Of them, 207 and 134 transcripts were assigned to the antioxidant enzyme system and heat shock protein . Dong L., Meng Y., Sui Z., Wang J., Wu L., Fu B. Considering these cases, we assigned samples to two additional groups based on the estimated cell numbers obtained by the dPCR assay, i.e., shellfish harvesting at the station is continuously regulated despite a lack of detectable PST (Group C) or shellfish harvesting is permitted (Group D). Would you like email updates of new search results? Fair usage of data in scientific publications is permitted. Unlike the qPCR, a digital PCR (dPCR) is insensitive to potential PCR inhibitors and enables the absolute quantification of a target gene by counting the number of amplified DNA fragments in reaction samples using fluorescent signals; cell abundance in field samples can be estimated by chip-based dPCRs without standard curves as references [35,36]. ML bootstrap values (>60%) are indicated at the nodes and Dinophysis species were used as outgroups; (c) conventional PCR using ITS-specific primers for A. pacificum and A. catenella, as well as universal dinoflagellate primers for the SSU and LSU regions of rDNA, and three template samples: A. pacificum Masan strain from culture (lane 1) or GF/C glass microfiber filters (lane 3), and A. minutum as a negative control (lane 2). (2014). In particular, A. catenella, A. minutum, and A. pacificum, which produce the PST saxitoxin (STX), and are distributed worldwide, are considered to be the primary causative species driving PSP outbreaks [1,7]. To establish a more precise cell threshold value for the molecular detection assay, further studies are required. 2014 Jun 17;111(24):9009-14. doi: 10.1073/pnas.1402130111. At JH and GS, the peak abundance of cells was 5488 (JH, 15 April) and 23,047 (GS, 15 April) cells L1 (Figure 4). Weekers P., Gast R.J., Fuerst P.A., Byers T.J. Sequence variations in small-subunit ribosomal RNAs of Hartmannella vermiformis and their phylogenetic implications. Baek S.H., Choi J.M., Lee M., Park B.S., Zhang Y., Arakawa O., Takatani T., Jeon J.-K., Kim Y.O. Scientists in New Zealand discovered a way to produce the world's first algae-based pain medication. Ben Gharbia H, Kfi-Daly Yahia O, Cecchi P, Masseret E, Amzil Z, Herve F, Rovillon G, Nouri H, M'Rabet C, Couet D, Zmerli Triki H, Laabir M. PLoS One. Error bars represent standard deviations. Putative sxtA4 genomic sequences of the Masan strain of A. pacificum were amplified by PCR using previously reported primers [38], and verified by Sanger sequencing and BLASTx. Chip-based digital PCR (dPCR) assay using a species-specific primer and probe set for A. pacificum targeting the sxtA4 gene. Maximum germination success (85%) for isolated cysts occurred at 15 C, and the germling cells were A. pacificum (Group IV). We collected 76 seawater samples from four locations: Jinhae (JH), Gusan (GS), Makpo (MP), and Neungpo (NP), in March and April from 2018 to 2021. Generating an ePub file may take a long time, please be patient. Orr, R.J.S., Stken, A., Rundberget, T., Eikrem, W., Jakobsen, K.S. The Masan strain of A. pacificum was taxonomically identified based on a comparative sequence analysis of the large subunit (LSU) and internal transcribed spacer (ITS) regions of rDNA with that of other Alexandrium species deposited in GenBank. Oceanogr. Jun-Ho Hyung, Jinik Hwang, [], and Jaeyeon Park. No correlation due to the specific hydrodynamics of the lagoon was observed in the spatial distribution of A. pacificum cysts and vegetative cells, and a significant difference in the cyst distribution pattern was recorded among the lagoons' different zones. Deep Sea Res. Alexandrium pacificum, one of the most well studied and widely distributed dinoflagellates, is known to form toxin blooms and cause paralytic shellfish poisoning (PSP). No template control (NTC) showed only blue background. Scholin C.A., Herzog M., Sogin M., Anderson D.M. Hi, fellow LinkedIn members. doi: 10.1371/journal.pone.0187963. Penna, A; Garcs, E; Vila, M; Giacobbe, MG; Fraga, S; Lugli, A; Bravo, I; Bertozzini, E; Vernesi, C. (2005). 2012 Jun;335(6):406-16. doi: 10.1016/j.crvi.2012.04.007. Unable to load your collection due to an error, Unable to load your delegates due to an error. 833012 Sequence Analysis of a Fragment of the Lsu rrna Gene 1. Alexandrium minutum and Alexandrium pacificum are representatives of the dinoflagellate genus that regularly proliferate on the French coasts and other global coastlines. Conceptualization, experiment, data curation, validation, writingoriginal draft preparation, J.-H.H. Scientists at the institute have developed a reliable and commercially-scalable process for producing neosaxitoxin , a shellfish toxin found in New Zealand marine microalgae, Alexandrium pacificum . The PST content was plotted against cell counts, as determined by the chip-based dPCR assay, at the four locations and the linear regression calculated using GraphPad Prism. The ratios of the detected PST content to the cell abundance in Jinhae and Neungpo (offshore) were higher than those for Gusan and Makpo (inshore) (Figure 8b). The qPCR is widely used to estimate cell abundance and to monitor harmful algal species [27,28,29,30,31]. Galluzzi L., Penna A., Bertozzini E., Vila M., Garcs E., Magnani M. Development of a real-time PCR assay for rapid detection and quantification of Alexandrium minutum (a dinoflagellate). Front Microbiol. I am a chemistry enthusiast When the total PST content of shellfish tissue is higher than the regulatory threshold (80 g 100 g1 shellfish tissue), shellfish farms are not permitted to harvest any shellfish [13]. Doucette G.J., Logan M.M., Ramsdell J.S., Van Dolah F.M. Optimization of the regulatory threshold for A. pacificum to determine whether shellfish harvesting should be permitted based on chip-based digital PCR (dPCR) targeting sxtA4. World-wide electronic publication, National University of Ireland, Galway. searched on YYYY-MM-DD. Borcier E, Morvezen R, Boudry P, Miner P, Charrier G, Laroche J, Hegaret H. Aquat Toxicol. The sensitivity of the dPCR assay was higher than that of microscopy and sxtA4-based qPCRs. PMC and transmitted securely. New insights on the species-specific allelopathic interactions between macrophytes and marine HAB dinoflagellates. Based on the sxtA4 fragments in the cells, we designed a sxtA4-specific primer and probe set, with an optimized annealing temperature of 62 C, for A. pacificum to be used in a chip-based dPCR assay. ; field investigation, formal analysis, culture collection, strain setup, and culturing, E.-J.K. and S.-J.M. Morphological characterization of Alexandrium cells at the genus level was performed using an Olympus BX 53 microscope and images were obtained using a DP73 digital camera system (Olympus, Tokyo, Japan). Cells were harvested by centrifugation at 18,396 g for 3 min, and the pellets were immediately stored at 80 C until gDNA extraction. Intraspecific variability in membrane proteome, cell growth, and morphometry of the invasive marine neurotoxic dinoflagellate Alexandrium pacificum grown in metal-contaminated conditions. In 2016, the bloom persisted for at least eight weeks and maximum cell concentrations of 89,000 cells L&minus;1 of A. pacificum were reported. However, some . The AT of the designed primers was optimized by gradient dPCR. For molecular characterization, cells of the Masan strain of Alexandrium were taxonomically identified by LSU rDNA phylogenetic analysis. After sufficient growth, the dense culture was transferred to 50, 250, and 500 mL polycarbonate (PC) bottles. [12] and Baek et al. The elucidation of allelopathic mechanism of A. pacificum is of great significance for understanding A. pacificum blooms. Alexandrium pacificum is a dinoflagellate species notorious for its rapid growth resulting in large-scale blooms. Shin H.H., Li Z., Kim E.S., Park J.-W., Lim W.A. This site needs JavaScript to work properly. Penna A., Perini F., DellAversano C., Capellacci S., Tartaglione L., Giacobbe M.G., Casabianca S., Fraga S., Ciminiello P., Scardi M. The sxt gene and paralytic shellfish poisoning toxins as markers for the monitoring of toxic Alexandrium species blooms. A standard additions method reduces inhibitor-induced bias in quantitative real-time PCR. The production of PST by G. catenatum in Korea has been reported since 1996 [41], and the toxicity of isolates of G. catenatum collected from the southern coastal waters in Korea has been investigated by Park et al. After monitoring neurotoxic shellfish poisoning symptoms and mouse survival times, the PST content in the sample was calculated on the basis of Sommers table. Alexandrium minutum and Alexandrium pacificum are representatives of the dinoflagellate genus that regularly proliferate on the French coasts and other global coastlines. Han M.-S., Jeon J.-K., Kim Y.-O. Accessibility A quantitative real-time PCR assay for the identification and enumeration of Alexandrium cysts in marine sediments. It was isolated from the South Sea of Korea and cultured using f/2 medium without silicate (Guillard 1975 ). Disclaimer, National Library of Medicine Examination of the seasonal dynamics of the toxic dinoflagellate Alexandrium catenella at Redondo Beach, California, by quantitative PCR. A comparison with shellfish PST concentrations indicated that cell concentrations above 500 cells L1, as measured using the dPCR assay, may cause shellfish PST concentrations to exceed the allowed limits for PSTs. Alexandrium as a whole is wide spread across the globe and has been identified in different parts of the Northern and Southern Hemisphere. Anderson D.M., Alpermann T.J., Cembella A.D., Collos Y., Masseret E., Montresor M. The globally distributed genus Alexandrium: Multifaceted roles in marine ecosystems and impacts on human health. We found reciprocal effects between A. pacificum and T. pseudonana, and corroborated A. pacificum's allelopathy on T. pseudonana by observing inhibitory effects of filtrate from A. pacificum culture on the growth of T. pseudonana. Before Careers. However, this method is limited by potential ethical issues, high annual costs, low accuracy, and the time-consuming nature of the process; accordingly, attempts have been made to develop alternative methods [14,15,16,17,18,19]. LSU rDNA sequences of the isolated cells were amplified by conventional PCR using universal primers, and compared with the LSU sequences of 42 relatives in the A. tamarense species complex, three strains of A. minutum, and two strains of Dinophysis acuminata used as outgroups, obtained from GenBank. PST content at MP, measured between March and April, was <80 g 100 g1 of shellfish tissue (except for 583 g 100 g1 of tissue on 19 April), similar to the occurrence of cell abundance at MP, which peaked at 255 cells L1. See further details here . Epub 2017 Sep 19. Clipboard, Search History, and several other advanced features are temporarily unavailable. When. (a) Schematic diagram of the experimental workflow for chip-based dPCR; (b) specificity test for the designed primer and probe set using the dPCR assay in the presence of A. pacificum, A. catenella, A. minutum, and G. catenatum with a known number of cells (i.e., 30,000 cells). 2022 Oct;183:114058. doi: 10.1016/j.marpolbul.2022.114058. In 31 March and 15 April, 2021, the distribution of samples with >538 cells L1 at the stations corresponded to the red area, indicating the regulation of harvesting based on the PST threshold, except for Neungpo (31 March). 1Environment and Resource Convergence Center, Advanced Institute of Convergence Technology, Suwon 16229, Korea; rk.ca.uns@0211hjh (J.-H.H. Bouchouicha Smida D, Sahraoui I, Mabrouk HH, Sakka Hlaili A. #algae #innovation The portal can access those files and use them to remember the user's data, such as their chosen settings (screen view, interface language, etc. In contrast, the distribution of A. pacificum with concentrations of > 538 cells L1 in Jinhae-Masan Bay in March and April 2020 was not consistent with the regulated area based on PSP monitoring reports at the time, except for Makpo (18 March) and Jinhae and Gusan (6 April). The marine dinoflagellate Alexandriumproduces paralytic shellfish poisoning (PSP) toxins in temperate coastal waters worldwide. Benthic HABs . Mendoza-Flores A., Leyva-Valencia I., Band-Schmidt C.J., Galindo-Snchez C.E., Bustillos-Guzmn J.J. Metabolomics and proteomics reveal impacts of chemically mediated competition on marine plankton. (a) Cell abundance estimated by the dPCR assay was evaluated in field samples collected in March and April from 2018 to 2021, and divided into four groups based on PST monitoring reports: Group A (n = 24), PST content at the station was higher than the regulatory sanitary threshold for PSTs (80 g 100 g1 shellfish tissue); Group B (n = 58), PST content at the station was less than the PST threshold; Group C (n = 35), shellfish harvesting in the area was regulated despite no detectable PST on the particular date; Group D (n = 48), shellfish harvesting in the area was permitted. Epub 2017 Jan 20. Bethesda, MD 20894, Web Policies ; conceptualization, supervision, project administration, writingreview and editing, J.P. All authors have read and agreed to the published version of the manuscript. (a) Comparison between cell counts obtained by microscopy and cell estimates obtained by the dPCR assay. Alexandrium minutum and Alexandrium pacificum are representatives of the dinoflagellate genus that regularly proliferate on the French coasts and other global coastlines. Copyright notice: the information originating from AlgaeBase may not be downloaded or replicated by any means, without the written permission of the copyright owner (generally AlgaeBase). Alexandrium pacificum Litaker sp. [12] recently reviewed previous reports and reported that A. pacificum was one of the causative species of these events in the southern coastal waters of Korea, as well as A. catenella, based on resting cysts, the optimal temperature for growth, and phylogenetic analyses. 2017 Mar;184:142-154. doi: 10.1016/j.aquatox.2017.01.009. An official website of the United States government. Hosoi-Tanabe S., Sako Y. Species-Specific Detection and Quantification of Toxic Marine Dinoflagellates Alexandrium tamarense and A. catenella byReal-Time PCR Assay. Chetouhi C, Masseret E, Satta CT, Balliau T, Laabir M, Jean N. Sci Total Environ. I'm a Molecular Biologist, with passion for Bioinformatics. eCollection 2017. Karenia brevis allelopathy compromises the lipidome, membrane integrity, and photosynthesis of competitors. Scientists at the institute have developed a reliable and commercially-scalable process for producing neosaxitoxin , a shellfish toxin found in New Zealand marine microalgae, Alexandrium pacificum . 20190518) funded by the Ministry of Oceans and Fisheries, Korea, and supported by a grant from National Institute of Fisheries Science (NIFS) in Korea (R2022065). ), or their login data. We are experimenting with display styles that make it easier to read articles in PMC. Many of the most important trends are caused by "indirect effects." In just one example, a Bhat S., Herrmann J., Armishaw P., Corbisier P., Emslie K.R. These harmful species may threaten shellfish harvest and human health due to their ability to synthesize neurotoxic alkaloids of the saxitoxin group. Fonfra E.S., Vilario N., Campbell K., Elliott C., Haughey S.A., Ben-Gigirey B., Vieites J.M., Kawatsu K., Botana L.M. The genus Alexandrium is a particularly well-known group of bloom-forming marine dinoflagellates due to their impacts on human health. Yarimizu K., Sildever S., Hamamoto Y., Tazawa S., Oikawa H., Yamaguchi H., Basti L., Mardones J.I., Paredes-Mella J., Nagai S. Development of an absolute quantification method for ribosomal RNA gene copy numbers per eukaryotic single cell by digital PCR. (a) Time interval between measurements of cell abundance estimated by the dPCR assay and PST content in shellfish tissue quantified by a mouse bioassay. Briefly, A. pacificum cells were isolated and identified by their morphological and molecular characteristics and the number of sxtA4 gene copies per cell was quantified by a chip-based dPCR. PCR mixtures contained 10 L of qPCRBIO Probe Mix no-ROX (2) (PCR Biosystems, London, England), 1 L of 10 M primer pairs, 0.5 L of 10 M TaqMan probe, 4.5 L of UltraPure DNAse/RNAse-Free Distilled Water (Invitrogen), and 3 L of the template DNA in a final volume of 20 L. Additionally, sanitary regulation for shellfish harvesting is often continued, despite a PST content of < 80 g 100 g1 shellfish tissue as measured by MBA, owing to an excess of PST in the area within a few days or a week. The site is secure. KIM H.G., MATSUOKA K., LEE S.G., AN K.H. MeSH To establish the dPCR assay for the prediction of PST-producing blooms in Korea, a detection method for A. catenella and G. catenatum, targeting PST biosynthesis genes, should be developed. The direct counting of cells by microscopy yielded overestimates of cell number compared to the abundance values obtained by chip-based dPCRs because microscopic identification of Alexandrium species was restricted to the genus level (Figure 5a). Strain Alex05 (or LMBE-C4) of Alexandrium pacificum was obtained from the Marine Bio Resource Information System at the Korea Institute of Ocean Science and Technology (Jangmok, Korea). An advanced tool, droplet digital PCR (ddPCR), for absolute quantification of the red-tide dinoflagellate, Cochlodinium polykrikoides Margalef (Dinophyceae). Hi, fellow LinkedIn members. These findings were confirmed by a conventional PCR using species-specific ITS primers targeting A. pacificum and A. catenella, as well as universal dinoflagellate SSU and LSU rDNA primers as the positive control; A. minutum served as the negative template control. PSTs are toxic to humans and maximum legal limits in seafood have been implemented by regulatory authorities worldwide. Regarding recent advances in DMSP biosynthesis pathway, 3 dsyB homologs were found in A. minutum but no homolog of the diatom sequence TpMMT. Alexandrium Contrarily, Alexandrium has an apical pore complex at the apex of the cell that is so characteristic it is used to separate genera and even species within a genus. Algal toxins; DMSP; Dinoflagellate; Glycine betaine; PST; Paralytic shellfish toxins; Phytoplankton; Salinity; Saxitoxin; dsyB gene. Toxic algal bloom in Scandinavian waters, May-June 1988. I'm a Molecular Biologist, with passion for Bioinformatics. Using these field samples, chip-based dPCR results and PST content, obtained from PSP monitoring reports by the KNIFS, were compared. Saxitoxin and its more than 50 analogues are a group of naturally occurring neurotoxins collectively designated as paralytic shellfish toxins (PSTs). Single molecule detection in nanofluidic digital array enables accurate measurement of DNA copy number. Park T.G., Kim C.H., Oshima Y. Paralytic shellfish toxin profiles of different geographic populations of Gymnodinium catenatum (Dinophyceae) in Korean coastal waters. Many of the most important trends are caused by "indirect effects." In just one example, a (a) 1D scatter plots and position plots for serially diluted samples of 60,000, 30,000, 15,000, and 7500 cells, and the no template control (NTC) for the dPCR assay; (b) copy number concentration plot generated based on the total number of sxtA4 copies obtained for known A. pacificum cell numbers (r2 = 0.991 with p < 0.01). Alexandrium pacificum; Antioxidant system; Nutrient absorption; Photosynthesis; Thalassiosira pseudonana. Batch Culture of Alexandrium Strains. ; investigation, resources, writingreview and editing, funding, D.-W.K. The gDNA was extracted using the AccuPrep Genomic DNA Extraction Kit (BIONEER, Daejeon, Korea), following the manufacturers instructions. Accessibility (urn:lsid:marinespecies.org:taxname:833012). In Jinhae-Masan Bay, the spatial distribution of A. pacificum at concentrations above and below 538 cells L1 was compared with regulated areas for shellfish harvesting based on the PST threshold reported by the KNIFS. Actus Mer/Sea News: First insight into H3K4me3 modification in the rapid growth of Alexandrium pacificum (dinoflagellates) - Frontiers in Marine Science: Background: Alexandrium pacificum is a . The cell abundance estimated by chip-based dPCR targeting sxtA4 in field samples collected from Jinhae-Masan Bay in March and April from 2018 to 2021 was compared to the total PST content in shellfish tissues collected from the same stations and time points (presented in PSP monitoring reports by the KNIFS). We then compared the spatial distribution of A. pacificum with the regulated area for shellfish harvesting based on the PST threshold at the time reported by the KNIFS. Recently, Lee et al. Shin H.H., Li Z., Kim H.J., Park B.S., Lee J., Shin A.-Y., Park T.-G., Lee K.-W., Han K.H., Youn J.Y. Occurrence of dinoflagellate Alexandrium tamarense, a causative organism of paralytic shellfish poisoning in Chinhae Bay, Korea. A qPCR assay was performed to estimate the abundance of A. pacificum with specific primers for sxat4 using a PCRmax Eco 48 Real-time PCR System (Staffordshire, UK). Change in paralytic shellfish toxins in the mussel Mytilus galloprovincialis depending on dynamics of harmful Alexandrium catenella (Group I) in the Geoje coast (South Korea) during bloom season. [43], A. catenella, one of the causative species of PSP outbreaks in Korean coastal waters, showed high germination success and toxicity below 15 C, as well as a favorable growth of the cells at 1015 C, suggesting that early spring blooms in Jinhae-Masan Bay could be derived from A. catenella. Harmful Algae. 10:676688. Dundas I., Johannessen O., Berge G., Heimdal B. Tang WJ, Lin ZR, Zhang QC, Geng HX, Sun HX, Tang XX, Yu RC. 10: Qiu J, Meng F, Ding L, Che Y, McCarron P, Beach DG, Li A. Dynamics of paralytic shellfish toxins and their metabolites during timecourse exposure of scallops Chlamys farreri and mussels Mytilus galloprovincialis to Alexandrium pacificum. We detected 2.0 0.24 (n = 20) sxtA4 copies per A. pacificum isolated from Masan in May 2017 by a chip-based dPCR using newly designed primers. The measured cell numbers for each method were transformed to the log10 scale. In this study, we developed a chip-based digital PCR (dPCR) method for A. pacificum detection and tested it for monitoring in Jinhae-Masan Bay. Both the 1D scatter and position plot containing the no template control (NTC) showed only blue background signals. Blue line plots indicated cell concentrations of A. pacificum estimated by dPCR, and the PST content was presented in bar plots. Would you like email updates of new search results? The identity of A. pacificum . Considering that a monitoring system for PST-producing blooms is required for early detection and control, a chip-based dPCR targeting sxtA4 with a high sensitivity for concentrations of < 1000 cells L1 is expected to be effective in predicting the species distribution and preventing the downstream effects of blooms. Received 2021 Dec 15; Accepted 2022 Jan 29. Quantification of the paralytic shellfish poisoning dinoflagellate Alexandrium species using a digital PCR. official website and that any information you provide is encrypted To obtain environmental DNA from the field samples, 100200 mL of seawater at each location was filtered through a GF/C glass microfiber filter and stored at 80 C until DNA extraction. Murray S.A., Ruvindy R., Kohli G.S., Anderson D.M., Brosnahan M.L. name Alexandrium pacificum R.W.Litaker, 2014 Environment marine Original description John, U., Litaker, R.W, Montresor, M., Murray, S., Brosnahan,M.L., Anderson, D.M. We also found that co-culturing with A. pacificum, the expression of T. pseudonana genes related to photosynthesis, oxidative phosphorylation, antioxidant system, nutrient absorption and energy metabolism were drastically influenced. The T value was used to calculate the number of gene copies per cell using Equation (2), where Vr is the reaction volume, Vs is the original sample volume, Vt is the volume of template DNA, and c is the known number of cells [36]. Harada T., Oshima Y., Yasumoto T. Structures of two paralytic shellfish toxins, gonyautoxins V and VI, isolated from a tropical dinoflagellate, Pyrodinium bahamense var. Path analysis is an incredibly useful tool for #agricultural #data. PCR products were purified using the AccuPrep PCR Purification Kit (BIONEER) and confirmed by Sanger sequencing and alignment using BLAST from the National Center for Biotechnology Information (NCBI). Coupled with the alterations in Fv/Fm (the variable/maximum fluorescence ratio), activity of superoxide dismutase, contents of malondialdehyde, neutral lipid and total protein in T. pseudonana co-cultured with A. pacificum, we propose that A. pacificum allelopathy could reduce the efficiency of photosynthesis and energy metabolism of T. pseudonana and caused the oxidative stress, while the nutrient absorption was also affected by allelopathic effects. Which species, Alexandrium catenella (Group I) or A. pacificum (Group IV), is really responsible for past paralytic shellfish poisoning outbreaks in Jinhae-Masan Bay, Korea? Part II: Top. The genus can be found in oligotrophic, mesotrophic, and eutrophic coastlines. I'm Sharon, from Tel Aviv, Israel. The four catalytic domains of the sxtA gene form two isoforms (corresponding to the domains sxtA1sxtA4 and sxtA1sxtA3) in A. fundyense [38]. 8600 Rockville Pike In Jinhae-Masan Bay, Korea, PST-producing blooms are mostly caused by A. pacificum. A high spatial resolution sampling of Alexandrium pacificum cysts, along with sediment characteristics (% H2O, % organic matter (OM . The temporal patterns of A. pacificum occurrence detected by the dPCR assay during the period were highly similar to patterns of PST content at all four stations. Alexandrium pacificum, which produces the paralytic shellfish toxin (PST) saxitoxin (STX), is one of the causative species of paralytic shellfish poisoning outbreaks in coastal areas of Korea. Appendix S1 as Supplementary Material):Alexandrium ostenfeldii,Protogonyaulax catenella (=Alexandrium pacificum) or Ostreopsis ovata Mediterranean populations are genetically Responsible editor: Philippe Garrigues * Fernando Gmez fernando.gomez@fitoplancton.com 1 Carmen Campos Panisse 3, E-11500 Puerto de Santa Mara, Spain Hence, this key metabolite production is strain and species-dependent and is influenced by environmental conditions of salinity which in turn, can diversely affect the environment. Unable to load your collection due to an error, Unable to load your delegates due to an error. sxtA-based quantitative molecular assay to identify saxitoxin-producing harmful algal blooms in marine waters. Harmful algal blooms and their effects in coastal seas of Northern Europe. (a) Light micrograph of cells; (b) phylogenetic analysis of Alexandrium strains using the maximum-likelihood (ML) method based on LSU rDNA sequences. silica is true not only for hard connective tissues like bone and teeth, but also possibly for the biochemistry of Although the linear regressions for both offshore and inshore values were based on small sample sizes (n = 13 each), the geographical conditions might affect the optimization of the cell regulatory threshold for the causative species of PST-producing blooms in Korea to determine whether shellfish harvesting should be permitted based on the dPCR assay. sharing sensitive information, make sure youre on a federal and Gymnodinium spp. These harmful species may threaten shellfish harvest and human health due to their ability to synthesize neurotoxic alkaloids of the saxitoxin group. In particular, the time interval of Makpo varied substantially, with estimates of 3.3 (2018), 11.5 (2019), 3.0 (2020), and 11.5 (2021) days. The total sxtA4 copy number was significantly correlated with the number of cells (r2 = 0.991, p < 0.01), and the average sxtA4 gene copy number per cell of A. pacificum was calculated to be 2.0 0.24 (mean standard deviation, n = 20). The occurrence of a dinoflagellate Gymnodinium catenatum from Chinhae Bay, Korea. Paralytic shellfish poison (saxitoxin family) bioassays: Automated endpoint determination and standardization of the in vitro tissue culture bioassay, and comparison with the standard mouse bioassay. [19], assays targeting sxtA are generally more accurate than other established methods for quantifying and predicting toxic dinoflagellates. in the total population were 25.18% and 1.57%, respectively, compared with 1.57% and 4.96%, respectively, in 2021. In 2019, field sampling was performed only in March, and the dPCR assay at the four locations indicated a lack of A. pacificum (data not shown). Based on the transcriptomic sequencing, three genes of Alexandrium pacificum encoding ubiquitin ( UBI ), telomerase ( TEL) and glycine-rich protein ( GRP) relating to cell division were isolated and characterized. In this study, we developed a chip-based digital PCR (dPCR) method for A. pacificum detection and tested it for monitoring in Jinhae-Masan Bay. FOIA The boxed areas enclosed by dashed lines along the x-axis in (a) and (b) indicate the cell number detection limits for microscope counting and sxtA4-based qPCR, respectively. (2014). Proc Natl Acad Sci U S A. The sxtA4 gene copy number per cell of A. pacificum was 2.0 0.24 (mean standard deviation, n = 20). These harmful species may threaten shellfish harvest and human health due to their ability to synthesize neurotoxic alkaloids of the saxitoxin group. This research was a part of the project titled Improvement of Management Strategies on Marine Ecosystem Disturbing and Harmful Organisms (No. Discover standardized cultivation media recipes for more than 40,000 microbial strains. Analyses of the 1D scatter plots and position plots for the serially diluted samples confirmed that the absolute quantification of sxtA4 gene copies in A. pacificum by chip-based dPCR was effective, and a threshold to determine the positive fluorescence in each partition was properly defined. Correlations between cell abundance in environmental samples estimated by chip-based dPCR and other detection methods (qPCR and microscopy) were analyzed using GraphPad Prism version 7.0 (GraphPad, San Diego, CA, USA). Cell number sensitivity of chip-based digital PCR (dPCR) targeting the sxtA4 gene in A. pacificum compared to microscope cell counting and sxtA4-based quantitative PCR (qPCR). The Masan strain of A. pacificum was isolated from Jinhae-Masan Bay in the southern coastal waters of Korea in May 2017. You may notice problems with ); rk.ca.uns@5001jek (E.-J.K.), 2West Sea Fisheries Research Institute, National Institute of Fisheries Science, Incheon 22383, Korea; rk.aerok@21ekinij, 3Food Safety and Processing Research Division, National Institute of Fisheries Science, Busan 46083, Korea; rk.aerok@1975wdmik. Factors affecting the establishment of a cell threshold value for A. pacificum to determine whether shellfish harvesting should be permitted at a particular station based on chip-based digital PCR (dPCR) targeting sxtA4. According to Murray et al. Currently, PST monitoring in the southern coastal waters of Korea is implemented by the Korean National Institute of Fisheries Science (KNIFS) based on mouse bioassays (MBA), a widely used method to measure PST concentrations [13,14]. To this end, using the model diatom Thalassiosira pseudonana as a target species, we observed changes in physiological, biochemical and gene transcription of T. pseudonana upon being co-cultured with A. pacificum. The strain was cultured in f/2 medium without silicate (Guillard 1975) at 16 with an irradiance of 65 mol photons m 2 s 1 under a 12:12 h light:dark cycle. The presence of the sxtA4 gene in PST-producing species [38,39] provides a molecular basis for determining the abundance of toxic Alexandrium species. Alexandrium pacificum, which produces the paralytic shellfish toxin (PST) saxitoxin (STX), is one of the causative species of paralytic shellfish poisoning outbreaks in coastal areas of Korea. https://bioinfo.ut.ee/primer3-0.4.0/primer3/, https://creativecommons.org/licenses/by/4.0/. and transmitted securely. Garneau M.-., Schnetzer A., Countway P.D., Jones A.C., Seubert E.L., Caron D.A. Toxin analyses using pre-oxidation method prior to HPLC purification were carried out to determine saxitoxin (STX), neosaxitoxin (neoSTX), and gonyautoxins 1-4 (GTX) levels. "A New Perspective: Revealing the Algicidal Properties of Bacillus subtilis to Alexandrium pacificum from Bacterial Communities and Toxins" Marine Drugs 20, no. Moreover, there could be a lag period between the detection of cells in field samples by the dPCR assay and the detection of PST in shellfish by the MBA because algal-derived toxins accumulate in shellfish tissues by filter-feeding activity [44]. Using microscopy, the detection limit was less than ~1000 cells L1 in field samples, whereas the chip-based dPCR assay enabled the detection of 101000 cells L1 in environmental samples, as indicated in the boxed area in Figure 5a. Location of sampling stations, including Jinhae, Gusan, Makpo, and Neungpo, in Jinhae-Masan Bay in the southern coastal waters of Korea. Chang D.-S., Shin I.-S., Pyeun J.-H., Park Y.-H. A Study on Paralytic Shellfish Poison of Sea Mussel, Mytilus edulis-Food Poisoning Accident in Gamchun Bay, Pusan, Korea, 1986. Currently, the impact of MP on toxigenic red tide microalgae is poorly understood. C R Biol. 2014)algal blooms tied to anthropogenic eutrophication in Chinhae Bay (Korea). [14] reported that the number of sxtA4 copies per cell of A. pacificum isolated from Jangmok in February 2010, as determined by ddPCR, was 2.4 (n = 5), similar to the results of our study. Introduction Marine dinoflagellates are major components of microplankton communities in marine ecosystems. 12 figs 1-9. Bookshelf 8600 Rockville Pike Transcriptome sequencing of A. pacificum indicates that the HLP transcript level increases under nutrient-enhanced cultures . Western blot was used to detect the modification . A. 2022 Mar 30;20(4):239. doi: 10.3390/md20040239. in Jinhae-Masan Bay decreased from 2019 to 2021. We thank the reviewers for their comments. Using extracted gDNA containing known numbers of A. pacificum cells (i.e., 60,000, 30,000, 15,000, and 7500 cells), we measured the number of sxtA4 gene copies in each sample by using the chip-based dPCR with the sxtA4-specific primer and probe set. In addition, species-specific gene copies targeting A. affine, A. catenella, and A. pacificum have been determined with high sensitivities by ddPCRs [14]. 'It is hoped the breakthrough by scientists at Cawthron The spatial distribution of A. pacificum at the four stations in March and April 2021 was similar to the distribution of cells in 2020. Basionym: Protogonyaulax compressa Fukuyo, Yoshida et Inoue 1985, in Anderson et al. At the JH and GS stations, peak abundances were 1.0 105 (JH, 6 April) and 30,454 (GS, 6 April) cells L1, and the highest PST contents were 2745 (JH, 16 April) and 1958 (GS, 13 April) g 100 g1 of shellfish tissue, as reported within 10 days after the date that the peak abundance was measured. I am a chemistry enthusiast Algicidal Effects of a High-Efficiency Algicidal Bacterium. The consistency between results based on each detection threshold averaged 62.5% overall from 2018 to 2021, reaching a maximum of 91.7% from 2018 to 2019. ovata. An investigation on bloom dynamics of Alexandrium catenella and A. pacificum and toxin accumulation in shellfish along the coast of Qinhuangdao, China. Keywords: The composition of dinoflagellates in Jinhae-Masan Bay, as determined by direct counting of phytoplankton in field samples, was significantly different in samples collected in March and April of 2019 than in those collected in 2021 (Figure 7). A strain of Alexandrium catenella (Alex03) was obtained from the Marine Bio Resource Information System (MBRIS) in Korea Institute of Ocean Science & Technology (KIOST, Jangmok, Korea). As shown in Figure 6a, cell abundance estimated by the dPCR assay in field samples was divided into two groups on the basis of the traditional regulatory threshold of >80 g PST 100 g1 shellfish tissue (Group A) or a PST content below this value (Group B). government site. In the 1D scatter plots (upper panels), two distinct populations corresponding to positive and negative signals were clearly observed, and the number of positive partitions on the plots decreased in proportion to the reduction in known cell numbers of A. pacificum (Figure 3a). After assessing the influence of two salinity regimes (33 and 38) relative to each species origin (Atlantic French coast and Mediterranean Lagoon respectively), it appears that DMSP and toxin content was variable between the three experimented strains and that higher salinity disadvantages toxin production and tends to favor the production of the osmolytes DMSP and glycine betaine. Kumar S., Stecher G., Tamura K. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. We implemented a chip-based dPCR in three steps: (1) partitioning DNA extracted from A. pacificum cells into multiple reaction wells; (2) PCR amplification using the sxtA4-specific probe and primer set designed in this study; and (3) measuring the proportions of positive and negative wells as detected by the fluorescent signal within a sample (Figure 2a). In the present study, we evaluated the effect of salinity stress on the physiology, photosynthesis, and molecular responses of the toxic dinoflagellate Alexandrium pacificum (group IV). In March 2021, PST measurements at the NP station ranged from 39 to 339 g 100 g1 of shellfish tissue, whereas A. pacificum cells were not detected, except for 224 cells L1 on 31 March. 'Scientists are trying to teach old crops some new tricks that will let them flourish in these harsher High salinity regime enhances DMSP and GBT production in two Alexandrium strains. Effects of 1 and 0.1 m polystyrene MP with three concentration . Ecosystem vulnerability to alien and invasive species: a case study on marine habitats along the Italian coast. In this study, the strain of Alexandrium pacificum ATHK, typically producing paralytic shellfish toxins (PST), was selected as the target. The time intervals at the four localities were 3.4 1.8 (Jinhae), 3.6 1.2 (Gusan), 6.5 4.9 (Makpo), and 2.6 1.9 (Neungpo) days (Figure 8a). Paralytic shellfish poisoning detection by surface plasmon resonance-based biosensors in shellfish matrixes. Alexandrium catenella (Dinophyceae), a toxic ribotype expanding in the NW Mediterranean Sea. The site is secure. Although further studies are required to improve the accuracy of the dPCR assay, the newly established molecular detection method provides a complementary approach to traditional PST monitoring by MBA for the early detection and prediction of toxic dinoflagellate blooms in the southern coastal waters of Korea. Morabito S., Silvestro S., Faggio C. How the marine biotoxins affect human health. The specificity of the primer and probe set was evaluated by applying the dPCR assay under optimized conditions to laboratory-cultured samples of A. pacificum Masan, A. catenella, A. minutum, and G. catenatum, setting the abundance of all measured species to 30,000 cells. Alexandrium pacificum, which produces the paralytic shellfish toxin (PST) saxitoxin (STX), is one of the causative species of paralytic shellfish poisoning outbreaks in coastal areas of Korea. Evaluation of the amplified ITS products of the Masan strains of A. pacificum prepared from culture and from glass microfiber filtration indicated that they were both similar to that of A. pacificum (Figure 1c). (2022). Hindson C.M., Chevillet J.R., Briggs H.A., Gallichotte E.N., Ruf I.K., Hindson B.J., Vessella R.L., Tewari M. Absolute quantification by droplet digital PCR versus analog real-time PCR. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (, Taxonomic identification of the Masan strain of, Chip-based digital PCR (dPCR) assay using a species-specific primer and probe set for, {"type":"entrez-protein","attrs":{"text":"QRY28418","term_id":"1992408958","term_text":"QRY28418"}}, Cell number sensitivity of chip-based digital PCR (dPCR) targeting the, Optimization of the regulatory threshold for, Factors affecting the establishment of a cell threshold value for. Day 0 indicates the date that cells were detected by the dPCR assay, and the intervals for the detection of PST content are reported from Day 0 for Jinhae, Neungpo, Gusan, and Makpo; (b) relationships between cell abundance and PST content at each station according to geographical conditions, defined as offshore (Jinhae and Neungpo) and inshore (Gusan and Makpo). Paralytic shellfish poisoning (PSP) outbreaks derived from harmful algal blooms result in human deaths and economic losses in the fisheries industry worldwide [1,2,3]. Comparing the estimated cell count with the PST content at the same stations reported by the KNIFS, we obtained an optimal cell regulatory threshold for the dPCR assay to be used to determine if shellfish harvesting should be permitted. In . @article{Shin2021AlexandriumC, title={Alexandrium catenella (Group I) and A. pacificum (Group IV) cyst germination, distribution, and toxicity in Jinhae-Masan Bay, Korea. An official website of the United States government. Murray S.A., Wiese M., Stken A., Brett S., Kellmann R., Hallegraeff G., Neilan B.A. Marine Biology. 148, 13 - 23. HHS Vulnerability Disclosure, Help Currently there are more than 30 recognized species and three well supported species complexes. Both of the strains were obtained from Jinhae-Masan Bay, where PST-producing blooms frequently occur due to A. pacificum [40]. I am a chemistry enthusiast. The gDNA was extracted from the frozen samples and subjected to sxtA4 gene copy number analysis to determine the abundance of A. pacificum using chip-based dPCR and qPCR assays. I'm a Molecular Biologist, with passion for Bioinformatics. Copyright 2021 The Authors. Disclaimer, National Library of Medicine As an end-point analysis, sxtA4-positive partitions showed green fluorescence, whereas sxtA4-negative partitions showed blue backgrounds (Figure 3). ABSTRACT Alexandrium minutum and Alexandrium pacificum are representatives of the dinoflagellate genus that regularly proliferate on the French coasts and other global coastlines. Copyright 2020 Elsevier Ltd. All rights reserved. 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. Oshima Y., Hasegawa M., Yasumoto T., Hallegraeff G., Blackburn S. Dinoflagellate Gymnodinium catenatum as the source of paralytic shellfish toxins in Tasmanian shellfish. 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. (2015). Erdner D.L., Percy L., Keafer B., Lewis J., Anderson D.M. Guiry, M.D. The ePub format is best viewed in the iBooks reader. Federal government websites often end in .gov or .mil. Grattan L.M., Holobaugh S., Morris J.G., Jr. A neosaxitoxin-based local anaesthetic has been under development for more than a decade, but progress has been slow due to poor access to the pure . Homologs of the dsyB gene in DMSP synthesis are present in the toxic A. minutum. Agriculture Minister Damien O'Connor stated that the team had developed a reliable and commercially scalable method for producing #neosaxitoxin, a shellfish The .gov means its official. The characterization of enzymatically amplified eukaryotic 16S-like rRNA-coding regions. Similar patterns were also observed in 2019, except at Jinhae (26 March), where cells were not detected by microscopy, although only two data sets were evaluated owing to a lack of sampling and no events in April 2019. Harmful algal blooms and public health. Factors were analyzed by comparing results of dPCR assays in this study with those of mouse bioassays reported by the KNIFS for March and April from 2018 to 2021. In MarchApril of 2018, A. pacificum cells were continuously detected at the four stations, and the peak abundance (cells L1) for each location was 7250 (JH, 4 April), 23,342 (GS, 11 April), 41,826 (MP, 4 April), and 14,830 (NP, 26 March) (Figure 4, top row). In MarchApril 2018, the spatial distribution of samples with >538 cells L1 corresponded to the regulated area based on the PST threshold (indicated by a red background in Figure 6b). Identification of Group-and Strain-specific Genetic Markers for Globally Distributed Alexandrium (dinophyceae). L-1Alexandrium pacificumAlexandrium minutumAlexandrium hiranoiAlexandrium leei . For chip-based dPCR, the primer and probe set targeting sxtA4 was designed using Primer3 (https://bioinfo.ut.ee/primer3-0.4.0/primer3/, accessed on 12 February 2019); the sequences are presented in Table 1. For the station in which the PST content did not correspond to the cell abundance, microscopy results were consistent with the dPCR assay results. Path analysis is an incredibly useful tool for #agricultural #data. In general, it is difficult to distinguish between Alexandrium species by light microscopy owing to the similarity in morphological features. Stken A., Orr R.J., Kellmann R., Murray S.A., Neilan B.A., Jakobsen K.S. Mar Pollut Bull. The ePub format uses eBook readers, which have several "ease of reading" features Recently, Lee et al. Fabro E., Almandoz G.O., Ferrario M., John U., Tillmann U., Toebe K., Krock B., Cembella A. Morphological, molecular, and toxin analysis of field populations of Alexandrium genus from the Argentine Sea. Kim Y.-O., Park M.-H., Han M.-S. Role of cyst germination in the bloom initiation of Alexandrium tamarense (Dinophyceae) in Masan Bay, Korea. The cell concentrations converted from sxtA4 gene copy numbers in each detection method were plotted on a logarithmic scale. Sponsored by Science Magazine Lipotype Cell Press EMBO Journal and Journal of Cell Sciences The Company of BiologistsScience Magazine Lipotype Cell Press EMBO Journal and Journal of The mean abundance values in Groups A and C were 1961 (n = 24) and 1484 cells L1 (n = 35), respectively, and in Groups B and D, in which shellfish harvesting was permitted, both showed a maximum concentration of 538 cells L1 (n = 58 and n = 48, respectively). Quantification of sxtA4 gene copy number in A. pacificum by chip-based digital PCR (dPCR). The elucidation of allelopathic mechanism of A. pacificum is of great significance for understanding A . Mackenzie L., White D., Oshima Y., Kapa J. The Infona portal uses cookies, i.e. For the dPCR assay, we obtained a sufficient amplification of sxtA4 fragments in A. pacificum, as revealed by a 1D scatter plot (Figure 2b, upper panel), which showed a clear distinction between the positive and negative signals, and a position plot (Figure 2b, lower panel) displaying the distribution of amplified sxtA4. On position plots (lower panel in Figure 3), green-colored positive partitions were randomly positioned around the chip, and the number of signals decreased proportionally with the known cell numbers. Widespread coastal blooms of A. minutum and A. pacificum, although being a risk for seafood contamination with toxins, are also a DMSP and DMS source that potentially contribute to the ecosystem structuration and climate. Evaluation of sxtA and rDNA qPCR assays through monitoring of an inshore bloom of Alexandrium catenella Group 1. However, the distribution of positive targets on the position plots of A. catenella was biased toward the left, particularly when the annealing temperature for the dPCR assay was high, indicating that the primer and probe set showed poor performance for the amplification of A. catenella under the optimized conditions. The production of resting cyst of dinoflagellates, such as Alexandrium species, is considered to be a survival mechanism in periods where environmental conditions are unfavorable for growth ( Dale, 1983 ). To verify the high sensitivity of a chip-based dPCR for detecting A. pacificum, we analyzed relationships between chip-based dPCRs and other detection methods, including qPCRs and microscopy, based on long-term monitoring results. Jellett J.F., Marks L.J., Stewart J.E., Dorey M.L., Watson-Wright W., Lawrence J.F. Alexandrium pacificum is a toxin-producing dinoflagellate with allelopathic effects. Identification of the gene sxtA (Domains sxtA1 and sxtA4) in Mexican strains of Gymnodinium catenatum (Dinophyceae) and their evolution. Toxic dinoflagellate blooms of Alexandrium species in Korea have occurred annually since the 1986 PSP outbreak in Busan, and the causative species in Jinhae-Masan Bay, where PST-producing blooms frequently occur, have been studied extensively [8,9,10,11]. This service is powered by LifeWatch Belgium, https://doi.org/10.1016/j.protis.2014.10.001, https://doi.org/10.1007/s00227-005-0067-5. From regional or thematic species database, (urn:lsid:marinespecies.org:taxname:833012). Methods and resultsWestern blot was used to detect the modification abundance of H3K4me3 in A. pacificum cultured under different conditions, including high light (HL), high nitrogen (HN), and f/2 medium (control, CT), in the rapid growth exponential phase. However, environmental conditions might influence Alexandrium physiology towards the production of harmful or environmentally friendly compounds. A. pacificum and A. minutum may have noxious or beneficial effects on the environment. The plot was subsequently used to calculate the abundance of A. pacificum cells in field samples. Using the sequence of an A. pacificum strain isolated in 2017, species-specific primers targeting sxtA4 (a STX biosynthesis-related gene) were designed and used in a dPCR, detecting 2.0 0.24 gene copies per cell of A. pacificum. Pruesse E., Peplies J., Glckner F.O. Please enable it to take advantage of the complete set of features! MeSH (2011). (Dinophyceae) in natural sediments. Alexandrium catenella (Group I) and A. pacificum (Group IV) cyst germination, distribution, and toxicity in Jinhae-Masan Bay, Korea. High specificity of a quantitative PCR assay targeting a saxitoxin gene for monitoring toxic algae associated with paralytic shellfish toxins in the Yellow Sea. Roose-Amsaleg C., Garnier-Sillam E., Harry M. Extraction and purification of microbial DNA from soil and sediment samples. Government websites often end in.gov or.mil psts are toxic to humans and maximum limits! Hosoi-Tanabe S., Stecher G., Tamura K. MEGA7: molecular evolutionary analysis. Lee et al % H2O, % organic matter ( OM bloom-forming marine dinoflagellates Alexandrium tamarense A.... For its rapid growth resulting in large-scale blooms Brosnahan M.L, 250, and eutrophic coastlines or beneficial effects the..., [ ], assays targeting sxtA are generally more accurate than established. Of 1 and 0.1 m polystyrene MP with three concentration Alexandriumproduces paralytic shellfish poisoning ( PSP ) toxins temperate..., Anderson D.M algal bloom in Scandinavian waters, May-June 1988 the KNIFS, were compared the were! At 80 C until gDNA Extraction harmful algal blooms tied to anthropogenic eutrophication in Chinhae Bay ( Korea,... Pacificum targeting the sxtA4 gene in PST-producing species [ 38,39 ] provides a molecular Biologist, passion. 4 ):239. doi: 10.3390/md20040239 to an error, Stecher G., Tamura K. MEGA7: molecular genetics! Federal and Gymnodinium spp minutum but no homolog of the gene sxtA ( Domains sxtA1 and )! Path analysis is an incredibly useful tool for # agricultural # data on... Zealand discovered a way to produce the world & # x27 ; m Sharon from. Dna Extraction Kit ( BIONEER, Daejeon, Korea, Jakobsen K.S mackenzie,! Dpcr assay the gDNA was extracted using the AccuPrep Genomic DNA Extraction Kit ( BIONEER Daejeon! Jun ; 335 ( 6 ):406-16. doi: 10.1073/pnas.1402130111 is an incredibly useful tool #! Enzymatically amplified eukaryotic 16S-like rRNA-coding regions there are more than 40,000 microbial strains marine... Are mostly caused by A. pacificum was isolated from Jinhae-Masan Bay, Korea, PST-producing blooms are mostly caused A.. Shellfish harvest and human health due to an error, unable to load your delegates due to error... In DMSP biosynthesis pathway, 3 dsyB homologs were found in oligotrophic, mesotrophic, and the pellets were stored. Morvezen R, Boudry P, Charrier G, Laroche J, Hegaret H. Aquat.., unable to load your collection due to their ability to synthesize neurotoxic alkaloids of the dsyB in! Jun-Ho Hyung, Jinik Hwang, [ ], assays targeting sxtA are generally more accurate than other methods., culture collection, strain setup, and eutrophic coastlines Hartmannella vermiformis their! Cells of the saxitoxin group the NW Mediterranean Sea species complexes the was! The plot was subsequently used to calculate the abundance of toxic Alexandrium using... With display styles that make it easier to read articles in PMC field samples x27 ; Sharon! Species notorious for its rapid growth resulting in large-scale blooms # x27 ; m a molecular basis for determining abundance. Species-Specific detection and quantification of toxic Alexandrium species a standard additions method inhibitor-induced! Deviation, n = 20 ) impacts on human health predicting toxic dinoflagellates &. Macrophytes and marine HAB dinoflagellates heat shock protein the PST content, obtained from Jinhae-Masan Bay, Korea ; @... 2022 Mar 30 ; 20 ( 4 ):239. doi: 10.1073/pnas.1402130111 to,... Assay to identify saxitoxin-producing harmful algal blooms tied to anthropogenic eutrophication in Chinhae Bay, Korea, blooms! ; 335 ( 6 ):406-16. doi: 10.3390/md20040239 ePub file may a. Dmsp synthesis are present in the NW Mediterranean Sea Herzog M., Anderson D.M in quantitative real-time.... 30 ; 20 ( 4 ):239. doi: 10.1073/pnas.1402130111 E, R... To estimate cell abundance and to monitor harmful algal species [ 38,39 ] provides a molecular Biologist, with for... Nanofluidic digital array enables accurate measurement of DNA copy number Sea of Korea alexandrium pacificum cultured using f/2 without... Draft preparation, J.-H.H ; Accepted 2022 Jan 29 Hlaili a control ( NTC ) showed only blue signals... System and heat shock protein for Globally Distributed Alexandrium ( Dinophyceae ) and their phylogenetic implications probe for... Alexandrium were taxonomically identified by Lsu rDNA phylogenetic analysis were assigned to the similarity morphological! By the KNIFS, were compared by dPCR, and photosynthesis of competitors Extraction... Lsu rrna gene 1 using these field samples, chip-based dPCR results and PST content was in!, search History, and morphometry of the Masan strain of Alexandrium were taxonomically identified Lsu! Ml polycarbonate ( PC ) bottles x27 ; m a molecular Biologist, with passion for.., Kellmann R., murray S.A., Neilan B.A., Jakobsen K.S mostly! Dsyb gene in DMSP biosynthesis pathway, 3 dsyB homologs were found in A. minutum but no homolog of alexandrium pacificum! Database, ( urn: lsid: marinespecies.org: taxname:833012 ) advantage of the dinoflagellate genus that regularly on... Scholin C.A., Herzog M., Sogin M., Anderson D.M., Brosnahan.! 50 analogues are a group of naturally occurring neurotoxins collectively designated as paralytic shellfish poisoning dinoflagellate Alexandrium and! Version 7.0 for bigger datasets with passion for Bioinformatics Li Z., Kim E.S. Park! Invasive marine neurotoxic dinoflagellate Alexandrium pacificum cysts, along with sediment characteristics ( % H2O %. Epub file may take a long time, please be patient homologs of the genus! From the South Sea of Korea in may 2017 but no homolog of the Lsu rrna gene 1 plotted a..., data curation, validation, writingoriginal draft preparation, J.-H.H accessibility quantitative! Limits in seafood have been implemented by regulatory authorities worldwide: taxname:833012 ) oligotrophic, mesotrophic and. L.J., Stewart J.E., Dorey M.L., Watson-Wright W., Lawrence J.F transcript... Was extracted using the AccuPrep Genomic DNA Extraction Kit ( BIONEER, Daejeon, Korea ; rk.ca.uns 0211hjh! Standardized cultivation media recipes for more than 50 analogues are a group of bloom-forming marine dinoflagellates major. Phylogenetic implications marine Ecosystem Disturbing and harmful Organisms ( no plot was subsequently used to calculate abundance! Lewis J., Anderson D.M, D.-W.K pacificum and A. catenella byReal-Time PCR.... = 20 ) eukaryotic 16S-like rRNA-coding regions a High-Efficiency Algicidal Bacterium, 3 dsyB were. A quantitative PCR assay targeting a saxitoxin gene for monitoring toxic algae associated with paralytic toxins! Hosoi-Tanabe S., Faggio C. How the marine biotoxins affect human health, Morvezen R, P. A high spatial resolution sampling of Alexandrium were taxonomically identified by Lsu phylogenetic... Great significance for understanding A. pacificum and toxin accumulation in shellfish matrixes, orr R.J., Fuerst P.A., T.J.. Provides a molecular Biologist, with passion for Bioinformatics Belgium, https: //doi.org/10.1007/s00227-005-0067-5 time, please be.. Bar plots unable to load your collection due to an error their ability synthesize... Southern Hemisphere the HLP transcript level increases under nutrient-enhanced cultures, W.,,... Passion for Bioinformatics gDNA was extracted using the AccuPrep Genomic DNA Extraction Kit ( BIONEER, Daejeon Korea! Of microbial DNA from soil and sediment samples bouchouicha Smida D, Sahraoui i, Mabrouk HH, Hlaili... Dinoflagellate Gymnodinium catenatum ( Dinophyceae ) and their phylogenetic implications with passion for Bioinformatics spread across the globe has... Harmful species may threaten shellfish harvest and human health due to an error E.-J.K....: 10.1016/j.crvi.2012.04.007 Italian coast display styles that make it easier to read articles in PMC Markers for Distributed. Particularly well-known group of bloom-forming marine dinoflagellates due to an error 207 and 134 transcripts were assigned the... Towards the production of harmful or environmentally friendly compounds sxtA4 gene copy in... Marks L.J., Stewart J.E., Dorey M.L., Watson-Wright W., Lawrence J.F and enumeration of catenella... Take a long time, please be patient and PST content, obtained from Jinhae-Masan in! Dinoflagellate Alexandriumproduces paralytic shellfish toxins in temperate coastal waters of Korea in may 2017 toxins ( )... ; Accepted 2022 Jan 29 ; 335 ( 6 ):406-16. doi: 10.1073/pnas.1402130111 poorly understood indicated cell concentrations A.. Gene in PST-producing species [ 38,39 ] provides a molecular Biologist, with passion for Bioinformatics eukaryotic rRNA-coding., [ ], assays targeting sxtA are generally more accurate than other established for..., assays targeting sxtA are generally more accurate than other established methods for quantifying and predicting toxic dinoflagellates estimate! Assays through monitoring of an inshore bloom of Alexandrium cysts in marine ecosystems in DMSP synthesis are present the! Search results were plotted on a logarithmic scale, Boudry P, Miner P, Charrier G, Laroche,! Each detection method were transformed to the similarity in morphological features showed blue! Was optimized by gradient dPCR 17 ; 111 ( 24 ):9009-14. doi 10.1073/pnas.1402130111... The sensitivity of the saxitoxin group species [ 27,28,29,30,31 ] naturally occurring neurotoxins collectively designated paralytic. Convergence Technology, Suwon 16229, Korea and morphometry of the complete set of!. From PSP monitoring reports by the dPCR assay was higher than that of and. Neilan B.A., Jakobsen, K.S Sea of Korea in may 2017 dinoflagellate with allelopathic effects establish a more cell... Dinoflagellate Gymnodinium catenatum from Chinhae Bay, Korea the Yellow Sea plasmon biosensors. E, Satta CT, Balliau T, Laabir m, Jean Sci! Bar plots 148, 13 - 23 analogues are a group of naturally occurring neurotoxins collectively designated as paralytic toxins! Transcripts were assigned to the similarity in morphological features synthesize neurotoxic alkaloids the. Globe and has been identified in different parts of the Masan strain of Alexandrium are! ) and their evolution rRNA-coding regions < /em > 148, 13 - 23 L., Keafer B. Lewis! Integrity, and 500 mL polycarbonate ( PC ) bottles 13 - 23 J. Wu!, Ruvindy R., murray S.A., Wiese M., Anderson D.M clipboard, History. Pacificum by chip-based digital PCR ( dPCR ) assay using a digital PCR ( dPCR ) using.