刺参南方度夏试验

选择刺参南方池塘养殖收获时的小规格个体,采用鲍鱼养殖池流水度夏,时间超过6个月。2年刺参度夏存活率分别为70.6%和89.1%;失重率为56.1%和61.8%。度夏后刺参再养殖约150天,成活率分别为94.9%和81.1%;增长率为10.5倍和13.6倍。     

Motility and fertility of cryopreserved spermatozoa of the Japanese sea cucumber Apostichopus japonicus

We developed both a cryopreservation method for Japanese sea cucumber spermatozoa and an artificial fertilization method using post‐thaw spermatozoa. Twenty per cent dimethyl sulfoxide (DMSO), 16% foetal bovine serum, and 64% artificial seawater were suitable cryodiluent, and the diluent was pre‐cooled to 0°C. Semen was diluted with the solution and enclosed in a 250 μl straw, cooled to ?50°C at 10.4 ± 0.4°C/min, and immediately immersed in liquid nitrogen. Although this method showed the highest post‐thaw motility in all the conditions we examined, its post‐thaw motility was still less than approximately 15%. Artificial fertilization was carried out by adding post‐thaw semen with a cryodiluent to the oocytes. The fertilization rate of 200 oocytes/ml seawater increased with the amount of post‐thaw semen from 1 to 5 μl but showed a significant decrease at 25 μl. This decrease was considered to be due to DMSO in the cryodiluent, because the fertilization rate of the fresh semen decreased sharply when the DMSO concentration around the oocytes was 1.0% or more. Further improvement in increasing post‐thaw motility and lowering the cryoprotectant concentration is necessary for commercial‐scale artificial fertilization.     

Effects of light intensity on daily activity rhythm of juvenile sea cucumber,Apostichopus japonicus (Selenka)

Eight light‐intensity treatments (natural light, continuous darkness, and 15, 30, 60, 125, 250 and 500 lx under LD 12:12 cycle) were used to investigate the effects of light intensity on the daily activity of 30.27±3.08 g sea cucumber Apostichopus japonicus (Selenka). Cyclic nocturnal activity patterns of behaviour were observed at different light intensities in the range of 15–500 lx under LD 12:12 cycle. And an ongoing nocturnal cycle persisted in DD cycle for up to 8 days, but longer feeding time and less marked rhythm occurred at continuous darkness. Under poor light conditions (I<5.18 lx), the daily activity rhythm of A. japonicus was governed by an innate biological clock and the effect of light intensity was not significant among different treatments. And more individuals tended to retreat to shelters (from 56.04% to 91.83%) with the increase of light intensity within the weak light condition (from 5.18 to 278 lx). However, the daily behaviours of A. japonicus were influenced under strong light conditions (>278 lx). Less than 8.17% individuals kept actively feeding and the proportion was not decreased with the increase of light intensity.     

A new system for the culture and stock enhancement of sea cucumber,Apostichopus japonicus (Selenka),in cofferdams

A new multilayer, plate‐type system for the culture and stock enhancement of sea cucumbers in cofferdam was developed. To optimize and evaluate the system, four experimental designs were implemented using polyethylene (PE)‐corrugated sheets of various colours, interval spacing and shapes/styles. Results showed that a system equipped with black PE‐corrugated sheets attracted more animals than either blue, green, transparent or a selection of mixed sheets (six transparent sheets in the upper layer and five black sheets in the lower layer) (P<0.05). Also, more animals gathered in the system with oblique‐angled sheets (30° to the base plate) than either a wavy (the bottom and every second sheet was at an angle of 10° to the base plate) or parallel arrangement (P<0.05), and more animals assembled in the system with 2 cm between sheets than spacings of 3, 4 or 5 cm (P<0.05). As expected, the upper layers of the systems attracted more animals than lower layers in most cases except for those with transparent and mixed oblique‐angled sheets with a 3 cm spacing (P<0.05). Thus, a system with black, oblique‐angled‐corrugated sheets and 2 cm spacing is recommended for Apostichopus japonicus (Selenka) culture and stock enhancement in cofferdams or ponds.     

Microsatellite genetic variation in wild and hatchery populations of the sea cucumber (Apostichopus japonicus Selenka) from northern China

Farming of the sea cucumber Apostichopus japonicus (Selenka) started 20 years ago and is still in rapid expansion in China. In order to assess the genetic status of both wild and cultivated stocks of this species, we used eight microsatellite markers to estimate the level of genetic diversity within five hatchery stocks and two wild populations of A. japonicus, and compared the degree of genetic differentiation between them. High levels of polymorphism were observed over all loci. The mean alleles and expected heterozygosities over the seven stocks were 10.4–12.3 and 0.735–0.783 respectively. The results of the microsatellite survey provide no evidence to show that hatchery practice of the sea cucumber in China to date has significantly affected the genetic variability of the cultured stocks. Significant differentiation was found between most pairs of the hatchery stocks and wild populations (F_st range: 0.008–0.036), and no obvious difference was detected between the wild populations (F_st=0.008). The information on the genetic variation and differentiation obtained in this study can be applied for future genetic monitoring of A. japonicus aquaculture stocks and will be useful for future genetic improvement by selective breeding, and for designing suitable management guidelines for these genetic materials.     

Influence of vibration caused by sound on migration of sea cucumber Apostichopus japonicus

Vibrations exist widely in the ocean, and the one caused by sound and biological movement plays an important role in the sensory system of marine organisms. The vibration caused by sound has many characteristics that make it a good candidate for modifying the movements of marine organisms. However, few studies have tested whether hydraulic vibration caused by sound has effects on the migration of sea cucumbers, and this has never been systematically studied previously in Apostichopus japonicus (Selenka). In order to understand whether hydraulic vibration caused by sound at different frequencies could affect the migration of sea cucumbers, in the present study, hydraulic vibration caused by sound at various frequencies was tested in the laboratory to determine their effects on the migration of A. japonicus at different sizes. The mean probability distribution was used as a statistical index to demonstrate the moving tendencies of the species. The experimental results showed that medium and small A. japonicus (<10 g ind.^?1) tend to move towards the low‐frequency vibration source caused by sound (100 Hz), whereas all sizes of A. japonicus tend to move backwards the high‐frequency vibration source caused by sound (10 000 and 28 000 Hz). This study provides basic data on the behavioural ecology of this species, as well as theoretical reference for the design of artificial reefs and harvesting equipment for A. japonicus.     

Effects of body size and water temperature on food consumption and growth in the sea cucumber Apostichopus japonicus (Selenka) with special reference to aestivation

To investigate the effects of body size and water temperature on feeding and growth in the sea cucumber Apostichopus japonicus (Selenka), the maximum rate of food consumption in terms of energy (C_maxe; J day^?1) and the specific growth rate in terms of energy (SGRe; % day^?1) in animals of three body sizes (mean±SE) – large (134.0±3.5 g), medium (73.6±2.2 g) and small (36.5±1.2 g) – were determined at water temperatures of 10, 15, 20, 25 and 30°C. Maximum rate of food consumption in terms of energy increased and SGRe decreased with increasing body weight at 10, 15 and 20°C. This trend, however, was not apparent at 25 and 30°C, which could be influenced by aestivation. High water temperatures (above 20°C) were disadvantageous to feeding and growth of this animal; SGRe of A. japonicus during aestivation was negative. The optimum temperatures for food consumption and for growth were similar and were between 14 and 15°C, and body size seemed to have a slight effect on the optimal temperature for food consumption or growth. Because aestivation of A. japonicus was temperature dependent, the present paper also documented the threshold temperatures to aestivation as indicated by feeding cessation. Deduced from daily food consumption of individuals, the threshold temperature to aestivation for large and medium animals (73.3–139.3 g) was 24.5?25.5°C, while that for small animals (28.9–40.7 g) was between 25.5 and 30.5°C. These values are higher than previous reports; differences in sign of aestivation, experimental condition and dwelling district of test animals could be the reasons.     

刺参一种囊膜病毒的分离及其超微结构观察

利用电镜负染技术检测发病的养殖刺参[Apostichopus japonicus（Selenka）]组织提取液。观察发现，提取液中存在大量病毒样粒子，该病毒粒子近似球形，具有囊膜。囊膜内可见高电子密度的核心结构。完整的病毒粒子直径为80～100nm，囊膜厚6～10nm，核心结构直径35-45nm，呈六边形。应用超薄切片技术对刺参的触手顶部、触手臂、疣足、呼吸树、背肠血管、肠等组织的病毒感染状况进行观察，发现该种病毒粒子大量存在于所检测各组织内。感染细胞超微结构表现为大量细胞器崩解形成空泡结构，并出现“髓袢样”结构等病理变化。根据观测结果，该病毒是一种无包涵体病毒。     

Effects of differently processed diets on growth,immunity and water quality of the sea cucumber,Apostichopus japonicus (Selenka, 1867)

The type of formulated diet used in aquaculture systems affects the feed efficiency and water quality. In this study, the water quality, growth performance and immunity of the sea cucumber, Apostichopus japonicus (Selenka, 1867), fed diets produced using four different processing methods (extruded pellet diet, cold‐bonded pellet diet, flake diet and mash diet) were quantified for 60 days to identify the optimal feed type for culturing of this species. Sea cucumbers grew faster, the feed conversion ratio was much lower, and immunity indicators were higher when fed the extruded pellet diet compared to the other diets. Sea cucumbers fed the extruded pellet diet had the highest specific growth rate (0.96), whereas A. japonicus fed the cold‐bonded pellet diet had the lowest specific growth rate (0.51). The lysozyme activity of sea cucumbers fed the extruded pellet diet was higher (57.52) than that of A. japonicus fed the cold‐bonded pellet diet (39.22), flake diet (37.32) and mash diet (32.68). When animals were fed the mash diet, the ammonia–nitrogen, nitrite–nitrogen and phosphate productions were higher than in animals fed the other diets. These results indicate that the extruded pellet diet is the optimum feed type for use in the culture of A. japonicus.     

不同光照强度对刺参幼参生长的影响

采用室内受控的实验方法研究了不同光照强度对刺参幼参生长的影响。实验设暗光、微强光、强光3个光照处理,同时每个光照处理组分别投喂两种饵料(人工配合饲料、人工配合饲料加底栖硅藻)。研究结果表明,不同光照强度下,幼参的生长差异显著(P<0.05),每种饵料处理下强光处理组幼参的特定生长率和日增重明显高于暗光处理组,而相同光照强度下两种饵料处理对幼参生长的影响差异不显著(P>0.05)。3个光照处理下幼参的特定生长率平均分别为0.26%、0.63%、0.98%/d,日增重为0.53、1.39、2.26 g/d。     

不同分子量壳聚糖对刺参(Apostichopus japonicus Selenka)生长和免疫功能的影响

以初始体重为(6.77±0.01)g的仿刺参(Apostichopus japonicus Selenka)为实验对象,在基础饲料中添加1％不同分子量的壳聚糖,分子量分别为35 kDa和400 kDa,进行为期56d的养殖实验,研究低分子壳聚糖(LMWC)和高分子壳聚糖(HMWC)对仿刺参生长和免疫相关酶活性的影响.结果显示,不同分子量的壳聚糖对仿刺参的生长均有促进作用,且LMWC能显著促进刺参的特定生长率(SGR)(P＜0.05).饲料中添加不同分子量壳聚糖,刺参体腔细胞中的总超氧化物歧化酶(T-SOD)活性比对照组均显著增高(P＜0.05).LMWC可显著增强刺参体腔细胞中酸性磷酸酶(ACP)活性(P＜0.05),并能提高碱性磷酸酶(AKP)和一氧化氮合酶(NOS)活性.HMWC对刺参体腔细胞中AKP和NOS活性均有显著增强作用(P＜0.05).研究表明,在刺参饲料中添加一定量的壳聚糖,对其生长和相关免疫酶活性有促进作用.     

5种海藻在刺参幼参饲料中的应用研究

本研究通过分析刺参(Apostichopus japonicus)幼参[(10.02±0.03) g]生长性能、体组成、肠道消化酶活性及非特异性免疫性能的变化,评价鼠尾藻(Sargassum thunbergii)、铜藻(Sargassaum horneri)、海带(Saccharina japonica)、海带渣、石莼(Ulva lactuca L.)及混合藻粉在刺参幼参养殖中的应用效果,在室内循环水系统中进行了为期56 d的养殖实验。结果显示,1) 鼠尾藻和混合藻粉组刺参的增重率(WGR)和特定生长率(SGR)极显著高于其他各藻粉组(P<0.01),海带渣组刺参的WGR和SGR最低,肠体比(IBR)和脏体比(VBR)均显著低于其他各组(P<0.05),藻粉对刺参存活率(SR)影响不显著(P>0.05);2) 不同藻粉对刺参体壁水分、粗灰分、粗蛋白和粗脂肪影响不显著(P>0.05);3) 鼠尾藻和混合藻粉组肠道胰蛋白酶活性极显著高于海带、海带渣和石莼组,海带渣组最低(P<0.01);藻粉对α-淀粉酶和脂肪酶活性无显著影响(P>0.05),但对刺参粪便的酸不溶性灰分具有极显著影响(P<0.01);4) 鼠尾藻组和混合藻粉组刺参肠道超氧化物歧化酶(T-SOD)、酸性磷酸酶(ACP)和碱性磷酸酶(AKP)活性极显著高于其他各组(P<0.01),海带渣组丙二醛(MDA)含量极显著高于其他各组(P<0.01)。研究表明,在本实验条件下,综合考虑刺参的生长性能及非特异性免疫性能,添加任一海藻均未达到替代鼠尾藻的饲喂效果,但多种海藻混合添加,其生长性能及非特异性免疫性能与单独添加鼠尾藻效果一致。     

仿刺参基因组大小的测定

仿刺参基因组大小测定采用流式细胞术,以鸡红血细胞DNA含量(2.5 pg/2C)为内标,用夏眠、正常和野生3个群体的仿刺参体腔细胞测定了52只刺参的单倍体基因组含量,其C-值为(0.90±0.06)pg,由此得出仿刺参基因组大小为(880.2±58.68)Mb。其中,夏眠、正常和野生群体C-值分别为(0.93±0.05)、(0.90±0.05)和(0.84±0.02)pg。运用独立样本t检验对比分析显示:(1)夏眠与正常的仿刺参基因组大小之间差异不显著,说明仿刺参基因组大小与夏眠这一生理因子不具有相关性;(2)池塘(围堰)养殖环境下与野生环境下的仿刺参基因组大小之间有显著差异,野生环境下的仿刺参基因组明显小于池塘(围堰)养殖环境下的基因组。据此推测,环境因子的改变可能对仿刺参乃至整个海参类群基因组DNA含量产生影响。实验还对C-值测定的发展和方法作了详尽的描述,同时在综合其他种类海参基因组大小研究的基础上,探讨了C-值与进化地位之间的关系。     

利用塑料大棚晒水进行刺参种苗越冬试验

2002年11月~2003年4月,在瓦房店市交流岛前哨海参育苗厂800 m3水体室内水泥池中,投放平均规格为5000头/kg,刺参种苗196 kg(98万头)。在没有锅炉进行水加温条件下,利用600 m3水体塑料大棚晒水进行刺参种苗越冬,水温2.0~13.2℃,平均盐度31.2,pH值8.2,DO>7 mg/L,经过近6个月的培育,共培育出规格为876头/kg的参苗836 kg(73.2万头),成活率74.7%,总重量增加了8.5倍。     

“胃萎缩症”仿刺参幼体及亲参组织中病毒观察

海参含有丰富的蛋白质和粘多糖,营养和药用价值极高.全世界海参约有1200种,在我国海域分布的有140多种,其中以黄、渤海的仿刺参(Apostichopus japonicus)品质最佳[1].近几年,仿刺参(俗称"刺参")人工养殖已成为辽宁、山东沿海水产业重要的养殖品种之一和新的经济增长点.     

Comparative study of water environment variation in the industrial aquaculture system of sea cucumber Apostichopus japonicus

This study investigated the microbial community, water quality, bacterial densities and growth performance in the industrial aquaculture system of sea cucumber Apostichopus japonicus without water exchange. Six treatments were set including, C (control treatment without sea cucumber culturing), S (small individual treatment), B (big individual treatment), Sd (small individual treatment with high density), Ss (small individual treatment adding carbohydrate source) and Sb (small individual treatment adding effective microorganisms). A total of 27916–32236 optimized reads and 564–742 operational taxonomic units (OTUs) were obtained from each samples. The phylum Proteobacteria, Actinobacteria, Bacteroidetes and Cyanobacteria predominated, representing 69.01–97.21% of the bacterial communities in the water samples. Cluster analysis and principal component analysis (PCA) showed that higher similarity was observed among S, Sd, Ss and Sb. The densities of TB and Vibrio in Ss were significantly higher than those in the other culture treatments after the 7th day. The concentrations of NH₄‐N, NO₂‐N, NO₃‐N and PO₄‐P in Ss and Sb were relatively lower than the other treatments. Conclusively, no deterioration was found in the water environment parameters during the 21‐day culture period without water exchange, indicating that low‐level water exchange protocol may be applied to the industrial aquaculture system. Based on the effects of different operations on culture system, industrial aquaculture is proved to be a viable way to rear sea cucumber.     

间歇投喂添加蛋氨酸硒饲料对仿刺参（Apostichopus japonicus Selenka）幼参生长和免疫指标的影响

在水温9℃～13℃下,将平均体质量0.745±0.3 g的仿刺参（Apostichopus japonicus Selenka）随机投放到50cm×40cm×30cm的塑料水槽内,每箱10头,共分为五组,每组3个平行,采用不同间隔时间投喂每kg添加0.5 mg蛋氨酸硒的饲料：第1、2、3、4组分别间隔0、5、10和15...     

Effects of a probiotic mixture (Bacillus subtilis YB‐1 and Bacillus cereus YB‐2) on disease resistance and non‐specific immunity of sea cucumber,Apostichopus japonicus (Selenka)

The effect of a commercially available compound probiotics product containing Bacillus subtilis YB‐1 (50%) and Bacillus cereus YB‐2 (50%) fed to sea cucumbers, Apostichopus japonicus (Selenka) on challenge infections and non‐specific immune responses was assessed. Sea cucumbers (were randomly allocated into nine aquariums at a density of 30 sea cucumbers per tank and triplicate groups) were fed diets containing 0 (control), 10⁷ and 10¹⁰ cfu (g diet)^?1 of the probiotics mixture for 32 days. The growth factors and immunological parameters were measured. In addition, the effects on resistance against Vibrio alginolyticus infection were also evaluated. The results indicate that all the immunological parameters (phagocytic activity, superoxide anion production, lysozyme activity, catalase activity and phenoloxidase activity) measured and the growth rate of sea cucumbers fed 10¹⁰ cfu of the probiotics mixture were significantly (P < 0.05) improved than control groups at 16 and 32 days. After challenging, the cumulative mortality for the control was 100%, whereas the cumulative mortality for sea cucumbers fed 10¹⁰ cfu of the probiotics mixture was 47% (P < 0.05). Although the total autochthonous intestinal heterotrophic bacterial counts were not affected by dietary treatment (P > 0.05), Bacillus sp. levels were significantly elevated in sea cucumbers fed the probiotics mixture (P < 0.05). These results confirmed that administration of the probiotics mixture in the diet stimulated non‐specific immune responses and enhanced the growth performance of sea cucumbers, and was effective in controlling infections caused by V. alginolyticus.