无公害刺参稳产健康养殖新技术

本文针对目前刺参养殖中存在的问题,简单明了叙述刺参无公害健康养殖一系列新技术和新方法,对推动海参养殖业持续、稳定、健康发展,起到积极作用和效果。     

Development strategies for the sea cucumber industry in China

The sea cucumber, Apostichopus japonicus, is an important marine aquaculture species in China. After nearly thirty years of development, the production of A. japonicus has become commercially lucrative and successful. In this report, current advances in sea cucumber industry are addressed in terms of the basic biology, culturing methods, and health care bene?ts. Next, the challenges restricting development of the sea cucumber industry are discussed, including weaknesses in the basic biological research, the problem of germplasm degradation, environmental stress caused by global climate change, and food safety problems. Finally, several strategies are presented that might contribute to sustainable development of the sea cucumber industry. These strategies include advances in genome studies, behavioral studies, selective breeding, ecological culture technologies, reforms in food safety management, and the development of health care functions based on contemporary medical practices. Thus, our work provides new insights into how to explore the sustainable development of the sea cucumber industry in the future.     

紫刺参早期生长发育与体色形成的研究

本研究对紫刺参胚胎和幼体发育过程进行了显微观测,比较分析了其幼体早期生长情况与体色形成过程。结果表明:(1)在水温21.0±0.2℃条件下,紫刺参受精卵在受精10～15min后释放第一极体,5h 30min～6h 30min进入囊胚期, 18～20h发育成原肠胚; 30～34h进入耳状幼体阶段, 8～10d变态发育为樽形幼体, 12～14d发育成稚参;紫刺参胚胎和幼体发育时序与普通刺参无显著差异。(2)紫刺参早期发育中的樽形幼体发生率和附着变态率分别为58.4±4.7%和45.1±2.7%,高于普通刺参。(3)紫刺参浮游阶段后期随日龄增长表现出较普通刺参发育更快的优势,在日龄6～7d表现出显著差异(P0.05)。(4)紫刺参稚参在日龄35d前后自背部开始着色,50d～60d腹部开始着色,110d体表紫色转变近乎成参体色,完成变色过程。     

四种重金属对刺参幼参的急性致毒效应

采用静水试验法,在水温18.5℃~20℃的条件下,用Cu2 ,Zn2 ,Cd2 和Cr6 四种重金属对刺参幼参进行了单一急性毒性试验。结果表明:Cu2 对刺参幼参的24h,48h,72h和96hLC50分别为0.299,0.176,0.133和0.120mg/L;Zn2 的24h,48h,72h和96hLC50分别为6.700,3.624,2.577和1.951mg/L;Cd2 的24h,48h和72hLC50分别为4.246,2.588和2.137mg/L;Cr6 的24h,48h和72hLC50分别为31.974,7.499和3.808mg/L;Cu2 ,Zn2 ,Cd2 和Cr6 对刺参幼参的安全浓度分别为0.018,0.362,0.259和0.750mg/L。四种重金属对刺参幼参的毒性大小依次为Cu2 >Cd2 >Zn2 >Cr6 。     

强光照和黑暗条件下刺参体壁的基因表达差异

参对光照变化非常敏感,研究刺参对光照的分子响应非常重要。本研究应用RNA测序获取了刺参暴露于强光（“强光”）、正常光照（“对照”）和完全黑暗（“黑暗”）环境下体壁的基因表达谱情况,通过“对照”与“黑暗”,“对照”与“强光”和“黑暗”与“强光”的比较,在|log₂ ratio|≥1和FDR≤0.001的标准下分别发现了1161、113和1705个差异表达基因（DEGs）。基因本体分析表明,“cellular process”和“binding”在“生物过程”和“分子功能”类别中的DEGs富集最多,而“cell”和“cell part”在“细胞组分”类别中的DEGs富集最多。将DEGs与Kyoto Encyclopedia基因和基因组数据库上的于214、41和229条通路进行比对,发现了51、2和57条通路分别显著富集。本研究发现的光特异性DEGs可作为深入研究刺参对光照变化的生化适应机制的重要目标基因。     

池塘养殖刺参安全度夏技术研究

文章概述夏季池塘养殖刺参死亡症状和灾情特点,分析刺参死亡原因主要包括池水盐度骤降、池水温度升高、池水pH值下降、池水分层、有机污染和刺参自身原因。在此基础上,提出池塘养殖刺参安全度夏的技术措施,主要包括科学合理选择养殖地址、规范池塘建设、做好度夏前准备工作、水质管理、及时清除水草和大型藻类、采取降温措施以及补充营养等,涉及大量具体操作方法,具有科学性和实用性。     

温度对刺参繁殖期消化酶和代谢酶活力的影响

为探究刺参(Apostichopus japonicus)繁殖期消化、代谢生理特点,作者以刺参亲参(240 g±15 g)为实验材料,设置了5个温度水平(6、10、14、18、21℃),研究了温度对刺参繁殖期内肠道消化酶和体壁肌肉代谢酶活力的影响。结果发现:温度对亲参繁殖期消化酶、代谢酶活力受培育水温影响显著(P0.05)。其中,胰蛋白酶与淀粉酶活力随水温升高呈先升高后降低的趋势,在10℃时活力最高且达到峰值;脂肪酶活力随着养殖水温升高呈下降趋势,在6℃时活力最高。己糖激酶、丙酮酸激酶、乳酸脱氢酶、苹果酸脱氢酶活力随水温升高出现先增高后降低的趋势,在10℃时达到峰值;而琥珀酸脱氢酶活力随温度升高呈下降趋势。研究表明,10~14℃温度下,亲参消化酶与代谢酶活力均处于较高水平,是理想的亲参室内促熟温度参数。     

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.     

Effect of Bacillus baekryungensis YD13 Supplemented in Diets on Growth Performance and Immune Response of Sea Cucumber（Apostichopus japonicus）

The effect of a potential probiotic on the growth performance and immune response of sea cucumber（Apostichopus japonicus） was investigated. Bacillus baekryungensis YD13 isolated from sea cucumber culturing ponds was added to sea cucumber basal feed as a probiotic in different doses（0, the control; 1×10^4（YD134）, 1×10^6（YD136） and 1×10^8（YD138） CFU g-1 of diet）, and administered orally to A. japonicus（initial mean wet weight 5.44 g ± 0.17 g）. The sea cucumbers were fed in 20 aquaria, 5 each treatment, for 60 d. At the end of growth trial, 20 sea cucumbers from each treatment were challenged with Vibrio splendidus. A. japonicus in YD134 and YD136 exhibited significantly better growth performance than control（P 〈0.05）. Five non-specific immune parameters including lysozyme, acid phosphatase, alkaline phosphatase, superoxide dismutase and catalase in coelomic fluid were measured to evaluate the immune response of A. japonicus to the probiotics. Results showed that all parameters were significantly improved when YD11 was supplemented in the dose of 1×10^6 CFU g^-1（P 〈0.05）. The cumulative incidence and mortality after the Vibrio splendidus challenge decreased significantly in sea cucumbers of YD136. Accordingly, 1×10^6 CFU g^-1 of YD13 in diet was recommended for the growth promotion and immune enhancement of A. japonicus.     

Identification of five sea cucumber species through PCR-RFLP analysis

Sea cucumbers are traditional marine food and Chinese medicine in Asia. The rapid expansion of sea cucumber market has resulted in various problems, such as commercial fraud and mislabeling. Conventionally, sea cucumber species could be distinguished by their morphological and anatomical characteristics; however, their identification becomes difficult when they are processed. The aim of this study was to develop a new convenient method of identifying and distinguishing sea cucumber species. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of mitochondrial cytochrome oxidase I gene (COI) was used to identifing five sea cucumber species (Apostichopus japonicus, Cucumaria frondosa, Thelenota ananas, Parastichopus californicus and Actinopyga lecanora). A 692 bp fragment of COI was searched for BamHI, KpnI, PstI, XbaI and Eco31I restriction sites with DNAMAN 6.0, which were then used to PCR-RFLP analysis. These five sea cucumber species can be discriminated from mixed sea cucumbers. The developed PCR-RFLP assay will facilitate the identification of sea cucumbers, making their source tracing and quality controlling feasible.