Trial grow‐out culture of sea cucumber Holothuria scabra in sea cages and pens

Field trials were conducted to determine the effects of size, grow‐out period and site on the growth and survival of Holothuria scabra. Juveniles were reared in sea cages in two sites for 2 months during two different times of the year. Juveniles from Experiment 1 were subsequently grown in sea pens for 8 months in one site. The significantly lower survival particularly of the small‐sized group (<0.1 g) in Pilar is likely due to reduced water and sediment quality because of proximity to fish‐farming activities. The importance of grow‐out period was evident in the significant differences in weight increments in all size groups and biomass in the small‐sized group (P < 0.10) in Lucero during Experiment 2 when water temperature range and incidence of rainfall were higher. The decrease in the average weight of adults during the second half of the grow‐out period may be due to the interplay of the reduction in salinity, onset of sexual maturity and biomass exceeding carrying capacity in the pens. The estimated biomass threshold in the sea cages and pens in Lucero were 300 g m^?2 and 380 g m^?2 respectively. Implications of the results on the viability of small‐scale pen grow‐out culture are discussed.     

糙海参胚胎和幼体发育的形态观察

为提高糙海参育苗技术,研究描述了糙海参从受精卵发育到稚参的形态变化,在显微镜下测定了受精卵、胚胎和幼体的大小,确定了糙海参胚胎发育的过程。结果表明,通过利用阴干、流水刺激法对成熟亲参进行人工催产,得到大量的受精卵,其受精率为90%以上。糙海参胚胎和幼体发育可分为受精卵、卵裂期、囊胚期、旋转囊胚期、原肠期、初耳状幼体、中耳状幼体、大耳状幼体、樽形幼体和稚参等阶段;在平均水温29℃,平均盐度34条件下,糙海参受精卵经3 h发育形成囊胚,4 h进入旋转囊胚期,5 h进入原肠期,19 h完成胚胎发育变态为耳状幼体;并经过7 d的生长与发育进入樽形幼体;第15天变态为稚参。观察发现,多精入卵现象则会导致胚胎发育不正常,最终使胚胎发育停止并死亡。在糙海参幼体发育过程中,大耳状幼体期幼虫臂的大小及其球状体的形成均可作为判断其幼体发育健康状况的重要指标:幼虫臂越大,球状体出现率越高,其幼虫的变态率和成活率也越高。对比发现,位于糙海参尾部的突出结构——尾突,为仿刺参和新西兰刺参所没有,这一结构差异同时导致了其骨片位置也不同;并且,糙海参胚胎和幼体与其他种类的海参在发育时间上存在一定差异。     

玉足海参与凡纳滨对虾的混养效果

在室内条件下进行了玉足海参与凡纳滨对虾的混养实验,分析了单养与混养两种条件下养殖水体营养盐结构以及底质成分的变化,测定了对虾与海参的存活率与生长性能。结果显示,混养海参可以明显改变养殖系统的营养盐结构,可使水体中的磷酸盐和硝酸盐浓度有所升高,同时也可有效地控制系统中氨氮浓度。混养海参也可以大幅度地降低沉积物中有机质和硫化物含量,实验结束时混养组硫化物含量为(7.71±1.33)mg/kg,仅相当于单养组浓度的1/3。混养海参对对虾生长及存活具有明显的促进作用,其中混养组对虾体长特异增长率为(0.69±0.13)%/d,显著优于单养组(0.45±0.06)%/d;混养组对虾成活率可达72.5%±22.9%,显著高于对照组55.0%±17.5%。在混养系统内,对虾不会对海参的生存造成负面影响,海参能够有效地选择摄食和利用沉积物中的营养物质(对食物中有机质的同化率可达36.36%±13.79%),并以较快的速度生长。结果表明,在对虾养殖系统中混养玉足海参具有明显的环境与经济效益。本研究可为我国海水养殖业的可持续发展提供一定的科学依据。     

Structural Characterization and Heparanase Inhibitory Activity of Fucosylated Glycosaminoglycan from Holothuria floridana

Unique fucosylated glycosaminoglycans (FG) have attracted increasing attention for various bioactivities. However, the precise structures of FGs usually vary in a species-specific manner. In this study, HfFG was isolated from Holothuria floridana and purified by anion exchange chromatography with the yield of ~0.9%. HfFG was composed of GlcA, GalNAc and Fuc, its molecular weight was 47.3 kDa, and the -OSO₃⁻/-COO⁻ molar ratio was 3.756. HfFG was depolymerized by a partial deacetylation–deaminative cleavage method to obtain the low-molecular-weight HfFG (dHfFG). Three oligosaccharide fragments (Fr-1, Fr-2, Fr-3) with different molecular weights were isolated from the dHfFG, and their structures were revealed by 1D and 2D NMR spectroscopy. HfFG should be composed of repeating trisaccharide units -{(L-FucS-α1,3-)d-GlcA-β1,3-d-GalNAc_4S6S-β1,4-}-, in which sulfated fucose (FucS) includes Fuc_2S4S, Fuc_3S4S and Fuc_4S residues linked to O-3 of GlcA in a ratio of 45:35:20. Furthermore, the heparanase inhibitory activities of native HfFG and oligosaccharide fragments (Fr-1, Fr-2, Fr-3) were evaluated. The native HfFG and its oligosaccharides exhibited heparanase inhibitory activities, and the activities increased with the increase of molecular weight. Additionally, structural characteristics such as sulfation patterns, the terminal structure of oligosaccharides and the presence of fucosyl branches may be important factors affecting heparanase inhibiting activity.     

Fucose-Rich Sulfated Polysaccharides from Two Vietnamese Sea Cucumbers Bohadschia argus and Holothuria (Theelothuria) spinifera: Structures and Anticoagulant Activity

Fucosylated chondroitin sulfates (FCSs) FCS-BA and FCS-HS, as well as fucan sulfates (FSs) FS-BA-AT and FS-HS-AT were isolated from the sea cucumbers Bohadschia argus and Holothuria (Theelothuria) spinifera, respectively. Purification of the polysaccharides was carried out by anion-exchange chromatography on DEAE-Sephacel column. Structural characterization of polysaccharides was performed in terms of monosaccharide and sulfate content, as well as using a series of non-destructive NMR spectroscopic methods. Both FCSs were shown to contain a chondroitin core [→3)-β-d-GalNAc-(1→4)-β-d-GlcA-(1→]_n bearing sulfated fucosyl branches at O-3 of every GlcA residue in the chain. These fucosyl residues were different in pattern of sulfation: FCS-BA contained Fuc2S4S, Fuc3S4S and Fuc4S at a ratio of 1:8:2, while FCS-HS contained these residues at a ratio of 2:2:1. Polysaccharides differed also in content of GalNAc4S6S and GalNAc4S units, the ratios being 14:1 for FCS-BA and 4:1 for FCS-HS. Both FCSs demonstrated significant anticoagulant activity in clotting time assay and potentiated inhibition of thrombin, but not of factor Xa. FS-BA-AT was shown to be a regular linear polymer of 4-linked α-L-fucopyranose 3-sulfate, the structure being confirmed by NMR spectra of desulfated polysaccharide. In spite of considerable sulfate content, FS-BA-AT was practically devoid of anticoagulant activity. FS-HS-AT cannot be purified completely from contamination of some FCS. Its structure was tentatively represented as a mixture of chains identical with FS-BA-AT and other chains built up of randomly sulfated alternating 4- and 3-linked α-L-fucopyranose residues.     

镉胁迫对银条开花期生理生态的影响

为了探讨镉胁迫对银条开花期生理生态的影响,采用盆栽试验方法,研究了镉胁迫对银条叶片生理特性的影响。结果表明:随着Cd浓度的提高,银条叶片叶绿素含量显著降低,Cd浓度与叶片叶绿素含量呈极显著负相关,相关系数r=-0.9852**(P0.01);细胞质膜透性、SOD活性均随镉处理浓度的增加显著升高、相关系数分别为r=0.9850**(P0.01)r、=0.9923*(P0.05);MDA含量、POD活性则先升高再降低,与镉浓度呈显著正相关,相关系数r分别为0.9845**(P0.01)0、.9018*(P0.05)。     

Amino Acid 129 of Cucumber mosaic virus Coat Protein Determines Local Symptom Expression and Systemic Movement in Tetragonia expansa, Momordica charantia and Physalis floridana

Local symptom expression and systemic movement of Cucumber mosaic virus (CMV) in Tetragonia expansa, Momordica charantia and Physalis floridana were mapped to the amino acid at position 129 of CMV coat protein (CP), using pseudorecombinants, chimeric RNAs, a site-directed mutant of RNA 3 and four strains of CMV : pepo-, SO-, MY17- and Y-CMV. Local and systemic symptoms caused by three strains, pepo-, SO- and MY17-CMV, and those by Y-CMV differed in the three host species. The three strains expressed local chlorotic spots at 24°C and systemic chlorotic spots and ringspots at 36°C, whereas Y-CMV developed local necrotic spots at 24°C but no systemic symptoms at 36°C in T. expansa. In M. charantia the three strains caused systemic chlorotic spots, whereas Y-CMV caused local necrotic spots. The three caused systemic mosaic and Y-CMV systemic necrosis in P. floridana. With pseudorecombinants combined with pepo- and Y-CMV RNAs, CMV RNA 3 was responsible for symptom expression and systemic infection. Inoculation with Y-CMV RNA 1, RNA 2 and chimeric RNA 3s exchanged CP gene fragments between pepo- and Y-CMV showed that NruI-XhoI fragment of CP was essential for symptom expression. Comparative analysis of the NruI-XhoI fragments revealed that only the amino acid at position 129 was common among the three strains but different from that of Y-CMV. Inoculation with a point mutant constructed by substituting one nucleotide resulting in an amino acid change from Ser to Pro at position 129 in Y-CMV CP verified the previous experiments. These results indicate that the amino acid at position 129 of CMV CP is the determinant for local symptom expression and systemic movement in the three host species. CMV CP containing Ser at position 129 may induce resistant responses in these plants. Received 29 June 2001/ Accepted in revised form 28 August 2001     

Improving the fitness of Holothuria arguinensis larvae through different microalgae diets

Development and design of adequate feeding protocols on sea cucumber larvae are necessary for improvements on larvae growth, survival and production of competent juveniles. This work assessed the Holothuria arguinensis larvae growth and survival, fed with three different single microalgae diets of Chaetoceros calcitrans, Isochrysis galbana and Tetraselmis chui, and two microalgae diets based on a combination of C. calcitrans and T. chuii (C:T; 1:1) and C. calcitrans, T. chuii and I. galbana (C:T:I; 1:1:1) during 2 months. Holothuria arguinensis larvae fed with T:C:I, showed higher survival and larger larval and stomach sizes than the ones fed with the other diets. Comparing the results obtained from the single diets, H. arguinensis larvae fed with I. galbana grew and survived better during the first 5 days. However, higher growth was obtained on the larvae fed with C. calcitrans from day 5 to day 12; since this day, larvae fed with T. chuii showed the largest size. These results could be related to the different microalgae size and morphology, in addition to the nutritional value of the different microalgae used. However, no competent larvae that metamorphosed into doliolaria were obtained during this experiment. The tanks used were transparent, which could be causing light stress to the larvae, since competent larvae were obtained in opaque production tanks in our aquaculture installations during the same time period. The combined diet of C. calcitrans, T. chuii and I. galbana (C:T:I) could improve the growth and larvae competency of H. arguinensis.     

Size measurement and nutritional condition evaluation methods in sandfish (Holothuria scabra Jaeger)

The aims of this study were to establish an accurate size measurement method and a nutritional condition evaluation method of Holothuria scabra (Jaeger). Although 0.5% KCl and 0.05% MgSO₄ did not induce anaesthesia, 2% menthol–ethanol for 20 min was found to be effective and harmless. The anaesthetization significantly reduced the coefficient of variation of the mean body length and weight by 68% and 43% respectively. During starvation, body size and weight decreased concomitantly, resulting in an unchanged condition factor (body weight/volume), suggesting that the condition factor cannot be used as an index of nutritional condition. Protein, cholesterol and carbohydrate concentrations in the body fluid were analysed to study the relationship with starvation. As the protein and cholesterol concentrations initially increased and then decreased during the starvation period, it is difficult to use them as an index of nutritional condition. The carbohydrate concentration showed a gradual one‐fold increase during 10 days of starvation, and it may be used as a proxy for nutritional condition; however, further physiological studies are needed. Body fluid density and volume relative to body size gradually increased and decreased, respectively, during starvation. These methods may be used to correctly monitor the conditions of H. scabra in studies for aquaculture and stock enhancement techniques.     

4种饵料对玉足海参幼体生长和发育的影响

为了解单一饵料或混合投喂对玉足海参(Holothuria leucospilota)幼体生长的影响,采用2种单胞藻(牟氏角毛藻、球等鞭金藻)和2种酵母(酿酒酵母、海洋红酵母)单一或组合搭配投喂玉足海参耳状幼体,测定并比较海参幼体的体长、胃宽、水体腔长、变态成活率等指标。试验结果显示：单一饵料组中,牟氏角毛藻对玉足海参幼体体长的生长及变态存活最有利,而球等鞭金藻对其脏器(胃、水体腔)的发育最有利;综合所有处理组的投喂效果看,混合投喂组比单一饵料组更有利于玉足海参幼体的生长和存活,其中酿酒酵母+海洋红酵母的混合投喂组合得到了最高的体长、胃宽和水体腔长特定生长率,牟氏角毛藻+海洋红酵母投喂组的变态成活率最高,变态发育速度最快。结果表明,在玉足海参幼体培养早期适合以混合酵母为主投喂,后期则可较多投喂牟氏角毛藻和海洋红酵母。