保幼激素拮抗物KK-42对南美白对虾Y-器官发育的影响

采用1.95×10-4mol.L-1的KK-42溶液浸润处理南美白对虾(Penaeus vannamei)幼虾(体长为4.2~5.0cm),从组织细胞学水平对分泌蜕皮激素的Y-器官结构进行初步研究,观察KK-42处理后幼虾Y-器官的形态结构变化。结果表明,伴随动物的生长,Y-器官腺细胞条索变粗,腺细胞核增大。KK-42处理后腺细胞核的结构、大小未见显著变化,腺细胞索的宽度明显高于同一时期对照组。     

制备K700bp成探针与美意和平湖两品系西蜂RAPD-PCR扩增产物及它们基因组DNA分子杂交的研究

Ｋ７００ｂｐ 是高产蜜西蜂引物Ｋ（５’－ ＣＧＧＣＣＣＣＴＧＣ－ ３’） ，通过ＲＡＰＤ－ ＰＣＲ 扩增出来的一个特有ＤＮＡ 片段。然后将Ｋ７００ｂｐ 制备成探针再与高、低产蜜西蜂的ＰＣＲ 产物及它们的基因组进行杂交。结果Ｋ７００探针只与高产蜜西蜂的ＰＣＲ 产物及其基因组杂交，证明Ｋ７００ｂｐ 确实是高产蜜西蜂的一个特有ＤＮＡ 片段。     

斑节对虾白斑综合症病毒部分基因组文库及核酸探针检测法

通过分离纯化白斑综合症病毒（ＷＳＳＶ）粒子，抽提病毒ＤＮＡ。用限制性内切酶ＥｃｏＲⅠ或ＳａｌⅠ酶切后，克隆入质粒ｐＢｌｕｅｓｃｒｉｐｔⅡＫＳ中，从而建立了ＷＳＳＶ部分基因组文库。估计ＷＳＳＶ基因组ＤＮＡ在１６５ｋｂ以上。将ＷＳＳＶ ＥｃｏＲⅠ克隆片段标记制备为探针。进行Ｓｏｕｔｈｅｒｎ杂交、打点杂交和原位杂交，其结果证明了克隆片段对ＷＳＳＶ特异，并为检测ＷＳＳＶ提供了方法。通过对部分基因组文库序列     

全封闭生态养殖斑节对虾Penaeus monodon Fabricius技术的研究

本文报道了深圳市宝安区水产科学研究所，于１９９８年和１９９９年连续二年，在珠江口低盐区，采用全封闭生态养殖斑节对是技术，并取得成功，这项新技术为预防虾病提供理论基础和实践经验，具有推广和应用价值。     

凡纳滨对虾引进群体和2个养殖群体遗传变异的微卫星分析

利用8个微卫星标记对引进的SPF凡纳滨对虾G0和两个养殖群体(G1,G2)进行遗传多样性分析。8个座位共获得64个等位基因,位点的等位基因数在5~l3之间。多态信息含量PIC在0.405 2~0.869 3之间,其中有6个位点为高度多态位点,适合于多态性分析。8个座位丢失的和新产生的等位基因共30个,占总数的47%。3个群体的平均观测杂合度分别为0.193 8、0.196 1、0.232 5,说明3个群体的遗传多样性较低。对近交系数Fis分析显示3个群体中存在近交,通过对哈迪温伯格平衡检验,显示所有座位均显著偏离平衡,存在杂合子缺失。通过配对Fst和Ne i遗传距离分析,显示3个群体之间有明显的遗传分化,说明种群结构发生了明显的遗传变异,变异可能来自于突变、随机漂变和选择的共同作用。实验结果能够很好地解释经过若干群体选育后,子代群体发生种质退化的现状,由此建议综合采用遗传育种的方法从引进的亲虾中筛选出性状优良稳定的仔虾作为虾苗。     

超高效液相色谱-串联质谱法测定凡纳滨对虾中的硝基呋喃代谢物残留

采用超高效液相色谱-串联质谱（UPLC-MS/MS）法,测定了凡纳滨对虾（Litopenaeus vannam ei）中氨基脲（SEM）、1-氨基-2-内酰脲（AHD）、3-氨基-2-唑烷基酮（AOZ）和5-甲基吗啉-3-氨基-2-唑烷基酮（AMOZ）4种硝基呋喃类代谢物。采用2-硝基苯甲醛作为衍生化试剂,氘、碳-13、氮-15同位素标记的SEM1-3C1-5N2、AHD1-3C3、AOZ-D4和AMOZ-D5为内标物,样品经衍生、乙酸乙酯提取、正己烷净化,以甲醇0.5 mmol.L-1乙酸铵水溶液（含0.1%甲酸）为流动相进行梯度洗脱,8 m in可将4种目标化合物完全分离并测定。结果表明,4种待测物在0.4～20.0 ng.mL-1范围内线性关系良好（R〉0.999）,检出限为0.03～0.15μg.kg-1。在0.5μg.kg-1、1.0μg.kg-1和2.0μg.kg-13个水平上进行加标回收试验,平均回收率为92.0%～107.3%,RSD为3.1%～8.7%。采用该法参加FAPAS国际水平测试,获得了满意的结果。     

斑节对虾C1q结合蛋白(PmC1qBP)的克隆及表达特征分析

Clq结合蛋白(ClqBP)是补体系统中的一种重要成分,它能与游离的补体分子Clq的胶原样区相互作用并启动多种细胞反应,包括增强吞噬作用、促进吞噬细胞对微生物的杀伤、诱导趋化作用,从而增强生物体的免疫性.本实验从构建的斑节对虾(Penaeus monodon)卵巢组织cDNA文库中筛选并克隆了PmClqBP基因.为研究PmClqBP在斑节对虾中的生物学功能,采用半定量和荧光定量的方法研究比较了PmClqBP基因在雌雄个体不同组织及卵巢不同发育阶段的差异表达情况.结果表明:雌雄个体的PmClqBP基因在性腺、胃、血细胞、肠中表达量存在极显著差异(P＜0.01).同时,PmClqBP在无节幼体和蚤状幼体时表达量低,从糠虾幼体至卵巢发育成熟过程中表达量有所增加并趋于稳定,揭示PmClqBP斑节对虾卵巢发育过程的重要功能基因.应激实验结果表明,经脂多糖(Lipopolysaccharide,LPS)刺激6h后PmClqBP在肝胰腺中的表达量极显著上调,提示PmClqBP参与了斑节对虾天然免疫应答,暗示其在斑节对虾先天性免疫调控中发挥着重要的作用.     

Growth performance of the white shrimp,Litopenaeus vannamei,fed on practical diets with increasing levels of the Antarctic krill meal,Euphausia superba,reared in clear‐ versus green‐water culture tanks

Litopenaeus vannamei were stocked in 25 clear‐water 500‐L tanks at 100 shrimp m^?2 and in 25 green‐water 1000‐L tanks at 60 animals m^?2. Four diets were formulated to include krill meal at 10, 50 or 110 g kg^?1; or krill oil at 25 g kg^?1 by replacing fish meal, fish oil, soybean lecithin and cholesterol. Diets had similar levels of crude protein, total energy and essential amino acids. After 72 days, shrimp reared in clear and green water showed no differences in performance among diets. In clear water, shrimp attained 13.1 ± 0.59 g body weight, 1.00 ± 0.06 g week^?1 growth, 81.4 ± 7.3% survival, 780 ± 118 g m^?2 yield, 16.9 ± 1.8 g shrimp^?1 apparent feed intake (AFI), and 2.18 ± 0.29 food conversion ratio (FCR). In green water, shrimp attained 14.3 ± 0.81 g body weight, 1.04 ± 0.09 g week^?1 growth, 91.4 ± 5.4% survival, 569 ± 69 g m^?2 yield, 20.9 ± 3.2 g shrimp^?1 AFI, and 2.22 ± 0.34 FCR. Diets containing krill meal or krill oil were able to fully replace the protein and lipid value of fish meal, fish oil, soybean lecithin and cholesterol at no cost to performance.     

Identification and characterization of Alix/AIP1 interacting proteins from the black tiger shrimp,Penaeus monodon

Apoptosis is proposed to be a major cause of death in shrimp viral infections. From our previous study, an apoptosis-related gene, Pm-Alix, was identified from the black tiger shrimp. Its expression was high in defence-related tissues including haemocytes and the lymphoid organ. To clarify its possible role in shrimp, we used Pm-Alix as bait in a yeast two-hybrid analysis to search for Alix interacting proteins in shrimp. Two cDNA sequences discovered had homology to a predicted ubiquitin C of the purple sea urchin, Strongylocentrotus purpuratus, and to a guanylyl cyclase of the red swamp crayfish, Procambarus clarkii. In vitro pull-down assays confirmed positive interaction between Pm-Alix and both proteins. Tissue distribution analysis revealed that Pm-Alix and the two binding partners were widely expressed in various tissues but more highly expressed in haemocytes. However, no significant positive or negative correlation was found in the expression of these genes as shrimp approached morbidity and death after challenge with white spot syndrome virus. Thus, the results suggested that Alix and its interacting partners did not play a direct role related to shrimp death.