海蜇毒素的提取及活性测定

利用液氮速冻、石英砂研磨破碎刺胞,用水溶液、醇溶液提取海蜇和沙蜇刺胞毒素。用Sepha-dex G-75柱层析分离得到2个峰,收集2个峰的液体冷冻干燥得到毒素。利用血平板快速检测毒素的活性。沙蜇毒素的溶血活性明显高于海蜇毒素。提取物中的活性成分的分子量约为10 000~60 000。这类毒素具有溶血、心脏毒性、神经毒性、肌肉毒性、皮肤坏死,影响离子转运以及胆碱酯酶活性等作用。     

Antibacterial and hemolytic effects of aqueous and organic extracts from different tissues of sea urchin <Emphasis Type="Italic">Echinometra mathaei</Emphasis> on pathogenic streptococci

In this study, the antibacterial and hemolytic effects of the gonadal tissue, test, spines and Aristotle’s lantern of the sea urchin (Echinometra mathaei) were investigated. The aerobic Gram-positive pathogenic bacteria Streptococcus mutans and Streptococcus sobrinus were selected because of their importance in oral diseases. The samples of sea urchin were collected from an intertidal zone in the Persian Gulf and then dissected. Finally, the crude extracts of the gonadal tissue, test, spines and Aristotle’s lantern of the samples were separately prepared by the solvents, including phosphate-buffered saline (PBS), 96 % ethanol (Et) and 80 % acetonitrile (ACN). The antibacterial activities of the extracts were evaluated using the well diffusion method in two concentrations of 1500 and 600 µg well^?1. The antibacterial effects against streptococcal species were observed in the organic extracts (Et and ACN) of the gonads and test while the extracts of spines and Aristotle’s lantern showed no antibacterial activity. The hemolytic activity of the PBS, 96 % Et, and 80 % ACN extracts from the sea urchin were evaluated using horse red blood cells. Hemolytic activity was observed only in the 80 % Et and ACN extracts of the gonad and test. In conclusion, the organic extracts of gonadal tissue showed the most inhibitory activity on the growth of S. mutans and S. sobrinus.     

Measurement of swimming speed of giant jellyfish <Emphasis Type="Italic">Nemopilema nomurai</Emphasis> using acoustics and visualization analysis

A species of giant jellyfish, Nemopilema nomurai, which has appeared only recently in the East China Sea, is an emerging nuisance in the northeastern region of Asia because of its extensive damage to fisheries. Until now, the biomass estimates of these jellyfish have mainly been obtained using trawl sampling and sighting survey methods. However, it is also necessary to determine the origin and diurnal migration patterns of these jellyfish. Drawbacks of the trawl sampling method are that it is effective only in estimating the density of jellyfish population distributed throughout the entire water column and requires considerable time. Another common analysis technique is the sighting method, which is effective only in the estimation of he density of jellyfish distributed in surface areas. The sighting method can determine distributions over wide areas in a short time. This method has limitations in investigating the vertical distribution and swimming behavior of jellyfishes. In our study, we utilized an echo sounding method extensively and effectively to overcome these limitations. Our method involved the use of a scientific echo sounder, acoustic camera, and conductivity-temperature-depth instrument during the drifting of a research vessel at various stations in the Yellow Sea. The acoustical method of particle tracking velocimetry (PTV) was used to analyze the swimming speed according to the vertical distribution of N. nomurai jellyfish. Results of the scientific echo sounder indicated that the jellyfish were mainly present in the water column from the surface up to a depth of 40 m. The mean swimming speed of the jellyfish was estimated as being 0.6 times the bell size (BS), with a tendency to maintain a certain speed. Further, results of a Monte Carlo simulation showed that the swimming speed was in the range of 0.46–0.89 BS. These results might be used as an index in a migration model, which may be useful to forecast the behavior and origin of the giant jellyfish entering inshore areas on a massive scale in northeastern Asia.     

安朵仙水母生活史的实验观察

安朵仙水母在固着的无性繁殖螅状体和浮游的有性繁殖水母体间交替,其生活史中绝大多数环节并不是自发进行的,而是受特定的物理、化学或生物因素的影响。静止水流和雌雄个体间的交互作用对于产卵和受精是非常重要的,受精卵在64h内可经浮浪幼虫阶段变态为4触手螅状体。螅状体能通过单碟型横裂产生水母体,但对温度和光照要求严格。出芽生殖是螅状体无性繁殖的唯一方式,出芽率与温度关系最为密切。大多数碟状体在横裂的过程中生长出口腕,并在脱离螅状体12～24h内翻转为口腕向上、外伞向下的倒立形态。水母体可通过食物和体内的共生藻获得能量。人工条件下,螅状体可长期保存,水母体可以饲养两年以上。     

Evaluation of antioxidant properties and biofunctions of polar,nonpolar, and water-soluble fractions extracted from gonad and body wall of the sea urchin Tripneustes gratilla

This study was performed to compare the various bioactivities of water-soluble, polar, and nonpolar extracts from the edible part (gonad) and waste (body wall) of the sea urchin Tripneustes gratilla. During antioxidant activity screening, these three extracts from both gonad and body wall showed various antioxidant activities in different tests. During human skin fibroblast (CCD966SK) viability and collagen screening, only water-soluble and polar extracts from gonad and body wall were observed to promote cell viability, while the water-soluble gonad extract promoted the collagen-generating activity of CCD966SK cells. The water-soluble and polar extracts from gonad and body wall showed proliferative activity towards different cell types. In contrast, the nonpolar extracts of gonad and body wall exerted antiproliferative effects on most tumor cells. These results indicate that the bioactivities of sea urchin extracts depend on the part of the urchin that the extract was taken from as well as the extraction method used.     

Food habits of fishes in the mangrove estuary of Urauchi River,Iriomote Island,southern Japan

The food habits of 67 fish species collected from the mangrove estuary of the Urauchi River, Iriomote Island, southern Japan were investigated using gut content analysis. Ontogenetic changes in food preference were recognized in nine species, including mugilids, gerreids, mullids, gobiids and tetraodontids. In most cases, juveniles of these species fed mostly on small crustaceans (e.g. calanoid and cyclopoid copepods and gammaridean amphipods) or detritus. With their subsequent growth, larger prey items (e.g. crabs and polychaetes) became dominant. A cluster analysis based on dietary overlaps showed that the mangrove fish assemblage comprised eight trophic groups (zooplankton, small benthic crustacean, large benthic crustacean, polychaete, fish, detritus, plant and insect feeders). Of these, large and small benthic crustacean feeders, which consumed mainly crabs and gammaridean amphipods, respectively, were the most abundantly represented in terms of species, whereas polychaete and insect feeders were each represented by only two species.     

4 种大型水母类ITS-5.8S rDNA 序列分析及其在钵水母类系统分析中的应用

由于钵水母类生物地理学研究的缺乏以及不同时期形态变异较大等原因,对其分类鉴定比较混乱和困难。为弥补形态学分类的缺陷,采用通用引物PCR扩增法,测定了分布于黄海北部和辽东湾海域海蜇(Rhopilema esculentum)、沙蜇(Nemopilema nomurai)、海月水母(Aurelia sp.)、白色霞水母(Cyanea nozakii)4种大型水母的ITS-5.8S rDNA序列,同时利用Gen Bank数据库中已有的钵水母纲(Scyphomedusae)ITS1(the Ribosomal First Internal Transcribed Spacer)同源序列对其进行序列分析并构建系统树,分析ITS1序列片段在大型水母种类鉴定方面的可行性及其在钵水母类系统及演化中的应用。结果显示,4种水母的ITS-5.8S rDNA序列变异较大且具有明显的序列长度多态性,序列长度范围675~833 bp。钵水母纲很多种类的ITS1序列具有种内长度多态性现象,这种长度多态性主要是由于微卫星重复次数不同所造成的。钵水母纲科间遗传距离为0.295~0.491,种间遗传距离为0.024~0.812;除白色霞水母和海蜇外,种内个体间遗传距离为0.000~0.099。采用ML法(maximum likelihood)和贝叶斯法(Bayesian)构建的分子系统树拓扑结构不完全相同且与形态分类学的观点不太一致。研究表明,ITS基因序列在钵水母纲不同阶元间变异较大,适合于钵水母纲种类鉴定和属内种间水平的系统进化研究。     

三峡库区26条支流浮游甲壳动物的群落结构

为了对三峡库区26条主要河流及长江干流浮游甲壳动物群落结构进行调查,2010年4月至2011年6月分四次对三峡库区主要河流进行浮游甲壳类进行定量和定性采集标本。152个采样点四次采样共鉴定出浮游甲壳动物50种,其中枝角类36种,桡足类14种。浮游甲壳类主要是以常见的广温性种类为主,四次调查中发现的种类均为长江流域常见种。三峡库区浮游甲壳类动物具有明显的季节性变化特征,四月份（春夏季）的存量和种类明显高于十月份（秋冬季）;也具有明显的纵向分布特征,相同支流上游的存量和种类一般低于回水区的数值。     

东海区夏、秋季大型水母分布区渔业资源特征分析

本文根据1999～2004年的夏季和秋季东海区大型水母和渔业资源的状况,分析了大型水母分布区内的渔业资源特征。结果表明大型水母夏季主要分布于30°30′～33°00′N间,秋季主要分布于32°00′N以北海域,并呈现由夏季到秋季逐渐往北转移的趋势;2003年是大型水母数量最高的年份,该年水母主要分布区内的渔业资源种类数最少,CPUE最低,Margalef种类丰富度指数也最低;2003年夏季水母高发区内渔获种类的优势度也较其它年份发生了显著变化,优势种变为黄;2003年水母高发区内带鱼、小黄鱼和银鲳的当龄鱼较少,银鲳的肥满度增加,小黄鱼的肥满度降低,而带鱼的肥满度未发生显著变化。本文对水母高发区内渔业资源变化的原因进行了初步解释,并对水母的防治措施提出了初步的建议。     

色素添加对咖啡金黄水母体色影响的初步研究

在人工繁殖水母的饵料中添加食用性人工合成色素、人工合成荧光色素、天然虾青素、铁元素和饲喂含有色素的海蜇等,测定水母采食饵料后感官指标、酪氨酸酶活力和总类胡萝卜素含量的变化,分析添加物质对水母生理机能的影响。试验结果显示,4种人工食用色素均能大大提升水母的观赏性,但颜色保持时间短,3 h后,所有水母均恢复原来的体色,水母状态未发现改变。水母注射水溶性荧光色素,10 h后才能通过水循环扩散到全身,荧光色维持时间可达30 d。水母饲喂天然虾青素、铁元素和含有色素的海蜇1个月后,肉眼观察,海蜇组的水母比对照组的水母颜色深,而虾青素组、FeSO_4组与对照组差异不明显。各组酪氨酸酶活力和总胡萝卜素含量均较低。组内个体间酪氨酸酶活力差异较大,但各组间差异均不显著(P>0.05)。总胡萝卜素在450 nm处吸收值最高。海蜇组水母的酪氨酸酶活力和总胡萝卜素含量均最高,分别为0.133 U/mL和251.01 mg/kg。虾青素组酪氨酸酶活力最低,为0.109 U/mL。食用色素组总胡萝卜素含量最低,仅为158.83 mg/kg,酪氨酸酶活力仅高于虾青素组,可能与色素并未在表皮和中胶层形成色素沉淀有关。试验结果表明,人工添加色素能获得颜色各异的水母,水溶性荧光色素能长期保存在动物体内,可作为体色调控色素。水母的色素合成与酪氨酸酶活力和总胡萝卜素含量有关。     

香菇多糖提取物体外抗IHNV的机制

本研究以虹鳟性腺细胞(rainbow trout gonad cell,RTG)为细胞模型,采用MTT法体外测定香菇多糖提取物的细胞毒性及其对传染性造血器官坏死病毒(IHNV)直接灭活、预防与生物合成的影响,探讨香菇多糖提取物抗IHNV的作用机制。结果显示:香菇多糖提取物对RTG细胞没有毒性作用。浓度为100μg/m L时,香菇多糖提取物对IHNV的直接灭活能力为52.48%,预防效果为50.38%,抑制IHNV生物合成的能力达84.41%。结果表明:香菇多糖提取物具有一定的抗IHNV作用,该作用主要与抑制IHNV在细胞内的生物合成有关。     

The vulnerability of fish and macroinvertebrate species with bioactive potential in a Mediterranean marine protected area

1. A high percentage of marine animals produce bioactive compounds that may play a leading role in the discovery of future compounds and drugs of marine origin. However, commercial fishing and other human activities leading to sea warming and pollution may affect these marine animals, even putting them in danger of extinction.
2. To date, no comprehensive studies have evaluated the conservation status of Mediterranean species with bioactive potential, which is crucial to better understanding of how these species cope with the impacts of human activity.
3. The study reviewed the bioactive potential and vulnerability of 833 fish and macro-invertebrate species inhabiting the marine protected area of Cap de Creus and surrounding areas. The most active taxa found were Porifera (49 out of 59 species; 83.0%) and Tunicata (17 out of 27 species; 63.0%). The most vulnerable species were Chondrichthyes (eight out of nine species) and Porifera (nine out of 12 species), which together account for over 75% of species classified as such.
4. Results emphasize the need to introduce specific management measures that protect vulnerable species with bioactive potential as this is a valuable component of marine ecosystems, as well as a potential source of molecules with pharmacological properties beneficial for human health.
5. Marine protected areas can contribute to preserving marine species of medical interest and achieving their sustainable use in the marine biotechnology industry.

     

Interaction of grammistins with lipids and their antibacterial activity

ABSTRACT: Grammistins are hemolytic and ichthyotoxic peptides in the skin secretion of soapfishes and are structurally characterized by their abundance in amphiphilic α-helicity. In the present study, their interaction with lipids and lipid vesicles as well as antibacterial activity were examined using four grammistins (Gs 1 and Gs 2 from Grammistes sexlineatus and Pp 1 and Pp 3 from Pogonoperca punctata ). The hemolytic activity of grammistins was inhibited by phospholipids but not by cholesterol. Moreover, grammistins released carboxyfluorescein entrapped within liposomes made of phosphatidylcholine. In contrast, grammistins were found to have antibacterial activity with a broad spectrum against nine species of bacteria, including both Gram-negative and Gram-positive groups. The potency of their antibacterial activity was not related to that of hemolytic activity, suggesting that grammistins bind to membrane phospholipids but lyse erythrocyte and bacterial membranes via different mechanisms. Conclusively, grammistins are new members of the family of cell non-selective membrane-lytic peptides with amphiphilic α-helices, being similar to pardaxins, which are secreted from the skin of soles, and to melittin, which is derived from bee venom.     

夏初辽东湾海蜇放流区大型水母和主要浮游动物

根据2005～2007年6月上旬和7月上旬在辽东湾海蜇放流区进行的大型水母调查及同期进行的浮游生物调查数据,分析了该区域大型水母及浮游动物的数量分布特点以及海区中浮游动物的生态类型和优势种,并且试图寻找浮游动物的数量分布与大型水母的数量分布的关系。结果表明,浮游动物的优势种为小型桡足类及桡足类幼体:双刺纺锤水蚤Acartia bifilosa、小拟哲水蚤Pa-racalanus parvus、强额拟哲水蚤Paracalanus crassirostris、桡足类幼体、无节幼体。大型水母主要有:海蜇Rhopilema esculentum、沙蜇Nemopilema nomurai、白色霞水母Cyanea nozakii。大型水母的高密集区往往位于浮游动物高丰度区的附近。     

乐清湾甲壳类群落结构特征及其影响因素

根据2016年5月份、8月份、11月份和2017年2月份在乐清湾海域的渔业资源调查数据,利用聚类分析、多元统计分析和冗余分析等方法,对该海域甲壳类的种类组成、生物多样性、群落结构特征及其与环境因子的关系进行了分析。结果显示,该海域共鉴定出甲壳类43种,隶属于2个目、18个科、28个属,全年优势种为三疣梭子蟹、日本蟳和哈氏仿对虾。甲壳类群落结构季节间变化差异显著,夏季甲壳类种类数最多,冬季最少,优势种种类季节变化较小。对不同水深带的甲壳类群落结构分析发现,5～10 m种类数最多,小于5 m和大于10 m种类数较少。单因素方差分析显示,不同水深甲壳类群落的丰富度指数(D)存在显著差异,均匀度指数(J')和多样性指数(H')均无显著差异。冗余分析结果显示,温度、水深和浑浊度是影响乐清湾甲壳类群落结构季节变化的重要环境因子,其中水深对甲壳类分布的影响较大。     

绿鳍马面鲀(Navodon septentrionalis)对4种大型水母的捕食行为

以绿鳍马面鲀为研究对象,通过室内受控实验,比较研究了其对我国沿海4种常见大型水母(海蜇、沙海蜇、海月水母和白色霞水母)的捕食差异。结果显示,体重为(215±20)g的绿鳍马面鲀对海月水母的捕食能力最强,日均最大摄食量为(150.7±18.6)g/fish,其次是海蜇和白色霞水母,日均摄食量分别为(129.7±11.6)和(120.0±19.3)g/fish,对沙海蜇的摄食量最少,为(92.5±11.3)g/fish;绿鳍马面鲀对海月水母与海蜇摄食量主要受投喂量影响,与规格无关,当投喂量小于其最大捕食量时,绿鳍马面鲀可捕食其周围所有水母,当投喂量超过其最大摄食量并继续增加时,绿鳍马面鲀摄食量保持不变,但残余水母的触手和伞部边缘均被啃食,继而导致水母摄食能力丧失,难以继续生存;在适口饵料冰鲜玉筋鱼充足的情况下,绿鳍马面鲀对水母具有明显的摄食偏向性,与仅投喂水母实验组相比,其对海月水母和海蜇的日均摄食量仅降低了20.2%和16.9%。研究结果表明,绿鳍马面鲀对上述4种水母皆能捕食。     

三种大型钵水母幼体对牙鲆和许氏平鲉仔稚鱼的捕食选择性

为探讨中国近海常见大型钵水母对鱼类资源补充的影响,本实验利用清除率法比较研究了伞径4 cm的3种大型水母幼体—海月水母(Aurelia aurita)、海蜇(Rhopilema esculentum)和沙海蜇(Nemopilema nomurai),对不同发育阶段的牙鲆(Paralichthys olivaceus)和许氏平鲉(Sebastes schlegeli)仔稚鱼的捕食量。结果表明,3种水母均可捕食牙鲆和许氏平鲉仔稚鱼,其捕食率随仔稚鱼生长而降低,其中海月水母可捕食规格15 mm牙鲆和20 mm许氏平鲉,海蜇能够捕食规格20 mm牙鲆和30 mm许氏平鲉,沙海蜇可捕食30 mm许氏平鲉;仔稚鱼密度对水母捕食率影响不显著,牙鲆的变态和底栖生活是其逃避被水母捕食的有效手段;不同水母物种对仔稚鱼的捕食效率主要与其活力相关,水母结构和毒性也是重要因素,相同规格3种水母对仔稚鱼捕食率由高到低为海蜇沙海蜇海月水母。本研究结果可为阐明水母与鱼类的动态关系,研究海洋鱼类的自然资源补充机制提供参考。     

Affinity isolation and mass spectral analysis of 1,10-phenanthroline (OP)-stimulated UV-damaged-DNA binding proteins expressed in zebrafish (Danio rerio) embryos

Our earlier studies indicated the high expression of a UV-damaged-DNA binding activity in zebrafish (Danio rerio) embryos at 12?h postfertilization (hpf). Two 30- to 35-kDa polypeptides homologous to the N-terminal lipovitellin 1 (Lv1) domain of the 150-kDa zebrafish vitellogenin 1 (zfVg1) were identified as the damage recognition factors in zebrafish extracts, and the metal-chelating agent 1,10-phenanthroline (OP) was found to inhibit the embryonic UV-damaged-DNA binding activity. This study further explored the DNA damage-sensing components in 12 hpf zebrafish extracts. UV-damaged-DNA binding proteins were enriched from zebrafish extracts by isoelectrofocusing. Both OP-sensitive and OP-stimulated, UV-damaged-DNA binding activities were detected in fractionated zebrafish extracts. Two-dimensional gel electrophoresis of proteins captured by an immobilized oligonucleotide carrying a UV-induced (6-4)photoproduct (6-4PP) revealed a 25-kDa polypeptide as the major 6-4PP-binding factor in an OP-stimulated fraction. Three 25-kDa factors that bound weakly to 6-4PPs were also isolated. The four polypeptides having pIs between 7.0 and 7.3 were unreactive to an anti-zfVg1 antibody targeting the Lv1 domain. Mass spectral analysis showed the appearance of amino acid sequences LPIIVTTYAK and IPEITMSK in all 25-kDa polypeptides and sequences exactly matching those contained in the four factors exist only in the C-terminal Lv2 domain of zfVg1, reflecting the origination of these factors from enzymatic cleavage of the Lv2 domain at slightly different positions. The OP-stimulated fraction produced a much stronger UV-dependent DNA incision activity in the presence than in the absence of OP, suggesting the association of these factors with DNA damage repair under metal-deficient conditions.     

条斑紫菜蛋白酶解液脱色脱腥技术研究

以条斑紫菜为原料采用木瓜蛋白酶制备紫菜蛋白酶解液,以活性回收率和脱色率为评价指标,研究酶解液的脱色脱腥技术。试验结果表明,紫菜盐酸提取液色素最大吸收峰为550nm;在单因素试验基础上,通过正交试验优化后的脱色工艺条件为活性炭用量10g/L、脱色温度60℃、脱色时间40min、pH 6.5,脱色后的酶解液呈浅黄色,活性回收率逾90%。     

Genome size distribution in phylum Cnidaria

Cnidarians are considered ancestral metazoans and, therefore, are important taxa for studying animal evolution. However, little is known about the group’s genome size (C value), which is an important parameter in whole-genome sequencing. To address this issue, we measured the C values of 27 cnidarian species from Japan, using flow cytometry, and found that they ranged from 0.26 to 3.56 pg. Excluding the results for Agalma elegans and Physalia physalis (order Siphonophorae), which had the highest C values among the species included in the present study, the C values for the cnidarians were 0.26–1.49 pg. In particular, we found that hydrozoans possessed relatively large and wide-ranging C values, indicating that evolution within the group involved considerable gains or losses of genomic content. Overall, the C values reported in the present study could be valuable for whole-genome sequencing, using next-generation sequencers, and for future research in cytogenetics.