Identification of Fucans from Four Species of Sea Cucumber by High Temperature ~1H NMR

Acidic polysaccharide, which has various biological activities, is one of the most important components of sea cucumber. In the present study, crude polysaccharide was extracted from four species of sea cucumber from three different geographical zones, Pearsonothuria graeffei（Pg） from Indo-Pacific, Holothuria vagabunda（Hv） from Norwegian Coast, Stichopus tremulu（St） from Western Indian Ocean, and Isostichopus badionotu（Ib） from Western Atlantic. The polysaccharide extract was separated and purified with a cellulose DEAE anion-exchange column to obtain corresponding sea cucumber fucans（SC-Fucs）. The chemical property of these SC-Fucs, including molecular weight, monosaccharide composition and sulfate content, was determined. Their structure was compared simply with fourier infrared spectrum analyzer and identified with high temperature 1H nuclear magnetic resonance spectrum analyzer（NMR） and room temperature 13 C NMR. The results indicated that Fuc-Pg obtained from the torrid zone mainly contained 2,4-O-disulfated and non-sulfated fucose residue, whereas Fuc-Ib from the temperate zone contained non-, 2-O- and 2,4-O-disulfated fucose residue; Fuc-St from the frigid zone and Fuc-Hv from the torrid zone contained mainly non-sulfated fucose residue. The proton of SC-Fucs was better resolved via high temperature 1H NMR than via room temperature 1H NMR. The fingerprint of sea cucumber in different sea regions was established based on the index of anomer hydrogen signal in SC-Fucs. Further work will help to understand whether there exists a close relationship between the geographical area of sea cucumber and the sulfation pattern of SC-Fucs.     

Analysis of fatty acid composition of sea cucumber Apostichopus japonicus using multivariate statistics

Fatty acids （FAs） provide energy and also can be used to trace trophic relationships among organisms. Sea cucumber Apostichopus japonicus goes into a state of aestivation during warm summer months. We examined fatty acid profiles in aestivated and non-aestivated A. japonicus using multivariate analyses （PERMANOVA, MDS, ANOSIM, and SIMPER）. The results indicate that the fatty acid profiles of aestivated and non-aestivated sea cucumbers differed significantly. The FAs that were produced by bacteria and brown kelp contributed the most to the differences in the fatty acid composition of aestivated and non- aestivated sea cucumbers. Aestivated sea cucumbers may synthesize FAs from heterotrophic bacteria during early aestivation, and long chain FAs such as eicosapentaenoic （EPA） and docosahexaenoic acid （DHA） that produced from intestinal degradation, are digested during deep aestivation. Specific changes in the fatty acid composition of A. japonicus during aestivation needs more detailed study in the future.     

Polyploidy induction by hydrostatic pressure shock and embryo development of sea cucumber Apostichopus japonicus

The manipulation of the chromosome set for commercially valuable marine animals is important for enhancing aquacultural production. In this study, triploid and tetraploid sea cucumber Apostichopus japonicus were induced by hydrostatic pressure shock, and the conditions of appropriate induction were tested with different starting times, and hydrostatic pressure intensities and durations. The highest rate of triploid induction reached 20% and that of tetraploid was 60%. In consideration of the survival rate and hatch rate, the appropriate treatment for triploid was 55 Mpa of hydrostatic pressure for 5 min at 55 min after fertilization (a.f.), while for tetraploid it was 60 Mpa for 5 min at 61 min a.f. The triploid of the sea cucumber could survive through the pelagic larval stage and attachment stage, and develop like the control group of the experiment. The tetraploid, however, could not survive the attachment stage.     

西沙群岛的棘皮动物 Ⅰ.海参纲

中国科学院海洋研究所曾多次到我国西沙群岛进行海洋生物的采集工作；作者参加过一次考察，并对该岛的棘皮动物进行过专门的采集与观察。 富饶的西沙群岛，盛产海参，有的地方，可谓俯拾皆是。全世界约40种海参可供食用，而西沙群岛即产有20种。世界上一些名贵的种类，例如梅花参、黑乳海参、蛇目白尼参和辐肛参等，该岛均有出产。 本文报道了经济意义较大的海参纲动物的研究结果，共检查了标本425个，鉴定为41种，分隶于3目，5科，16属，内有一新种，有14种在我国系首次记录。     

热带珊瑚礁区红腹海参及绿刺参食物来源分析——脂肪酸标志法

沉积食性海参通过大量摄取沉积物在热带珊瑚礁生态系统中发挥重要的生态功能,但其食物来源组成尚不清楚.本文研究了海南三亚近岸典型珊瑚礁海域红腹海参(Holothuria edulis)及绿刺参(Stichopus chloronotus)脂肪酸组成,利用脂肪酸标志法探讨了夏季和冬季两种海参的食物来源异同.结果表明,红腹海参...     

室内养殖不同月龄糙海参营养成分分析

【目的】分析室内养殖的4、7和10月龄干、鲜糙海参的营养成分。【方法】用国标统一方法分析。【结果】随着生长发育,糙海参干体粗脂肪含量呈增加趋势,而粗蛋白和总多糖含量则呈下降趋势(P0.05);不同月龄糙海参鲜体内水分、粗蛋白、总多糖、总皂苷和粗脂肪含量差异有统计学意义(P0.05),粗脂肪含量低于检出限。在糙海参体内共检出17种氨基酸,其中8种必需氨基酸、1种半必需氨基酸和8种非必需氨基酸。4月龄糙海参干体各氨基酸含量显著低于7月龄和10月龄糙海参(P0.05),各月龄糙海参氨基酸含量由大到小顺序较为一致,为谷氨酸、甘氨酸、天冬氨酸、精氨酸、丙氨酸、脯氨酸,但鲜体蛋氨酸、亮氨酸,赖氨酸和组氨酸含量差异有统计学意义(P0.05),而其他氨基酸含量差异无统计学意义(P0.05)。不同月龄糙海参干体必需、非必需氨基酸比值和必需氨基酸占总氨基酸比例分别为0.36~0.38,26.58%~27.55%,鲜品的必需、非必需氨基酸比值和必需氨基酸占总氨基酸比例分别为0.33~0.36、26.06%~26.34%;不同月龄糙海参干体的鲜味、药用氨基酸占总氨基酸比例分别为60.74%~61.46%、66.29%~66.21%,鲜品的鲜味、药用氨基酸占总氨基酸比例分别为62.86%~64.03%、66.13%~66.46%。【结论】室内养殖糙海参品质和营养价值较高。     

糙海参卵子发生及卵黄发生的超微结构

通过扫描与透射电镜观察了糙海参(Holothuria scabra)卵子发生及卵黄发生的过程。结果表明,糙海参卵子属于均黄卵,其卵子发生经历卵原细胞、卵黄发生前卵母细胞、卵黄发生期卵母细胞3个阶段。其中,卵黄发生期卵母细胞分别包括卵黄发生早期、中期、后期卵母细胞3个阶段。卵原细胞核大且呈椭圆形,胞质中只分布少量的线粒体;卵黄发生前卵母细胞内有一个空泡状的生发泡(卵核);卵黄发生早期,细胞器种类丰富,数量多;卵黄发生中期,卵黄合成旺盛,是内源性卵黄大量形成的阶段,多种细胞器活跃参与形成卵黄;卵黄发生后期,细胞体积最大,细胞核明显,且位于胞质中间或者靠近动物极突。高尔基体、线粒体及核糖体是形成内源性卵黄最主要的细胞器。卵质膜通过胞饮作用参与外源性卵黄的形成。本结果可为糙海参繁殖生物学和人工繁殖提供参考。     

Generation of Reactive Oxygen Species in Different Fractions of the Coelomocytes of Holothurian Eupentacta fraudatrix in Response to the Thermostable Toxin of Yersinia pseudotuberculosis in Vitro

Pure fraction (92% - 95%) of phagocytes (FP) and a mixture of amoebocytes (62%) and morula cells (38 % )-FPMC- of the holothurian Eupentacta fraudatrix ( Holothuroidea,Dendrochirota ) were obtained by using ficoll-verographine step gradient. Basal production of reactive oxygen species (ROS) in FP quantified by using reduction of nitroblue tetrazolium (NBT) was more than twice that in FPMC. Thermostable toxin of Yersinia pseudotuberculosis (TST) at differentconcentrations ( 0.2; 0.5; 2.5 μg/ml, but not 0.1 μg/ml) stimulated NBT reduction in FPMC after 24 h incubation. In FP, TST at concentrations of 0.1 and 0.2 μg/ml inhibited and at concentrations of 0.5 and 2.5 μg/ml stimulated NBT reduction after 24 h incubation. Maximal effect was observed in FP and FPMC at TST concentrations of 0.5 and 0.2 μg/ml, respectively. Addition of catalase (0.7 μg/ml) to the cells treated with TST (2.5 μg/ml) was followed by a decrease in NBT reduction compared to that under toxin treatment alone. TST stimulated superoxide dismutase activity in ccncentration-dependent manner (maximum at 0.5 μg/ml concentration in FP) after 24 htreatment, and this stimulation was prevented by a commercial catalase. Plant lectin concanavalin A stimulated NBT reduction more than 5-fold in FPMC compared to the control. With addition of TST, lectin stimulated ROS to lesser extent than that with lectin alone. When catalase, TST, and lectin were added into the FPMC simultaneously, ROS increase was similar to that under lectin treatment alone. On the whole, data obtained indicated that ROS generation in holothurian coelomocytes especially occurs in both stimulated and not stimulated phagocytes, and that changes in ROS production by these cells may be one of the mechanisms of antibacterial protection of holothurians.     

糙海参精子的发生及超微形态结构

为了解糙海参(Holothuria scabra)精子发生的过程及其形态结构,作者于2013年11月~2014年4月每月采集2头海南岛的野生雄性糙海参,取精巢组织固定后分别制作投射电镜、扫描电镜切片和石蜡包埋切片,在电镜和光镜下连续观察。观察结果表明,糙海参精子发生经历了精原细胞、初级精母细胞、次级精母细胞、精细胞和成熟精子5个阶段。精细胞分化过程可分为3个时期,早期:细胞核前端出现前顶体囊;中期:顶体腔内出现围顶体物质;后期:胞质往尾部方向收缩,形成精子中部。糙海参成熟精子全长约60μm,由头部、中部和尾部3部分组成,为典型的鞭毛型精子。精子头部呈球形,精子核中顶体形态为"U"型内陷囊状体,围顶体物质呈"月牙"形,核染色质凝集成团块状。精子中部由一个巨大的线粒体和中心粒复合体融合而成。精子尾部鞭毛横切面为典型的"9+2"微管结构。本研究可为糙海参的繁殖生物学和人工育苗提供理论参考。