• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.128-128
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• 2003

The annually outbreak of paralytic shellfish poisoning (PSP) were caused by toxic dinolagellate A. tamarense and A. catenella in Korea. The purpose of this study were to investigate the distribution of PSP-causative organisms, A. tamarense and A. catenella and their species classification. Sediment (Saemangeum, the south open sea) and water samples (southeastern coast) were sampled to establish clonal isolates in 2003. After isolation and purification, strains were cultured under $17^{\circ}C$, f/2 media, 14:10=L:D cycle. PST analysis and species identification were performed by HPLC-FD method and specific DNA probe, respectively. Thirty-ons strains were isolated from the Saemangeum reclamation, southeastern coast including Jinhae Bay and south open sea. PSTs were detected in all cultured strains. In eight strains from south offshore, major toxin components are GTX5, C1/2 and minors are GTX3/4, dcGTX3, neoSTX. Sixteen strains from south coastal area have GTX1/4, neoSTX, C1/2 as major toxin components and GTX2/3 as minors. Seven strains from the Saemangeum reclamation have GTX5, C1/2 as major toxin components and GTX1/2/3/4 as minors. Thus, among eight south offshore isolates, four A. tamarense have more toxic (38.31~l19.16 fmol.$cell^{-1}$) than A. catenella (3.78~13.13 fmol.$cell^{-1}$). With the previous results of different toxin composition, toxin components and toxin contents, .it is toxin profile that could used to diagnosis of regional toxic population and geographical distribution of both A. tamarense and A. catenella and their toxigenic strains.

• Journal of Microbiology and Biotechnology
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• v.14 no.5
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• pp.959-966
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• 2004

Twenty-three cultures of harmful algal bloom (HAB)-(causing dinoflagellates were isolated from the coastal waters of Korea. For each of the 14 morphospecies, the nuclearencoded small subunit (SSU) rDNA was analyzed to determine the phylogenetic relatedness of the species. Despite temporal and spatial isolation, 3-4 clonal cultures of Alexandrium catenella, Cochlodinium polykrikoides, and Gymnodinium catenatum had 100% identical SSU rDNA sequences. In contrast, heterogeneities in the SSU rDNA sequences were observed in Akashiwo sanguinea and Lingulodinium polyedrum strains. Extreme sequence polymorphism was shown within the SSU rRNA genes of an Al. tamarense clonal culture. A homology search in GenBank revealed that 11 dinoflagellate species were located in clusters corresponding to their morphological classification. The SSU rDNA sequences of C. polykrikoides, Gyrodinium instriatum, and Pheopolykrikos hartmannii, which were determined for the first time in this study, showed the following phylogenetic relationships: C. polykrikoides formed an independent branch separated from other dinoflagellates; Gyr. instriatum was placed in a monophyletic group with Gyr. dorsum and Gyr. uncatenum; and Ph. hartmanii, which forms a distinct two-celled pseudocolony, belonged to Gymnodinium sensu Hansen and Moestrup.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.6
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• pp.887-893
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• 2007

Influences of forage DM and addition of cane molasses on silage characteristics of berseem (Trifolium alexandrium) and lucerne (Medicago sativa) and their ruminal digestion kinetics in Nili buffaloes were studied. Berseem and lucerne fodders (at one tenth bloom) were ensiled with wheat straw in laboratory silos to achieve 20, 30, 40% forage DM and without wheat straw (control); each forage DM level was supplemented with 2, 4 and 6% of cane molasses at ensiling. The pH and lactic acid contents of berseem and lucerne silages were affected by both forage DM and addition of molasses. Dry matter, CP and true protein (TP) of berseem and lucerne silages were affected by forage DM at ensiling but were not affected by the addition of cane molasses. Higher DM, CP and TP losses were observed when berseem and lucerne fodders were ensiled either without wheat straw or with wheat straw to achieve 20% and 40% forage DM at ensiling compared with 30% DM at ensiling. Fiber fractions (NDF, ADF, hemicellulose and cellulose) of berseem silage and lucerne silage were significantly increased with increasing forage DM at ensiling. Addition of cane molasses did not affect the DM, CP, TP and fiber fractions of both berseem and lucerne silages. Berseem and lucerne ensiled at 30% DM with 2% cane molasses were screened for comparative ruminal digestion kinetics with their respective fodders. Addition of wheat straw to berseem or lucerne fodder at ensiling depressed DM and NDF ruminal degradability. However, ruminal lag time, rate of degradation and extent of digestion of silages were similar to their respective fodders. In conclusion, berseem and lucerne could be ensiled with wheat straw to increase their DM to 30% along with 2% molasses for buffaloes.

• ALGAE
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• v.31 no.4
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• pp.391-401
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• 2016

To develop an easy and rapid method of quantifying lipid contents of marine dinoflagellates, we quantified lipid contents of common dinoflagellate species using a colorimetric method based on the sulpho-phospho-vanillin reaction. In this method, the optical density measured using a spectrophotometer was significantly positively correlated with the known lipid content of a standard oil (Canola oil). When using this method, the lipid content of each of the dinoflagellates Alexandrium minutum, Prorocentrum micans, P. minimum, and Lingulodinium polyedrum was also significantly positively correlated with the optical density and equivalent intensity of color. Thus, when comparing the color intensity or the optical density of a sample of a microalgal species with known color intensities or optical density, the lipid content of the target species could be rapidly quantified. Furthermore, the results of the sensitivity tests showed that only $1-3{\times}10^5cells$ of P. minimum and A. minutum, $10^4cells$ of P. micans, and $10^3cells$ of L. polyedrum (approximately 1-5 mL of dense cultures) were needed to determine the lipid content per cell. When the lipid content per cell of 9 dinoflagellates, a diatom, and a chlorophyte was analyzed using this method, the lipid content per cell of these microalgae, with the exception of the diatom, were significantly positively correlated with cell size, however, volume specific lipid content per cell was negatively correlated with cell size. Thus, this sulpho-phospho-vanillin method is an easy and rapid method of quantifying the lipid content of autotrophic, mixotrophic, and heterotrophic dinoflagellate species.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.6
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• pp.669-674
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• 2017

Paralytic shellfish toxins (PSTs) are produced by marine dinoflagellate phytoplankton Alexandrium spp. and Gymnodinium spp. These toxins accumulate in filter feeding organisms such as bivalves and the ingestion of contaminated shellfish can cause illness in humans. The mouse bioassay (MBA) has been the preferred PST testing method worldwide for more than 50 years. However, this assay has several disadvantages, such as detection limits, non-toxic-profiles, and the ethical issues of using animals. The aim of this study was to establish an alternative to the MBA method for testing for PSTs. We optimized the analysis conditions of a post-column oxidation-high performance liquid chromatography (PCOX-HPLC) method and the Scotia Rapid Test Kit, and then compared the accuracy of these methods to the MBA method. The results demonstrated a strong correlation between the PCOX-HPLC method and the MBA, although the PCOX-HPLC method required expensive equipment and standard material, and was time consuming. The Scotia Rapid Test Kit promises to be a useful tool, as it provided rapid and qualitative results, although the method sometimes gave a false positive result that could not be explained by toxin profiles.

• Journal of Life Science
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• v.14 no.4
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• pp.589-592
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• 2004

The growth response of the fish-killing dinoflagellate Cochlodinium polykrikoides was studied in cultures, using the treatment of Ceratium extracts by a methanol, a water-soluble, and a cell-free medium. The cell-free medium had the most increasing on the growth of C. polykrikoides cultures, enriched with $\geq$ 25% Ceratium, whereas the methanol and water-souble fractions did not affect the growth of C. polykrikoides exposed to even higher concentration. In particular, the cell-free medium also increased the growth of Gyrodinium impudicum and Chaetoceros sp., similar species to C. polykrikoides. In contrast to C. polykrikoides, G. impudicum and Chaetoceros sp., the growth of Alexandrium tamarense was inhibited significantly, and there was no great effect on the growth of Prorocentrum minimum. These results imply that Ceratium extracts may play an important role in the stimulatory effect of C. polykrikoides, and they have to affect the interaction between C. polykrikoides and Ceratium when co-existing.

• Journal of Life Science
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• v.11 no.2
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• pp.74-80
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• 2001

The nucleotide sequences of the internal transcribed spacer regions (ITS1 and ITS2) of ribosomal DNA (rDNA), and the 5.85 rRNA gene, have been determined for 13 strains of dinoflagellates in order to analyze the phylo-genetic relationship. The DNA sequences contained considerable variation in the ITS regions, but little in the 5.85 rDNA. In addition, the ITS1 was more variable than the ITS2 in all species examined. The nucleotide length of this region varied from 519 bp to 596 bp depending on the taxa. The investigated taxa were divided into three large groups based on the ITS length, i. e., a group with short ITS region (A. fraterculus and Alexandrium sp.), a with ITS region group (P. micans, P. minimum and P. triestinum) and a with ITS region group (G. impudicum, C. polykrikoides, G. sanguineum, G. catenatum and H. triquetra). The relationship between nucleotide length of ITS1 and that of ITS2 was negative, whereas G+C content and nucleotide length showed positive correlation. In phylogenetic analyses producing NJ trees, the topology was similar cluster and clearly divided the taxa into three groups based on 5.8S rDNA that were similar to those based on morphological characteristics. In particular, G. impudicum was more closely related to G. catenatum than to C. polykrikoides using phylogenetic analysis. From this study, we chew that the length of ITS region contributes to discriminate Korean harmful algal species and ITS analysis is a useful method for resolving the systematic relationships of dinoflagellates.

• ALGAE
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• v.33 no.3
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• pp.279-290
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• 2018

Microalgae have been utilized in diverse industries including aquaculture. Among the microalgae, dinoflagellates are known to have various bioactive compounds, and thus the interest in their application to industry has increased. In order to test their potential as food materials for aquaculture animals, the crude protein contents and compositions of amino acids of six dinoflagellates Heterocapsa rotundata (family Heterocapsaceae), Ansanella granifera (Suessiaceae), Alexandrium andersonii (Ostreopsidaceae), Takayama tasmanica (Brachidiniaceae), Takayama helix, and Gymnodinium smaydae (Gymnodiniaceae) belonging to diverse families were analyzed. The percentage of the amount of the crude protein relative to dry weight of T. tasmanica was the highest (65%) and that of A. andersonii was the lowest (26%). However, the highest percentage of total detected amino acids in crude protein was found in A. andersonii (98.2%). In all six dinoflagellates, glutamic acid was the most dominant amino acid in crude protein. However, the second main amino acid was aspartic acid for H. rotundata, A. granifera, T. helix, and G. smaydae, but were arginine and leucine for A. andersonii and T. tasmanica, respectively. Furthermore, T. tasmanica and T. helix did not have taurine and gamma-aminobutyric acid, whereas the other dinoflagellates possessed them. The percentages of essential amino acid contents of the dinoflagellates met the requirement levels for juvenile shrimps. In addition, the dinoflagellates were not toxic to the brine shrimp Artemia salina. Compared with the other microalgae reported so far, H. rotundata and A. andersonii can be used for arginine-rich diets, T. tasmanica for valine and leucine-rich diets, A. granifera for histidine-rich diets, T. helix for threonine-rich diets, and G. smaydae for lysine-rich diets. Therefore, based on their biochemical composition and toxicity to Artemia, the dinoflagellates could be used as essential amino acid sources for cultivating animals in the aquaculture industry.

• Fisheries and Aquatic Sciences
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• v.26 no.8
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• pp.470-481
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• 2023

Algal overgrowth in shrimp culture ponds can affect the quality of the aquatic environment, thereby adversely affecting the shrimp and causing economic losses. The objective of this study was to evaluate the variation in phytoplankton composition in intensive shrimp ponds in Bac Lieu province, Vietnam. Phytoplankton samples were collected in three black tiger shrimp (Penaeus monodon) ponds and three whiteleg shrimp (Litopenaeus vannamei) ponds. The collected data were analyzed using SPSS and canonical correlation analysis softwares. In total, 75 species of phytoplankton were recorded in black tiger shrimp ponds and 64 species in whiteleg shrimp ponds. Diatoms had the highest species diversity with 29-30 species (39%-47%), followed by green algae with 9-19 species (14%-25%); species numbers of other phyla varied from 5-12 (8%-16%). The total number of phytoplankton species throughout the study varied from 34-50 species. Algal density was relatively high and ranged from 497,091-2,229,500 ind./L and 1,301,134-2,237,758 ind./L in black tiger shrimp and whiteleg shrimp ponds, respectively. The diatom density tended to increase during the final stage of the production cycle in black tiger shrimp ponds. Blue-green algae and dinoflagellates also increased in abundance at the end of the cycle, which can affect shrimp growth. Diatoms were significantly positively correlated with pH, salinity, total ammonia nitrogen, and nitrate (NO₃^-) concentrations (p < 0.05). Blue-green algae and dinoflagellates were positively correlated with salinity, phosphate (PO₄^3-), and NO₃^-. Algal species diversity was lower in the whiteleg shrimp ponds than in the black tiger shrimp ponds. Several dominant algal genera were recorded in the shrimp ponds, including Nannochloropsis, Gyrosigma, Chaetoceros, Alexandrium, and Microcystis. The results of this study provide basic data for further investigations, and they contribute to the management of algae in brackish-water shrimp ponds.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.15 no.11
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• pp.6958-6965
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• 2014

This study examined the occurring pattern of potential risky species and the related abiotic factors for port-specific environmental management considering the control of ballast water-induced foreign species at Incheon seaport. From a total of 62 species observed during the study, 13 red-tide and 7 toxic phytoplankton, normally occurring species in Korean waters, occurred from the seasonal investigation at the inner and outer sites of the Incheon seaport from 2007 to 2009. The number of potential risky phytoplankton was relatively high at the outer site of the port during summer and winter. Red-tide species, such as Skeletonema spp., Thalassiosira nordenskioldii, and Paralia sulcata, dominated the total standing crops at the inner site (avg. 72.4%) and outer site (avg. 77.6%) in spring and summer, being positively correlated with the concentrations of total suspended solids (TSS) and pH (p<0.05). In summer, the red-tide species (Skeletonema spp.) and toxic species (Alexandrium catenella, A. tamarense, Dinophysis acuminata and Pseudo-nitzschia spp.) co-dominated (avg. 74.2%) at the inner site, while Skeletonema spp. and P. sulcata predominated (avg. 67.2%) at the outer site. During the study periods, the toxic species were significantly and positively correlated with the chemical oxygen demand (COD), dissolved inorganic nitrogen, silicate and phosphate (p < 0.05). The chlorophyll-a (chl-a) concentration of phytoplankton at the outer site ranged from 1.49 to $5.46{\mu}g/L$ on average, which was 3-5 times higher than that at the inner site in spring, summer and autumn, whereas there was no difference in the concentration between inner (avg. $0.94{\mu}g/L$) and outer (avg. $0.95{\mu}g/L$) sites in winter. In summary, diverse red-tide species dominated and a relatively high chl-a concentration existed at the outer site, whereas a relatively high number of toxic species and low chl-a concentration was observed at the inner site in summer. The potential risky species can outbreak in association with the concentration of nutrients, COD and TSS, suggesting that distinctive management of potential risky species is needed considering the environmental characteristics of Incheon seaport.