Jellyfish Cyanea Nozakii Research Articles

Identification and characterization of the key lethal toxin from jellyfish Cyanea nozakii

Jellyfish Cyanea nozakii venom is a complex mixture of various toxins, most of which are proteinous biological macromolecules and are considered to be responsible for clinical symptoms or even death after a severe sting. Previous transcriptome and proteome analysis identified hundreds of toxins in the venom, including hemolysins, C-type lectin, phospholipase A2, potassium channel inhibitor, metalloprotease, etc. However, it is not clear which toxin in the venom plays the most important role in lethality. Herein, we isolated the key lethal toxin (Letoxcn) from jellyfish Cyanea nozakii using anion exchange chromatography, size-exclusion chromatography, and cation exchange chromatography. The molecular weight of Letoxcn is ∼50 kDa with the N-terminal sequences of QADAEKVNLPVGVCV. Peptide mass fingerprinting analysis of Letoxcn shows that it may have some motifs of phospholipase, metalloproteinase, thrombin-like enzyme, potassium channel toxin, etc. However, only metalloproteinase activity but no hemolytic, PLA2, or blood coagulation activity was observed from in vitro toxicity analysis. Overall, this study uncovered and characterized the key lethal toxin in the venom of jellyfish Cyanea nozakii, which will not only help to reveal the molecule mechanism of the lethality, but also develop effective treatment like antivenom for this jellyfish sting in the future.

Evaluation of the complete nuclear rDNA unit sequence of the jellyfish Cyanea nozakii Kishinouye (Scyphozoa: Semaeostomeae) for molecular discrimination

ABSTRACT The harmful jellyfish Cyanea nozakii Kishinouye has frequently occurred on Korean coasts, and its blooms have caused serious ecological and economic damages. DNA sequences of the C. nozakii for molecular detection and discrimination are relatively scarce. In this study, we determined the complete sequence of a single unit of tandemly repeated ribosomal DNA (rDNA) of the Korean C. nozakii and characterized the molecular features of the rDNA. The complete rDNA contained 8,003 bp (48.4% GC) with the same gene arrangement (18S, ITS1, 5.8S, ITS2, 28S, and IGS) to the typical eukaryotes. Dot plot analysis showed that the coding regions (18S, 5.8S, and 28S) were highly conserved, while the non-coding regions (ITS1, ITS2, and IGS) were more variable and parsimony-informative. The IGS contained a putative transcription termination signal (poly(T) tract) and four repeats of block minisatellites. Phylogenetic analyses using 18S and 28S rDNA revealed well-resolved relationships of C. nozakii within the order Semaeostomeae, separating it from other Cyanea species. The complete rDNA sequence provides various options for the selection of jellyfish taxonomic markers and may be useful for discriminating between species of C. nozakii and phylogeny reconstruction with close relatives.

Updated descriptions of the nematocysts of the scyphozoan jellyfish Cyanea nozakii Kishinouye, 1891 (Cnidaria, Scyphozoa)

Nematocysts are typical cnidarian organelles that can discharge and release venom under physicochemical stimuli for predation and defense. Cyanea nozakii Kishinouye, 1891, a dominant jellyfish-blooming species in Chinese coastal waters, possesses numerous stinging nematocysts on its tentacles, and causes many envenomations every year. However, detailed taxonomic information of nematocysts in C. nozakii is elusive. In the present study, the morphological characteristics of nematocysts from C. nozakii were examined by combined light and scanning-electron microscopy. Five nematocyst types were revealed in the cnidome of C. nozakii, including common nematocyst types identified in Scyphozoa: atrichous isorhizas (a-atrich/O-atrich), microbasic euryteles, and microbasic birhopaloids type II. Importantly, two seldom reported types, microbasic b-mastigophores and microbasic birhopaloids type I, were also found, for the first time, in the cnidome of C. nozakii. This study contributes to understanding of the cnidome of C. nozakii, and the present study strongly suggests that the nematocysts of Scyphozoa are more diverse and complex than previously reported, which sheds new light on the nematocyst types in Scyphozoa species.

Isotopic evidence for size-based dietary shifts in the jellyfish Cyanea nozakii in the northern East China Sea

Abstract Cyanea nozakii is a common bloom-forming Scyphomedusa in coastal waters of China. To understand the diet of this jellyfish and its trophic relationship with other zooplankton groups, stable isotope δ13C and δ15N values of C. nozakii in the bloom area within the northern East China Sea (ECS) in August 2017 were analyzed. We examined: first, the size-based variation in C. nozakii stable isotope values, second, the contributions of different food sources to the C. nozakii diet, and possible size-based dietary shifts in individual C. nozakii and, third, the contribution of gelatinous prey to the C. nozakii diet. Isotope values of differently sized C. nozakii ranged from −18.81 to −16.88‰ for δ13C and from 8.23 to 10.46‰ for δ15N. A significant change in δ15N values implies that the trophic position and the diet of C. nozakii changed as body size increased. The MixSIAR model indicated that C. nozakii underwent a dietary shift during growth, from a seston-dominant diet in smaller medusae to a zooplankton-dominant diet in larger medusae. Copepods were an important food source for C. nozakii, irrespective of size. The mean proportions of gelatinous zooplankton increased significantly in the C. nozakii diet with increasing C. nozakii diameter, suggesting that larger C. nozakii consumed more gelatinous prey. The diet of larger C. nozakii included 9.54% Nemopilema nomurai and 9.53% small medusae. The results suggest that intraguild predation among these jellyfish exist in the northern ECS.

Proteomics, transcriptomics and toxicity analysis of venom proteins from jellyfish Cyanea nozakii

Proteomics, transcriptomics and toxicity analysis of venom proteins from jellyfish Cyanea nozakii

Combined Proteome and Toxicology Approach Reveals the Lethality of Venom Toxins from Jellyfish Cyanea nozakii.

Jellyfish are a type of poisonous cnidarian invertebrate that secrete lethal venom for predation or defense. Human beings often become victims of jellyfish stings accidentally while swimming or fishing and suffer severe pain, itching, swelling, inflammation, shock, and even death. Jellyfish venom is composed of various toxins, and the lethal toxin is the most toxic and hazardous component of the venom, which is responsible for deaths caused by jellyfish stings and envenomation. Our previous study revealed many toxins in jellyfish venom, including phospholipase A2, metalloproteinase, and protease inhibitors. However, it is still unknown which type of toxin is lethal and how it works. Herein a combined toxicology analysis, proteome strategy, and purification approach was employed to investigate the lethality of the venom of the jellyfish Cyanea nozakii. Toxicity analysis revealed that cardiotoxicity including acute myocardial infarction and a significant decrease in both heart rate and blood pressure is the primary cause of death. Purified lethal toxin containing a fraction of jellyfish venom was subsequently subjected to proteome analysis and bioinformation analysis. A total of 316 and 374 homologous proteins were identified, including phospholipase A2-like toxins and metalloprotease-like toxins. Furthermore, we confirmed that the lethality of the jellyfish venom is related to metalloproteinase activity but without any phospholipase A2 activity or hemolytic activity. Altogether, this study not only provides a comprehensive understanding of the lethal mechanism of jellyfish venom but also provides very useful information for the therapeutic or rescue strategy for severe jellyfish stings.

Comparative study of the hemolytic and cytotoxic activities of nematocyst venoms from the jellyfish Cyanea nozakii Kishinouye and Nemopilema nomurai Kishinouye

Two species of jellyfish, Cyanea nozakii Kishinouye and Nemopilema nomurai Kishinouye, have occurred offcoastal areas of the northeastern China Sea, Yellow Sea, and Bohai Sea in recent years. They influence marine ecosystem safety and fishery production, and also pose a risk to human health. The current study examined the hemolytic and cytotoxic activities of crude venoms extracted from the nematocysts of C. nozakii and N. nomurai. The results showed that there were more nematocysts on tentacles from C. nozakii than on tentacles of the same length from N. nomurai. The protein concentration per nematocyst extracted from N. nomurai was higher than that from C. nozakii. Both nematocyst venoms showed dose- and timedependent hemolytic activity on erythrocytes from chicken, pigeon, and sheep, with sheep erythrocytes being the most sensitive, with EC50 values of 69.69 and 63.62 μg/mL over a 30-min exposure with N. nomurai and C. nozakii nematocyst venoms, respectively. A cytotoxic assay of both jellyfish venoms on A431 human epidermal carcinoma cells resulted in IC50 values of 68.6 and 40.9 μg/mL after 24-h incubation, respectively, with venom from C. nozakii showing stronger cytotoxic activity than that from N. nomurai. The results of current study indicate that nematocyst venom from C. nozakii had stronger hemolytic and cytotoxic activities than that from N. nomurai and, thus, C. nozakii might be more harmful to the health of humans and other species than are N. nomurai when they appear in coastal waters.

The complete mitogenome of ghost jellyfish Cyanea nozakii (Cnidaria, Semaeostomeae, Cyaneidae)

The cnidarian jellyfishes are impressive organisms to show animal mitochondrial genomic diversities. Their mitogenome structure is linear and tRNA content has one or two in numbers, which is highly different than other metazoans. In this study, a complete mitogenome of the ghost jellyfish Cyanea nozakii (Cnidaria, Semaeostomeae, Cyaneidae) was sequenced and analyzed. The mitgenome is 17,381 bp long with 38.5% A, 16.0% C, 13.9% G, and 31.6% T nucleotide distributions. In addition, phylogenetic relationship of C. nozakii in the class Scyphozoa was investigated by using mitochondrial protein coding genes. Due to results, C. nozakii was positioned in the paraphyletic order Semaeostomeae. This is the first complete mitogenome from the genus Cyanea.

Investigation into the hemolytic activity of tentacle venom from jellyfish Cyanea nozakii Kishinouye

Cyanea nozakii Kishinouy e (C. nozakii), a giant cnidarian of the class Scyphomedusae, order Semaeostomeae and family Cyaneidae, is widely distributed in the East China Sea, the Yellow Sea and the Bohai Sea, and is abundant from late summer to early autumn. Venom produced by C. nozakii during mass agglomerations can contaminate seawater resulting in death of the halobios and seriously damage commercial fisheries. Swimmers and fishermen commonly suff er painful stings from this jellyfish, resulting in local edema, tingling, breathing difficulties, depressed blood pressure and even death. Such effects arise from the complex mixture of biologically active molecules that make up jellyfish venom. In the present study, the hemolytic activity of venom from tentacles of C. nozakii and factors aff ecting its activity were assayed. The HU50 ( defined as the amount of protein required to lyse 50 % of erythrocytes) of the venom against dove and chicken erythrocytes was 34 and 59 μg/mL, respectively. Carboxylmethyl chitosan and glycerol could increase hemolytic activity at concentrations greater than 0.06% and 0.2 mol/L, respectively.

Development of a rapid assay to detect the jellyfish Cyanea nozakii using a loop-mediated isothermal amplification method

Blooms of the harmful jellyfish Cyanea nozakii, which are a severe nuisance to fisheries and tourisms, frequently occur in the northern East China Sea, Yellow Sea, and Bohai Sea. To provide early warning of this species, a simple and effective molecular method for identifying C. nozakii was developed using the loop-mediated isothermal amplification method (LAMP). The LAMP assay is highly specific and uses a set of four primers that target six different regions on the mitochondrial cytochrome c oxidase subunit I (COI) gene of C. nozakii. The amplification conditions, including the dNTP and betaine concentrations, the inner primer to outer primer concentration ratio, reaction time and temperature, were optimized. The LAMP assay amplified DNA extracted from tissue samples of C. nozakii but did not amplify DNA from other common scyphozoans and hydrozoans collected in the same region. In addition, the LAMP assay was more sensitive than conventional PCR. Therefore, the established LAMP assay is a sensitive, specific, fast, and easily performed method for detection of C. nozakii at different stages in their life cycle.

Cytotoxicity of the venom from the nematocysts of jellyfish Cyanea nozakii Kishinouye

In this article, the cytotoxicity of the venom from the nematocysts of jellyfish Cyanea nozakii Kishinouye on human hepatoma cells (Bel-7402, SMMC-7721) and human colon cancer cells (H630) was investigated first. Of the three kinds of cells, the venom had the strongest cytotoxicity on H630 cells with the 50% lethal concentration (IC(50)) of 5.1 μg/ml. However, the IC(50) on Bel-7402 and SMMC-7721 was approximate 17.9 and 24.3 μg/ml, respectively. The cytotoxicity of the venom was affected by pH, temperature and storage conditions. At the pH 4.5-8.5, the venom displayed obvious cytotoxicity and the percentage of survival was about 50%. When pre-incubated at temperatures over 60°C for as short as 10 min, the percentage of survival sharply improved from 4.6% up to 80%. The venom was stored in a more stable condition at -80°C and in lyophilized state compared to other storage conditions used in this study. Lactate dehydrogenase release assay performed on H630 cells indicated that exposure to the venom could result in damage to the cell membrane.

Two-step purification and in vitro characterization of a hemolysin from the venom of jellyfish Cyanea nozakii Kishinouye

Hemolysin is one of the most hazardous components in the venom of Cyanea nozakii Kishinouye. Here we describe the purification and in vitro characterization of the hemolysin, which we named CnPH. The CnPH was isolated by anion-exchange and size-exclusion chromatography from the nematocyst venom. Two protein bands with molecular masses of 20kDa, 60kDa respectively were shown in the reducing SDS–PAGE analysis of the CnPH. And Approximately 5μg/mL of the CnPH resulted in 50% hemolysis of the erythrocyte suspension. The hemolytic activity of the CnPH was both temperature and pH dependent. Moreover, it was significantly inhibited in the presence of divalent metal cations, including Cu2+, Mg2+, Mn2+, Zn2+ and Ca2+, but enhanced in the presence of EDTA. However, how CnPH performs its hemolytic activity is not yet clear, therefore the mechanism of the hemolytic activity of the CnPH is under research.

Partial characterization of the hemolytic activity of the nematocyst venom from the jellyfish Cyanea nozakii Kishinouye

Using a recently developed technique to extract jellyfish venom from nematocysts, the present study investigated the hemolytic activity of Cyanea nozakii Kishinouye nematocyst venom on chicken erythrocytes. Venom extract caused a significant concentration-dependent hemolytic effect. The extract could retain its activity at −80 °C but was unstable when kept at 4 °C and −20 °C for 2 days. The hemolytic activity was inhibited by heating within the range of 37–100 °C. The extract was active over a pH range of 5.0–8.63 and the pH optima for the extract was 7.8. Incubation of the venom with sphingomyelin specially inhibited hemolytic activity by up to 70%. Cu 2+ and Mn 2+ greatly reduced the hemolytic activity while Mg 2+, Sr 2+ and Ba 2+ produced a relatively low inhibiting effect on the hemolytic activity. Treatment with Ca 2+ induced a concentration-dependent increase in the hemolytic activity. In the presence of 5 mM EDTA, all the hemolytic activity was lost, however, the venom containing 1.5 mM EDTA was stable in the long-term storage. PLA 2 activity was also found in the nematocyst venom of C. nozakii. These characteristics provide us a fundamental knowledge in the C. nozakii nematocyst venom which would benefit future research.

Isolation and characterization of lethal proteins in nematocyst venom of the jellyfish Cyanea nozakii Kishinouye

Cyanea nozakii Kishinouye, a jellyfish widely distributed in coastal areas of China, has garnered attention because of its stinging capacity and the resulting public health hazard. We used a recently developed technique to extract jellyfish venom from nematocysts; the present study investigates the lethality of C. nozakii venom. The nematocyst contents were extremely toxic to the grass carp, Ctenopharyngodon idellus, producing typical neurotoxin toxicity. The ID 50 was about 0.6 μg protein/g fish. Toxin samples were stable when kept at −80 °C, but after 48 h, an 80% decline in lethality occurred at −20 °C. Poor stability of the venom was observed within the range of 65–80 °C and at pH 3.5. The venom was hydrolyzed by a proteolytic enzyme, trypsin. Fractionation of the venom yielded two protein bands with molecular weights of 60 kDa and 50 kDa. Our results provide the first evidence that C. nozakii produces lethal toxins. These characteristics highlight the need for the isolation and molecular characterization of new active toxins in C. nozakii.

Comparison of life cycles and morphology of Cyanea nozakii and other scyphozoans

Mass aggregations of the giant jellyfish Cyanea nozakii have occurred in China since the end of the 20th century. In particular, C. nozakii bloomed abnormally in Liaodong Bay during July and August of 2004 and the catch of edible Rhopilema esculentum was greatly reduced. In order to clarify the causes leading to mass occurrences of C. nozakii, the reproductive cycle and morphological characters of this species were compared with other common jellyfish species using data from both laboratory–rearing experiments and field investigations. The results showed that, in particular, settling of planulae and asexual reproductive strategies of scyphystomae made C. nozakii more capable of thriving under unfavorable physical conditions than R. esculentum. The ephyrae of Nemopilema nomurai, R. esculentum, C. nozakii, Aurelia aurita, Rhopilema hispidum and Rhopilema asamushi were differentiated by different shapes of lappets, rhopalar clefts, gastric filaments and nematocyst batteries. In addition, the life cycle of C. nozakii was compared with earlier reports on the life cycle of Cyanea capillata, R. esculentum, Rhizostoma pulmo, Stomolophus meleagris and N. nomurai, and the morphology of adult C. nozakii was compared and contrasted with species of the genus Cyanea (C. capillata, C. ferruginea and C. purpurea).