浙江沿海三疣梭子蟹的养殖模式

为开发三疣梭子蟹低盐耐受性相关甲基化标记,采用甲基化敏感性高分辨率熔解曲线法(methylation sensitive high resolution melting curve, MS-HRM)进行了三疣梭子蟹低盐耐受性相关甲基化位点的筛选和应用。结果显示,从三疣梭子蟹转录组数据库中共开发了8个甲基化标记,其中6个标记与低盐度耐受性显著相关,在低盐度条件下表现出显著的去甲基化或甲基化,这些标记分别位于V-ATP酶(V-type proton ATPase)、CLC型氯离子通道、琥珀酸脱氢酶、NAD(P)转氢酶(NAD(P)、磷酸乙醇酸性磷酸酶、NADH还原酶基因,为三疣梭子蟹耐低盐新品种的分子标记辅助选育提供候选工具。研究表明,“宁象1号”耐低盐能力显著高于野生群体,且选育群体中位点Pt-M1、Pt-M2的去甲基化率达到100%,可能有利于低盐条件下保持体内Na⁺和Cl⁻的平衡。     

菲律宾蛤仔二种I型溶菌酶的重组表达和抑菌活性

为研究病原微生物刺激后,菲律宾蛤仔I型溶菌酶的免疫应答反应,实验采用重组表达技术,从菲律宾蛤仔中得到2种I型溶菌酶的重组蛋白rVpILYZ-1和rVpILYZ-2,发现其均具有较为广谱的抑菌活性;其中,rVpILYZ-1对副溶血弧菌、灿烂弧菌和藤黄微球菌等具较强的抑菌活性,而rVpILYZ-2对副溶血弧菌抑菌活性较强,且二者均通过非酶抑菌活性和胞壁酸酶活性来发挥抑菌作用。结果显示,rVpILYZ-1最适pH和温度分别为6.5和20 ℃,而rVpILYZ-2最适pH和温度则分别为4.5和10 ℃。此外,rVpILYZ-1和rVpILYZ-2均具免疫调理活性,能够促进血细胞对病原菌的吞噬作用。研究表明,VpILYZ-1和VpILYZ-2在菲律宾蛤仔清除病原微生物的免疫应答反应中发挥重要作用。     

基于图像识别技术研究不同海区三疣梭子蟹甲壳白色斑纹特征及蜕壳前后斑纹特征的变化

为进行三疣梭子蟹外观特征的研究,本研究通过计算机视觉技术建立三疣梭子蟹背部白色斑纹评估方法,经人工对比验证其准确性达100%。在此基础上,对来自海域的30只三疣梭子蟹[平均体质量(2.8±0.56)g]进行跟踪观察,发现蜕壳前后白色斑纹数量和位置保持不变,而白色斑纹面积和斑纹区域面积则会随着蜕壳后蟹壳面积的增加而扩大;蜕壳后,白色斑纹面积增加89.33%±8.61%,斑纹区域面积增加90.51%±7.95%,蟹壳面积增加94.66%±8.26%,均显著正相关。为进一步研究不同海区梭子蟹白色斑纹分布特征,对来自朝鲜、中国辽宁和浙江等不同海区的三疣梭子蟹进行白色斑纹个数(N)、白色斑纹面积(S1)、斑纹区域面积(S2)、白色斑纹面积百分比(R1)、斑纹区域面积百分比(R2)和密度(D)等6项参数分析,结果发现,不同海区三疣梭子蟹斑纹性状存在显著特点,朝鲜海区呈现斑纹小、分布面积少、占蟹壳总面积比例低的特点;辽宁海区呈现斑纹小、分布面积多、占蟹壳总面积比例高的特点;浙江海区呈现斑纹大、分布面积少、占蟹壳总面积比例低的特点。本研究将计算机视觉技术应用于水产养殖研究,通过建立评估参数阐明三疣梭子蟹白色斑纹的生长规律,揭示了不同群体的三疣梭子蟹白色斑纹具有地理多样性的特征。     

三疣梭子蟹低盐耐受性相关甲基化标记的开发及验证

为开发三疣梭子蟹低盐耐受性相关甲基化标记,采用甲基化敏感性高分辨率熔解曲线法(methylation sensitive high resolution melting curve, MS-HRM)进行了三疣梭子蟹低盐耐受性相关甲基化位点的筛选和应用。结果显示,从三疣梭子蟹转录组数据库中共开发了8个甲基化标记,其中6个标记与低盐度耐受性显著相关,在低盐度条件下表现出显著的去甲基化或甲基化,这些标记分别位于V-ATP酶(V-type proton ATPase)、CLC型氯离子通道、琥珀酸脱氢酶、NAD(P)转氢酶(NAD(P)、磷酸乙醇酸性磷酸酶、NADH还原酶基因,为三疣梭子蟹耐低盐新品种的分子标记辅助选育提供候选工具。研究表明,“宁象1号”耐低盐能力显著高于野生群体,且选育群体中位点Pt-M1、Pt-M2的去甲基化率达到100%,可能有利于低盐条件下保持体内Na⁺和Cl^-的平衡。     

Effects of salinity on growth,nutrient composition,fatty acid composition and energy metabolism of Scylla paramamosain during indoor overwintering

Three salinities (4‰, 12‰ and 25‰) were selected to determine the effects of different salinities on the growth, survival, nutrition and energy metabolism of Scylla paramamosain during indoor overwintering. Growth performance (survival rate, condition factor, hepatopancreatic index, weight gain rate and specific growth rate), nutrient composition (ash, moisture, crude fat, crude protein and total nitrogen), fatty acids, energy metabolites (glycogen, glucose, triglycerides, cholesterol and lactic acid) and energy metabolic enzymes (hexokinase, pyruvate kinase, lactate dehydrogenase and succinate dehydrogenase) were measured and calculated. The results showed that S. paramamosain exhibited the best growth and survival at 25‰ and worst at 4‰ after overwintering. In addition to consuming crude fat, excess protein was also broken down to provide energy in the 4‰ and 12‰ groups. The content of unsaturated fatty acids in the 25‰ group was higher after overwintering. Glycogen and glucose consumption and triglycerides, cholesterol and lactic acid production were lowest in the 25‰ group. The activity of hexokinase, pyruvate kinase, lactate dehydrogenase and succinate dehydrogenase was highest in the 4‰ group and lowest in the 25‰ group. The activity of hexokinase, pyruvate kinase, lactate dehydrogenase and succinate dehydrogenase in the hepatopancreas was higher than in the muscle. In conclusion, S. paramamosain uses less energy, has more unsaturated fatty acids and has a higher survival rate at a salinity of 25‰ after overwintering. The results of this study provide helpful information the indoor overwintering S. paramamosain in aquaculture production systems.     

拟穴青蟹软壳蟹与硬壳蟹营养品质和风味差异综合比较分析

为明确在最佳收集时间段内不同时间点的软壳蟹和硬壳蟹营养与风味物质的差异,本试验采用顶空固相微萃取(SPME)、气质联用(GS-MS)方法比较了拟穴青蟹硬壳蟹与蜕壳后0～48 h软壳蟹肌肉和肝胰腺2种组织中水分、灰分、氨基酸、矿物质元素、脂肪酸及挥发性物质含量.结果 表明,软壳蟹2种组织在蜕壳0h水分含量最高,硬壳蟹灰分...     

丙酸睾酮对三疣梭子蟹氨基酸和脂肪酸组成的影响

为研究丙酸睾酮(TP)对三疣梭子蟹氨基酸和脂肪酸组成的影响,试验以三疣梭子蟹I期幼蟹为材料,随机分为对照组(饲料中无丙酸睾酮)、TP 200处理组(饲料含200 mg·kg^-1TP)和TP 400处理组(饲料含400 mg·kg^-1TP),养殖90 d后,采用液相色谱质谱联用法检测血清TP、睾酮(T)和雌二醇(E₂)含量,并测定各组生长指标及存活率,检测肌肉和肝胰腺的基本营养成分、氨基酸及脂肪酸组成。结果显示,三疣梭子蟹血清中性激素含量较高,表明其具有较强的性激素富集能力。添加TP对三疣梭子蟹肝胰腺指数及蜕壳频率无显著影响,但成活率显著降低(P<0.05),TP 200组三疣梭子蟹的增重率、特定生长率及全甲宽等指标显著高于对照组及TP 400组(P<0.05)。TP 200组和TP 400组的总氨基酸、必需氨基酸含量与对照组相比无显著差异,但精氨酸含量显著降低(P<0.05)。TP 200组和TP 400组肌肉脂肪酸组成和含量与对照组相比无显著差异,但肝胰腺中部分脂肪酸特别是高度不饱和脂肪酸(C20∶2...     

Overwintering behaviour affects distinctive flavour of mud crab Scylla paramamosain in commercial farms

In order to reveal the influence of overwintering behaviour on the flavour of Scylla paramamosain, this study selected crabs of before and after overwintering in SM (Sanmen County, Zhejiang Province), HP (Hepu County, Guangxi Province) and WN (Wanning city, Hainan Province). The contents of flavouring‐related substances in hepatopancreas, muscles and gonads were determined by high‐performance liquid chromatography (HPLC); taste activity value (TAV) and equivalent umami concentration (EUC) were calculated. The results showed that sweetish amino acids were mainly accumulated in muscles, umami amino acids were mainly accumulated in hepatopancreas, free nucleotides were mainly accumulated in gonads, and the EUC value in gonads was the highest. After overwintering, the content of sweetish, umami amino acids and free nucleotides in muscle and hepatopancreas decreased significantly, while the content of flavour‐tasting amino acids in gonads did not change significantly, but the content of umami nucleotides increased significantly. Therefore, overwintering behaviour decreased the flavour of muscle and hepatopancreas, slightly enhanced gonad and the overall flavour of S. paramamosain before overwintering was better than that after overwintering.     

管角螺受精过程的荧光显微观察及保育卵形成分析

为了研究管角螺受精卵在受精和早期卵裂过程中的核相变化以及保育卵的形成,实验通过Hoechst 33258染色、荧光显微观察方法,对管角螺受精和早期卵裂过程中核行为的细胞学变化进行了详细观察,并对其胚胎发育过程中的数量变化进行统计分析。结果显示,管角螺的成熟未受精卵呈卵圆形,平均卵径(0.33±0.03)mm,核相处于第一次成熟分裂中期,在水温为(26±2)°C条件下,1 h精子入卵,2~3 h受精卵先后排出第1、第2极体,完成第一次和第二次成熟分裂,6 h雌、雄性原核形成,靠近融合,第一次卵裂形成2细胞,12~24 h时2细胞继续分裂生成4细胞。研究表明,管角螺受精卵在成熟分裂和卵裂过程中表现出发育的不同步性,同个卵荚内多个发育阶段共存,受精率为43.14%,而受精卵正常发育概率仅5.70%。卵荚内多数卵子不受精或者受精卵停止发育,成为营养卵(保育卵),为正常可育卵消化利用,这两种来源的保育卵比率约为3∶2。管角螺胚胎发育过程中,1~6 d内胚胎数减少明显,担轮幼虫发育率为0.48%,之后数量稳定,直至稚螺出膜。     

贮藏在海藻酸钙胶丸中的荔枝种子活力

从健康果实中取出的荔枝种子分别放入加有0．1％代森锌的海藻酸钙溶液中，然后在2％氯化钙溶液中胶丸化，形成胶丸种子。胶丸种子和非胶丸种子在10℃下贮存80天，非胶丸种子在10℃中贮藏40天发芽率是11．7％，胶丸种子（4％海藻酸钙）在贮藏80天后仍有53．3％的发芽率。结果表明用海藻酸钙制成的胶丸可延长荔枝这样的“顽拗型”种子的贮藏寿命。