牡蛎糖原的研究 (II)——牡蛎糖原的分子结构

该研究以新鲜近江牡蛎 (Crassostrea ariakensis) 软体组织为实验材料,通过比较分析不同蒽酮硫酸比例、不同反应时间的吸光值确定最优反应条件,并对方法的最低检出限、重复性、稳定性和精确性进行评价,最终建立了牡蛎糖原含量的微量检测体系。建立的微量反应体系体积为300 μL,主要包括0.2%蒽酮硫酸溶液200 μL,样品待测液100 μL;沸水浴反应时间为10 min。该方法的葡萄糖最低检出限为0.001 5 mg·mL⁻¹,标准曲线变异系数小于4%,证实该方法具有较高的检测灵敏度;显色反应完成后,室温条件下120 min内其吸光值基本不变,稳定性较高;测定牡蛎外套膜、鳃、唇瓣、性腺、肝胰腺、闭壳肌的加标回收率介于95.3%~105.8%,说明该法具有较高的准确性。因此,该研究建立的微量反应体系测定牡蛎糖原的方法具有试剂用量小、操作简单、单样品成本低等优势,同时重复性、稳定性、精确性均较高,适用于大批样品批量测定。该研究为快速、高效完成牡蛎样品糖原的检测提供了有效的技术方法。     

健跳港上游湾区牡蛎潮间带分带格局和资源补充研究#br#

为了解牡蛎在潮间带的生态特性及资源分布情况,调查了浙江三门健跳港上游湾区牡蛎潮间带分带格局,并检验附苗器类型和地点对牡蛎资源补充的影响。结果显示:潮间带定量调查共记录到3种牡蛎,即熊本牡蛎(Crassostrea sikamea)、近江牡蛎(C.ariakensis)和香港牡蛎(C.hongkongensis),首次在浙江海域记录到香港牡蛎的自然分布。熊本牡蛎在调查的5个潮区(1.4 m、2.2 m、3.0 m、3.8m、4.6 m)均有分布,其平均密度沿增加的高程梯度呈现上升趋势,其中4.6 m潮区熊本牡蛎密度显著高于1.4 m和2.2 m潮区(P<0.05)。近江牡蛎和香港牡蛎仅分布于最底层的1.4 m和2.2 m潮区。牡蛎资源补充实验结果表明,该湾区具有很高的熊本牡蛎资源补充量,采苗地点(P<0.05)和附苗器类型(P<0.05)均显著影响牡蛎苗的密度和壳高,总体上水泥片的附苗效果要好于塑胶轮胎,上游S1点附苗效果优于下游S2点,推荐上游S1点作为熊本牡蛎附苗场。     

Assessment of the performance of Pacific oyster (Crassostrea gigas) larvae fed with fresh and preserved Pavlova lutheri concentrates

The Prymnesiophycea Pavlova lutheriwas produced using a traditional technique (batch mode in carboys) and by an innovative technique (semi-continuous mode in alveolarphotobioreactor). Two experimental feedingtrials were carried out with Pacific oyster(Crassostrea gigas) larvae with freshand preserved (4 °C and 1 °C)P. lutheri concentrates used inmonospecific and bispecific diets, incombination with fresh Chaetoceroscalcitrans forma pumilum. Nodifferences in terms of gross composition andlarval performance were noticed between thefresh algae biomass produced by the twotechniques. The low temperatures allowedpreservation of the nutritionalcharacteristics of the P. lutheri freshculture for the time period considered (27days). The P. lutheri concentratespreserved at 4 °C or 1 °C didnot differ substantially in terms ofnutritional value. Larvae fed a monospecificdiet of P. lutheri, both fresh andpreserved, exhibited a modest growth rate (<2.1 µm day^–1), in some cases notsignificantly different from that observed inthe unfed larvae (the control), and a lowsurvival rate. In contrast, when Chaetoceros calcitrans forma pumilumwas used as a monospecific diet, good growthperformance (>4.5 µm day^–1) and ahigh survival rate (>86%) were observed inlarvae. A substitution of 50% (trial 1) or80% (trial 2) of fresh C. calcitransforma pumilum with the preserved P. lutheri concentrates did not adverselyaffect growth rate or survival of C.gigas larvae.     

In vitro modulation of reactive oxygen and nitrogen intermediate (ROI/RNI) production in Crassostrea gigas hemocytes

Bivalve hemocyte competence has been measured by quantifying functional characteristics, including reactive oxygen intermediate (ROI) production after activation with zymosan or phorbol myristate acetate (PMA). However, untreated oyster hemocytes also produce ROI and RNI (reactive nitrogen intermediates) after bleeding even if not stimulated by zymosan or PMA. Extensive investigation of this parameter by flow cytometry showed that, in vitro, ROI/RNI production by untreated hemocytes maintained in seawater appeared to be independent of both bacterial burden in the serum and non-self particle phagocytosis. ROI/RNI production in granulocytes was higher than in hyalinocytes and could be intensified when activated by zymosan but not by PMA. Both cell types used NADPH-oxidase- and NO-synthase-like pathways to produce these molecules; the NO-synthase pathway seemed relatively more dominant in hyalinocytes and NADPH-oxidase appeared more effective in granulocytes. These results provide new insights for interpreting the modulation of ROI/RNI production by untreated hemocytes shown by other studies, relative to environmental conditions or physiological status of the oysters.     

The Negative Relationship between Fouling Organisms and the Content of Eicosapentaenoic Acid and Docosahexaenoic Acid in Cultivated Pacific Oysters,Crassostrea gigas

Bivalves serve as an important aquaculture product, as they are the source of essential fatty acids, such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), in our diet. However, their cultivation in the wild can be affected by fouling organisms that, in turn, affect their EPA and DHA content. The effects of fouling organisms on the EPA and DHA contents of cultivated bivalves have not been well documented. We examined the effects of fouling organisms on the EPA and DHA contents and condition index of cultured oysters, Crassostrea gigas, in an aquaculture system. We sampled two-year-old oysters from five sites in Shizugawa Bay, Japan, in August 2014. Most of the fouling organisms were sponges, macroalgae, and Mytilus galloprovincialis. A significant negative relationship existed between the DHA content in C. gigas and the presence of sponges and macroalgae. A lower C. gigas EPA content corresponded to a higher M. galloprovincialis fouling mass and a lower C. gigas condition index. This can be explained by dietary competition between C. gigas and M. galloprovincialis for diatoms, which were the main producer of EPA in our study sites. Our findings indicate that fouling organisms likely reduce the EPA and DHA content in cultivated oysters. Therefore, our results suggest that the current efforts to remove fouling organisms from oyster clusters is an effective strategy to enhance the content of EPA and DHA in oysters.     

Electrophysiological Evaluation of Pacific Oyster (Crassostrea gigas) Sensitivity to Saxitoxin and Tetrodotoxin

Pacific oysters (Crassostrea gigas) may bio-accumulate high levels of paralytic shellfish toxins (PST) during harmful algal blooms of the genus Alexandrium. These blooms regularly occur in coastal waters, affecting oyster health and marketability. The aim of our study was to analyse the PST-sensitivity of nerves of Pacific oysters in relation with toxin bio-accumulation. The results show that C. gigas nerves have micromolar range of saxitoxin (STX) sensitivity, thus providing intermediate STX sensitivity compared to other bivalve species. However, theses nerves were much less sensitive to tetrodotoxin. The STX-sensitivity of compound nerve action potential (CNAP) recorded from oysters experimentally fed with Alexandrium minutum (toxic-alga-exposed oysters), or Tisochrysis lutea, a non-toxic microalga (control oysters), revealed that oysters could be separated into STX-resistant and STX-sensitive categories, regardless of the diet. Moreover, the percentage of toxin-sensitive nerves was lower, and the STX concentration necessary to inhibit 50% of CNAP higher, in recently toxic-alga-exposed oysters than in control bivalves. However, no obvious correlation was observed between nerve sensitivity to STX and the STX content in oyster digestive glands. None of the nerves isolated from wild and farmed oysters was detected to be sensitive to tetrodotoxin. In conclusion, this study highlights the good potential of cerebrovisceral nerves of Pacific oysters for electrophysiological and pharmacological studies. In addition, this study shows, for the first time, that C. gigas nerves have micromolar range of STX sensitivity. The STX sensitivity decreases, at least temporary, upon recent oyster exposure to dinoflagellates producing PST under natural, but not experimental environment.     

太平洋牡蛎卵母细胞发育及卵黄发生的超微结构

利用透射电镜观察了太平洋牡蛎（ Ｃｒａｓｓｏｓｔｒｅａ ｇｉｇａｓ） 卵母细胞的发育及卵黄发生。卵母细胞的发育可分为卵黄合成前卵母细胞与卵黄合成期卵母细胞２ 个阶段。卵黄颗粒来源于线粒体、内质网、高尔基体、吞饮小泡等多种细胞器。