三种乌贼内骨骼生化成分研究

曼氏无针乌贼（Sepiella maindroni）和金乌贼（Sepia esculenta）的内骨骼称为海螵蛸,可用于制酸、止血等。通过对曼氏无针乌贼、金乌贼和虎斑乌贼（Sepia pharaonis）内骨骼生化成分的比较,有望实现传统中药海螵蛸原料质量的提升和替代。分别使用烘干干燥法、高温灼烧法、凯氏定氮法、索氏提取法、酸碱浸泡法、茚三酮柱后衍生离子交换色谱法、离子发光色谱法检测水分、灰分、粗蛋白、粗脂肪、粗甲壳素、氨基酸及矿物质。结果显示：三种内骨骼营养成分中,虎斑乌贼水分和粗蛋白含量最高,分别为（2.52±0.16）%和（3.43±0.10）%;金乌贼灰分和粗脂肪含量最高,分别为（94.7±0.26）%和（0.24±0.04）%;甲壳素含量最高的为曼氏无针乌贼内骨骼,为（9.48±0.36）%,约为其他两种的1.50倍。三种乌贼内骨骼共检出15种氨基酸,其中必需氨基酸（EAA）7种,半必需氨基酸（HEAA）1种,非必需氨基酸（NEAA）7种,三种乌贼骨的EAA/TAA为44.64%~47.09%,符合FAO/WHO中对于蛋白质EAA/TAA的理想评定标准。金乌贼内骨骼的钙含量最高（3.93×10⁵ mg/kg）、曼氏无针乌贼的钠含量最高（1.02×10⁴ mg/kg）、虎斑乌贼的钾含量最高（589.5 mg/kg）。可见三种乌贼内骨骼均是低脂肪、高甲壳素、高矿物质的中药。虎斑乌贼内骨骼蛋白和灰分营养成分含量较为丰富,是否可以作为传统海螵蛸的替代品,值得进一步探讨。     

头足类腕上吸盘和内壳扫描电镜观察

本文采用扫描电子显微镜对7种头足类腕上的吸盘及乌贼的内壳进行观察。结果表明,蛸科（Octopodidae）的短蛸（Octopus ocellatus）、真蛸（O. vulgaris）和长蛸（O. variabilis）腕的吸盘均无齿,乌贼科（Sepiidae）的日本无针乌贼（Sepia japonica）和金乌贼（S.esculenta）的腕和触腕吸盘上的角质环均有齿,且齿的形状不一样;虎斑乌贼（S. pharoanis）触腕的吸盘角质环光滑无齿,枪乌贼科(Loliginidae)的中国枪乌贼（Uroteuthis chinensis）腕上吸盘的角质环无齿。对３种乌贼的内壳的横切面进行扫描电镜观察,乌贼的内壳的背表面和腹表面的结构均是由文石和铺在上面的球型颗粒构成,但文石的形状和小球的排列存在种间差异,这些差异可以作为头足类的分类辅助特征。     

浙南近海头足类种类组成及生态位分析

在浙南海域布设42个调查站位, 于2015年11月、2016年2月、5月和8月进行了4个航次的系统调查。针对获得的头足类样品, 运用相对重要性指数、生态位宽度和生态位重叠等生态指标, 结合非度量多维标度法(nMDS)和典范对应分析(CCA)对生态位的重叠和分化进行分析。结果表明: 调查海域有头足类14种, 优势种为剑尖枪乌贼、多钩钩腕乌贼、曼氏无针乌贼和火枪乌贼; 生态位变化范围为0.09—7.01, 多钩钩腕乌贼、剑尖枪乌贼、柏氏四盘耳乌贼和火枪乌贼为广生态位种, 四盘耳乌贼、曼氏无针乌贼和田乡枪乌贼为中生态位种, 其余为窄生态位种; 生态位重叠值范围为0.00—0.70, 种间变化较大; 影响头足类分布的最主要因素是悬浮物, 其次为叶绿素和盐度, 温度和溶氧影响相对较小, 这些环境因素综合解释了此区域头足类群落结构83%的总变异。     

金乌贼生活习性的初步观察

金乌贼 Sepia esculenta Hoyle俗称墨鱼或乌鱼,属软体动物、头足纲、二鳃目、十腕亚目,是我国北方沿海习见的一种乌贼,产量颇多,肉味鲜美,可供食用,鲜食或腌渍皆可,贝壳(海螵蛸)可做药用。     

金乌贼墨汁营养成分分析及评价

采用氨基酸自动分析仪、原子吸收仪、液相色谱及常规化学分析法对金乌贼(Sepia esculenta)墨汁的基本营养组分以及氨基酸、矿物质元素、维生素的组成和含量进行了测定。在所检测的17种氨基酸中,含有7种必需氨基酸和2种半必需氨基酸。同时富含Mg、Ca、K、Na等常量矿物元素以及Fe、sr、Al等微量元素和脂溶性、水溶性维生素。     

拟目乌贼卵子发生与卵巢发育

为了丰富拟目乌贼(Sepia lycidas)生物学资料, 为人工育苗与养殖提供理论依据, 采用解剖学和组织学的方法, 对水泥池养殖条件下拟目乌贼卵子发生和卵巢发育进行了研究。结果表明: 经过6个月水泥池养殖, 平均体重为256.34 g, 最大体重达到457.08 g, 个别发育成熟, 绝大部分未达性成熟。卵子发生不同步, 根据细胞形态、细胞大小、滤泡细胞形态和卵黄形成情况可分为卵原细胞阶段(卵原细胞期)、原生质生长阶段(无滤泡期、单层滤泡期和双层滤泡期)、间质生长阶段(滤泡内折早期、滤泡内折中期和滤泡内折晚期)和营养质生长阶段(卵黄发生早期、卵黄发生晚期和成熟期), 共4个阶段10个时期。卵巢发育根据外观形态、性腺指数变化和切面上各期细胞所占的比例, 可分为形成前期、形成期、小生长期、大生长期、成熟前期和成熟期6个时期。拟目乌贼繁殖周期为一年。       

东北羊草草地钾,钠含量特征的研究

东北羊草(Aneurolepidium chinense)各器官地上部分的钾含量在生长初期最高,以后逐渐降低;钠含量的变化规律与钾相似,仅在8月份有明显增高。群落地上部分的钾、钠含量高于地下部分。羊草地上部分的钾、钠积累量在生长季中的变化为单峰曲线,峰值分别出现于7月和8月,寸草苔(Carex duriuscula)和针蔺(Heleocharis acicularis)地上部分的钾、钠积累量变化与生物量相似,为双峰型。群落地下部分的钾、钠积累量明显高于地上部分,茎、叶中钾、钠积累量相近。群落的钾、钠积累量分别占根层土壤钾、钠贮量的0.25%和0.17%,占交换性及水溶性钾、钠贮量之和的2.31%和0.93%。     

盐度对不同日龄日本无针乌贼幼体生长及存活的影响

对不同日龄(1日龄、7日龄、15日龄)的日本无针乌贼在不同盐度(10、15、20、25、30、35、40)条件下的生长及存活状况进行了研究。结果表明,各日龄组日本无针乌贼的存活率及增重率均随盐度的升高而升高,但当盐度超过一定值后,其存活率及增重率随盐度的升高而降低。日本无针乌贼的适宜盐度范围为20～30,不同日龄乌贼幼体的适宜盐度范围略有不同,其中15日龄幼体对盐度的耐受性要优于1日龄及7日龄乌贼幼体。乌贼幼体的最适盐度约为25,其生长和存活均较高。     

曼氏无针乌贼神经肽sCAP基因克隆与组织表达定位

为研究曼氏无针乌贼(Sepiella japonica)小心激肽(small cardioactive peptides, sCAP)生理功能, 通过RACE技术克隆曼氏无针乌贼sCAP基因(简称SjsCAP, GenBank登录号:MG779491), 得到总长度为696 bp的cDNA序列, 包括111 bp的5′非编码区(UTR)和324 bp的3′UTR, 预测的开放阅读框(ORF)共261 bp, 编码86个氨基酸, 相对分子量(MW)为9.331 kD, 等电点(pI)为8.52。信号肽以及跨膜区预测结果表明, sCAP中含有明显的信号肽序列和跨膜区结构。因此, 推测该蛋白可能由细胞内分泌到细胞外发挥作用。亲水性分析显示该蛋白为亲水性蛋白。基于sCAP氨基酸序列进行的系统进化分析表明曼氏无针乌贼与商乌贼(Sepia officinalis)亲缘关系最近, 相似性达到90%。通过荧光定量PCR (Quantitative real-time PCR, qRT-PCR)技术对SjsCAP基因在成熟雄性和雌性曼氏无针乌贼不同组织中的表达量进行分析, 结果显示sCAP主要在视叶中显著表达, 在脑中也有较高的表达量, 其中雄性个体表达量显著高于雌性个体。原位杂交实验结果显示sCAP基因在曼氏无针乌贼的脑组织的视叶、食道上神经团的垂直叶、亚垂直、脑脚叶、背外侧叶和视腺均可以观察到明显的阳性杂交信号。sCAP基因的成功克隆以及组织表达定位分析为sCAP的亚细胞定位以及生物学功能研究奠定一定的基础, 同时为曼氏无针乌贼的种质资源保护与开发提供一定的理论支持。     

盐度和温度对不同规格虎斑乌贼幼体的耗氧率、排氨率和窒息点的影响

为探究体重、盐度和温度对不同规格的虎斑乌贼(Sepia pharaonis)幼体耗氧率、排氨率以及其窒息点的影响。采用单因子试验设计和密闭静水法, 对不同体重下(0.212、0.385、0.476、0.597、0.754和0.946 g)虎斑乌贼幼体的耗氧率和排氨率, 以及不同体重(0.476、0.673、1.341、3.873和4.205 g)幼体的窒息点进行了测定, 同时研究了不同盐度(19‰、22‰、25‰、28‰和31‰)和温度(18、21、24、27和30℃)对不同规格[A: 体重(0.366±0.042) g, B: 体重(0.556±0.038) g, C: 体重(0.844±0.051) g]的虎斑乌贼幼体耗氧率(R_O)和排氨率(R_N)的影响。结果表明: (1)虎斑乌贼幼体体重对耗氧率和排氨率均影响显著(P<0.05)。随着幼体体重的增长, 耗氧率和排氨率显著下降, 个体越小耗氧率和排氨率越大; (2)盐度对幼体的耗氧率和排氨率有显著影响(P<0.05), 均随着盐度的增加呈先下降后上升的趋势, 其中, A规格和B规格的幼体在盐度25‰时, 耗氧率和排氨率显著低于盐度19‰、22‰和31‰时的3个试验组(P<0.05), 而与盐度28‰时无显著差异(P>0.05); C规格幼体的耗氧率在盐度28‰时显著低于盐度19‰组(P<0.05), 而排氨率在盐度25‰时显著低于盐度19‰和31‰两组(P<0.05)。盐度对A规格幼体的氧氮比(O/N)值有显著影响(P<0.05), 而对B规格和C规格的幼体无显著影响(P>0.05)。(3)温度对不同规格幼体的耗氧率和排氨率有显著影响(P<0.05), 均随温度的增长, 呈先上升后下降的趋势。其中, A规格和C规格幼体的耗氧率在27℃时, 显著高于18和30℃两组(P<0.05), B规格的在水温24℃时显著高于18和30℃(P<0.05); A规格和B规格幼体的排氨率分别在24和27℃时, 显著高于其他温度组(P<0.05), C规格的在温度27℃时, 显著高于18和30℃两组(P<0.05); 温度对A规格幼体的O/N值有显著影响(P<0.05), 而对B规格和C规格的幼体无显著影响(P>0.05)。(4)虎斑乌贼幼体的窒息点为0.84—1.62 mg/L, 随着体重的增加而逐渐降低。     

Phylogeny of selected Sepiidae (Mollusca, Cephalopoda) based on 12S, 16S, and COI sequences, with comments on the taxonomic reliability of several morphological characters

Phylogenetic relationships among 11 species of sepiids from Japanese waters and Sepia officinalis from Mediterranean were studied using partial sequences of the mitochondrial 12S rRNA, 16S rRNA, and cytochrome c oxidase subunit I genes. These three genes had been analyzed in an Atlantic species S. elagans and was obtained from database. In the two-gene set analysis (16S+COI), sequence data of another 4 species were added from database. We also studied morphological characters of radulae, tentacular clubs, and cuttlebones. The molecular phylogeny was not congruent with relationships detected by the number of rows in radulae and the arrangement of suckers on the tentacular club. As to the cuttlebone shape, the molecular phylogeny suggests the separation of two groups, Doratosepion species with a lanceolate cuttlebone and the others with a broad cuttlebone. Our molecular phylogenetic study revealed these sepiids are separated into four clades. The first clade includes Sepia officinalis, S. hierrendda, S. bertheloti, S. pharaonis and Sepiella japonica. The second clade consists of S. latimanus and Metasepia tullbergi from sub-tropical waters. The third clade includes Sepia esculenta, S. madokai, S. aculeata and S. lycidas, which have a cuttlebone with a prominent spine. The fourth clade consists of Doratosepion species complex, S. kobiensis, S. lorigera, S. pardex, S. peterseni, and S. sp., which are characterized by a narrow cuttlebone with a distinct outer cone at the posterior end. The lack of membranous structures in the cuttlebone is a synapomorphy for this clade. S. elegans did not clearly belong to any of these clades and might represent the fifth clade.     

Structural Comparison of Cephalopod Hemocyanins: Phylogenetic Significance

Hemocyanins, the respiratory molecules of cephalopod mollusks, are hollow cylinders with five internal arches. Three hemocyanins representative of three orders of cephalopods (Benthoctopus species, Octopoda; Vampyroteuthis infernalis, Vampyromorpha; Sepia officinalis, Sepioidea) were subjected to cryoelectron microscopy and three-dimensional (3D) reconstruction. The structure of Benthoctopus hemocyanin, solved at 26.4-A resolution, possesses arches comprising two identical functional units. The similarity between these functional units and the structure recently observed in X-ray crystallography for Octopus by Cuff et al. (J. Mol. Biol., 1998, 232, 522-529) allows the identification of their N- and C-terminal domains in the 3D reconstruction volume. Conversely, arches present in the 3D reconstruction volume of Sepia hemocyanin (21.8 A resolution) contain four functional units that are disposed differently. The strong resemblance between the reconstruction volumes of Vampyroteuthis (21.4-A resolution) and Benthoctopus hemocyanins suggests that Sepioidea diverged from a group containing Octopoda and Vampyromorpha.     

Phylogenetic relationships among major species of japanese coleoid cephalopods (Mollusca: Cephalopoda) using three mitochondrial DNA sequences

Phylogenetic relationships among 36 species of major coleoid cephalopods from Japanese waters were studied using partial sequences of three mitochondrial genes, 16S rDNA, 12S rDNA, and cytochrome c oxidase subunit I gene. Octopoda and Decapoda were monophylic groups. Within Sepioidea, Sepiadariidae and Sepiolidae were not closely related to Sepiidae, but rather related to Teuthoidea. Sepiidae with a distinct calcareous shell formed a single cluster. Myopsida was closely related to Oegopsida. Within Octopoda, Opisthoteuthis depressa and Argonauta argo diverged earlier than Octopodiidae. The common octopuses in Japanese waters were separated into three clusters. The first cluster occupied a basal position, and includes large-sized octopuses, such as Enteroctopus dofleini and Octopus (Paroctopus) conispadiceus from the continental shelf and upper slope. The second cluster consisted of long-armed octopuses, such as O. ornatus, O. minor, and O. sasakii. The third cluster contained small- to medium-sized octopus, such as Amphioctopus fangsiao, A. areolatus, O. cyaneus, and O. vulgaris, in which several species possess ocelli on the web. The second cluster formed the sister group to the third cluster.     

温度对拟目乌贼幼体日生长率和存活率的影响

以温度为单因素变量,在人工饲养环境中,分析水温对拟目乌贼幼体日生长率与存活率的影响。实验结果表明,当水温处于28.1～32℃之间时,拟目乌贼幼体的日生长率与水温呈正相关关系;当水温处于32～32.67℃时,拟目乌贼幼体的日生长率与水温呈负相关关系;当水温处于29.5～32.6℃时,拟目乌贼幼体不易出现幼体死亡(死亡率<6.25%),适宜拟目乌贼幼体的存活;而当水温处于32.3～34℃之间时,拟目乌贼幼体出现了13.16%～61.16%的幼体死亡,不适宜拟目乌贼幼体的存活。实验结果表明,当水温处于29.5～32℃之间时,拟目乌贼幼体的存活率高(存活率>93.75%),且日生长率处于0.24%～1.76%,适宜拟目乌贼幼体的存活。     

Actin gene family evolution and the phylogeny of coleoid cephalopods (Mollusca: Cephalopoda)

Phylogenetic analysis conducted on a 784-bp fragment of 82 actin gene sequences of 44 coleoid cephalopod taxa, along with results obtained from genomic Southern blot analysis, confirmed the presence of at least three distinct actin loci in coleoids. Actin isoforms were characteri zed through phylogenetic analysis of representative cephalopod sequences from each of the three isoforms, along with translated actin cDNA sequences from a diverse array of metazoan taxa downloaded from GenBank. One of the three isoforms found in cephalopods was closely related to actin sequences expressed in the muscular tissues of other molluscs. A second isoform was most similar to cytoplasmic-specific actin amino acid sequences. The muscle type actins of molluscs were found to be distinct from those of arthropods, suggesting at least two independent derivations of muscle actins in the protostome lineage, although statistical support for this conclusion was lacking. Parsimony and maximum-likelihood analyses of two of the isoforms from which >30 orthologous coleoid sequences had been obtained (one of the cytoplasmic actins and the muscle actin) supported the monophyly of several higher-level coleoid taxa. These included the superorders Octopodiformes and Decapodiformes, the order Octopoda, the octopod suborder Incirrata, and the teuthoid suborder Myopsida. The monophyly of several taxonomic groups within the Decapodiformes was not supported, including the orders Teuthoidea and Sepioidea and the teuthoid suborder Oegopsida. Parametric bootstrap analysis conducted on the simulated cytoplasmic actin data set provided statistical support to reject the monophyly of the Sepioidea. Although parametric bootstrap analysis of the muscle actin isoform did not reject sepioid monophyly at the 5% level, the results (rejection at P: = 0.068) were certainly suggestive of sepioid nonmonophyly.     

Cuttlefish camouflage: a quantitative study of patterning

To investigate camouflage design, we compared the responses of two species of cuttlefish ( Sepia officinalis and Sepia pharaonis ) with controlled but naturalistic backgrounds, consisting of mixtures of 1-mm and 9-mm diameter coloured pebbles. Quantitative analysis of image data using methods adapted from functional imaging research found differences in how the two species camouflage themselves. Whereas S. officinalis switches from background resemblance to a disruptive pattern as it moves from a fine to a coarsely patterned background particle, S. pharaonis blends the two types of pattern. We suggest that the differences may arise because S. pharaonis needs to produce camouflage that is effective when viewed over a relatively wide range of distances.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 92 , 335–345.     

First report on the stomach contents of long‐finned pilot whales,Globicephala melas,stranded in New Zealand

Abstract

Stomach contents of the long‐finned pilot whale, Globicephala melas, are reported for the first time from New Zealand waters. Analyses based on two male and three female whales (2.5–5.3 m in length) that stranded on Farewell Spit, Golden Bay, South Island in December 2005 revealed a diet comprised exclusively of cephalopods (2-33 lower cephalopod beaks per stomach). Two genera of cephalopod from two orders; arrow squid, Nototodarus spp. (Teuthoidea: Ommastrephidae), and common octopus, Pinnoctopus cordiformis (Octopoda: Octopodidae) were represented. A further five pilot whale stomachs were examined and found to be empty.     

Mitochondrial genome structure and evolution in the living fossil vampire squid, Vampyroteuthis infernalis, and extant cephalopods

Complete nucleotide sequences of mitochondrial (mt) genomes of the "living fossil" cephalopod Vampyroteuthis infernalis (Vampyromorpha) and the cuttlefish Sepia esculenta (Sepiida) were determined. The V. infernalis mt genome structure is identical to the incirrate octopod Octopus vulgaris mt genome structure, and is therefore more similar to that of the polyplacophoran Katharina tunicata, than to that of the other "living fossil" cephalopod Nautilus macromphalus. The mt genome structure of S. esculenta is identical to that of Sepia officinalis. Molecular phylogenetic analyses based on the mt protein genes from the completely sequenced cephalopod mt genomes suggested the monophyletic relationship of two myopsid squids Loligo bleekeri and Sepiotheuthis lessoniana, and the monophyletic relationship of two oegopsid squids Watasenia scintillans, and Todarodes pacificus. Sepiida appeared as the sister group of Teuthida (Myopsida + Oegopsida). The phylogenetic position of Vampyromorpha appeared as the sister group of Octopoda, although the monophyly of Vampyromorpha and Decapodiformes cannot be rejected outright by our phylogenetic analyses. The hypothesis that Vampyromorpha is basal among the coleoid cephalopods can be rejected because of low statistical support. Therefore, it is reasonable to recognize three major groups in Coleoidea--Vampyromorpha, Octopoda, and Decapodiformes.