Effects of dietary β-glucan and glycyrrhizin on non-specific immunity and disease resistance of the sea cucumber (<Emphasis Type="Italic">Apostichopus japonicus</Emphasis> Selenka) challenged with <Emphasis Type="Italic">Vibrio splendidus</Emphasis>

Sea cucumbers, Apostichopus japonicus Selenka, were fed diets containing non-immunostimulant (basal diet), 0.2% β-glucan and 0.02% glycyrrhizin in a recirculatory water system for 45 days, and subsequently challenged with Vibrio splendidus by injection at 1.0×10⁸ cfu / sea cucumber for 15 days. Phagocytic capacity (PC), intracellular superoxide anion production (ISAP), lysozyme (LSZ) activity and superoxide dismutase (SOD) activity in the coelomic fluid were analyzed on the 0^th, 5^th, 10^th and 15^th days after injection. Results showed that after the 45-day feeding period, PC, ISAP, LSZ activity and SOD activity in sea cucumbers fed with dietary β-glucan or glycyrrhizin were significantly higher than in those fed with the basal diet. On the 5th day after infection, all the immune parameters examined in the sea cucumbers injected with V. splendidus decreased in value significantly. On the 15^th day, PC, ISAP and LSZ activity returned to levels similar to those on the 0^th day. For the sea cucumbers injected with saline, there were no significant differences in all the immune parameters examined and in the cumulative morbidity during the 15-day challenging trial. After injecting with V. splendidus, the cumulative morbidity of sea cucumbers fed with the basal diet was significantly higher than those fed with dietary β-glucan or glycyrrhizin when challenged with V. splendidus challenged sea cucumber fed with the basal diet was significantly higher than those fed with dietary β-glucan or glycyrrhizin. There was no significant difference in cumulative morbidity between the dietary β-glucan and glycyrrhizin treatments over time.     

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.     

不同温度的有机溶剂分提对鯷鱼鱼油得率及脂肪酸组成的影响

讨论了不同温度的有机溶剂分级提取对鳀鱼鱼油得率、脂肪酸组成及EPA、DHA含量变化的影响。经15℃干法分级提取的液态鳀鱼鱼油,在设定的4种温度下,液态鳀鱼鱼油得率均随溶剂量的增加而提高;在相同溶剂量的情况下,随着分级提取温度的降低,液态鳀鱼鱼油得率呈下降的趋势。有机溶剂分级提取对14碳短碳链的饱和与不饱和脂肪酸及16碳、18碳的饱和脂肪酸的脱除效果是明显的,但对脱除20碳、22碳的长碳链饱和脂肪酸的效果不明显。在4种试验温度下,EPA和DHA的总量分别为29.88%、31.74%、35.27%和41.80%,高出美国阿拉斯佳深海鱼油对应总量(28.76%)的1.12%～13.04%。     

温度变化对刺参同工酶的影响

采用同工酶电泳方法,对在不同温度处理下刺参(Apostichopus japonicus)的乳酸脱氢酶(LDH)、酯酶(EST)和超氧化歧化酶(SOD)的表达情况进行分析。本文结果显示刺参组织内无LDH同工酶的表达。在10℃到20℃的温度突变处理中,12 h内EST和SOD均有新增酶带,在20℃到10℃的温度突变1 h内EST表达有新增酶带。3 h内SOD表达有新增酶带。在不同变温幅度的处理中,EST和SOD只有1条酶带。这表明,刺参对温度突变可产生积极的响应,对于其适应环境温度的变化具有重要意义。     

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 。     

鲈鱼苗烂鳃、烂尾病病原菌的研究

１９９７年４～５月,对花鲈（Ｌａｔｅｏｌａｂｒａｘｊａｐｏｎｉｃｕｓ）鱼苗期细菌性烂鳃和烂尾病的病原菌进行了研究。从具有明显症状的病鱼的病灶组织分离到２株致病菌Ｃ－１和Ｗ－１,经人工感染及从人工感染发病鱼体再分离的Ｗ－４和Ｗ－５菌株的再感染试验结果表明,所分离的细菌为鲈鱼苗烂尾,烂鳃病的致病菌。经６４项生物学特性,生理生化特征测试结果表明,这４株菌可归为一类,同属于鳗弧菌（Ｖｉｂｒｉｏａｎｇｕｉｌａｒｕｍ）。药敏试验结果表明,痢特灵、ＣＩＮ１００、ＭＥＬ２５、呋喃妥因、氯霉素、磺胺药＋ＴＭＰ和美洛西林等７种药物对病原菌有显著的抑制作用。     

Nitrogen and Phosphorus Budget of a Polyculture System of Sea Cucumber （Apostichopus japonicus）, Jellyfish （Rhopilema esculenta） and Shrimp （Fenneropenaeus chinensis）

The nitrogen(N) and phosphorus(P) budget and the ecological efficiency of a polyculture system of sea cucumber(Apostichopus japonicus), jellyfish(Rhopilema esculenta) and shrimp(Fenneropenaeus chinensis) were studied in a cofferdam, 120.2 ha in size. The nutrients were supplied by spring tide inflow. In total, 139600 kg N yr-1 and 9730 kg P yr-1 input to the system; while 118900 kg N yr-1 and 2840 kg P yr-1 outflowed from the system concurrently, thus the outflow was 85.7%(N) and 29.2%(P) of inflow. The production of N and P was 889.5 kg yr-1 and 49.28 kg yr-1(sea cucumber) and 204 kg yr-1 and 18.03 kg yr-1(jellyfish and shrimp), respectively. The utilization rate of N and P by polycultured animals was 7.8‰ and 6.9‰, respectively, 21.9% and 38% higher than that of monocultured sea cucumber. Our results indicated that the polyculture system was an efficient culture system of animals and a remediation system of coastal environment as well; it scavenged 14.3% and 70.8% of N and P, respectively. Such an ecological efficiency may be improved further by increasing either the stocking density or the size of sea cucumber or both.     

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.     

Influence of dietary probiotic Bacillus TC22 and Prebiotic fructooligosaccharide on growth, immune responses and disease resistance against Vibrio splendidus infection in sea cucumber Apostichopus japonicus

The effects of probiotic Bacillus TC22 (isolated from intestine of infected sea cucumber) and prebiotic fructooligosaccharide (FOS) on growth, immunity and disease resistance in sea cucumber Apostichopus japonicus were studied. Six experimental diets were formulated with combinations of three levels of TC22 (0, 107 and 109 CFU g-1 diet) and two levels of FOS (0 and 0.5%) in a 3 × 2 factorial experiment. At the end of the 8-week feeding trial,animals were challenged by injecting Vibrio splendidus. The results revealed that the specific growth rates (SGR) of sea cucumbers were not affected by TC22 and FOS, or the interaction between TC22 and FOS (P > 0.05). However,there were significant interactions between TC22 and FOS for immune response and disease resistance in sea cucumbers (P < 0.05). When sea cucumbers were fed with TC22 at 109 CFU g?1 feed and 0.5% FOS alone or in combination, the phagocytosis, respiratory burst and phenoloxidase activity of sea cucumber coelomocytes were significantly enhanced;the disease resistance against V. splendidus infection was also increased significantly (P < 0.05).However, dietary combination of TC22 at 109 CFU g-1 diet and 0.5% FOS did not yield significantly higher efficiency than either TC22 at 109 CFU g-1 diet or 0.5% FOS used alone (P > 0.05). Therefore, further studies should examine the effects of combinations of other levels of FOS(> 0.5% or < 0.5%) and TC22 on the immunity and disease resistance of sea cucumbers.     

盐度调控对花鲈(Lateolabrax maculatus)生长相关基因表达的影响

生长激素受体(GHR)作为GH/IGF轴的中心环节,在内分泌调控中发挥重要作用。本实验采用c DNA末端快速扩增法(RACE)技术克隆出花鲈GHR1和GHR2的c DNA全长序列。急性低盐度调控实验设为海水组、半海水组和淡水组。测定了急性低盐度调控24、48、96、144、192h后,花鲈肝脏中GHRs、IGF-1及垂体GH表达情况。结果表明,GHR1 c DNA全长序列2436bp,编码637个氨基酸;GHR2 c DNA全长序列2940bp,编码582个氨基酸。GHR1与GHR2由信号肽、胞外区、跨膜区、胞内区组成,且结构存在差异。脑、肾、鳃中GHR1表达明显高于GHR2;而在肌肉、垂体、肝脏、盲肠、胸腺、心脏中,GHR2表达明显高于GHR1。24h时,各组GHR1表达不变,GHR2、GH、IGF-1显著下降。之后,相对于海水组,淡水组和半海水组GHRs和IGF-1表达升高,而GH下降,GH与GHR负相关。据结果推测,花鲈GHR2可能为SL受体,GH/IGF轴参与低渗调控可能是通过增加GHRs,进而激活下游IGF-1表达而实现。