Three Loxocaudinae species (Ostracoda, Podocopida) from South Korea.

For many ostracod groups in Korea, published records are missing or are very limited. Loxocaudinae is one such subfamily, with only one named species, Loxocaudaorientalis Schornikov, 2011 reported from Korea. Having fewer than 50 species, this subfamily can be considered a small ostracod group, with most of the species known only by their shell morphology. The diagnoses of genera are based on the shell characters that are often homoplastic, and soft body appendages that are difficult to observe, such as the mandibular exopodite. Because of this, the validity of the entire subfamily and some of its genera have been questioned. Here three Loxocaudinae species were collected from the marine macrobenthic assemblages from Korea. Two are new and belong to the genus Glacioloxoconcha Hartmann, 1990, previously known only from Antarctica: Glacioloxoconchajeongokensis sp. nov. and Glacioloxoconchajisepoensis sp. nov. Loxocaudaorientalis is briefly redescribed, with some of the populations having unusual morphological features. COI and 18S rRNA sequences of all three species are provided and the latter marker used to assess the position of the subfamily within the family Loxoconchidae and the superfamily Cytheroidea. The resulting tree shows that within the family Loxoconchidae, the genera Glacioloxoconcha and Loxocauda Schornikov, 1969 are the most closely related, with very shallow but well-supported branches. Polyphyletic and paraphyletic natures of several Cytheroidea families are discussed, inferred from the reconstructed phylogeny.

Population Genetic Structures and Demographic Expansion of the Exotic Jellyfish Carybdea brevipedalia in Korean Coasts Inferred from Mitochondrial COI Analysis.

Carybdea brevipedalia Kishinouye, 1891 is a poisonous jellyfish that usually occurs only in Japanese coastal regions. However, it was recently found on the Korean coast, thus expanding its known geographical range. In this study, we analyzed the population genetics and demographic histories of 113 C. brevipedalia specimens from the southern and eastern coastal regions of Korea by sequencing mitochondrial DNA cytochrome c oxidase subunit I (COI). We identified 42 C. brevipedalia COI haplotypes with high genetic diversity and a significant genetic structure. Populations were highly differentiated based on geographic location and distinctly divided into A and B clades. The results of Mantel tests indicated that geographic distance influenced the genetic distance between the two clades. Moreover, demographic analyses (neutrality tests) and the star-like profile of the Templeton, Crandall, and Sing (TCS) haplotype network indicated that C. brevipedalia had recently expanded into the southern and eastern coastal regions of Korea. These findings suggest that C. brevipedalia populations along the Korean coast have significant genetic differentiation that could be influenced by geographic isolation and subsequent adaptation to regional ecological conditions.

Comparative Study of Toxic Effects and Pathophysiology of Envenomations Induced by Carybdea brevipedalia (Cnidaria: Cubozoa) and Nemopilema nomurai (Cnidaria: Scyphozoa) Jellyfish Venoms.

Jellyfish stings can result in local tissue damage and systemic pathophysiological sequelae. Despite constant occurrences of jellyfish stings in oceans throughout the world, the toxinological assessment of these jellyfish envenomations has not been adequately reported in quantitative as well as in qualitative measurements. Herein, we have examined and compared the in vivo toxic effects and pathophysiologic alterations using experimental animal models for two representative stinging jellyfish classes, i.e., Cubozoa and Scyphozoa. For this study, mice were administered with venom extracts of either Carybdea brevipedalia (Cnidaria: Cubozoa) or Nemopilema nomurai (Cnidaria: Scyphozoa). From the intraperitoneal (IP) administration study, the median lethal doses leading to the deaths of mice 24 h post-treatment after (LD50) for C. brevipedalia venom (CbV) and N. nomurai venom (NnV) were 0.905 and 4.4697 mg/kg, respectively. The acute toxicity (i.e., lethality) of CbV was much higher with a significantly accelerated time to death value compared with those of NnV. The edematogenic activity induced by CbV was considerably (83.57/25 = 3.343-fold) greater than NnV. For the evaluation of their dermal toxicities, the epidermis, dermis, subcutaneous tissues, and skeletal muscles were evaluated toxinologically/histopathologically following the intradermal administration of the venoms. The minimal hemorrhagic doses (MHD) of the venoms were found to be 55.6 and 83.4 µg/mouse for CbV and NnV, respectively. Furthermore, the CbV injection resulted in extensive alterations of mouse dermal tissues, including severe edema, and hemorrhagic/necrotic lesions, with the minimum necrotizing dose (MND) of 95.42 µg/kg body weight. The skin damaging effects of CbV appeared to be considerably greater, compared with those of NnV (MND = 177.99 µg/kg). The present results indicate that the toxicities and pathophysiologic effects of jellyfish venom extracts may vary from species to species. As predicted from the previous reports on these jellyfish envenomations, the crude venom extracts of C. brevipedalia exhibit much more potent toxicity than that of N. nomurai in the present study. These observations may contribute to our understanding of the toxicities of jellyfish venoms, as well as their mode of toxinological actions, which might be helpful for establishing the therapeutic strategies of jellyfish stings.

The complete mitochondrial genome of the hydrozoan jellyfish Turritopsis lata Lendenfeld, 1885 (Cnidaria; Hydrozoa; Anthoathecata) with molecular phylogenetic analysis.

In this study, we sequenced and analyzed the complete mitochondrial genome (mtgenome) of the hydrozoan jellyfish Turritopsis lata. The mtgenome was a complete linear form (15,047 bp in length, 30.9% A, 42.1% T, 12.5% C, and 14.5% G), including 13 protein coding genes (PCGs) (cox1, cox2, cox3, atp6, atp8, nad1, nad2, nad3, nad4, nad4L, nad5, nad6, and cytb), 2 tRNAs (tRNAMet and tRNATrp), and 2 rRNAs (12S and 16S rRNA). The genome structure of the T. lata was completely identical to those of other species within the subclass Hydroidolina. In addition, our molecular phylogenetic analysis using 13 PCGs within hydrozoans showed that T. lata was the closest to Turritopsis dohrnii.

Complete mitochondrial genome of the hydrozoan jellyfish Blackfordia virginica Mayer, 1910 (Cnidaria; Hydrozoa; Leptothecata) with phylogenetic analysis.

In this study, we analyzed the complete mitochondrial genome of the hydrozoan jellyfish Blackfordia virginica. The genome was a linear form (15,109 bp long, 73.6% AT), including 13 protein-coding genes (cox2, atp8, atp6, cox3, nad2, nad5, nad6, nad3, nad4L, nad1, nad4, cytB, and cox1), 2 tRNAs (tRNA-Met and tRNA-Trp), and 2 rRNAs (12S and 16S RNA). The genome structure of the B. virginica was completely identical to mitochondrial genomes of other hydrozoans that belonged to Leptothecata and Anthoathecata. Molecular phylogenetic analysis within hydrozoan species showed that B. virginica was the closest to the hydrozoan Laomedea flexuosa.

Proteomic Changes during the Dermal Toxicity Induced by Nemopilema nomurai Jellyfish Venom in HaCaT Human Keratinocyte.

Jellyfish venom is well known for its local skin toxicities and various lethal accidents. The main symptoms of local jellyfish envenomation include skin lesions, burning, prickling, stinging pain, red, brown, or purplish tracks on the skin, itching, and swelling, leading to dermonecrosis and scar formation. However, the molecular mechanism behind the action of jellyfish venom on human skin cells is rarely understood. In the present study, we have treated the human HaCaT keratinocyte with Nemopilema nomurai jellyfish venom (NnV) to study detailed mechanisms of actions behind the skin symptoms after jellyfish envenomation. Using two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF/MS), cellular changes at proteome level were examined. The treatment of NnV resulted in the decrease of HaCaT cell viability in a concentration-dependent manner. Using NnV (at IC50), the proteome level alterations were determined at 12 h and 24 h after the venom treatment. Briefly, 70 protein spots with significant quantitative changes were picked from the gels for MALDI-TOF/MS. In total, 44 differentially abundant proteins were successfully identified, among which 19 proteins were increased, whereas 25 proteins were decreased in the abundance levels comparing with their respective control spots. DAPs involved in cell survival and development (e.g., Plasminogen, Vinculin, EMILIN-1, Basonuclin2, Focal adhesion kinase 1, FAM83B, Peroxisome proliferator-activated receptor-gamma co-activator 1-alpha) decreased their expression, whereas stress or immune response-related proteins (e.g., Toll-like receptor 4, Aminopeptidase N, MKL/Myocardin-like protein 1, hypoxia up-regulated protein 1, Heat shock protein 105 kDa, Ephrin type-A receptor 1, with some protease (or peptidase) enzymes) were up-regulated. In conclusion, the present findings may exhibit some possible key players during skin damage and suggest therapeutic strategies for preventing jellyfish envenomation.

Dataset of Swimming behavioral alterations in Danio rerio by Nemopilema nomurai jellyfish venom.

This article reports data associated with Prakash et al. [1]. Nemopilema nomurai jellyfish venom (NnV) can lead to neurotoxicity in zebrafish (Danio rerio) model. In the present study, zebrafish were treated with NnV by intraperitoneal injection and the swimming behavior of each fish was evaluated using a score scale. The dose of NnV in each treatment group was based on the protein concentration of NnV. Swimming is the main locomotory movements in the fishes. NnV modulated the swimming behavior of Danio rerio in a dose-dependent manner. In this article provided data are directly related to the previously published research article - "Danio rerio as an alternative vertebrate model for jellyfish venom study: the toxinological aspects of Nemopilema nomurai venom" [1] where the downregulation of acetylcholinesterase activity as well as histopathological alterations were observed from the brain of Danio rerio treated with NnV. Here we provide datasets, including mortality rate table, swimming behavior graph, and videos of zebrafish after NnV envenomation.

Population Genetic Structures of the Jellyfish Aurelia coerulea Polyps along Korean Coasts and Implications as Revealed by Mitochondrial COI.

The moon jellyfish, Aurelia coerulea, is is globally distributed, and its blooms have been responsible for severe environmental impacts. Benthic polyp populations are important for forming and maintaining medusa populations; however, their genetic structures are mostly unknown. Here, we analysed the genetic structure and phylogeographic pattern of A. coerulea polyps using the mitochondrial COI of 229 specimens collected from four different coastal regions of Korea. Molecular discrimination by COI assigned all polyps to A. coruelea. Population genetics revealed 53 haplotypes with high diversity and significant genetic structure, distinguishing two haplogroups (A and B) that coexist in all regions. Haplogroup A exhibited a star-like haplotype network pattern, while haplogroup B demonstrated a branched haplotype network pattern. Our results suggest that, the two haplogroups detected have existed in sympatry along Korean coasts. However, haplogroup A may have been established by a recent population expansion, while haplogroup B may have been established a long time ago. The strong genetic structure found within the polyp population of A. coerulea may have an effect on the moon jellyfish blooms on Korean coasts.

Danio rerio as an alternative vertebrate model for jellyfish venom study: The toxinological aspects of Nemopilema nomurai venom.

Nemopilema nomurai venom (NnV) is severely toxic to many organisms. However, the mechanism of its poisoning has not been properly understood yet. The present work demonstrates that zebrafish (Danio rerio) is an alternative vertebrate model for studying NnV jellyfish venom for the first time. In this model, NnV appears to cause severe hemorrhage and inflammation in cardiopulmonary regions of zebrafish. NnV also altered the swimming behavior of zebrafish accompanied by a significant downregulation of acetylcholinesterase (AChE) activity in brain tissues. Histopathological changes observed for various organs of D. rerio caused by NnV corresponded to an increase in lactate dehydrogenase (LDH) activity in tissues. NnV also significantly altered glutathione S-transferase (GST) activity in cardiopulmonary and brain tissues of D. rerio. SDS-PAGE revealed many protein bands of NnV of various sizes after silver staining. Taken together, these results indicate that Danio rerio can be a useful alternative animal model for jellyfish venom toxicology studies. Findings of the present study also suggest that Danio rerio could be used to develop an effective treatment strategy and discover the mechanism of action of jellyfish venom envenomation.

Protective effect of epigallocatechin-3-gallate (EGCG) on toxic metalloproteinases-mediated skin damage induced by Scyphozoan jellyfish envenomation.

Jellyfish stingings are currently raising serious public health concerns around the world. Hence, the search for an effective first aid reagent for the envenomation has been the goal of many investigators in the field. There have been a few previous reports of in vivo as well as in vivo studies suggesting the metalloproteinase activity of scyphozoan jellyfish venom, such as N. nomurai venom (NnV), plays a major role in the pathogenesis. These results have inspired us to develop a metalloproteinase inhibitor as a candidate for the treatment of Scyphozoan jellyfish envenomation. It has been previously demonstrated that the major polyphenol component in green tea, epigallocatechin-3-gallate (EGCG), can inhibit metalloproteinase activity of snake venoms. In fact, plant polyphenols as potential therapeutics have been shown to exert positive effects on neutralizing snake venoms and toxins. In the present study, we found that EGCG significantly inhibits the toxic proteases of NnV in a concentration-dependent manner. Human keratinocyte (HaCaT) and Human dermal fibroblast (HDF) cell culture studies showed that EGCG treatment can protect the cells from NnV-induced cytotoxicity which has been accompanied by the down-regulation of human matrix metalloproteinase (MMP)-2 and -9. Simulated rat NnV envenomation study disclosed that topical treatments with EGCG considerably ameliorated the progression of the dermonecrotic lesions caused by NnV. EGCG also reduced the activitions of tissue MMP-2 and MMP-9, which seem to be crucial players in the dermal toxic responses induced by NnV. Therefore, we propose that EGCG might be an effective therapeutic agent for the treatment of cutaneoous jellyfish symptoms.

The complete mitochondrial genome of the hydrozoan jellyfish Spirocodon saltatrix (Cnidaria; Hydrozoa; Anthoathecata) with phylogeny analysis.

In the present study, we sequenced and analyzed the complete mitochondrial genome of the hydrozoan jellyfish Spirocodon saltatrix. The mitochondrial genome was a linear form (15,752 bp long, 70.4% AT), consisting of 13 protein coding genes (cox1, cox2, atp8, atp6, cox3, nad2, nad5, nad6, nad3, nad4L, nad1, nad4, and cytB), two tRNAs (tRNA-Met and tRNA-Trp), and two rRNAs (12S and 16S). Mitochondrial gene arrangement of the S. saltatrix was completely identical to already-known mitochondrial genomes of hydrozoans. Molecular phylogenetic analysis using 13 protein-coding genes showed that S. saltatrix was closely related to the hydrozoan Clava multicornis.

Proteomic Analysis of Novel Components of Nemopilema nomurai Jellyfish Venom: Deciphering the Mode of Action.

Nowadays, proliferation of jellyfish has become a severe matter in many coastal areas around the world. Jellyfish Nemopilema nomurai is one of the most perilous organisms and leads to significant deleterious outcomes such as harm to the fishery, damage the coastal equipment, and moreover, its envenomation can be hazardous to the victims. Till now, the components of Nemopilema nomurai venom (NnV) are unknown owing to scant transcriptomics and genomic data. In the current research, we have explored a proteomic approach to identify NnV components and their interrelation with pathological effects caused by the jellyfish sting. Altogether, 150 proteins were identified, comprising toxins and other distinct proteins that are substantial in nematocyst genesis and nematocyte growth by employing two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI/TOF/MS). The identified toxins are phospholipase A2, phospholipase D Li Sic Tox beta IDI, a serine protease, putative Kunitz-type serine protease inhibitor, disintegrin and metalloproteinase, hemolysin, leukotoxin, three finger toxin MALT0044C, allergens, venom prothrombin activator trocarin D, tripeptide Gsp 9.1, and along with other toxin proteins. These toxins are relatively well characterized in the venoms of other poisonous species to induce pathogenesis, hemolysis, inflammation, proteolysis, blood coagulation, cytolysis, hemorrhagic activity, and type 1 hypersensitivity, suggesting that these toxins in NnV can also cause similar deleterious consequences. Our proteomic works indicate that NnV protein profile represents valuable source which leads to better understanding the clinical features of the jellyfish stings. As one of the largest jellyfish in the world, Nemopilema nomurai sting is considered to be harmful to humans due to its potent toxicity. The identification and functional characterization of its venom components have been poorly described and are beyond our knowledge. Here is the first report demonstrating the methodical overview of NnV proteomics research, providing significant information to understand the mechanism of NnV envenomation. Our proteomics findings can provide a platform for novel protein discovery and development of practical ways to deal with jellyfish stings on human beings.

Proteomic Investigation to Identify Anticancer Targets of Nemopilema nomurai Jellyfish Venom in Human Hepatocarcinoma HepG2 Cells.

Nemopilema nomurai is a giant jellyfish that blooms in East Asian seas. Recently, N. nomurai venom (NnV) was characterized from a toxicological and pharmacological point of view. A mild dose of NnV inhibits the growth of various kinds of cancer cells, mainly hepatic cancer cells. The present study aims to identify the potential therapeutic targets and mechanism of NnV in the growth inhibition of cancer cells. Human hepatocellular carcinoma (HepG2) cells were treated with NnV, and its proteome was analyzed using two-dimensional gel electrophoresis, followed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI/TOF/MS). The quantity of twenty four proteins in NnV-treated HepG2 cells varied compared to non-treated control cells. Among them, the amounts of fourteen proteins decreased and ten proteins showed elevated levels. We also found that the amounts of several cancer biomarkers and oncoproteins, which usually increase in various types of cancer cells, decreased after NnV treatment. The representative proteins included proliferating cell nuclear antigen (PCNA), glucose-regulated protein 78 (GRP78), glucose-6-phosphate dehydrogenase (G6PD), elongation factor 1γ (EF1γ), nucleolar and spindle-associated protein (NuSAP), and activator of 90 kDa heat shock protein ATPase homolog 1 (AHSA1). Western blotting also confirmed altered levels of PCNA, GRP78, and G6PD in NnV-treated HepG2 cells. In summary, the proteomic approach explains the mode of action of NnV as an anticancer agent. Further characterization of NnV may help to unveil novel therapeutic agents in cancer treatment.

Nemopilema nomurai jellyfish venom exerts an anti-metastatic effect by inhibiting Smad- and NF-κB-mediated epithelial-mesenchymal transition in HepG2 cells.

Epithelial-mesenchymal transition (EMT) is a key initial step in metastasis for malignant cancer cells to obtain invasive and motile properties. Inhibiting EMT has become a new strategy for cancer therapy. In our previous in vivo study, Nemopilema nomurai jellyfish venom (NnV) -treated HepG2 xenograft mice group showed that E-cadherin expression was strongly detected compared with non-treated groups. Therefore, this study aimed to determine whether NnV could inhibit the invasive and migratory abilities of HepG2 human hepatocellular carcinoma cells and to examine its effect on EMT. Our results revealed that transforming growth factor (TGF)-ß1 induced cell morphological changes and downregulated E-cadherin and ß-catenin expression, but upregulated N-cadherin and vimentin expression through the Smad and NF-κB pathways in HepG2 cells. Treatment of TGF-ß1-stimulated HepG2 cells with NnV reversed the EMT-related marker expression, thereby inhibiting cell migration and invasion. NnV also significantly suppressed the activation of p-Smad3, Smad4, and p-NF-κB in a dose-dependent manner. These data indicated that NnV can significantly suppress cell migration and invasion by inhibiting EMT in HepG2 cells, and therefore might be a promising target for hepatocellular carcinoma therapeutics.

Comprehensive Analysis of the Jellyfish Chrysaora pacifica (Goette, 1886) (Semaeostomeae: Pelagiidae) with Description of the Complete rDNA Sequence.

Jinho Chae, Yoseph Seo, Won Bae Yu, Won Duk Yoon, Hye Eun Lee, Soo-Jung Chang, and Jang-Seu Ki (2018) The Scyphomedusae genus Chrysaora consists of highly diversified jellyfishes. Although morphological systematics of the genus has been documented over the past century, characterization of molecular taxonomy has been attempted only recently. In the present study, we sequenced an 8,167 bp region, encompassing a single ribosomal DNA (rDNA) repeat unit, from Chrysaora pacifica, and used it for phylogenetic analyses. The tandemly repeated rDNA units turned out to consist of both coding and noncoding regions, whose arrangement was found to be the same as that of a typical eukaryote. None of the 5S rRNA sequences were found among the repeat units. Comparative analyses of jellyfish rDNA sequences showed that the 28S locus is highly informative and divergent compared to the 18S locus. Phylogenetic analyses of the 18S and 28S loci revealed that the Semaeostomeae order of jellyfish is separated into taxonomic groups by families and genera, with a few exceptions. The family Pelagiidae was in a clade separate from other groups, thus forming a monophyletic lineage. All Chrysaora included here formed a strongly supported clade within the family Pelagiidae, and Pelagiidae manifested a sister relationship with Cyanea. Nonetheless, Chrysaora was found to be paraphyletic in both 18S and 28S phylogenies. Chrysaora pacifica was clearly distinct from close relatives C. melanaster and C. quinquecirrha. These results provide a special reference for the DNA taxonomy of Pelagiidae jellyfishes in terms of nuclear cistron rDNA sequences and improve our understanding of the molecular phylogenetic relationships among Semaeostomeae jellyfishes.

A short note on a euphausiid, Euphausia pacifica, an important food source to demersal fishes in Uljin area, the eastern coast of Korea.

Gut contents of two species of demersal fishes, Pleurogrammus azonus and Dasycottus steiger were examined to understand the importance of a euphausiid, Euphausia pacifica as prey of demersal fishes at coastal waters off Uljin, the eastem Korea where aggregations of large amounts of the euphausiids were found. Euphausiids were significantly important food source to the species of fishes examined. All of the stomachs of P. azonus examined contained euphausiids (% of a food item frequency occurred, F = 100%) in March and June 2003. Individual numbers of euphusiids in the guts ranged from 10 to 540 individuals per gut. Percents of individual number of food item were 99.60 and 99.96 for euphausiids in March and June 2003, respectively The guts of D. steiger also frequently contained euphausiids (F = 86.67 and 79.20% in October 2002 and June 2003, respectively), while amphipods (F = 36.67% and 16.17%,) and other fishes (F = 33.33% and 45.80%) were also often observed. Pleurogrammus azonus preferred larger euphausiids with size range 16.9-28.4 mm, while the range of euphausiids distributed in the ambient water column was much broader (from eggs to adults).

Distribution of a pelagic tunicate, Salpa fusiformis in warm surface current of the eastern Korean waters and its impingement on cooling water intakes of Uljin nuclear power plant.

Impingement of a large amount of gelatinous plankton, Salpa fusiformis on the seawater intake system-screens in a nuclear power plant at Uljin was firstly recorded on 18th June 2003. Whole amount of the clogged animals was estimated were presumptively at 295 tons and the shortage of cooling seawater supply by the animal clogging caused 38% of decrease in generation capability of the power plant. Zooplankton collection with a multiple towing net during the day and at night from 5 to 6 June 2003 included various gelatinous zooplanktons known to be warm water species such as salps and siphonophores. Comparatively larger species, Salpa fusiformis occupied 25.4% in individual density among the gelatinous plankton and showed surface distribution in the depth shallower than thermocline, performing little diel vertical migration. Temperature, salinity and satellite data also showed warm surface current predominated over the southern coastal region near the power plant in June. The results suggested that warm surface current occasionally extended into the neritic region may transfer S. fusiformis, to the waters off the power plant. The environmental factors and their relation to ecobiology of the large quantity of salpa population that are being sucked into the intake channel of the power plant are discussed.

Comparing the distribution of ciliate plankton in inner and outer areas of a harbor divided by an artificial breakwater.

The distribution of ciliate plankton was compared between inner and outer areas of a harbor divided by an artificial breakwater in Kuryongpo, on the eastern coast of Korea, from February 2001 to October 2003. Less dissolved oxygen and higher concentrations of nitrogenous nutrients and phosphate were observed in the inner area. The abundance of oligotrich ciliates peaked in February 2001, when nanoflagellates bloomed in the inner area. The photosynthetic ciliate Mesodinium rubrum showed differing population dynamics annually, with blooming peaks in October 2001 in the inner area and in February 2003 in the outer area. The tintinnid species Tintinnopsis beroidea and Helicostomella subulata were generally more abundant in the outer area. Total ciliates were significantly related to oligotrich abundance in the inner area, and to tintinnid abundances in the outer area. Ciliate distribution showed quantitative and qualitative differences between the inner and outer areas in connection with the distribution of other plankton communities: oligotrich ciliate abundance increased with nanoflagellate blooms; dominance of M. rubrum was consistent with blooming of micro-sized phytoplankton (diatoms and dinoflagellates); large-sized tintinnids concurred with small dinoflagellates; and ciliate abundances decreased with mesozooplankton increases. The results indicate that the breakwater induces eutrophication in the inner area and provides suitable conditions for nanoflagellate blooms, which serially trigger opportunistic increases in oligotrich ciliates.