects
fungi, 0603 Evolutionary Biology, 0605 Microbiology, Microbiology
Abstract

Determining the adaptive capacity of marine phytoplankton is important in predicting changes in phytoplankton responses to ocean warming. Phytoplankton may consist of high levels of standing phenotypic and genetic variability, the basis of rapid evolution; however, few studies have quantified trait variability within and amongst closely related diatom species. Using 35 clonal cultures of the ubiquitous marine diatom Leptocylindrus isolated from six locations, spanning 2000 km of the south-eastern Australian coastline, we found evidence of significant intraspecific morphological and metabolic trait variability, which for 8 of 9 traits (growth rate, biovolume, C:N, silica deposition, silica incorporation rate, chl-a, and photosynthetic efficiency under dark adapted, growth irradiance, and high-light adaptation) were greater within a species than between species. Moreover, only two traits revealed a latitudinal trend with strains isolated from lower latitudes showing significantly higher silicification rates and protein:lipid content compared to their higher latitude counterparts. These data mirror recent studies on diatom intraspecific genetic diversity, which has found comparable levels of genetic diversity at a single site to those thousands of kilometres apart, and provide evidence of a functional role of diatom diversity that will allow for rapid adaptation via ecological selection on standing variation in response to changing conditions.

Published
2020

19. Sexual reproduction and genetic polymorphism within the cosmopolitan marine diatom Pseudo-nitzschia pungens

Author

Kim JH, Ajani P, Murray SA, Kim J-H, Lim HC, Teng ST, Lim PT, Han M-S, and Park BS

Abstract

Different clades belonging to the cosmopolitan marine diatom Pseudo-nitzschia pungens appear to be present in different oceanic environments, however, a 'hybrid zone', where populations of different clades interbreed, has also been reported. Many studies have investigated the sexual reproduction of P. pungens, focused on morphology and life cycle, rather than the role of sexual reproduction in mixing the genomes of their parents. We carried out crossing experiments to determine the sexual compatibility/incompatibility between different clades of P. pungens, and examined the genetic polymorphism in the ITS2 region. Sexual reproduction did not occur only between clades II and III under any of experimental temperature conditions. Four offspring strains were established between clade I and III successfully. Strains established from offspring were found interbreed with other offspring strains as well as viable with their parental strains. We confirmed the hybrid sequence patterns between clades I and III and found novel sequence types including polymorphic single nucleotide polymorphisms (SNPs) in the offspring strains. Our results implicate that gene exchange and mixing between different clades are still possible, and that sexual reproduction is a significant ecological strategy to maintain the genetic diversity within this diatom species.

Published
2020

20. Functional significance of phylogeographic structure in a toxic benthic marine microbial eukaryote over a latitudinal gradient along the East Australian Current

Author

Verma A, Hughes DJ, Harwood DT, Suggett DJ, Ralph PJ, and Murray SA

Subjects
fungi, 0602 Ecology, 0603 Evolutionary Biology
Abstract

Genetic diversity in marine microbial eukaryotic populations (protists) drives their ecological success by enabling diverse phenotypes to respond rapidly to changing environmental conditions. Despite enormous population sizes and lack of barriers to gene flow, genetic differentiation that is associated with geographic distance, currents, and environmental gradients has been reported from planktonic protists. However, for benthic protists, which have reduced dispersal opportunities, phylogeography and its phenotypic significance are little known. In recent years, the East Australian Current (EAC) has intensified its southward flow, associated with the tropicalization of temperate waters. Benthic harmful algal species have been increasingly found in south-eastern Australia. Yet little is known about the potential of these species to adapt or extend their range in relation to changing conditions. Here, we examine genetic diversity and functional niche divergence in a toxic benthic dinoflagellate, Ostreopsis cf. siamensis, along a 1,500 km north-south gradient in southeastern Australia. Sixty-eight strains were established from eight sampling sites. The study revealed long-standing genetic diversity among strains established from the northern-most sites, along with large phenotypic variation in observed physiological traits such as growth rates, cell volume, production of palytoxin-like compounds, and photophysiological parameters. Strains from the southern populations were more uniform in both genetic and functional traits, and have possibly colonized their habitats more recently. Our study reports significant genetic and functional trait variability in a benthic harmful algal species, indicative of high adaptability, and a possible climate-driven range extension. The observed high trait variation may facilitate development of harmful algal blooms under dynamic coastal environmental conditions.

Published
2020

21. First Detection of Paralytic Shellfish Toxins from Alexandrium pacificum above the Regulatory Limit in Blue Mussels (Mytilus galloprovincialis) in New South Wales, Australia.

Author

Barua, A, Ajani, PA, Ruvindy, R, Farrell, H, Zammit, A, Brett, S, Hill, D, Sarowar, C, Hoppenrath, M, Murray, SA, Barua, A, Ajani, PA, Ruvindy, R, Farrell, H, Zammit, A, Brett, S, Hill, D, Sarowar, C, Hoppenrath, M, and Murray, SA

Abstract

In 2016, 2017 and 2018, elevated levels of the species Alexandrium pacificum were detected within a blue mussel (Mytilus galloprovincialis) aquaculture area at Twofold Bay on the south coast of New South Wales, Australia. In 2016, the bloom persisted for at least eight weeks and maximum cell concentrations of 89,000 cells L-1 of A. pacificum were reported. The identity of A. pacificum was confirmed using molecular genetic tools (qPCR and amplicon sequencing) and complemented by light and scanning electron microscopy of cultured strains. Maximum reported concentrations of paralytic shellfish toxins (PSTs) in mussel tissue was 7.2 mg/kg PST STX equivalent. Elevated cell concentrations of A. pacificum were reported along the adjacent coastal shelf areas, and positive PST results were reported from nearby oyster producing estuaries during 2016. This is the first record of PSTs above the regulatory limit (0.8 mg/kg) in commercial aquaculture in New South Wales since the establishment of routine biotoxin monitoring in 2005. The intensity and duration of the 2016 A. pacificum bloom were unusual given the relatively low abundances of A. pacificum in estuarine and coastal waters of the region found in the prior 10 years.

Published
2020

22. First report of the potentially toxic marine diatom Pseudo-nitzschia simulans (Bacillariophyceae) from the East Australian Current

Author

Ajani, PA, Lim, HC, Verma, A, Lassudrie, M, McBean, K, Doblin, MA, Murray, SA, Ajani, PA, Lim, HC, Verma, A, Lassudrie, M, McBean, K, Doblin, MA, and Murray, SA

Abstract

© 2020 Japanese Society of Phycology Certain species of the marine diatom genus Pseudo-nitzschia are responsible for the production of the domoic acid (DA), a neurotoxin that can bioaccumulate in the food chain and cause amnesic shellfish poisoning (ASP) in animals and humans. This study extends our knowledge by reporting on the first observation of the potentially toxic species Pseudo-nitzschia simulans from this region. One clonal strain of P. simulans was isolated from the East Australian Current and characterized using light and transmission electron microscopy, and phylogenetic analyses based on regions of the internal transcribed spacer (ITS) and the D1–D3 region of the large subunit (LSU) of the nuclear-encoded ribosomal deoxyribonucleic acid (rDNA), as well as examined for DA production as measured by liquid chromatography–mass spectrometry. Although this strain was non-toxic under the defined growth conditions, the results unambiguously confirmed that this isolate is the potentially toxic species P. simulans – the first report of this species from the Southern Hemisphere.

Published
2020

23. Fifteen years of Pseudo-nitzschia in an Australian estuary, including the first potentially toxic P. delicatissima bloom in the southern hemisphere

Author

Ajani, PA, Larsson, ME, Woodcock, S, Rubio, A, Farrell, H, Brett, S, Murray, SA, Ajani, PA, Larsson, ME, Woodcock, S, Rubio, A, Farrell, H, Brett, S, and Murray, SA

Abstract

© 2020 Elsevier Ltd In March 2018, an unprecedented bloom of a species of Pseudo-nitzschia occurred in Berowra Creek, a major tributary of the Hawkesbury River estuary, New South Wales, Australia (max cell abundance 5.7 × 106 cells L−1). Morphological and phylogenetic analysis showed the bloom was dominated by the species P. delicatissima, with toxin characterization by LC-MS confirming one of the seven strains isolated produced domoic acid (DA), at a concentration of 0.34 pg DA per cell. In response to this unprecedented bloom, we used a fifteen year series of phytoplankton and physico-chemical data to relate environmental factors to spatial and temporal variability of Pseudo-nitzschia species throughout the estuary. Species in the Pseudo-nitzschia delicatissima group consistently dominated all locations monitored throughout the estuary, with peak cell densities observed in the austral autumn (lowest in winter). Overall, Generalised Additive Models (GAMs) suggested that high cell concentrations of Pseudo-nitzschia were linked to an increase in soluble reactive phosphorus and a decrease in total nitrogen at all sites, with up to 55% of the deviance explained. Twenty species of Pseudo-nitzschia have now been reported in Australian coastal waters, four of which are confirmed DA producers. We hypothesise that increased urbanization, nutrient input and warmer waters could combine to increase Pseudo-nitzschia blooms in south-eastern Australia.

Published
2020

24. Transcriptomic investigation into polyketide toxin synthesis in Ostreopsis (Dinophyceae) species

Author

Verma, A, Kohli, GS, Harwood, DT, Ralph, PJ, and Murray, SA

Subjects
Microbiology
Abstract

© 2019 Society for Applied Microbiology and John Wiley & Sons Ltd. In marine ecosystems, dinoflagellates can become highly abundant and even dominant at times, despite their comparatively slow growth. Their ecological success may be related to their production of complex toxic polyketide compounds. Ostreopsis species produce potent palytoxin-like compounds (PLTX), which are associated with human skin and eye irritations, and illnesses through the consumption of contaminated seafood. To investigate the genetic basis of PLTX-like compounds, we sequenced and annotated transcriptomes from two PLTX-producing Ostreopsis species; O. cf. ovata, O. cf. siamensis, one non-PLTX producing species, O. rhodesae and compared them to a close phylogenetic relative and non-PLTX producer, Coolia malayensis. We found no clear differences in the presence or diversity of ketosynthase and ketoreductase transcripts between PLTX producing and non-producing Ostreopsis and Coolia species, as both groups contained >90 and > 10 phylogenetically diverse ketosynthase and ketoreductase transcripts, respectively. We report for the first-time type I single-, multi-domain polyketide synthases (PKSs) and hybrid non-ribosomal peptide synthase/PKS transcripts from all species. The long multi-modular PKSs were insufficient by themselves to synthesize the large complex polyether backbone of PLTX-like compounds. This implies that numerous PKS domains, including both single and multi-, work together on the biosynthesis of PLTX-like and other related polyketide compounds.

Published
2019

25. Morphology and phylogenetics of benthic prorocentrum species (Dinophyceae) from tropical Northwestern Australia

Author

Verma, A, Kazandjian, A, Sarowar, C, Harwood, DT, Murray, JS, Pargmann, I, Hoppenrath, M, Murray, SA, Verma, A, Kazandjian, A, Sarowar, C, Harwood, DT, Murray, JS, Pargmann, I, Hoppenrath, M, and Murray, SA

Abstract

© 2019 by the authors. Approximately 70 species of Prorocentrum are known, of which around 30 species are associated with benthic habitats. Some produce okadaic acid (OA), dinophysistoxin (DTX) and their derivatives, which are involved in diarrhetic shellfish poisoning. In this study, we isolated and characterized Prorocentrum concavum and P. malayense from Broome in north Western Australia using light and scanning electron microscopy as well as molecular sequences of large subunit regions of ribosomal DNA, marking the first record of these species from Australian waters. The morphology of the motile cells of P. malayense was similar to P. concavum in the light microscopy, but differed by the smooth thecal surface, the pore pattern and the production of mucous stalk-like structures and a hyaline sheath around the non-motile cells. P. malayense could also be differentiated from other closely related species, P. leve and P. foraminosum, despite the similarity in thecal surface and pore pattern, by its platelet formula and morphologies. We tested the production of OA and DTXs from both species, but found that they did not produce detectable levels of these toxins in the given culturing conditions. This study aids in establishing more effective monitoring of potential harmful algal taxa in Australian waters for aquaculture and recreational purposes.

Published
2019

26. Large expert-curated database for benchmarking document similarity detection in biomedical literature search

Author

Brown, P, Tan, A-C, El-Esawi, MA, Liehr, T, Blanck, O, Gladue, DP, Almeida, GMF, Cernava, T, Sorzano, CO, Yeung, AWK, Engel, MS, Chandrasekaran, AR, Muth, T, Staege, MS, Daulatabad, SV, Widera, D, Zhang, J, Meule, A, Honjo, K, Pourret, O, Yin, C-C, Zhang, Z, Cascella, M, Flegel, WA, Goodyear, CS, van Raaij, MJ, Bukowy-Bieryllo, Z, Campana, LG, Kurniawan, NA, Lalaouna, D, Huttner, FJ, Ammerman, BA, Ehret, F, Cobine, PA, Tan, E-C, Han, H, Xia, W, McCrum, C, Dings, RPM, Marinello, F, Nilsson, H, Nixon, B, Voskarides, K, Yang, L, Costa, VD, Bengtsson-Palme, J, Bradshaw, W, Grimm, DG, Kumar, N, Martis, E, Prieto, D, Sabnis, SC, Amer, SEDR, Liew, AWC, Perco, P, Rahimi, F, Riva, G, Zhang, C, Devkota, HP, Ogami, K, Basharat, Z, Fierz, W, Siebers, R, Tan, K-H, Boehme, KA, Brenneisen, P, Brown, JAL, Dalrymple, BP, Harvey, DJ, Ng, G, Werten, S, Bleackley, M, Dai, Z, Dhariwal, R, Gelfer, Y, Hartmann, MD, Miotla, P, Tamaian, R, Govender, P, Gurney-Champion, OJ, Kauppila, JH, Zhang, X, Echeverria, N, Subhash, S, Sallmon, H, Tofani, M, Bae, T, Bosch, O, Cuiv, PO, Danchin, A, Diouf, B, Eerola, T, Evangelou, E, Filipp, FV, Klump, H, Kurgan, L, Smith, SS, Terrier, O, Tuttle, N, Ascher, DB, Janga, SC, Schulte, LN, Becker, D, Browngardt, C, Bush, SJ, Gaullier, G, Ide, K, Meseko, C, Werner, GDA, Zaucha, J, Al-Farha, AA, Greenwald, NF, Popoola, SI, Rahman, MS, Xu, J, Yang, SY, Hiroi, N, Alper, OM, Baker, CI, Bitzer, M, Chacko, G, Debrabant, B, Dixon, R, Forano, E, Gilliham, M, Kelly, S, Klempnauer, K-H, Lidbury, BA, Lin, MZ, Lynch, I, Ma, W, Maibach, EW, Mather, DE, Nandakumar, KS, Ohgami, RS, Parchi, P, Tressoldi, P, Xue, Y, Armitage, C, Barraud, P, Chatzitheochari, S, Coelho, LP, Diao, J, Doxey, AC, Gobet, A, Hu, P, Kaiser, S, Mitchell, KM, Salama, MF, Shabalin, IG, Song, H, Stevanovic, D, Yadollahpour, A, Zeng, E, Zinke, K, Alimba, CG, Beyene, TJ, Cao, Z, Chan, SS, Gatchell, M, Kleppe, A, Piotrowski, M, Torga, G, Woldesemayat, AA, Cosacak, MI, Haston, S, Ross, SA, Williams, R, Wong, A, Abramowitz, MK, Effiong, A, Lee, S, Abid, MB, Agarabi, C, Alaux, C, Albrecht, DR, Atkins, GJ, Beck, CR, Bonvin, AMJJ, Bourke, E, Brand, T, Braun, RJ, Bull, JA, Cardoso, P, Carter, D, Delahay, RM, Ducommun, B, Duijf, PHG, Epp, T, Eskelinen, E-L, Fallah, M, Farber, DB, Fernandez-Triana, J, Feyerabend, F, Florio, T, Friebe, M, Furuta, S, Gabrielsen, M, Gruber, J, Grybos, M, Han, Q, Heinrich, M, Helantera, H, Huber, M, Jeltsch, A, Jiang, F, Josse, C, Jurman, G, Kamiya, H, de Keersmaecker, K, Kristiansson, E, de Leeuw, F-E, Li, J, Liang, S, Lopez-Escamez, JA, Lopez-Ruiz, FJ, Marchbank, KJ, Marschalek, R, Martin, CS, Miele, AE, Montagutelli, X, Morcillo, E, Nicoletti, R, Niehof, M, O'Toole, R, Ohtomo, T, Oster, H, Palma, J-A, Paterson, R, Peifer, M, Portilla, M, Portillo, MC, Pritchard, AL, Pusch, S, Raghava, GPS, Roberts, NJ, Ross, K, Schuele, B, Sergeant, K, Shen, J, Stella, A, Sukocheva, O, Uversky, VN, Vanneste, S, Villet, MH, Viveiros, M, Vorholt, JA, Weinstock, C, Yamato, M, Zabetakis, I, Zhao, X, Ziegler, A, Aizat, WM, Atlas, L, Bridges, KM, Chakraborty, S, Deschodt, M, Domingues, HS, Esfahlani, SS, Falk, S, Guisado, JL, Kane, NC, Kueberuwa, G, Lau, CL, Liang, D, Liu, E, Luu, AM, Ma, C, Ma, L, Moyer, R, Norris, AD, Panthee, S, Parsons, JR, Peng, Y, Pinto, IM, Reschke, CR, Sillanpaa, E, Stewart, CJ, Uhle, F, Yang, H, Zhou, K, Zhu, S, Ashry, M, Bergsland, N, Berthold, M, Chen, C-E, Colella, V, Cuypers, M, Eskew, EA, Fan, X, Gajda, M, Gonzalezlez-Prendes, R, Goodin, A, Graham, EB, Groen, EJN, Gutierrez-Sacristan, A, Habes, M, Heffler, E, Higginbottom, DB, Janzen, T, Jayaraman, J, Jibb, LA, Jongen, S, Kinyanjui, T, Koleva-Kolarova, RG, Li, Z, Liu, Y-P, Lund, BA, Lussier, AA, Mier, P, Moore, MD, Nagler, K, Orme, MW, Pearson, JA, Prajapati, AS, Saito, Y, Troder, SE, Uchendu, F, Verloh, N, Voutchkova, DD, Abu-Zaid, A, Bakkach, J, Baumert, P, Dono, M, Hanson, J, Herbelet, S, Hobbs, E, Kulkarni, A, Liu, S, Loft, ND, Reddan, T, Senghore, T, Vindin, H, Xu, H, Bannon, R, Chen, B, Cheung, JTK, Cooper, J, Esnakul, AK, Feghali, KA, Ghelardi, E, Gnasso, A, Horbar, J, Lai, HM, Ma, R, Pan, Z, Peres, MA, Pranata, R, Seow, E, Sydes, M, Testoni, I, Westermair, AL, Yang, Y, Afnan, M, Albiol, J, Albuquerque, LG, Amiya, E, Amorim, RM, An, Q, Andersen, SU, Aplin, JD, Argyropoulos, C, Asmann, YW, Assaeed, AM, Atanasov, AG, Atchison, DA, Avery, SV, Avillach, P, Baade, PD, Backman, L, Badie, C, Baldi, A, Ball, E, Bardot, O, Barnett, AG, Basner, M, Batra, J, Bazanova, OM, Beale, A, Beddoe, T, Bell, ML, Berezikov, E, Berners-Price, S, Bernhardt, P, Berry, E, Bessa, TB, Billington, C, Birch, J, Blakely, RD, Blaskovich, MAT, Blum, R, Boelaert, M, Bogdanos, D, Bosch, C, Bourgoin, T, Bouvard, D, Boykin, LM, Bradley, G, Braun, D, Brownlie, J, Bruhl, A, Burt, A, Butler, LM, Byrareddy, SN, Byrne, HJ, Cabantous, S, Calatayud, S, Candal, E, Carlson, K, Casillas, S, Castelvetro, V, Caswell, PT, Cavalli, G, Cerovsky, V, Chagoyen, M, Chen, C-S, Chen, DF, Chen, H, Chen, J-T, Chen, Y, Cheng, C, Cheng, J, Chinapaw, M, Chinopoulos, C, Cho, WCS, Chong, L, Chowdhury, D, Chwalibog, A, Ciresi, A, Cockcroft, S, Conesa, A, Cook, PA, Cooper, DN, Coqueret, O, Corea, EM, Costa, E, Coupland, C, Crawford, SY, Cruz, AD, Cui, H, Cui, Q, Culver, DC, D'Angiulli, A, Dahms, TES, Daigle, F, Dalgleish, R, Danielsen, HE, Darras, S, Davidson, SM, Day, DA, Degirmenci, V, Demaison, L, Devriendt, K, Ding, J, Dogan, Y, Dong, XC, Donner, CF, Dressick, W, Drevon, CA, Duan, H, Ducho, C, Dumaz, N, Dwarakanath, BS, Ebell, MH, Eisenhardt, S, Elkum, N, Engel, N, Erickson, TB, Fairhead, M, Faville, MJ, Fejzo, MS, Festa, F, Feteira, A, Flood-Page, P, Forsayeth, J, Fox, SA, Franks, SJ, Frentiu, FD, Frilander, MJ, Fu, X, Fujita, S, Galea, I, Galluzzi, L, Gani, F, Ganpule, AP, Garcia-Alix, A, Gedye, K, Giordano, M, Giunta, C, Gleeson, PA, Goarant, C, Gong, H, Gora, D, Gough, MJ, Goyal, R, Graham, KE, Grande-Perez, A, Graves, PM, Greidanus, H, Grice, D, Grunau, C, Gumulya, Y, Guo, Y, Gurevich, VV, Gusev, O, Hacker, E, Hage, SR, Hagen, G, Hahn, S, Haller, DM, Hammerschmidt, S, Han, J, Han, R, Handfield, M, Hapuarachchi, HC, Harder, T, Hardingham, JE, Heck, M, Heers, M, Hew, KF, Higuchi, Y, St Hilaire, C, Hilton, R, Hodzic, E, Hone, A, Hongoh, Y, Hu, G, Huber, HP, Hueso, LE, Huirne, J, Hurt, L, Idborg, H, Ikeo, K, Ingley, E, Jakeman, PM, Jensen, A, Jia, H, Jia, S, Jiang, J, Jiang, X, Jin, Y, Jo, D, Johnson, AM, Johnston, M, Jonscher, KR, Jorens, PG, Jorgensen, JOL, Joubert, JW, Jung, S-H, Junior, AM, Kahan, T, Kamboj, SK, Kang, Y-K, Karamanos, Y, Karp, NA, Kelly, R, Kenna, R, Kennedy, J, Kersten, B, Khalaf, RA, Khalid, JM, Khatlani, T, Khider, T, Kijanka, GS, King, SRB, Kluz, T, Knox, P, Kobayashi, T, Koch, K-W, Kohonen-Corish, MRJ, Kong, X, Konkle-Parker, D, Korpela, KM, Kostrikis, LG, Kraiczy, P, Kratz, H, Krause, G, Krebsbach, PH, Kristensen, SR, Kumari, P, Kunimatsu, A, Kurdak, H, Kwon, YD, Lachat, C, Lagisz, M, Laky, B, Lammerding, J, Lange, M, Larrosa, M, Laslett, AL, LeClair, EE, Lee, K-W, Lee, M-Y, Lee, M-S, Li, G, Lieb, K, Lim, YY, Lindsey, ML, Line, P-D, Liu, D, Liu, F, Liu, H, Lloyd, VK, Lo, T-W, Locci, E, Loidl, J, Lorenzen, J, Lorkowski, S, Lovell, NH, Lu, H, Lu, W, Lu, Z, Luengo, GS, Lundh, L-G, Lysy, PA, Mabb, A, Mack, HG, Mackey, DA, Mahdavi, SR, Maher, P, Maher, T, Maity, SN, Malgrange, B, Mamoulakis, C, Mangoni, AA, Manke, T, Manstead, ASR, Mantalaris, A, Marsal, J, Marschall, H-U, Martin, FL, Martinez-Raga, J, Martinez-Salas, E, Mathieu, D, Matsui, Y, Maza, E, McCutcheon, JE, Mckay, GJ, McMillan, B, McMillan, N, Meads, C, Medina, L, Merrick, BA, Metzger, DW, Meunier, FA, Michaelis, M, Micheau, O, Mihara, H, Mintz, EM, Mizukami, T, Moalic, Y, Mohapatra, DP, Monteiro, A, Montes, M, Moran, JV, Morozov, SY, Mort, M, Murai, N, Murphy, DJ, Murphy, SK, Murray, SA, Naganawa, S, Nammi, S, Nasios, G, Natoli, RM, Nguyen, F, Nicol, C, van Nieuwerburgh, F, Nilsen, EB, Nobile, CJ, O'Mahony, M, Ohlsson, S, Olatunbosun, O, Olofsson, P, Ortiz, A, Ostrikov, K, Otto, S, Outeiro, TF, Ouyang, S, Paganoni, S, Page, A, Palm, C, Paradies, Y, Parsons, MH, Parsons, N, Pascal, P, Paul, E, Peckham, M, Pedemonte, N, Pellizzon, MA, Petrelli, M, Pichugin, A, Pinto, CJC, Plevris, JN, Pollesello, P, Polz, M, Ponti, G, Porcelli, P, Prince, M, Quinn, GP, Quinn, TJ, Ramula, S, Rappsilber, J, Rehfeldt, F, Reiling, JH, Remacle, C, Rezaei, M, Riddick, EW, Ritter, U, Roach, NW, Roberts, DD, Robles, G, Rodrigues, T, Rodriguez, C, Roislien, J, Roobol, MJ, Rowe, JA, Ruepp, A, van Ruitenbeek, J, Rust, P, Saad, S, Sack, GH, Santos, M, Saudemont, A, Sava, G, Schrading, S, Schramm, A, Schreiber, M, Schuler, S, Schymkowitz, J, Sczyrba, A, Seib, KL, Shi, H-P, Shimada, T, Shin, J-S, Shortt, C, Silveyra, P, Skinner, D, Small, I, Smeets, PAM, So, P-W, Solano, F, Sonenshine, DE, Song, J, Southall, T, Speakman, JR, Srinivasan, MV, Stabile, LP, Stasiak, A, Steadman, KJ, Stein, N, Stephens, AW, Stewart, DI, Stine, K, Storlazzi, C, Stoynova, NV, Strzalka, W, Suarez, OM, Sultana, T, Sumant, AV, Summers, MJ, Sun, G, Tacon, P, Tanaka, K, Tang, H, Tanino, Y, Targett-Adams, P, Tayebi, M, Tayyem, R, Tebbe, CC, Telfer, EE, Tempel, W, Teodorczyk-Injeyan, JA, Thijs, G, Thorne, S, Thrift, AG, Tiffon, C, Tinnefeld, P, Tjahjono, DH, Tolle, F, Toth, E, del Tredici, AL, Tsapas, A, Tsirigotis, K, Turak, A, Tzotzos, G, Udo, EE, Utsumi, T, Vaidyanathan, S, Vaillant, M, Valsesia, A, Vandenbroucke, RE, Veiga, FH, Vendrell, M, Vesk, PA, Vickers, P, Victor, VM, Villemur, R, Vohl, M-C, Voolstra, CR, Vuillemin, A, Wakelin, S, Waldron, L, Walsh, LJ, Wang, AY, Wang, F, Wang, Y, Watanabe, Y, Weigert, A, Wen, J-C, Wham, C, White, EP, Wiener, J, Wilharm, G, Wilkinson, S, Willmann, R, Wilson, C, Wirth, B, Wojan, TR, Wolff, M, Wong, BM, Wu, T-W, Wuerbel, H, Xiao, X, Xu, D, Xu, JW, Xue, B, Yalcin, S, Yan, H, Yang, E-C, Yang, S, Yang, W, Ye, Y, Ye, Z-Q, Yli-Kauhaluoma, J, Yoneyama, H, Yu, Y, Yuan, G-C, Yuh, C-H, Zaccolo, M, Zeng, C, Zevnik, B, Zhang, L, Zhang, Y, 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W, Suarez, OM, Sultana, T, Sumant, AV, Summers, MJ, Sun, G, Tacon, P, Tanaka, K, Tang, H, Tanino, Y, Targett-Adams, P, Tayebi, M, Tayyem, R, Tebbe, CC, Telfer, EE, Tempel, W, Teodorczyk-Injeyan, JA, Thijs, G, Thorne, S, Thrift, AG, Tiffon, C, Tinnefeld, P, Tjahjono, DH, Tolle, F, Toth, E, del Tredici, AL, Tsapas, A, Tsirigotis, K, Turak, A, Tzotzos, G, Udo, EE, Utsumi, T, Vaidyanathan, S, Vaillant, M, Valsesia, A, Vandenbroucke, RE, Veiga, FH, Vendrell, M, Vesk, PA, Vickers, P, Victor, VM, Villemur, R, Vohl, M-C, Voolstra, CR, Vuillemin, A, Wakelin, S, Waldron, L, Walsh, LJ, Wang, AY, Wang, F, Wang, Y, Watanabe, Y, Weigert, A, Wen, J-C, Wham, C, White, EP, Wiener, J, Wilharm, G, Wilkinson, S, Willmann, R, Wilson, C, Wirth, B, Wojan, TR, Wolff, M, Wong, BM, Wu, T-W, Wuerbel, H, Xiao, X, Xu, D, Xu, JW, Xue, B, Yalcin, S, Yan, H, Yang, E-C, Yang, S, Yang, W, Ye, Y, Ye, Z-Q, Yli-Kauhaluoma, J, Yoneyama, H, Yu, Y, Yuan, G-C, Yuh, C-H, Zaccolo, M, Zeng, C, Zevnik, B, Zhang, L, Zhang, Y, Zhang, Z-Y, Zhao, Y, Zhou, M, Zuberbier, T, Aanei, CM, Ahmad, R, Al-Lawama, M, Alanio, A, Allardyce, J, Alonso-Caneiro, D, Atack, JM, Baier, D, Bansal, A, Benezeth, Y, Berbesque, C, Berrevoet, F, Biedermann, PHW, Bijleveld, E, Bittner, F, Blombach, F, Van den Bos, W, Boudreau, SA, Bramoweth, AD, Braubach, O, Cai, Y, Campbell, M, Catry, T, Chen, X, Cheng, S, Chung, H-J, Chavez-Fumagalli, MA, Conway, A, Costa, BM, Cyr, N, Dean, LT, Denzel, MS, Dlamini, SV, Dudley, KJ, Dufies, M, Ecke, T, Eckweiler, D, Eixarch, E, El-Adawy, H, Emmrich, JV, Eustace, AJ, Falter-Wagner, CM, Fuss, J, Gao, J, Gill, MR, Gloyn, L, Goggs, R, Govinden, U, Greene, G, Greiff, V, Grundle, DS, Gruneberg, P, Gumede, N, Haore, G, Harrison, P, Hoenner, X, Hojsgaard, D, Hori, H, Ikonomopoulou, MP, Jeurissen, P, Johnson, DM, Kabra, D, Kamagata, K, Karmakar, C, Kasian, O, Kaye, LK, Khan, MM, Kim, Y-M, Kish, JK, Kobold, S, Kohanbash, G, Kohls, G, Kugler, J-M, Kumar, G, Lacy-Colson, J, Latif, A, Lauschke, VM, Li, B, Lim, CJ, Liu, X, Lu, J-J, Lu, Q, Mahavadi, P, Marzocchi, U, McGarrigle, CA, van Meerten, T, Min, R, Moal, I, Molari, M, Molleman, L, Mondal, SR, Van de Mortel, T, Moss, WN, Moultos, OA, Mukherjee, M, Nakayama, K, Narayan, E, Navaratnarajah, Neumann, P-A, Nie, J, Nie, Y, Niemeyer, F, Fiona, Nwaiwu, O, Oldenmenger, WH, Olumayede, E, Ou, J, Pallebage-Gamarallage, M, Pearce, SP, Pelkonen, T, Pelleri, MC, Pereira, JL, Pheko, M, Pinto, KA, Piovesan, A, Pluess, M, Podolsky, IM, Prescott, J, Qi, D, Qi, X, Raikou, VD, Ranft, A, Rhodes, J, Rotge, J-Y, Rowe, AD, Saggar, M, Schuon, RA, Shahid, S, Shalchyan, V, Shirvalkar, P, Shiryayev, O, Singh, J, Smout, MJ, Soares, A, Song, C, Srivastava, K, Srivastava, RK, Sun, J, Szabo, A, Szymanski, W, Tai, CNP, Takeuchi, H, Tanadini-Lang, S, Tang, F, Tao, W, Theron, G, Tian, CF, Tian, Y-S, Tuttle, LM, Valenti, A, Verlot, P, Walker, M, Wang, J, Welter, D, Winslade, M, Wu, D, Wu, Y-R, Xiao, H, Xu, B, Xu, Z, Yang, D, Yang, M, Yankilevich, P, You, Y, Yu, C, Zhan, J, Zhang, G, Zhang, K, Zhang, T, Zhao, G, Zhao, J, Zhou, X, Zhu, Z, Ajani, PA, Anazodo, UC, Bagloee, SA, Bail, K, Bar, I, Bathelt, J, Benkeser, D, Bernier, ML, Blanchard, AM, Boakye, DW, Bonatsos, V, Boon, MH, Bouboulis, G, Bromfield, E, Brown, J, Bul, KCM, Burton, KJ, Butkowski, EG, Carroll, G, Chao, F, Charrier, EE, Chen, Y-C, Chenguang, Choi, JR, Christoffersen, T, Comel, JC, Cosse, C, Cui, Y, van Dessel, P, Dhaval, Diodato, D, Duffey, M, Dutt, A, Egea, LG, El-Said, M, Faye, M, Fernandez-Fernandez, B, Foley, KG, Founou, LL, Fu, F, Gadelkareem, RA, Galimov, E, Garip, G, Gemmill, A, Gouil, Q, Grey, J, Gridneva, Z, Grothe, MJ, Grebert, T, Guerrero, F, Guignard, L, Haenssgen, MJ, Hasler, D, Holgate, JY, Huang, A, Hulse-Kemp, AM, Jean-Quartier, C, Jeon, S-M, Jia, Y, Jutzeler, C, Kalatzis, P, Karim, M, Karsay, K, Keitel, A, Kempe, A, Keown, JR, Khoo, CM, Khwaja, N, Kievit, RA, Kosanic, A, Koutoukidis, DA, Kramer, P, Kumar, D, Kirag, N, Lanza, G, Le, TD, Leem, JW, Leightley, D, Leite, A, Lercher, L, Li, Y, Lim, R, Lima, LRA, Lin, L, Ling, T, Liu, Y, Liu, Z, Lu, Y, Lum, FM, Luo, H, Machhi, J, Macleod, A, Macwan, I, Madala, HR, Madani, N, de Maio, N, Makowiecki, K, Mallinson, DJ, Margelyte, R, Maria, C, Markonis, Y, Marsili, L, Mavoa, S, McWilliams, L, Megersa, M, Mendes, CSM, Menichetti, J, Mercieca-Bebber, R, Miller, JJ, Minde, D-PM, Minges, A, Mishra, E, Mishra, VR, Moores, C, Morrice, N, Moskalensky, AE, Navarin, N, Negera, E, Nolet, P, Nordberg, A, Norden, R, Nowicki, JP, Olova, N, Olszewski, P, Onzima, R, Pan, C-L, Park, C, Park, DI, Park, S, Patil, CD, Pedro, SA, Perry, SR, Peter, J, Peterson, BM, Pezzuolo, A, Pozdnyakov, I, Qian, S, Qin, L, Rafe, A, Raote, I, Raza, A, Rebl, H, Refai, O, Regan, T, Richa, T, Richardson, MF, Robinson, KR, Rossoni, L, Rouet, R, Safaei, S, Schneeberger, PHH, Schwotzer, D, Sebastian, A, Selinski, J, Seltmann, S, Sha, F, Shalev, N, Shang, J-L, Singer, J, Singh, M, Smith, T, Solomon-Moore, E, Song, L, Soraggi, S, Stanley, R, Steckhan, N, Strobl, F, Subissi, L, Supriyanto, I, Surve, CR, Suzuki, T, Syme, C, Sorelius, K, Tang, Y, Tantawy, M, Tennakoon, S, Teseo, S, Toelzer, C, Tomov, N, Tovar, M, Tran, L, Tripathi, S, Tuladhar, AM, Ukubuiwe, AC, Ung, COL, Valgepea, K, Vatanparast, H, Vidal, A, Wang, Q, Watari, R, Webster, R, Wei, J, Wibowo, D, Wingenbach, TSH, Xavier, RM, Xiao, S, Xiong, P, Xu, S, Yao, R, Yao, W, Yin, Q, Zaitsu, M, Zeineb, Z, Zhan, X-Y, Zhang, R, Zhang, W, Zheng, S, Zhou, B, Ahmad, H, Akinwumi, SA, Albery, GF, Alhowimel, A, Ali, J, Alshehri, M, Alsuhaibani, M, Anikin, A, Azubuike, SO, Bach-Mortensen, A, Baltiansky, L, Bartas, M, Belachew, KY, Bhardwaj, V, Binder, K, Bland, NS, Boah, M, Bullen, B, Calabro, GE, Callahan, TJ, Cao, B, Chalmers, K, Chang, W, Che, Z, Chen, ATY, Chen, Z, Choi, Y, Chowdhury, MAK, Christensen, MR, Cooke, RSC, Cottini, M, Covington, NV, Cunningham, C, Delarocque, J, Devos, L, Dhar, AR, Ding, K-F, Dong, K, Dong, Z, Dreyer, N, Ekstrand, C, Fardet, T, Feleke, BE, Feurer, T, Freitas, A, Gao, T, Asefa, NG, Giganti, F, Grabowski, P, Guerra-Mora, JR, Guo, C, Guo, X, Gupta, H, He, S, Heijne, M, Heinemann, S, Hogrebe, A, Huang, Z, Iskander-Rizk, S, Iyer, LM, Jahan, Y, James, AS, Joel, E, Joffroy, B, Jegousse, C, Kambondo, G, Karnati, P, Kaya, C, Ke, A, Kelly, D, Kickert, R, Kidibule, PE, Kieselmann, JP, Kim, HJ, Kitazawa, T, Lamberts, A, Liang, H, Linn, SN, Litfin, T, Liusuo, W, Lygirou, V, Mahato, AK, Mai, Z-M, Major, RW, Mali, S, Mallis, P, Mao, W, Marvin-Dowle, K, Mason, LD, Merideth, B, Merino-Plaza, MJ, Merlaen, B, Messina, R, Mishra, AK, Muhammad, J, Musinguzi, C, Nanou, A, Naqash, A, Nguyen, JT, Nguyen, TTH, Ni, D, Nida, Notcovich, S, Ohst, B, Ollivier, QR, Osses, DF, Peng, X, Plantinga, A, Pulia, M, Rafiq, M, Raman, A, Raucher-Chene, D, Rawski, R, Ray, A, Razak, LA, Rudolf, K, Rusch, P, Sadoine, ML, Schmidt, A, Schurr, R, Searles, S, Sharma, S, Sheehan, B, Shi, C, Shohayeb, B, Sommerlad, A, Strehlow, J, Sun, X, Sundar, R, Taherzadeh, G, Tahir, NDM, Tang, J, Testa, J, Tian, Z, Tingting, Q, Verheijen, GP, Vickstrom, C, Wang, T, Wang, X, Wang, Z, Wei, P, Wilson, A, Wyart, Yassine, A-A, Yousefzadeh, A, Zare, A, Zeng, Z, Zhang, H, Zhou, J, Zhu, D, Adamo, V, Adeyemo, AA, Aggelidou, M, Al-Owaifeer, AM, Al-Riyami, AZ, Alzghari, SK, Andersen, V, Angus, K, Asaduzzaman, M, Asady, H, Ato, D, Bai, X, Baines, RL, Ballantyne, M, Ban, B, Beck, J, Ben-Nafa, W, Black, E, Blancher, A, Blankstein, R, Bodagh, N, Borges, PAV, Brooks, A, Brox-Ponce, J, Brunetti, A, Canham, CD, Carninci, P, Carvajal, R, Chang, SC, Chao, J, Chatterjee, P, Chhatriwalla, AK, Chikowe, I, Chuang, T-J, Collevatti, RG, Valera-Cornejo, DA, Cuenda, A, Dao, M, Dauga, D, Deng, Z, Devkota, K, Doan, LV, Elewa, YHA, Fan, D, Faruk, M, Feifei, S, Ferguson, TS, Fleres, F, Foster, EJ, Foster, CS, Furer, T, Gao, Y, Garcia-Rivera, EJ, Gazdar, A, George, RB, Ghosh, S, Gianchecchi, E, Gleason, JM, Hackshaw, A, Hall, A, Hall, R, Harper, P, Hogg, WE, Huang, G, Hunter, KE, IJzerman, AP, Jesus, C, Jian, G, Lewis, JS, Kanj, SS, Kaur, H, Kheir, F, Kichatova, VS, Kiyani, M, Klein, R, Kovesi, T, Kraschnewski, JL, Kumar, AP, Labutin, D, Lazo-Langner, A, Leclercq, G, Li, M, Li, Q, Li, T, Liao, W-T, Liao, Z-Y, Lin, J, Lizer, J, Lobreglio, G, Lowies, C, Lu, C, Majeed, H, Martin, A, Martinez-Sobrido, L, Meresh, E, Middelveen, M, Mohebbi, A, Mota, J, Mozaheb, Z, Muyaya, L, Nandhakumar, A, Ng, SHX, Obeidat, M, Oh, D-H, Owais, M, Pace-Asciak, P, Panwar, A, Patterson, C, Penagos-Tabaree, F, Pianosi, PT, Pinzi, V, Pridans, C, Psaroulaki, A, Pujala, RK, Pulido-Arjona, L, Qi, P-F, Rahman, P, Rai, NK, Rassaf, T, Refardt, J, Ricciardi, W, Riess, O, Rovas, A, Sacks, FM, Saleh, S, Sampson, C, Schmutz, A, Sepanski, R, Sharma, N, Spearman, P, Subramaniapillai, M, Swali, R, Tan, CM, Tellechea, JI, Thomas, L-M, Tong, X, Veys, R, Vitriol, V, Wang, H-D, Waugh, J, Webb, SA, Williams, BA, Workman, AD, Xiang, T, Xie, L-X, Xu, T, Yang, C, Yoon, JG, Yuan, CM, Zaritsky, A, Zhao, H, Zuckerman, H, Lyu, R, Pullan, W, and Zhou, Y

Published
2019

28. Harmful Algae Introductions: Vectors of Transfer, Mitigation, and Management

Author

Shumway, S, Burkholder, JA, Morton, S, Murray, SA, Hallegraeff, G, Shumway, S, Burkholder, JA, Morton, S, Murray, SA, and Hallegraeff, G

Abstract

Chapters in this volume will provide readers with information on causes of HAB, successful management and monitoring programs, control, prevention, and mitigation strategies, economic consequences of HAB, associated risks to human health, ...

Published
2019

29. Characterisation of Two Toxic Gambierdiscus spp. (Gonyaulacales, Dinophyceae) from the Great Barrier Reef (Australia): G. lewisii sp. nov. and G. holmesii sp. nov.

Author

Kretzschmar, AL, Larsson, ME, Hoppenrath, M, Doblin, MA, Murray, SA, Kretzschmar, AL, Larsson, ME, Hoppenrath, M, Doblin, MA, and Murray, SA

Abstract

© 2019 Elsevier GmbH Ciguatera fish poisoning (CFP) is a human illness caused via consumption of seafood contaminated with neurotoxins produced by some species from the epiphytic dinoflagellate genus Gambierdiscus. In this study, we describe two new species of Gambierdiscus isolated from Heron Island in the Southern Great Barrier Reef, Queensland, Australia. These new species were analysed using light microscopy, scanning electron microscopy, and phylogenetic analyses of nuclear encoded ribosomal ITS, SSU as well as D1-D3 and D8-D10 of the LSU gene regions. Gambierdiscus lewisii sp. nov. (Po, 3′, 0a, 7″, 6c,? s, 5‴, 0p, 2′‴) is distinguished by its strong reticulate-foveate ornamentation and is genetically distinct from its sister species, G. pacificus. Gambierdiscus holmesii sp. nov. (Po, 3′, 0a, 7″, 6c, 6s?, 5‴, 0p, 2′‴) is morphologically distinct from the genetically similar species G. silvae because of a strongly ventrally displaced apical pore complex and a characteristic fold at the anterior edge of the sulcus. Both G. lewisii and G. holmesii produce putative Maitotoxin-(44-Methylgambierone) and compounds which show ciguatoxin and maitotoxin-like activities. Identification of two new Gambierdiscus species will enable us to more accurately assess the risk of CFP in Australia and internationally.

Published
2019

31. Evaluation of sxtA and rDNA qPCR assays through monitoring of an inshore bloom of Alexandrium catenella Group 1

Author

Murray, SA, Ruvindy, R, Kohli, GS, Anderson, DM, Brosnahan, ML, Murray, SA, Ruvindy, R, Kohli, GS, Anderson, DM, and Brosnahan, ML

Abstract

© 2019, The Author(s). Alexandrium catenella (formerly A. tamarense Group 1, or A. fundyense) is the leading cause of Paralytic Shellfish Poisoning in North and South America, Europe, Africa, Australia and Asia. The quantification of A.catenella via sxtA, a gene involved in Paralytic Shellfish Toxin synthesis, may be a promising approach, but has not been evaluated in situ on blooms of A. catenella, in which cell abundances may vary from not detectable to in the order of 106 cells L−1. In this study, we compared sxtA assay performance to a qPCR assay targeted to a species-specific region of ribosomal DNA (rDNA) and an established fluorescent in situ hybridization (FISH) microscopy method. Passing-Bablok regression analyses revealed the sxtA assay to overestimate abundances when <5 cell equivalents A. catenella DNA were analysed, but otherwise was closer to microscopy estimates than the rDNA assay, which overestimated abundance across the full range of concentrations analysed, indicative of a copy number difference between the bloom population and a culture used for assay calibration a priori. In contrast, the sxtA assay performed more consistently, indicating less copy number variation. The sxtA assay was generally reliable, fast and effective in quantifying A. catenella and was predictive of PST contamination of shellfish.

Published
2019

32. The genetic basis of toxin biosynthesis in dinoflagellates

Author

Verma, A, Barua, A, Ruvindy, R, Savela, H, Ajani, PA, Murray, SA, Verma, A, Barua, A, Ruvindy, R, Savela, H, Ajani, PA, and Murray, SA

Abstract

© 2019 by the authors. Licensee MDPI, Basel, Switzerland. In marine ecosystems, dinoflagellates can become highly abundant and even dominant at times, despite their comparatively slow growth rates. One factor that may play a role in their ecological success is the production of complex secondary metabolite compounds that can have anti-predator, allelopathic, or other toxic effects on marine organisms, and also cause seafood poisoning in humans. Our knowledge about the genes involved in toxin biosynthesis in dinoflagellates is currently limited due to the complex genomic features of these organisms. Most recently, the sequencing of dinoflagellate transcriptomes has provided us with valuable insights into the biosynthesis of polyketide and alkaloid-based toxin molecules in dinoflagellate species. This review synthesizes the recent progress that has been made in understanding the evolution, biosynthetic pathways, and gene regulation in dinoflagellates with the aid of transcriptomic and other molecular genetic tools, and provides a pathway for future studies of dinoflagellates in this exciting omics era.

Published
2019

33. Bloom drivers of the potentially harmful dinoflagellate Prorocentrum minimum (Pavillard) Schiller in a south eastern temperate Australian estuary

Author

Ajani, PA, Larsson, ME, Woodcock, S, Rubio, A, Farrell, H, Brett, S, Murray, SA, Ajani, PA, Larsson, ME, Woodcock, S, Rubio, A, Farrell, H, Brett, S, and Murray, SA

Abstract

© 2018 Elsevier Ltd Harmful algal blooms are an increasing concern in the estuarine reaches of the Hawkesbury-Nepean River, one of the largest coastal rivers systems in south eastern Australia. In the austral spring of 2016, an unprecedented bloom of the harmful mixotrophic dinoflagellate Prorocentrum minimum occurred in Berowra Creek (maximum cell abundance 1.9E+06 cells L−1, 89% of the total phytoplankton community), a major tributary of this river system. In response to this bloom, our study utilises an estuary-wide, thirteen-year time series of phytoplankton abundance and environmental data to examine the spatial and temporal patterns of this harmful alga and its potential bloom drivers in this system. P. minimum cell densities and environmental parameters varied over large spatial scales, with sites located in the main channel of the estuary significantly differing from those in the more urbanized tributary of Berowra Creek. Generalised additive modelling outputs suggested that blooms of P. minimum are complex, but generally corresponded to a spatial gradient of eutrophication and salinity, whereby P. minimum growth and concomitant high chlorophyll-a concentrations were enhanced at sites that were generally less saline and more eutrophic than others. Furthermore, temporal patterns suggested that blooms occurred abruptly and lasted up to three weeks, most often during the austral autumn to spring. While significant correlations were observed between rainfall and nutrients at all other sites, suggesting a pathway for nutrient availability, the association between rainfall and nutrient delivery was generally not observed in Berowra Creek (a 15-m deep site) suggesting that a continual supply of nutrients, coupled with unique bathymetry and water residence time at this site, are the most likely contributing factors to phytoplankton growth. This study presents the most comprehensive examination of P. minimum in any southern hemisphere estuary to date and highlights t

Published
2018

34. Toxicology of gambierdiscus spp. (dinophyceae) from tropical and temperate Australian waters

Author

Larsson, ME, Laczka, OF, Tim Harwood, D, Lewis, RJ, Himaya, SWA, Murray, SA, Doblin, MA, Larsson, ME, Laczka, OF, Tim Harwood, D, Lewis, RJ, Himaya, SWA, Murray, SA, and Doblin, MA

Abstract

© 2018 by the authors. Ciguatera Fish Poisoning (CFP) is a human illness caused by the consumption of marine fish contaminated with ciguatoxins (CTX) and possibly maitotoxins (MTX), produced by species from the benthic dinoflagellate genus Gambierdiscus. Here, we describe the identity and toxicology of Gambierdiscus spp. isolated from the tropical and temperate waters of eastern Australia. Based on newly cultured strains, we found that four Gambierdiscus species were present at the tropical location, including G. carpenteri, G. lapillus and two others which were not genetically identical to other currently described species within the genus, and may represent new species. Only G. carpenteri was identified from the temperate location. Using LC-MS/MS analysis we did not find any characterized microalgal CTXs (P-CTX-3B, P-CTX-3C, P-CTX-4A and P-CTX-4B) or MTX-1; however, putative maitotoxin-3 (MTX-3) was detected in all species except for the temperate population of G. carpenteri. Using the Ca2+ influx SH-SY5Y cell Fluorescent Imaging Plate Reader (FLIPR) bioassay we found CTX-like activity in extracts of the unidentified Gambierdiscus strains and trace level activity in strains of G. lapillus. While no detectable CTX-like activity was observed in tropical or temperate strains of G. carpenteri, all species showed strong maitotoxin-like activity. This study, which represents the most comprehensive analyses of the toxicology of Gambierdiscus strains isolated from Australia to date, suggests that CFP in this region may be caused by currently undescribed ciguatoxins and maitotoxins.

Published
2018

35. The microbiome of the cosmopolitan diatom leptocylindrus reveals significant spatial and temporal variability

Author

Ajani, PA, Kahlke, T, Siboni, N, Carney, R, Murray, SA, Seymour, JR, Ajani, PA, Kahlke, T, Siboni, N, Carney, R, Murray, SA, and Seymour, JR

Abstract

Copyright © 2018 Ajani, Kahlke, Siboni, Carney, Murray and Seymour. The ecological interactions between phytoplankton and marine bacteria have important implications for the productivity and biogeochemistry of ocean ecosystems. In this study we characterized the microbial assemblages associated with multiple isolates of the ecologically important diatom Leptocylindrus using amplicon sequencing of the 16S rRNA gene, to examine levels of conservation of the microbiome across closely related species or strains. We also assessed if the microbiome structure of a given diatom strain was dependent on the location from which it was isolated and if the microbiome of cultured isolates significantly changed overtime from the seawater in which they were isolated. The bacterial assemblages from 36 strains belonging to three species (Leptocylindrus danicus, Leptocylindrus convexus, and Leptocylindrus aporus) isolated from six locations spanning > 1000 km of south east Australian coastline over 1 year, were dominated by the Rhodobacteraceae (∼60%) and the Flavobacteriaceae (∼10%). Across all strains, only one 'core OTU' (Roseovarius sp.) was identified across all samples. We observed no significant differences in bacterial community composition between diatom species. Significant differences in microbiome structure were, however, observed between diatom strains collected at different sampling times and from differing locations, albeit these two factors were coupled. Moreover, while bacterial communities under domestication varied from the seawater in which they were isolated, they remained specific to the location/month of origin, i.e., different regions and time points harbored distinct bacterial communities. Our study delivers new knowledge in relation to diatom-bacterial associations, revealing that the location/time from which a diatom is isolated plays an important role in shaping its microbiome.

Published
2018

36. A new diatom species P. Hallegraeffii sp. Nov. Belonging to the toxic genus Pseudo-nitzschia (Bacillariophyceae) from the East Australian Current

Author

Ajani, PA, Verma, A, Lassudrie, M, Doblin, MA, Murray, SA, Ajani, PA, Verma, A, Lassudrie, M, Doblin, MA, and Murray, SA

Abstract

© 2018 Ajani et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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.

Published
2018

39. A new species of Gambierdiscus (Dinophyceae) from the south-west Pacific: Gambierdiscus honu sp. nov.

Author

Rhodes, L, Smith, KF, Verma, A, Curley, BG, Harwood, DT, Murray, S, Kohli, GS, Solomona, D, Rongo, T, Munday, R, Murray, SA, Rhodes, L, Smith, KF, Verma, A, Curley, BG, Harwood, DT, Murray, S, Kohli, GS, Solomona, D, Rongo, T, Munday, R, and Murray, SA

Abstract

© 2017 Elsevier B.V. Two isolates of a new tropical, epiphytic dinoflagellate species, Gambierdiscus honu sp. nov., were obtained from macroalgae sampled in Rarotonga, Cook Islands, and from North Meyer Island, Kermadec Islands. Gambierdiscus honu sp. nov. had the common Gambierdiscus Kofoidian plate formula: Po, 3′, 6″, 6C?, 6 or 7S, 5‴, 1p and 2⁗. The characteristic morphological features of this species were its relatively small short dorsoventral length and width and the shape of individual plates, in particular the combination of the hatchet-shaped 2′ and pentagonal 3′ plates and the length to width ratio of the antapical 1p plate. The combination of these characteristics plus the smooth thecal surface and equal sized 1⁗ and 2⁗ plates differentiated this species from other Gambierdiscus species. The phylogenetic analyses supported the unique description. Both isolates of G. honu produced the putative maitotoxin (MTX)-3 analogue, but neither produced ciguatoxin (CTX) or MTX. Extracts of G. honu were shown to be highly toxic to mice by intraperitoneal injection (0.2 mg/kg), although less toxic by gavage. It is possible that toxins other than putative MTX-3 are produced.

Published
2017

40. Role of Modular Polyketide Synthases in the Production of Polyether Ladder Compounds in Ciguatoxin-Producing Gambierdiscus polynesiensis and G. excentricus (Dinophyceae)

Author

Kohli, GS, Campbell, K, John, U, Smith, KF, Fraga, S, Rhodes, LL, Murray, SA, Kohli, GS, Campbell, K, John, U, Smith, KF, Fraga, S, Rhodes, LL, and Murray, SA

Abstract

© 2017 The Author(s) Journal of Eukaryotic Microbiology © 2017 International Society of Protistologists Gambierdiscus, a benthic dinoflagellate, produces ciguatoxins that cause the human illness Ciguatera. Ciguatoxins are polyether ladder compounds that have a polyketide origin, indicating that polyketide synthases (PKS) are involved in their production. We sequenced transcriptomes of Gambierdiscus excentricus and Gambierdiscus polynesiensis and found 264 contigs encoding single domain ketoacyl synthases (KS; G. excentricus: 106, G. polynesiensis: 143) and ketoreductases (KR; G. excentricus: 7, G. polynesiensis: 8) with sequence similarity to type I PKSs, as reported in other dinoflagellates. In addition, 24 contigs (G. excentricus: 3, G. polynesiensis: 21) encoding multiple PKS domains (forming typical type I PKSs modules) were found. The proposed structure produced by one of these megasynthases resembles a partial carbon backbone of a polyether ladder compound. Seventeen contigs encoding single domain KS, KR, s-malonyltransacylase, dehydratase and enoyl reductase with sequence similarity to type II fatty acid synthases (FAS) in plants were found. Type I PKS and type II FAS genes were distinguished based on the arrangement of domains on the contigs and their sequence similarity and phylogenetic clustering with known PKS/FAS genes in other organisms. This differentiation of PKS and FAS pathways in Gambierdiscus is important, as it will facilitate approaches to investigating toxin biosynthesis pathways in dinoflagellates.

Published
2017

42. Unprecedented Alexandrium blooms in a previously low biotoxin risk area of Tasmania, Australia.

Author

Proenca, LA, Hallegraeff, G, Bolch, C, Condie, S, Dorantes-Aranda, J, Murray, SA, Quinlan, R, Ruvindy, R, Turnbull, A, Ugalde, S, Wilson, K, Proenca, LA, Hallegraeff, G, Bolch, C, Condie, S, Dorantes-Aranda, J, Murray, SA, Quinlan, R, Ruvindy, R, Turnbull, A, Ugalde, S, and Wilson, K

Abstract

During October 2012, a shipment of blue mussels (Mytilus galloprovincialis) from the poorly monitored east coast of Tasmania, Australia, was tested by Japanese import authorities and found to be contaminated with unacceptable levels of Paralytic Shellfish Toxins (PSTs; 10 mg/kg). Subsequently local oysters, scallops, clams, the viscera of abalone and rock lobsters were also found to be contaminated. This led to a global product recall and loss to the local economy of AUD 23M. Following low toxicity during 2013 and 2014 and implementation of minimal shellfish farm closures, a more severe bloom event occurred during July-November 2015 and again June-September 2016 (up to 300,000 Alexandrium cells/L; 24 mg/kg PST in mussels, 6 mg/kg in Crassostrea gigas oysters), also causing 4 human illnesses resulting in hospitalization after consumption of wild shellfish. While Alexandrium tamarense had been detected in low concentrations in southeastern Australia since 1987, all cultured strains belonged to the mostly non-toxic group 5 (now designated A. australiense; detected since 1987) and weakly toxic group 4 (A. pacificum; detected in 1997). In contrast, the 2012 to 2016 outbreaks were dominated by highly toxic group 1 (A. fundyense) never detected previously in the Australian region. Molecular analyses suggest that A. fundyense may have been a cryptic ribotype previously present in Tasmania, but newly stimulated by altered water column stratification conditions driven by changing rainfall and temperature patterns. Increased seafood and plankton monitoring of the area now include the implementation of Alexandrium qPCR, routine Neogen™ immunological and HPLC PST tests, but ultimately may also drive change in harvesting strategies and aquaculture species selection by the local seafood industry.

Published
2017

43. Both modular and single-domain Type I polyketide synthases are expressed in the brevetoxin-producing dinoflagellate, Karenia brevis (Dinophyceae)

Author

Van Dolah, FM, Kohli, GS, Morey, JS, Murray, SA, Van Dolah, FM, Kohli, GS, Morey, JS, and Murray, SA

Abstract

© 2017 The Authors Journal of Phycology published by Wiley Periodicals, Inc. on behalf of Phycological Society of America Dinoflagellates are prolific producers of polyketide compounds, many of which are potent toxins with adverse impacts on human and marine animal health. To identify polyketide synthase (PKS) genes in the brevetoxin-producing dinoflagellate, Karenia brevis, we assembled a transcriptome from 595 million Illumina reads, sampled under different growth conditions. The assembly included 125,687 transcripts greater than 300 nt in length, with over half having >100× coverage. We found 121 transcripts encoding Type I ketosynthase (KS) domains, of which 99 encoded single KS domains, while 22 contained multiple KS domains arranged in 1–3 protein modules. Phylogenetic analysis placed all single domain and a majority of multidomain KSs within a monophyletic clade of protist PKSs. In contrast with the highly amplified single-domain KSs, only eight single-domain ketoreductase transcripts were found in the assembly, suggesting that they are more evolutionarily conserved. The multidomain PKSs were dominated by trans-acyltransferase architectures, which were recently shown to be prevalent in other algal protists. Karenia brevis also expressed several hybrid nonribosomal peptide synthetase (NRPS)/PKS sequences, including a burA-like sequence previously reported in a wide variety of dinoflagellates. This contrasts with a similarly deep transcriptome of Gambierdiscus polynesiensis, which lacked NRPS/PKS other than the burA-like transcript, and may reflect the presence of amide-containing polyketides in K. brevis and their absence from G. polynesiensis. In concert with other recent transcriptome analyses, this study provides evidence for both single domain and multidomain PKSs in the synthesis of polyketide compounds in dinoflagellates.

Published
2017

44. Management of ciguatoxin risk in eastern Australia

Author

Farrell, H, Murray, SA, Zammit, A, Edwards, AW, Farrell, H, Murray, SA, Zammit, A, and Edwards, AW

Abstract

© 2017 by the authors. Licensee MDPI, Basel, Switzerland. Between 2014 and 2016, five cases of ciguatera fish poisoning (CFP), involving twenty four individuals, were linked to Spanish Mackerel (Scomberomorus commerson) caught in the coastal waters of the state of New South Wales (NSW) on the east coast of Australia. Previously, documented cases of CFP in NSW were few, and primarily linked to fish imported from other regions. Since 2015, thirteen individuals were affected across four additional CFP cases in NSW, linked to fish imported from tropical locations. The apparent increase in CFP in NSW from locally sourced catch, combined with the risk of CFP from imported fish, has highlighted several considerations that should be incorporated into risk management strategies to minimize CFP exposure for seafood consumers.

Published
2017

45. Assessment of the metabarcoding approach for community analysis of benthic-epiphytic dinoflagellates using mock communities

Author

Smith, KF, Kohli, GS, Murray, SA, Rhodes, LL, Smith, KF, Kohli, GS, Murray, SA, and Rhodes, LL

Abstract

© 2017 The Royal Society of New Zealand. In this study, we assessed the use of DNA metabarcoding as a method for biodiversity assessment of benthic-epiphytic dinoflagellate communities and for detecting rare, toxin-producing taxa. Mock communities and three primer pairs were used to establish the recovery of species signal and quantitative representation of species in the samples, as well as to determine primer biases, bioinformatic filtering steps, and threshold levels. Samples were analysed using high-throughput sequencing Illumina™ MiSeq technology. We did not find a relationship between read number and cell abundance for all treatments. However, the method was extremely sensitive, with two of the primer pairs detecting a single cell representing less than 0.001% of the cells in the sample. Benthic and epiphytic dinoflagellate communities were also collected from the Bay of Islands (Northland, New Zealand). Dinophyceae species richness was much higher when samples were analysed using metabarcoding than when analysed by microscopy, and we detected several new taxonomic records for New Zealand.

Published
2017

46. Comparative performance of four immunological test kits for the detection of Paralytic Shellfish Toxins in Tasmanian shellfish

Author

Dorantes-Aranda, JJ, Campbell, K, Bradbury, A, Elliott, CT, Harwood, DT, Murray, SA, Ugalde, SC, Wilson, K, Burgoyne, M, Hallegraeff, GM, Dorantes-Aranda, JJ, Campbell, K, Bradbury, A, Elliott, CT, Harwood, DT, Murray, SA, Ugalde, SC, Wilson, K, Burgoyne, M, and Hallegraeff, GM

Abstract

© 2016 Elsevier Ltd Blooms of the toxic dinoflagellate Alexandrium tamarense (Group 1) seriously impacted the Tasmanian shellfish industry during 2012 and 2015, necessitating product recalls and intensive paralytic shellfish toxin (PST) product testing. The performance of four commercial PST test kits, Abraxis™, Europroxima™, Scotia™ and Neogen™, was compared with the official AOAC LC-FLD method for contaminated mussels and oysters. Abraxis and Europroxima kits underestimated PST in 35–100% of samples when using standard protocols but quantification improved when concentrated extracts were further diluted (underestimation ≤18%). The Scotia kit (cut off 0.2–0.7 mg STX-diHCl eq/kg) delivered 0% false negatives, but 27% false positives. Neogen produced 5% false negatives and 13% false positives when the cut off was altered to 0.5–0.6 mg STX-diHCl eq/kg, the introduction of a conversion step eliminated false negatives. Based on their sensitivity, ease of use and performance, the Neogen kit proved the most suitable kit for use with Tasmanian mussels and oysters. Once formally validated for regulatory purposes, the Neogen kit could provide shellfish growers with a rapid tool for harvesting decisions at the farm gate. Effective rapid screening preventing compliant samples undergoing testing using the more expensive and time consuming LC-FLD method will result in significant savings in analytical costs.

Published
2017

48. Diversity, temporal distribution and physiology of the centric diatom Leptocylindrus Cleve (Bacillariophyta) from a southern hemisphere upwelling system

Author

Ajani, PA, Armbrecht, LH, Kersten, O, Kohli, GS, and Murray, SA

Subjects
fungi, Marine Biology & Hydrobiology
Abstract

© 2016 The International Society for Diatom Research. The marine diatom Leptocylindrus is a major component of phytoplankton blooms in coastal ecosystems and upwelling regions worldwide, however, little is known about this genus in the southern hemisphere. Whilst Leptocylindrus danicus has been reported from south-eastern (SE) Australia since the 1930s, there has been neither unequivocal species identification nor focused examination of the temporal abundance of Leptocylindrus in this region. Such investigations are crucial in the context of climate change and the strengthening of the East Australian Current, which is expected to result in alterations to the seasonal abundance and distribution of Leptocylindrus along the east Australian coast. Thus we also describe the temporal distribution of Leptocylindrus based on 50 years of records, revealing that this diatom is a key component of the seasonal phytoplankton cycle, with greatest abundance in the austral spring and summer. Using light and transmission electron microscopy and molecular phylogenetics based on the nuclear-encoded ITS1–5.8S–ITS2 rDNA region, our study unambiguously revealed three species, L. danicus, Leptocylindrus convexus and Leptocylindrus aporus from 34 clonal isolates from SE Australia, with the majority (82%) of strains identified as L. danicus. Furthermore, we investigated the growth, auxospore and resting spore formation of the most commonly occurring species, L. danicus, under four temperature and irradiance scenarios. The diatom reached maximum growth rates (µMax, 1.71 divisions day−1) under relatively high temperatures (25°C) and light conditions (100 µmol photons m−2 s−1) between days 2 and 7 of the experiment. When temperature and light regimes were reduced (18°C, 50 µmol photons m−2 s−1) auxospores and resting spores were formed. The rapid growth rate and potential of L. danicus to form auxospores are important survival mechanisms in coastal upwelling systems and likely to result in the continued success of this species in Eastern Australia. The ecological, physiological and evolutionary response of this significant diatom group to further ocean warming should be the focus of future investigations.

Published
2016

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