Factors affecting the protease activity of venom from jellyfish Rhopilema esculentum Kishinouye

In this paper, the effects of some chemical and physical factors such as temperature, pH values, glycerol, and divalent metal cations on the protease activity of venom from jellyfish, Rhopilema esculentum Kishinouye, were assayed. Protease activity was dependent on temperature and pH values. Zn²⁺, Mg²⁺, and Mn²⁺ in sodium phosphate buffer (0.02 M, pH 8.0) could increase protease activity. Mn²⁺ had the best effects among the three metal cations and the effect was about 20 times of that of Zn²⁺ or Mg²⁺ and its maximal protease activity was 2.3 × 10⁵ U/mL. EDTA could increase protease activity. PMSF had hardly affected protease activity. O-Phenanthroline and glycerol played an important part in inhibiting protease activity and their maximal inhibiting rates were 87.5% and 82.1%, respectively.     

Studies on the hemolytic activity of tentacle extracts of jellyfish Rhopilema esculentum Kishinouye: application of orthogonal test

The present work is first reporting the hemolytic activity of venom from jellyfish Rhopilema esculentum Kishinouye extracted by different phosphate buffer solutions and incubated at different temperature according to the orthogonal test L6(1) x 3(6). Of the seven controllable independent variables, incubated temperature and phenylmethylsulfonyl fluoride (PMSF) had strongest effect on the hemolytic activity.     

Radical scavenging activity of protein from tentacles of jellyfish Rhopilema esculentum

In this study, radical scavenging activity of protein from tentacles of jellyfish Rhopilema esculentum (R. esculentum) was assayed including superoxide anion radical and hydroxyl radical scavenging. The protein samples showed strong scavenging activity on superoxide anion radical and values EC50 of full protein (FP), first fraction (FF), second fraction (SF), and 30% (NH4)2 SO4 precipitate (Fr-1) were 2.65, 7.28, 1.10, and 22.51 microg/mL, respectively, while values EC50 of BHA, BHT, and alpha-tocopherol were 31, 61, and 88 microg/mL, respectively. Also, the protein samples had strong scavenging effect on hydroxyl radical and the values EC50 of FP, FF, SF, Fr-1, and Fr-2 were 48.91, 27.72, 1.82, 16.36, and 160.93 microg/mL, but values EC50 of Vc and mannitol were 1907 and 4536 microg/mL, respectively. Of the five protein samples, SF had the strongest radical scavenging activity and may have a use as a possible supplement in the food and pharmaceutical industries. The radical scavenging activity was stable at high temperature so that R. esculentum may be used as a kind of natural functional food.     

从海蜇中筛选抗真菌活性的海洋细菌菌株

从中国北部湾海域采集的海蜇样品中分离到577株海洋细菌,以条件致病真菌白色念珠菌(Candida albicans)作为敏感测试菌株,采用琼脂块法测定了577株海洋细菌的抗真菌活性,结果表明有119株具有抗白色念珠菌活性,占总数的20.6%。对其中的40株高活性菌株进行抗真菌和抗细菌活性研究,结果表明,有21株海洋细菌对3种或3种以上的真菌有抑制活性,23株对细菌有不同程度的抑制作用,其中1株海洋细菌I0s4-18具有广谱的抗菌作用,对革兰氏阴性细菌、革兰氏阳性细菌、酵母菌及丝状真菌均具有抑制作用。海蜇共附生的微生物中能产生抗真菌活性的微生物的比例是很高的,I0s4-18菌株具有良好的开发前景。     

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 20 kDa, 60 kDa respectively were shown in the reducing SDS-PAGE analysis of the CnPH. And Approximately 5 μg/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.     

基于碳氮稳定同位素技术研究辽东湾海蜇的食性和营养级

应用稳定同位素技术分析了渤海辽东湾海蜇及其可能摄食饵料生物的碳、氮稳定同位素比值,对其食物组成和营养级进行了研究.结果表明： 辽东湾海蜇的δ¹³C值范围是-20.27‰～-23.06‰,跨度为2.79‰,平均值为-21.33‰;δ¹⁵N值范围是6.82‰～10.03‰,跨度为3.21‰,平均值为8.25‰.海蜇的饵料生物主要包括悬浮物、浮游植物、鱼卵、≤1000 μm 浮游动物、1000～1500 μm 浮游动物和＞1500 μm 浮游动物,其中以≤1000 μm 浮游动物为主,贡献率高达71%～88%;其他饵料生物的重要性由高到低依次为>1500 μm浮游动物、1000～1500 μm浮游动物、悬浮物、浮游植物和鱼卵,其平均贡献率分别为6%～19%、0%～22%、0%～10%、0%～8%、0%～2%.经检验发现,海蜇伞径长与δ¹⁵N的相关性不显著(P >0.05),而与δ¹³C呈显著负相关(P<0.05).以悬浮物为基准的辽东湾不同伞径组海蜇的营养级范围为2.79～3.88,平均营养级为3.28.本研究结果说明,海蜇作为大型浮游动物在辽东湾生态系统中对小型浮游动物的能量传递起着关键的调控作用,研究结果将为深入研究辽东湾生物群落的营养结构及整个生态系统的物质循环与能量流动提供基础资料.     

Monoclonal antibodies neutralizing the haemolytic activity of box jellyfish (Chironex fleckeri) tentacle extracts

• 1.1. Three monoclonal antibodies have been produced which neutralize in vitro the haemolytic activity present in tentacle extracts of the box jellyfish (Chironex fleckeri).
• 2.2. Two of these monoclonal antibodies bound specifically to a component of relative molecular mass 50,000 in tentacle extract on Western blots.
• 3.3. This binding only occurred when the extracts were electrophoresed under non-reducing conditions.
• 4.4. The third monoclonal antibody did not display binding to Western blots of tentacle extract under any of our experimental conditions.

     

海蜇胶原蛋白的提取及纯化(英文)

通过胃蛋白酶的限制性酶切、选择性盐析及阳离子交换柱的层析作用,低温从海蜇中胶层提取可得到胃蛋白酶促溶且去除端肽(telopeptide)的胶原蛋白(PSC)。紫外-可见扫描测得PSC的最大吸收峰位于235nm。凝胶电泳及氨基酸分析显示这些都符合Ⅰ型胶原的特征,同水母S.nomurai和Rhopilema asamushi来源的胶原蛋白基本一致,作为一个低等动物,其氨基酸分析显示了脊椎动物和非脊椎动物的微小差异。     

Glycaemic effects of tentacle extract of the jellyfish Acromitus rabanchatu (Annandale)

Tentacle extract of the common jellyfish A. rabanchatu, caused glycaemic alteration in fasting rabbits. Intravenous administration of the extract produced a significant rise followed by a significant fall in blood sugar level. Glucose tolerance in rabbits was also significantly increased. Extract-mediated hypoglycaemic response was fully abolished in alloxan diabetic rabbits. Preliminary separation on Sephadex G 50 indicated the hypoglycaemic factor to be a non-lethal protein of molecular weight less than 20 kDa. Heat treatment of extract and G 50 separated fraction P2 demonstrated total loss of hypoglycaemic activity.     

水母胶原蛋白抗氧化活性研究

提取水母胶原蛋白,观察其体内外抗氧化作用。体外采用邻苯三酚自氧化法观察水母胶原蛋白对超氧阴离子(·O2-)的清除作用,采用Fenton反应体系观察对羟自由基(·OH)的清除作用;体内观察水母胶原蛋白对小鼠血清、肝脏、大脑中超氧化物歧比酶(SOD)、过氧化氢酶(CAT),丙二醛(MDA)含量的影响。结果表明:水母胶原蛋白对·O2-、·OH具有良好的清除作用,呈一定的剂量依赖关系,能明显提高小鼠血清、肝脏、大脑中的SOD、CAT含量,能在一定程度上降低血清、肝脏、大脑中的MDA水平。因此,水母胶原蛋白具有显著的抗氧化生物活性。     

Stock enhancement of the edible jellyfish (<Emphasis Type="Italic">Rhopilema esculentum</Emphasis> Kishinouye) in Liaodong Bay,China: a review

Among the edible species, jellyfish Rhopilema esculentum, is one of the most abundant and important fishery species in China. The jellyfish fishery is characterized by considerable fluctuations in catch and a very short fishing season. In this article, we first review the research results on the biology of R. esculentum, which previously were published in Chinese, as related to the jellyfish enhancement and fishery. Next, we review results from enhancement experiments conducted from 1984 to 2004, with the aims of stabilizing and increasing catch. During 2005 and 2006, stock enhancement of R. esculentum was carried out on a large scale for the first time in Liaodong Bay, China, where 414 million juvenile jellyfish (umbrella diameter > 1 cm) were released. We present results of these enhancements, including the survey methods, catch prediction, enhancement assessment, and fishery management. In 2005 and 2006, the recapture rate of released jellyfish was 3.0 and 3.2%, respectively. The fishermen earned ￥ 159 million during the 2 years. The ratio of the input (cost of culturing juvenile jellyfish) to the output (value of the sales) was about 1:18. The high commercial value of R. esculentum enhancement in Liaodong Bay makes this a very successful enterprise. Guest editors: K. A. Pitt & J. E. Purcell Jellyfish Blooms: Causes, Consequences, and Recent Advances     

海蜇(Rhopilema esculentum)even-skipped基因的克隆及生物信息学分析

Even-skipped(eve)是一个含有同源框结构域的转录调控因子,在体节形成、神经分化、尾芽发育等过程中发挥着重要作用.为探究eve在钵水母变态发育中的作用,本研究以海蜇转录组测序获得的eve基因类似序列为基础,通过RACE技术获得了海蜇eve基因(ReEve)的cDNA全长,并已提交GenBank数据库,登录号...     

Nematocysts of Rhopilema nomadica (Scyphozoa: Rhizostomeae), an immigrant jellyfish in the eastern mediterranean

Rhopilema nomadica—a recently discovered scyphomedusa in the eastern Mediterranean—is considered a lessepsian migrant. Its nematocysts were extracted from the scapular and mouth-arm tentacles and examined using light and electron microscopy techniques. The morphometric parameters of the nematocysts were measured before and after complete discharge. Three categories of nematocysts were identified: heterotrichous isorhiza haploneme, holotrichous isorhiza haploneme, and heterotrichous microbasic eurytele. The relative abundance of the nematocysts and their occurrence in tissues of the jellyfish were noted. A brief discussion concerning the classification of certain types of nematocysts is given. A comparison with the available data on other Rhopilema species revealed that the nematocyst categories of R. nomadica are more similar to those of the Atlantic R. verrilli than to those of the Western Pacific R. esculentum. A brief comparison of the injuries caused by these species is given. © 1995 Wiley-Liss, Inc.     

骆驼蓬醇提取物杀虫活性初步研究

骆驼蓬全草醇粗提物分为石油醚、氯仿、乙酸乙酯、正丁醇、水溶性5部分,提取得率分别为4.6%、1.7%、0.3%、4.8%、7.6%.分别配成2.0g·L-1至0.1g·L-1一系列梯度浓度,采用浸叶法进行小麦蚜虫的杀灭活性试验,48h后,结果表明,浓度越高,活性越大,其中在2.0g·L-1处理时水部分和氯仿部分活性非常明显,小麦蚜虫校正死亡率可达95.46%、90.83%.另外化学成分系统预试结果表明茎叶含有丰富的生物碱,黄酮,酚性成分等.     

灰毛豆甲醇提取物的杀虫活性

研究灰毛豆(Tephrosia purpurea)各部位提取物的杀虫活性及其作用方式。结果表明,灰毛豆对白纹伊蚊Aedes albopictus Skuse幼虫、菜青虫Pieris rapae(L.)、斜纹夜蛾Prodenialitura(Fabricius)幼虫和黄曲条跳甲Phyllotreta striolata(Fabricius)成虫都有杀虫活性。灰毛豆种子、树皮、根皮、豆荚、枝条和树叶的甲醇提取物对白纹伊蚊4龄幼虫24h的LC50值分别是22.1,97.7,36.6,142.6,165.6和618.3mg.L-1,树干木质部没有杀虫活性;灰毛豆种子、树皮和根皮甲醇提取物对3龄菜青虫24h的触杀毒力LC50值分别是232.1,206.3和236.7mg.L-1;种子、树皮、根皮、枝条、树叶和豆荚对3龄菜青虫的24h胃毒毒力LC50值分别是192.6,168.4,249.7,524.5,1001.0,和510.7mg.L-1。在取食或接触有提取物的叶碟后,5龄菜青虫会出现取食量减少和生长发育变慢的亚致死现象。这些研究结果表明,灰毛豆除茎干木质部以外的其它各部位均含有杀虫活性成分,其作用方式为胃毒和触杀。     

Insecticidal activity of recombinant baculovirus co-expressing Bacillus thuringiensis crystal protein and Kunitz-type toxin isolated from the venom of bumblebee Bombus ignitus

To improve the insecticidal activity of Autographa californica nucleopolyhedrovirus (AcMNPV), using co-expression of Bacillus thuringiensis crystal protein and a Kunitz-type toxin isolated from bumblebee Bombus ignitus venom, a recombinant AcMNPV, ApPolh5-3006BiKTI, expressing Bi-KTI under the control of early promoter from Cotesia plutellae bracovirus (CpBV) was constructed. In this recombinant virus, B. thuringiensis cry1-5 crystal protein gene was introduced into the genome by the fusion of polyhedrin-cry1-5 under the control of polyhedrin gene promoter. RT-PCR analysis indicated that both Bi-KTI and polyhedrin-cry1-5 fusion protein were successfully expressed from the infected cells. In addition, SDS-PAGE revealed that polyhedrin-cry1-5 fusion protein expressed by recombinant viruses was occluded into the polyhedra. ApPolh5-3006BiKTI showed an improved insecticidal activity against larvae of Plutella xylostella and Spodoptera exigua. At low dosage rates, it was more effective against S. exigua than on P. xylostella, but more rapid insecticidal activity was shown in P. xylostella. These results strongly suggest that co-expression of Bt toxin and Kunitz-type toxins could be successfully applied to improve the insecticidal activity of baculoviruses.     

Pharmacological actions of tentacle extract of the jellyfish, Acromitus rabanchatu, occurring in the Bay of Bengal

Tentacle extract of A.rabanchatu, produced a fall of blood pressure in cat, rat and guinea pig. Hypotension produced in cat remained unantagonized by blockers of acetylcholine, histamine and 5-HT. On isolated guinea pig heart, the extract significantly reduced the rate and amplitude of contraction leading to irreversible cardiac arrest. In cats and rats, the respiratory rate and amplitude was decreased significantly and resulted in temporary apnoea. The extract also produced vasoconstriction in perfused rat hindquarter preparation and increased cutaneous capillary permeability. The extract produced contraction in several isolated smooth muscle preparations. Contraction on guinea pig ileum was partly antagonized by atropine and cyproheptadine. On isolated rat phrenic nerve diaphragm and chick biventer cervicis, the extract produced irreversible blockade of the electrical stimulation-induced twitch responses. Haemolytic and myonecrotic activity was exhibited by the extract. LD50 was found to be 7.7 mg/kg (iv, mice).     

海蜇养殖对池塘底泥营养盐和大型底栖动物群落结构的影响

2009年5-9月,对荣成靖海湾大型养殖池塘海蜇(Rhopilema esculentum Kishinouye)养殖期间与养殖期前后底泥营养盐及大型底栖动物群落结构变化进行了研究。结果表明,7月份海蜇养殖期间,由于海蜇的避光性而产生的上下浮动的行为特征造成了对水体的扰动作用, 与海蜇的生物沉积作用共同导致养殖海区(实验点)与邻近非养殖海区(对照点)之间各项底泥营养盐指标均存在显著性差异,其中,实验点氨氮(NH₄-N)、硝氮(NO₃-N)和沉降速率(sedimentation rate, SR)显著高于对照点,实验点叶绿素a(Chla)、总有机物(TOM)和总有机碳(TOC)含量显著低于对照点。多变量聚类分析结果表明,海蜇养殖对养殖池塘的大型底栖动物群落结构产生显著影响,并且7月份实验点大型底栖动物生物多样性指数(H')和均匀度指数(J) 随海蜇放养显著增大,并显著高于对照点。大型底栖动物群落多样性指数与沉降速率(SR)和底泥TOM含量分别表现出显著正相关和负相关,而与其他营养盐指标无显著相关性。