Preliminary Study of the Optimum Dietary Riboflavin Level in Sea Cucumber,Apostichopus japonicus (Selenka)

A 14‐wk feeding trial was carried out to evaluate the optimum dietary riboflavin (vitamin B₂) level in juvenile sea cucumber, Apostichopus japonicus. A total of 360 sea cucumbers averaging 1.49 ± 0.07 g (mean ± SD) were randomly distributed into 18 rectangular plastic tanks, and each tank was then randomly assigned to one of three replicates of six diets containing 0.29 (B₂0), 4.22 (B₂4), 8.93 (B₂9), 17.9 (B₂18), 56.7 (B₂57), and 577 (B₂577) mg riboflavin/kg diet. At the end of 14 wk of feeding trial, average weight gain (WG) of sea cucumbers fed B₂9, B₂18, B₂57, and B₂577 diets were significantly (P < 0.05) higher than that of animals fed B₂0 diet. WG of sea cucumbers fed B₂18, B₂57, and B₂577 diets were significantly higher than those of animals fed B₂0 and B₂4 diets. Specific growth rate of sea cucumbers fed B₂9, B₂18, B₂57, and B₂577 diets were significantly higher than those of animals fed B₂0 and B₂4 diets. Feed efficiency of sea cucumbers fed B₂9, B₂57, and B₂577 diets were significantly higher than those of animals fed B₂0 and B₂4 diets. Although significant differences were recorded in proximate composition of sea cucumbers fed the experimental diets, no clear trends were observed. The ANOVA test suggested that the optimum dietary riboflavin level in sea cucumber could be 17.9 mg/kg diet but broken‐line analysis of WG indicated a level of 9.73 mg riboflavin/kg diet. Therefore, these results may indicate that the optimum dietary riboflavin level in sea cucumber could be higher than 9.73 mg/kg diet but lower than 17.9 mg/kg diet.     

Preliminary Study of the Optimum Dietary Ascorbic Acid Level in Sea Cucumber,Apostichopus japonicus (Selenka)

A 14‐wk feeding trial was carried out to evaluate the optimum dietary ascorbic acid (AA) level in juvenile sea cucumber, Apostichopus japonicus. Sea cucumbers averaging 1.49 ± 0.07 g (mean ± SD) were randomly distributed into 18 rectangular plastic tanks of 20 L capacity in a recirculating system (20 animals per tank). Six semipurified experimental diets were formulated to contain 0 (l ‐ascorbyl‐2‐monophosphate [AMP]; AMP0), 30 (AMP24), 60 (AMP48), 120 (AMP100), 240 (AMP206), and 1200 (AMP1045) mg AA/kg diet in the form of AMP using casein as the main protein source. Sea cucumbers were fed each of the six experimental diets in triplicate groups. At the end of 14 wk of feeding trial, weight gain (WG), specific growth rate (SGR), and feed efficiency (FE) of sea cucumbers fed AMP100, AMP206, and AMP1045 were significantly (P < 0.05) higher than those of animals fed AMP0, AMP24, and AMP48. However, there were no significant differences in WG, SGR, and FE among sea cucumbers fed AMP100, AMP206, and AMP1045 and among animals fed AMP0, AMP24, and AMP48. Whole‐body vitamin C concentration increased with AA content of the diets. Broken‐line analysis of WG showed an optimum dietary AA level of 105.3 mg AA/kg diet in sea cucumber. These results indicated that the optimum dietary vitamin C level in sea cucumber in the form of AMP could be greater than 100 mg AA/kg diet but less than 105.3 mg AA/kg diet.     

Optimum Level of Dietary n‐3 Highly Unsaturated Fatty Acids for Juvenile Sea Cucumber,Apostichopus japonicus

A feeding trial was conducted to estimate the optimum level of dietary n‐3 highly unsaturated fatty acids (HUFAs) for juvenile sea cucumber, Apostichopus japonicas, based on growth performance and fatty acid compositions. Diets with five n‐3 HUFAs levels (0.15, 0.22, 0.33, 0.38, and 0.46%) were fed to sea cucumber juveniles (1.97 ± 0.01 g) once a day for 60 d. The sea cucumbers fed diets containing 0.22% n‐3 HUFAs showed significantly (P < 0.05) higher body weight gain, feed efficiency, and protein efficiency ratio than the sea cucumbers fed diets containing 0.15% n‐3 HUFAs, but not significantly different (P > 0.05) from those of sea cucumbers fed diets containing 0.33, 0.38, and 0.46% n‐3 HUFAs. The sea cucumbers fed diets containing 0.46% n‐3 HUFAs showed significantly (P < 0.05) higher eicosapentaenoic acid and saturated fatty acid than the sea cucumber fed diets containing 0.15% n‐3 HUFAs, but not significantly different (P > 0.05) from those of sea cucumbers fed diets containing 0.22, 0.33, and 0.38% n‐3 HUFAs. The results of growth performance and n‐3 HUFA compositions of body wall indicated that the optimum level of dietary n‐3 HUFAs for juvenile sea cucumber is between 0.22 and 0.46%.     

Using Phage PSM‐1 to Control Shewanella marisflavi Infection in Juvenile Sea Cucumber,Apostichopus japonicus

Shewanella marisflavi is considered responsible for infection in juvenile sea cucumber, Apostichopus japonicas. A bacteriophage specific to S. marisflavi was isolated from seafood wholesale market sewage and designated as vB_SmaM_PSM (PSM‐1). The phage was classified as a member of the Myoviridae family by negative staining transmission electronic microscopy. The stability of phage PSM‐1 was observed after treatment with different temperature gradients and pH values. The results showed that the lytic activity of phage PSM‐1 was optimum at 28 C with a pH of 7.0 and ceased at temperatures higher than 60 C. A one‐step growth curve analysis of the phage revealed eclipse and latent periods of 20 and 40 min, respectively, with a burst size of 219 plaque forming units per infected cell. The ability of phage PSM‐1 to inhibit the growth of S. marisflavi AP629 was tested in vitro using the phage with three multiplicity of infection (MOI) values of 0.1, 1, and 10. All treated cultures that produced an inhibition of the growth of S. marisflavi AP629 were compared with the untreated culture, and the treatment group with a MOI value of 10 showed the highest inhibition effect among the three doses. In an in vivo performance experiment, the sea cucumbers (15–20 g) were randomly assigned to five groups (one negative control, one antibiotic treatment group, and three phage treatment groups) in a phage injection trial as well as a phage immersion trial. All groups were infected with S. marisflavi strain AP629 (3.3 × 10⁶ colony forming units/g, LD₅₀ [lethal dose 50%] level) by intraperitoneal injection. After 8 d of observation, the survival rates ranged from 40 to 80% in all phage treatment groups, while 10% in the negative control group. The optimal therapeutic time points of the two trials were also determined. The results showed that using phage prior to infection could be a good choice to protect sea cucumbers. The overall results of this experiment indicate that using phage may be a feasible biological control agent to prevent S. marisflavi infection in sea cucumbers.     

复合蛋白酶在即食海参加工中的应用

用复合蛋白酶处理盐渍海参,作为加工冻干即食海参的预处理样品,研究其酶处理最适工艺。试验结果表明,酶处理温度40℃,处理50 m in,水浸泡13 h,酶用量为肉重0.075%时,处理后的海参胶原蛋白溶解度(DD%)为(38.14±0.52)%,与水发海参接近[(39.34±0.47)%],口感达到最佳。从酶处理海参和盐渍海参的红外光谱分析可知,二者的营养成分无明显差异,达到生产冻干即食海参的要求。     

低聚果糖对仿刺参生长及生长免疫因子的影响

在基础饲料中分别添加0(对照组)、0.20%、0.40%、0.60%、0.80%、1.00%的低聚果糖,制作6组等氮、等能的试验饲料,投喂初始体质量约20 g的仿刺参14周,研究低聚果糖对仿刺参生长、体成分、机体生长免疫因子及肠道糖代谢酶活力的影响。试验结果表明,低聚果糖添加量对仿刺参的成活率无显著影响(P>0.05),但显著影响了仿刺参的质量增加率和特定生长率(P<0.05);仿刺参体壁基本成分不受低聚果糖添加量的影响(P>0.05),总糖含量随低聚果糖的升高而升高(P<0.05);体壁中甘氨酸、半胱氨酸及精氨酸含量随低聚果糖含量的增加呈先升后降的趋势,分别在0.40%、0.50%、0.80%组达到最高,显著高于0%组(P<0.05);体腔液中生长激素、补体C3、补体C4及一氧化氮合酶含量均随低聚果糖的升高呈先升后降的趋势,但肠道丙酮酸激酶和己糖激酶活力均随低聚果糖的升高而升高(P<0.05)。以质量增加率为评价指标,经二次曲线回归分析,仿刺参饲料中低聚果糖的适宜添加量为0.55%。     

海参疾病学研究进展

海参具有极高的营养和药用价值,历来被视为海珍奇品。近年来,海参价格不断上涨,市场需求量迅速增加,随着采捕强度的增大,导致自然资源量急剧减少。为解决市场膨胀和资源缩减问题,海     

仿刺参SARM基因的克隆、表达和功能研究

采用RACE方法克隆了仿刺参TLR信号通路MyD88非依赖途径接头分子SARM,命名为AjSARM.AjSARM基因的序列全长3874 bp,开放阅读框为2412 bp,编码803个氨基酸.通过SMART软件对Aj SARM蛋白进行蛋白结构预测,发现Aj SARM蛋白由2个SAM结构域和1个C端TIR结构域组成.采用实...     

Effects of Dietary Taurine on the Growth,Digestive Enzymes,and Antioxidant Capacity in Juvenile Sea Cucumber,Apostichopus japonicus

Taurine has been widely used as a growth‐ or health‐promoting additive in aquatic animals because of its multiple functions, while little work has been done on its effects on sea cucumbers, in spite of the occurrence of serious diseases. In this study, juvenile sea cucumbers (4.68 ± 0.04 g) were fed diets supplemented with taurine at 0% (control), 0.1, 0.25, 0.5, 1, and 2% for 8 wk. At the end of an 8‐wk feeding trial, growth performance of sea cucumbers was not significantly affected by dietary taurine (P > 0.05). However, dietary taurine significantly elevated intestinal lipase activities of sea cucumbers (P < 0.05). Intestinal amylase activity and trypsin activity in sea cucumbers did not show significant changes after animals were fed diets supplemented with taurine (P > 0.05). Dietary taurine at all five dosages significantly increased total antioxidant capacity in sea cucumbers, while superoxide dismutase activity in groups with dietary taurine at 0.25 and 0.5% was significantly higher than that of the control group (P < 0.05). Therefore, it appears that taurine could be used as a potential feed additive to confer better health of farmed sea cucumbers.     

A3α‐peptidoglycan extracted from Bifidobacterium sp. could enhance immunological ability of Apostichopus japonicus

To assess the effects of A3α‐peptidoglycan (A3α‐PG) extracted from Bifidobacterium sp. on the immune response and disease resistance of sea cucumber, different concentrations (0, 0.5, 5 and 50 mg mL^?1) of A3α‐PG suspensions were used to perform hypodermic injection on Apostichopus japonicus, followed by a Vibrio splendidus challenge. Total coelomocyte count (TCC), phagocytosis activity and activities of four immunological enzymes in both cell‐free coelomic fluid (extra‐cellular, EC) and coelomocyte lysate supernatant (intracellular, IC), including acid phosphatase (ACP), alkaline phosphatase (ALP), superoxide dismutase (SOD) and peroxidase (POD), were measured at 2, 6, 14 and 24 h post injection (hpi). The TCC was not significantly affected (P > 0.05) by A3α‐PG, ranging from 1.84 × 10⁶ to 3.53 × 10⁶ cells mL^?1. The coelomocyte phagocytosis activity was significantly activated (P < 0.05) in all the A3α‐PG treatments, whereas no significant difference was observed between them except 24 hpi (P > 0.05). The EC‐ACP activity in the 5.0 mg mL^?1 treatment increased significantly (P < 0.05) at all sampling times, while the IC‐ACP activity in the 50 mg mL^?1 treatment increased significantly (P < 0.05) at 2 hpi. Also, the 5.0 mg mL^?1 treatment had significant (P < 0.05) increase in the EC‐ALP activity within 14 hpi and the EC‐POD activity at 2 hpi, respectively, while significantly (P < 0.05) enhanced IC‐ALP and IC‐POD activities were observed in the 50 mg mL^?1 treatment within 6 hpi and at 2 hpi, respectively. Only the 5.0 mg mL^?1 treatment showed significant (P < 0.05) increase in the EC‐SOD activity at 2 hpi and IC‐SOD activity within 14 hpi, respectively. The challenge test showed that the animals treated with 50 mg mL^?1 of A3α‐PG had notably lower cumulative mortality after 14 days following V. splendidus exposure. All together, these results suggest that A3α‐PG could positively enhance immune response that effectively promotes the health status of A. japonicus against V. splendidus infection.     

养殖仿刺参溃烂病病因初探

从患病仿刺参分离获得的两株优势菌2004—A1和2004—A2,经人工感染回接健康参试验,均获得与原发病相同症状,证实此两株菌是病原菌。另通过人工创伤感染试验确认海参受外伤后,容易感染发生溃烂病。     

刺参的增养殖技术

刺参隶属于棘皮动物门 ,海参纲 ,木盾手目 ,刺参科。产于黄、渤海的刺参 ,其蛋白质含量高 ,糖类丰富而不含胆固醇 ,可滋补强身。据有关资料记载 ,刺参可治阳痿、肠燥便秘、肺结核、再生障碍性贫血、糖尿病等 ;其体壁结缔组织、体腔膜和真皮内腺管所含的多种酸性粘多糖 ,对人体生长、愈创、抗炎、成骨和预防组织老化、动脉硬化等有着特殊功能 ;刺参肠子和性腺的加工品在日本被视为美味而备受欢迎 ,价格昂贵。刺参可谓全身是宝 ,因其经济价值极高而被称之为“参”中之冠。近几年 ,刺参的价格一路攀升 ,居高不下 ,吸引了众多的投资者在北方寻找…     

玉米蛋白含量对仿刺参幼参生长和消化的影响

在水温12.0～16.0 ℃下,将平均体质量8.56 g 的仿刺参幼参放养于容量50 L(50 cm× 40 cm ×30 cm)的水槽中,饲料以玉米蛋白为蛋白源,按饲料中粗蛋白含量12.0%、15.0%、18.0%、21.0%和24.0%,分别添加11.5%(F1)、16.1%(F2)、20.8%(F3)、25.7%(F4)和30.7 %(F5)玉米蛋白,以不同数量的海泥平衡.饲养60 d,各组幼参的存活率均为100%,但特殊生长率却随饲料中玉米蛋白含量的增加而下降.饲料中玉米蛋白含量由11.5%升至20.8%时,幼参的特殊生长率差异不显著(P＞0.05),当玉米蛋白含量达25.7%和30.7%时,幼参的特殊生长率显著低于F1、F2和F3组(P＜0.05).摄食不同试验饲料的幼参对饲料中蛋白质的消化率随饲料中玉米蛋白含量的增加而降低,消化道中蛋白酶和脂肪酶的活力差异不显著,而当饲料玉米蛋白含量超过20.8 %时,淀粉酶的活力明显下降.摄食试验饲料的幼参体内蛋白、脂肪、灰分和水分含量差异不显著(P＞0.05).     

辛硫磷对刺参幼参的急性毒性效应

研究了静水条件下辛硫磷对刺参幼参(0.8±0.1)g的急性毒性试验,以期为海参养殖环境监测提供理论依据。利用Bliss法,得出辛硫磷对刺参幼参在24、48、72 h半致死质量浓度分别为0.329、0.1970、.141 mg/L,并计算出安全质量浓度为0.021 mg/L。     

仿刺参纤维胶凝蛋白基因的分子特征及表达分析

纤维胶凝蛋白是非特异性免疫系统中的重要分子。通过转录组测序及cDNA末端快速克隆技术得到一条仿刺参纤维胶凝蛋白基因的全长cDNA序列,并将其编码的蛋白命名为仿刺参纤维胶凝蛋白-1。获得的基因cDNA全长为1951bp,其中5′-末端非翻译区为397bp,3′-末端非翻译区为666bp,开放阅读框为888bp,编码295个氨基酸,N端16个氨基酸为信号肽,信号肽后面有两个G-X-Y重复序列,C端为纤维蛋白素原结构域。采用实时荧光定量PCR方法分析了仿刺参纤维胶凝蛋白-1基因在仿刺参幼参不同组织及细菌脂多糖刺激后的时序表达规律,结果显示仿刺参纤维胶凝蛋白-1基因在仿刺参的肠道、呼吸树、体腔细胞和体壁均有表达,且肠道的表达量最高;脂多糖刺激后,4种组织的仿刺参纤维胶凝蛋白-1基因表达量均有变化,且以肠道和体壁表达量的变化最为显著;此外,仿刺参纤维胶凝蛋白-1基因在仿刺参4种组织中的表达量变化具有不同的时序性,表明仿刺参纤维胶凝蛋白-1可能在仿刺参免疫应答中起重要作用。     

仿刺参线粒体全基因组序列结构及比较研究

利用已构建的仿刺参cDNA文库得到的线粒体DNA(mtDNA)相关基因序列设计扩增引物,测定了大连仿刺参线粒体基因组全序列,并对其进行了基因构成和进化分析。仿刺参线粒体基因组序列长16 109bp,其基因构成与其他后口动物基本一致,包括37个基因(2个rRNA基因、22个tRNA基因和13个蛋白质编码基因)和3个主要的非编码区。在其37个基因中,ND6、tRNASer(AGN)、tRNAGln、tRNAAla、tRNAVal、TrnaAsp位于L链上,其余均位于H链上。在13个蛋白质编码基因中,除ND1的起始密码子为GTG外,其余均以ATG作为起始密码子;除Cytb以"T"作为终止密码子外,其他蛋白质基因均具有完全的终止密码子,且在已知的棘皮动物线粒体蛋白质基因中,部分基因的起始和终止密码子表现出一定的纲内特异性。比较分析了大连、青岛、威海仿刺参线粒体基因组,三者的基因组成和排列相同,碱基组成相近,蛋白质编码基因的起始和终止密码子完全一致,但存在核苷酸和氨基酸序列的差异。三者的控制区序列存在多个插入/缺失和SNP位点。根据COI、Cytb和ND4计算了三者之间的遗传距离为0.006～0.018,遗传距离分析和系统进化关系分析都显示青岛仿刺参和威海仿刺参的关系较大连仿刺参更近。     

仿刺参对盐度的耐受能力研究

设置不同的盐度梯度(10～45),根据仿刺参的存活、溃烂、着壁状态、活力、排便和疣刺伸展情况,进行仿刺参的耐盐范围和耐盐能力分析.结果表明,仿刺参适宜生长的盐度为20～35,最适生长盐度为25～30.研究发现,仿刺参在盐度为15和40下生长一段时间后,就会逐步适应其所生长的环境.     

Effects of dietary supplementation of A3α‐peptidoglycan on the growth,immune response and defence of sea cucumber Apostichopus japonicus

To determine the effects of A3α‐peptidoglycan (A3α‐PG) extracted from Bifidobacterium sp. on the growth, immune response and disease resistance of sea cucumber Apostichopus japonicus, a 70‐day feeding trial was conducted in this study. A total of 216 sea cucumbers were fed with four practical diets prepared from a commercial feed with different contents (0, 1.5, 2.5 and 4.0 g kg^?1) of A3α‐PG. The specific growth rate (SGR), total coelomocyte count (TCC), phagocytotic activity and activities of four immunological enzymes in both cell‐free coelomic fluid (extracellular, EC) and coelomocyte lysate supernatant (intracellular, IC), including acid phosphatase (ACP), alkaline phosphatase (ALP), peroxidase (POD) and superoxide dismutase (SOD), were measured at the end of the feeding trial. Finally, the animals were administered a 16‐day Vibrio splendidus challenge via intraperitoneal injection to test the potency of A3α‐PG on disease resistance. Compared with the control (0 g kg^?1 A3α‐PG), a significant increase (P < 0.05) in SGR was observed in the groups fed with 1.5 and 2.5 g kg^?1 A3a‐PG. The TCC, ranging from 7.25 × 10⁶ to 1.05 × 10⁷ cells mL^?1, was not significantly affected (P > 0.05) by A3α‐PG,. Coelomocyte phagocytotic activities in all of the A3α‐PG‐supplemented groups were significantly activated (P < 0.05), but no significant difference (P > 0.05) was observed. Sea cucumbers fed with 1.5 and 2.5 g kg^?1 A3α‐PG exhibited significant activation (P < 0.05) of EC/IC‐ACP, EC/IC‐ALP, and EC/IC‐POD activities. A significant increase in EC‐SOD activities (P < 0.05) was exhibited by all groups with A3α‐PG supplementation. The challenge test showed that animals fed with diets containing 2.5 and 4.0 g kg^?1 A3α‐PG had significantly lower cumulative mortalities compared with the control 16 days after exposure. All of the results presented here show that A3α‐PG can positively enhance the growth, immune response and disease resistance of sea cucumber, suggesting that dietary supplementation of A3α‐PG has potential applications in the health management of economic species of sea cucumber.